BKM-120

• Catalog No.: S2247
• CAS Number: 1202777-78-3
• M. F.: C18H21F3N6O2
• M. W.: 410.3935496
• Storage: -20°C

SYNONYMS

• IUPAC name: 5-[2,6-bis(morpholin-4-yl)pyrimidin-4-yl]-4-(trifluoromethyl)pyridin-2-amine
• IUPAC Traditional name: 5-[2,6-bis(morpholin-4-yl)pyrimidin-4-yl]-4-(trifluoromethyl)pyridin-2-amine
• Synonyms: BKM120

DATABASE IDS

• CAS Number: 1202777-78-3

PROPERTIES

• Target: PI3K
• Salt Data: Free Base
• Storage Condition: -20°C

DETAILS

Description (English)

Research Area

• Description: Solid tumours, Endometrial cancer , Breast cancer

Biological Activity

• Description: 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.
• Targets: p110α; p110β; p110δ; p110γ
• IC50: 52-99 nM; 166 nM; 116 nM; 262 nM ^[1]
• In Vitro: 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]
• In Vivo: 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]
• Clinical Trials: Current under Phase II in men with metastatic castration-resistant prostate cancer.

Protocol

• Protocol: Kinase Assay ^[1]
• 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 MaxiSorp^TM 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 [γ^33P]-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 TopCount^TM counter.
• Protocol: Cell Assay ^[1]
• 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 10^3 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.
• Protocol: Animal Study ^[1]
• 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.).

References

• References: [1] Burger MT, et al. ACS Med Chem Lett, 2011, 2 (10), 774–779.
• References: [2] Zheng Y, et al. J Mol Med (Berl), 2011 Dec 30.
• References: [3] Park E, et al. Int J Oncol, 2012, 40(4), 1259-1266.
• References: [4] Koul D, et al. Clin Cancer Res, 2012, 18(1), 184-195.

REFERENCES

• Park E, et al. Int J Oncol, 2012, 40(4), 1259-1266.
• Koul D, et al. Clin Cancer Res, 2012, 18(1), 184-195.
• Burger MT, et al. ACS Med Chem Lett, 2011, 2 (10), 774–779.
• Zheng Y, et al. J Mol Med (Berl), 2011 Dec 30.