Ipatasertib

SKU: orb1300935

Description

Ipatasertib (GDC-0068) is a potent, ATP-competitive pan-Akt inhibitor with low nanomolar IC50 values against Akt1, Akt2, and Akt3. It induces p53-independent apoptosis by inhibiting Akt to activate FoxO3a and NF-κB, which co-transcribe PUMA, leading to Bax-mediated mitochondrial apoptosis. This mechanism has been investigated in vitro and in vivo for cancer research, particularly in tumors with PTEN deficiency or Akt hyperactivation.

Research Area

Cell Biology

Key Properties

• CAS Number: 1001264-89-6
• MW: 458.0
• Purity: >98%
• Formula: C₂₄H₃₂ClN₅O₄₂
• SMILES: C[C@H]1C2=C(N=CN=C2[C@H](O)C1)N3CCN(C([C@H](CNC(C)C)C4=CC=C(Cl)C=C4)=O)CC3
• Target: Kinase
• Solubility: Soluble in DMSO (up to 25 mg/ml) or ethanol (up to 20 mg/ml)

Bioactivity

• Target IC50:
  Akt1:5 nM|PKA:3100 nM|Akt2:18 nM|Akt3:8 nM
• In Vivo:
  METHODS: Nude mice were subcutaneously injected with HCT116 WT or PUMA−/−, and model mice were treated with Ipatasertib (GDC-0068) (30 mg/kg, oral, 15 days). Representative tumors at the end of the experiment, tumor weights, and c tumor volumes at specified time points after treatment were calculated to investigate whether PUMA-mediated apoptosis is essential for the anti-tumor activity of ipatasertib. RESULTS Ipatasertib (GDC-0068) significantly inhibited the growth of WT tumors; immunohistochemical staining showed that the expression of P-Akt was reduced in both WT and PUMA; Ki67 was significantly reduced in WT tumors, but there was no significant change in PUMA; C-Caspase3 was significantly increased in WT tumors and slightly increased in PUMA; Ipatasertib (GDC-0068) has a PUMA-dependent anti-tumor effect in colon cancer.
• In Vitro:
  METHODS: At 0, 3, 6, 12, and 24 hours after HCT116 cells were treated with Ipatasertib (GDC-0068) (1-20 μM), cell viability was detected in HCT116 by CCK-8 to study how ipatasertib affects tumor progression. RESULTS HCT116 cell viability decreased significantly with increasing dose or time, and Ipatasertib (GDC-0068) could inhibit cell proliferation in a dose- and time-dependent manner. METHODS: HCT116 cells were treated with ipatasertib (GDC-0068) (10 μM), and the expression of p53 or PUMA in HCT116 WT and p53−/− was analyzed by Western blotting; ipatasertib (GDC-0068)-induced PUMA mRNA in WT, p53−/−HCT116 and DLD1 was analyzed by real-time qPCR and normalized to the housekeeping gene β-actin. RESULTS Ipatasertib (GDC-0068) treatment increased the expression level of PUMA with increasing doses; this upregulation was observed in WT (HCT116, RKO), p53 mutants (DLD1, HT29), and p53; Ipatasertib (GDC-0068) can lead to p53-independent transcriptional activation of PUMA and inhibit cell proliferation.
• Cell Research:
  GDC-0068 is prepared in DMSO and stored, and then diluted with appropriate medium before use. The 384-well plates are seeded with 2,000 cells per well in a volume of 54 μL per well followed by incubation at 37°C under 5% CO2 overnight (~16 hours). Compounds (e.g., GDC-0068) are diluted in DMSO to generate the desired stock concentrations then added in a volume of 6 μL per well. All treatments are tested in quadruplicates. 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 IC50 is calculated from a 4-parameter curve analysis (XLfit) and is determined from a minimum of 3 experiments. For cell lines that failed to achieve an IC50, the highest concentration tested (10 μM) is listed.

Storage & Handling

• Storage: -20°C
• Expiration Date: 12 months from date of receipt.
• Disclaimer: For research use only

Alternative Names

GDC-0068; RG7440

Compound Identifiers

PubChem CID

24788740