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Research Area
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Description
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Solid tumours, Endometrial cancer , Breast cancer |
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Biological Activity
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Description
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BKM120 is a selective PI3K inhibitor of p110α, p110β, p110δ and p110γ with IC50 of 52-99 nM, 166 nM, 116 nM and 262 nM, respectively. |
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Targets
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p110α |
p110β |
p110δ |
p110γ |
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IC50 |
52-99 nM |
166 nM |
116 nM |
262 nM [1] |
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In Vitro
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BKM120 is not sensitive to Class III and Class IV PI3K's or PI4K. NVP-BKM120 shows great antiproliferation activity to PI3K deregulated cell lines including A2780, U87MG, MCF7 and DU145 with GI50 of 0.1-0.7 nM. [1] BKM120 induces multiple myeloma cells (ARP1, ARK, MM.1S, MM1.R and U266) apoptosis, which results in increased G1-phase cells and decreased S-phase cells. BKM120 induced CD138+ primary MM cell apoptosis and has significant lower cytotoxicity toward CD138? stromal cells. BKM120 exposure could cause upregulation of BimS and downregulation of XIAP. [2] BKM120 demonstrates antiproliferative activity in human gastric cancer cell lines by decreasing mTOR downstream signaling. BKM120 could increase either p-ERK or p-STAT3 in KRAS mutant gastric cancer cells. Combination with STAT3 blockade, BKM120 shows a synergism in cells harboring mutated KRAS by inducing apoptosis, but not in KRAS wild-type cells. [3] A recent study shows that BKM120 shows differential forms of cell death on the basis of p53 status of the cells with p53 wild-type cells undergoing apoptotic cell death and p53 mutant/deleted cells having a mitotic catastrophe cell death. BKM120 mediates mitotic catastrophe mainly through Aurora B kinase. [4] |
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In Vivo
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BKM120 completely inhibits pAktser473 in A2780 xenograft tumors at doses of 30, 60, or 100 mg/kg, respectively. BKM120 also shows antitumor activity against U87MG glioma model at doses of 30 and 60 mg/kg. [1] BKM120 treatment results in significantly reduced tumor volume and level of circulating human kappa chain at 5 μM/kg/day?1in ARP1 SCID mouse model, with prolonged survival. [2] |
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Clinical Trials
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Current under Phase II in men with metastatic castration-resistant prostate cancer. |
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Features
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Protocol
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Kinase Assay
[1]
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| PI3K biochemical assay (ATP depletion assay) |
BKM120 is dissolved in DMSO and directly distributed into a black 384-well plate at 1.25 μL per well. To start the reaction, 25 μL of 10 nM PI3 kinase and 5 μg/mL 1-α-phosphatidylinositol (PI) in assay buffer (10 mM Tris pH 7.5, 5 mM MgCl2, 20 mM NaCl, 1 mM DTT and 0.05% CHAPS) are added into each well followed by 25 μL of 2 μM ATP in assay buffer. The reaction is performed until approx 50% of the ATP is depleted and then stopped by the addition of 25 μL of KinaseGlo solution. The stopped reaction is incubated for 5 minutes and the remaining ATP is then detected via luminescence. |
| PI3K biochemical assay (filter binding assay) |
50 μL/well of a 1:1 mixture of 100 μL/mL L-α-phosphatidylinositol and L-α-phosphatidylserine dissolved in chloroform:ethanol (2.2:7.8) is pipetted into 96-well MaxiSorpTM plates. The solvents are evaporated at room temperature and plates are washed with Tris-buffered saline (TBS, pH7.4). PI3Kα is incubated for 60 min at room temperature in coated plates in 50 μL medium containing [γ33P]-ATP (~6 kBq/well), 0.5 μM ATP (or higher), 5 mM MgCl2, 150 mM NaCl, 25 mM Tris-HCl pH7.4, and 1% DMSO. The reaction is started by adding PI3Kα (0.4 μg/ml, <2 nm)="" and="" stopped="" by="" adding="" 50="" μl="" of="" 50="" mm="" edta.="" plates="" are="" washed="" twice="" with="" tbs="" and="" dried;="" 100="" μl/well="">TM PS is added, and bound radioactivity is determined using a TopCountTM counter. |
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Cell Assay
[1]
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| Cell Lines |
A2780 cells. |
| Concentrations |
0-6.6 μM |
| Incubation Time |
3 days. |
| Methods |
A2780 cells are cultured in DMEM supplemented with 10% FBS, L-glutamine, sodium pyruvate, and antibiotics. Cells are plated in the same medium at a density of 103 cells per well, 100 μL per well into black-walled-clear-bottom plates and incubated for 3-5 hours. BKM120 supplied in DMSO (20 mM) is diluted. The diluted BKM120 solution (2 μL), is then added to cell medium (500 μL) cell medium (concentration from 0-6.6 μM). Equal volumes of this solution (100 μL) are added to the cells in 96 well plates and incubated at 37 °C for 3 days and developed using Cell Titer Glo. Inhibition of cell proliferation is determined by luminescence read using Trilux. |
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Animal Study
[1]
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| Animal Models |
U87MG and A2780 xenografts are established in female nu/nu mice. |
| Formulation |
In 15% Captisol. |
| Doses |
~60 mg/kg. |
| Administration |
Dosed orally daily (q.d.). |
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References |
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[1] Burger MT, et al. ACS Med Chem Lett, 2011, 2 (10), 774–779.
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[2] Zheng Y, et al. J Mol Med (Berl), 2011 Dec 30.
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[3] Park E, et al. Int J Oncol, 2012, 40(4), 1259-1266.
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[4] Koul D, et al. Clin Cancer Res, 2012, 18(1), 184-195.
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