Table 1.
Biochemical and Cellular Activity of GSK2110183 and GSK2141795.
Figure 1.
Effect of GSK2141795 on AKT signaling and growth inhibition in human cancer cell lines.
A, BT474 (left) and LNCaP (right) cell lines were treated with DMSO or GSK2141795 for 1 h. Western analysis was performed to assay levels of phosphorylated and total GSK-3, PRAS40, AKT and ERK, phosphorylated FOXO, Caspase 9, and MEK. Tubulin was used as a loading control. B, Scatter plot of EC50's for anti-proliferative effect of GSK2141795 against various haematological cancer cell lines. Cell lines are grouped according to their disease classification and sub-divided into cellular origin of B cell (black diamond), pre-B cell (red diamond), T cell (blue diamond) or other (grey diamond). C, Scatter plot of EC50's for various solid cancer cell lines treated with GSK2141795. Cells were treated as described above. Cell lines are grouped by tissue of origin then further divided by genetic status. KRAS/BRAFV600E mutation status is represented by color with mutant (red), wild type (black) or other (blue); whereas PIK3CA or PTEN status represented with shape, mutant (diamond) and wild type (circle).
Figure 2.
Effect of GSK2110183 on AKT signaling and growth inhibition in human cancer cell lines.
A, BT474 (left) and LNCaP (right) cell lines were treated with DMSO or GSK2110183 for 1 h. Western analysis was performed to assay levels of phosphorylated and total GSK-3, PRAS40, AKT and ERK, phosphorylated Foxo, Caspase 9, and MEK. Tubulin was used as a loading control. B, Scatter plot of EC50's for anti-proliferative effect of GSK2110183 against various haematological cancer cell lines. Cell lines are grouped according to their disease classification and sub-divided into cellular origin of B cell (black diamond), pre-B cell (red diamond), T cell (blue diamond) or other (grey diamond). C, Scatter plot of EC50's for various solid cancer cell lines treated with GSK2110183. Cells were treated as described above. Cell lines are grouped by tissue of origin then further divided by genetic status. KRAS/BRAFV600E mutation status is represented by color with mutant (red), wild type (black) or other (blue); whereas PIK3CA or PTEN status represented with shape, mutant (diamond) and wild type (circle).
Figure 3.
Effect of GSK2141795 on cell cycle.
LNCaP, BT474, A3 and I9.2 cells lines were treated with GSK2141795 for 24; bars, SD.
Figure 4.
Dose responsive PD/PK relationship of GSK2110183 and GSK2141795 in BT474 tumor xenografts.
Mice bearing BT474 tumors were treated with vehicle or GSK2110183 at 10, 30 or 100/kg (A) or GSK2141795 at 3, 10 and 30 mg/kg (B) daily for 7 days (QDx7; n = 3/group). Tumors were harvested and analysed by ELISA for phosphorylated and total PRAS40 levels. The concentration of compound in the tumor (black triangles; ng/g) and blood (grey squares; ng/mL) was quantified by LC/MS-MS. Data represents mean ± s.d. *p<0.01.
Figure 5.
The impact of GSK2110183 and GSK2141795 on glucose homeostasis in vivo.
Blood glucose (black squares) and plasma insulin (black circles) levels were assayed over time in mice treated with a single efficacious dose of GSK2110183 (30 mg/kg) or GSK2141795 (100 mg/kg). Data represents mean ± s.d.
Figure 6.
Anti-tumor activity of GSK2141795 in vivo.
Mice bearing either BT474 (A) or SKOV3 (B) tumors were treated with vehicle (black line) or GSK2141795 at 10 (red line), 20 (blue line) or 30 (gold line) mg/kg, once daily for 21 d (QDx21). Duration of treatment is shown by the horizontal grey line. Tumor volume was measured twice per week. Data represents mean ± SEM. #p<0.05, *p<0.01.
Figure 7.
Combination anti-tumor effect of AKT and MEK inhibitors in mouse models of pancreatic cancer.
Mice bearing HPAC (A) or CAPAN-2 (B) tumor xenografts were treated with either vehicle (black line), 0.3 mg/kg GSK1120212 (purple line), 30 mg/kg GSK2141795 (green line) or the combination of both (blue line). Data represents mean ± SEM. #p<0.05, *p<0.01. C, Immunohistochemical evaluation of HPAC tumor xenografts treated with vehicle, GSK1120212 at 0.3 mg/kg QDx3 (MEK inh), GSK2141795 at 30 mg/kg QDx3 (AKT inh) or the combination of GSK1120212 plus GSK2141795 at 0.3 and 30 mg/kg, respectively (combo). Tumor tissues were stained with markers of proliferation (Ki67), apoptosis (Cleaved Caspase 3; CC3), MAPK pathway activation (phospho-ERK, dually phospho-ERK and total ERK) and AKT pathway activation (phospho-PRAS40, total PRAS40, phospho-S6 kinase, total S6K, phospho-AKT and total AKT). Quantitative image analysis of various immune-staining is represented as a percentage of the vehicle control (100%). Data represents mean ± s.d.