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Biological Activity
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Description
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AS-252424 is a novel, potent and selective inhibitor of PI3Kγ with IC50 of 33 nM. |
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Targets
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PI3Kγ |
CK2 (casein kinase 2) |
PI3Kα |
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IC50 |
33 nM |
20 nM |
935 nM [1] |
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In Vitro
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AS-252424 is a furan-2-ylmethylene thiazolidinedione as a selective ATP-competitive PI3Kγ inhibitor with IC50 with 33 nM. AS-252424 shows reduced potency on PI3Kα with an IC50 with 935 nM. When screening against 80 different Ser/Thr and Tyr kinases, AS-252424 doesn’t show significant inhibit towards any of them at 10 μM except for CK2. AS-252424 inhibits C5a-mediated PKB/Akt phosphorylation in a concentration- dependent manner with submicromolar or low-micromolarIC50 value. AS-252424 inhibits MCP-1-mediated chemotaxis in wild-type primary monocytes in a concentration-dependent manner with an IC50 value of 52 μM, as well as in the monocytic cell line THP-1 with an IC50 value of 53 μM. [1] AS252424 specifically blocks proliferation in the pancreatic cancer cell lines HPAF and Capan1, as assessed by cell counting. [2] A recent research indicates 100 nM of AS-252424 significantly reduces [Ca2+]i, ICa and Ca2+ transients in HL-1 cardiomyocytes. [3] |
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In Vivo
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Oral administration of AS-252424 at 10 mg/kg results in moderate reduction of neutrophil recruitment (35%), almost matching the result observed in PI3Kγ-deficient mice. [1] |
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Clinical Trials
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Features
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Protocol
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Kinase Assay
[1]
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| In vitro PI3Kγ Kinase Assay |
Human PI3Kγ (100 ng) is incubated at RT with kinase buffer (10 mM MgCl2, 1 mM β-glycerophosphate, 1 mM DTT, 0.1 mM Na3VO4, 0.1% Na Cholate and 15 M ATP/100 nCi γ[33]ATP, final concentrations) and lipid vesicles containing 18 M PtdIns and 250 M of PtdSer (final concentrations), in the presence of AS-252424 or DMSO. Kinase reaction is stopped by adding 250 g of Neomycin-coated Scintillation Proximity Assay (SPA) beads and preceded. |
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Cell Assay
[1]
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| Cell Lines |
Raw-264 macrophage |
| Concentrations |
0.01 nM to 0.1 μM |
| Incubation Time |
5 minutes |
| Methods |
After 3 hours of starvation in serum-free medium, Raw-264 macrophages are pretreated with AS-252424 or DMSO for 30 minutes and stimulated for 5 minutes with 50 nM C5a. PKB/Akt phosphorylation is monitored using phospho-Ser-473 Akt specific antibody and standard ELISA protocols. |
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Animal Study
[1]
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| Animal Models |
Intraperitoneally injecting female C3H mice with thioglycollate (40 ml/kg) to induced a peritonitis mouse model |
| Formulation |
AS-252424 is dissolved in vehicle (0.5% carboxymethylcellulose/0.25% Tween-20) and adjusted the solution to 10 mL/kg of body weight. |
| Doses |
10 mg/kg |
| Administration |
Oral administration 15 minutes before injection of thioglycollate |
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References |
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[1] Pomel V, et al, J Med Chem, 2006, 49(13), 3857-3871.
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[2] Edling CE, et al, Clin Cancer Res, 2010, 16(20), 4928-4937.
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[3] Graves BM, et al, J Biomed Sci, 2012, 19(1), 59 [Epub ahead of print]
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