Molecular Weight (MW)

307.34

Formula

C19H17NO3

CAS No.

154447-36-6

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 36 mg/mL (117.1 mM)

Water:<1 mg/mL (slightly soluble or insoluble)

Ethanol: 21 mg/mL (68.32 mM)

Solubility (In vivo)

4% DMSO+30% PEG 300+5% Tween 80+ddH2O: 5 mg/mL

Chemical Name

2-morpholino-8-phenyl-4H-chromen-4-one

Synonym

LY294002; LY-294002; LY 294002; NSC 697286; SF 1101

SMILES

O=C1C=C(N2CCOCC2)OC3=C(C4=CC=CC=C4)C=CC=C13

In Vitro

In vitro activity: LY294002 inactivates Akt/PKB, consequently inhibiting cell proliferation and inducing apoptosis. LY294002 demonstrates a remarkable growth-inhibitory and apoptosis-inducing effect in these colon cancer cell lines, with decreased expression of phosphorylated Akt (Ser473). LY294002 induces marked nuclear pyknosis and diminished cytoplasmic volume in the tumor cells. Thus, LY294002 markedly inhibits ovarian cancer cell proliferation in vitro. LY294002 induces specific G1 arrest in cell growth, leading to almost complete inhibition of melanoma cell proliferation and partial inhibition of MG-63 (osteosarcoma cell line) proliferation. The effect of LY294002 on cell cycle progression may provide insights into a possible link between the PI3K activation pathway and cancer cell cycle regulation.

Kinase Assay: PI3K inhibition by LY294002 is determined in a radiometric assay using purified, recombinant enzymes with 1 μM ATP. The kinase reaction is carried out for 1 hour at room temperature (24oC) and is terminated by addition of PBS. IC50 values are subsequently determined using a sigmoidal dose–response curve fit (variable slope). CK2 and GSK3β (glycogen synthase kinase 3β) inhibition is established by kinase selectivity screening. LY294002 is tested against the Upstate panel of kinases in 10 μM ATP.

Cell Assay: 1.0×105 cells (100 μL volume/well, Colon cancer cell lines DLD-1, LoVo, HCT15, and Colo205 ) are inoculated into 96-well microtiter plates. LY294002 is added to triplicate wells and cultured at 37oC for 0–48 hours. After treatment, 10 μL of Premix WST-1 are added to each microculture well, and the plates are incubated for 60 minutes at 37oC, after which absorbance at 450 nm is measured with a microplate reader.

In Vivo

Administration of GDC-0941 at 75 mg/kg/day displays significant inhibitory effect against established human U87MG glioblastoma xenografts in female NCr athymic mice, with tumor growth inhibition of 83%. Oral administration of GDC-0941 at 150 mg/kg/day inhibits the growth of HER2-amplified, trastuzumab-resistant MDA-MB-361.1 xenografts in mice, and significantly delays the tumor progression, in association with potent induced apoptosis in tumors. GDC-0941 (75 mg/kg/day) treatment for 2 weeks induces ~40% reduction in tumor volume of spontaneous B-cell follicular lymphomas developed in PTEN+/-LKB1+/hypo mice, accompanied by ablation of phosphorylation of Akt, S6K and SGK (serum and glucocorticoid protein kinase) protein kinases.

Animal model

Two groups of athymic nude mice (5–7 weeks) are inoculated i.p. with OVCAR-3 cells

Formulation & Dosage

DMSO + 0.25 ml of PBS; 100 mg/kg ; i.p.

References

[1] Chaussade C, et al. Biochem J, 2007, 404(3), 449-58.; [2] Semba S, et al. Clin Cancer Res, 2002, 8(6), 1957-63.