| 包装 | 价格(元) |
| 10mM (in 1mL DMSO) | 电议 |
| 10mg | 电议 |
| 50mg | 电议 |
| 100mg | 电议 |
Kinase experiment: | Ten centimeter square dishes are seeded with 2 million cells in a volume of 10 mL followed by incubation at 37℃ under 5% CO2 overnight (appr 16 hours). Cells are treated with the indicated concentration of GDC-0941, Apitolisib (GDC-0980), or mTOR1/2 inhibitor for the time indicated. Following treatment, cells are washed with cold PBS and lysed in 1X Cell Extraction Buffer, 1 mM PMSF, and Phosphatase Inhibitor Cocktails 1 and 2 are all needed. Protein concentration is determined using the Pierce BCA Protein Assay Kit. For immunoblots, equal protein amounts are separated by electrophoresis through NuPage Bis-Tris 10% gradient gels; proteins are transferred onto polyvinylidene difluoride membranes using the Criterion system and protocol. |
Cell experiment: | Three hundred and eighty-four-well plates are seeded with 2,000 cells/well in a volume of 54 μL per well followed by incubation at 37℃ under 5% CO2 overnight (appr 16 hours). Compounds are diluted in dimethyl sulfoxide to generate the desired stock concentrations then added in a volume of 6 μL per well. All treatments are tested in quadruplicate. After 4 days incubation, relative numbers of viable cells are estimated using CellTiter-Glo and total luminescence is measured on a Wallac Multilabel Reader. The concentration of drug resulting in 50% inhibition of cell viability (IC50) or 50% maximal effective concentration (EC50) is determined using Prism software. For cell lines that failed to achieve an IC50 the highest concentration tested (20 μM) is listed. |
Animal experiment: | Human prostate cancer PC3 cells are resuspended in Hank’s Balanced Salt Solution and 3×106 cells implanted subcutaneously into the right hind flank of athymic nu/nu (nude) mice. Tumors are monitored until they reach a mean tumor volume of 150-200 mm3 prior to the initiation of dosing. MCF7.1 cells resuspended in a 1:1 mixture of Hank’s Buffered Salt Solution and Matrigel Basement Membrane Matrix are 5×106 subcutaneously implanted into the right hind flank of athymic nu/nu (nude) mice. Prior to cell inoculation, 17β-estradiol (0.36 mg/pellet, 60-day release, no. SE-121) are implanted into the dorsal shoulder blade area of each nude mouse. After implantation of cells, tumors are monitored until they reach a mean tumor volume of 250-350 mm3 prior to initiating dosing. Compound 2 is dissolved in 0.5% methylcellulose with 0.2% Tween-80 (MCT). Female nude (nu/nu) mice that are 6-8 weeks old and weighed 20-30 g are obtained from Charles River Laboratories. Tumor bearing mice are dosed daily for 14-21 days depending on the xenograft model with 100 μL of vehicle (MCT) or test agent orally. |
| 产品描述 | GDC-0980 (RG7422) is a selective, novel, potent and orally bioavailable inhibitor of class 1 PI3K/mTOR kinase with the Ki value of 17nmol/L for mTOR kinase [1]. GDC-0980 (RG7422) has shown the effective inhibition with the IC50 values of ＜200nmol/L in prostate, breast and NSCLC lines, respectinely. In addition, GDC-0980 (RG7422) has been reported to reduce cell viability in KPL4 cells by cell-cycle inhibition and induction of apoptosis with the IC50 value of 109nM and the EC50 value of 78nM. Furthermore, GDC-0980 (RG7422) has been revealed to significantly inhibit tumor responses in xenograft models in oral dose of the compound. Besides, GDC-0980 (RG7422) has been noted to enhance the antitumor activity in combination with antitumor agents ( docetaxel) in vivo [1]. References: |
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