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Research Area
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Description
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Myocardial infarction |
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Biological Activity
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Description
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TG100-115 is a potent and dual selective inhibitor for PI3Kγ and PI3Kδ with IC50 of 83 nM and 235 nM, respectively. |
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Targets
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PI3Kγ |
PI3Kδ |
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IC50 |
83 nM |
235 nM [1] |
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In Vitro
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TG100-115 inhibits PI3Kγ and -δ, with IC50 of 83 and 235 nM, respectively. TG100-115 is not active for PI3Kα and -β, with IC50 of 1.2 and 1.3 mM. In human umbilical vein endothelial cells (HUVECs), TG100-115 (up to 10 μM) has no effects on cell proliferation and VEGF-stimulated ERK phosphorylation. However, TG100-115 (10 μM) interrupts other VEGF signaling pathways, such as those that culminate in VE-cadherin phosphorylation. [1] In HUVECs, TG100-115 (10 μM) inhibits the VEGF-induced increase of total level of VE-cadherin. TG100-115 inhibits VEGF mediated phosphorylation of mTOR and p70S6 kinase, both of which are downstream of PI3K. TG100-115 (125 nM to 10 μM) also inhibits FGF-stimulated phosphorylation of Akt. [2] |
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In Vivo
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In Miles assay models, TG100-115 (1–5 mg/kg) reduces edema formation and inflammation in rats. In rigorous rodent and porcine models of myocardial ischemia (MI), TG100-115 (0.5–5 mg/kg) provides potent cardioprotection, limits infarct development, and preserves myocardial function. [1] In mice, TG100-115 (5 mg/kg) markedly diminishes vascular permeability (VP) in response to either Sema3A or VEGF, indicating that both factors may depend on PI3Kγ/δ to induce VP. [3] In a mouse asthma model, aerosolized TG100-115 markedly reduces the pulmonary eosinophilia, inhibits interleukin-13 and mucin accumulation. [4] |
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Clinical Trials
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TG100-115 is currently under a Phase I/II clinical trial in myocardial infarction. |
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Features
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TG100-115 is a potent and dual selective PI3Kγ/δ inhibitor. |
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Protocol
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Kinase Assay
[1]
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| PI3K assays |
Forty mL of reaction buffer (20 mM Tris/4 mM MgCl2/10 mM NaCl, pH 7.4) containing 50 mM D-myo-phosphatidylinositol 4,5-bisphosphate substrate and the desired PI3K isoform are aliquoted to 96-well plates; kinase concentrations are 250-500 ng/well, such that linear kinetics are achieved over 90 min. TG100-115 is then added as 2.5 mL of a DMSO stock to final concentration range of 100 mM to 1 nM. Reactions are initiated by addition of 10 mL of ATP to a final concentration of 3 mM, and after 90 min, 50 mL of Kinase-Glo reagent added to quantify residual ATP levels; luminosity is measured using an Ultra 384 instrument. Control reactions omitting either TG100-115 or substrate are also performed. IC50 values are derived from experimental data by nonlinear curve fitting using Prism Version 4. |
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Cell Assay
[1]
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| Cell Lines |
Human umbilical vein endothelial cells (HUVECs) |
| Concentrations |
10 μM, dissolved in DMSO as stock solution |
| Incubation Time |
24, 48, and 72 hours |
| Methods |
Cells plated in 96-well cluster plates (5 × 103 cells/well) are cultured in assay medium (containing 0.5% serum and 50 ng/ml VEGF) in the presence or absence of TG100-115, and cell numbers are quantified by XTT assay 24, 48, or 72 hours later. |
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Animal Study
[1]
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| Animal Models |
Rat (Sprague-Dawley) myocardial ischemia (MI) model |
| Formulation |
Dissolved in PEG or sulfobutyl ether β-cyclodextrin |
| Doses |
0.5–5 mg/kg |
| Administration |
By intravenous injection. |
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References |
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[1] Doukas J, et al. Proc Natl Acad Sci U S A, 2006, 103(52), 19866-19871.
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[2] Palanki MS, et al. J Med Chem, 2007, 50(18), 4279-4294.
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[3] Acevedo LM, et al. Blood, 2008, 111(5), 2674-2680.
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[4] Doukas J, et al. J Pharmacol Exp Ther, 2009, 328(3), 758-765.
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