Human non-small-cell lung carcinoma wild type p53-containing H460 and A549, human non-small-cell lung carcinoma p53-null H1299, and human colon cancer HCT116 (p53^+/+ and p53^-/-) cells are used. Cells (1.5×10⁵) are plated into 6-well plates, and incubated at 37°C overnight. After treatment of Inauhzin and Nutlin 3 at the indicated concentrations for 48 h, cells are harvested, fixed in 70% ice-cold ethanol overnight at -20°C, resuspended in propidium iodide-solution (50 µg/mL PI, 0.1 mg/mL RNase A, 0.05% Tritin X-100 in PBS) for 40 min at 37°C, then analyzed for DNA content using a flow cytometer and proprietary software^[2].

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

Mice^[2]
Five-week-old female SCID mice are used. Mice are subcutaneously inoculated with 3×10⁶ HCT116^p53+/+ cells in the right flank and tumor growth is monitored with calipers. After the mean tumor volume reaches 50-100 mm³, animals are administered Inauhzin intraperitoneally (IP), Nutlin 3 orally, or vehicles (4% DMSO for Inauhzin, EtOH: Tween: 5% Glucose=5:5:90 for Nutlin 3). Tumor volume is measured every other day, and inhibition of tumor growth (T/C) is calculated on the last day of treatment.

MCE has not independently confirmed the accuracy of these methods. They are for reference only.

• [1]. Yu Z, et al. Design, synthesis and biological evaluation of sulfamide and triazole benzodiazepines as novel p53-MDM2 inhibitors. Int J Mol Sci. 2014 Sep 5;15(9):15741-53.

  [2]. Zhang Y, et al. Inauhzin and Nutlin3 synergistically activate p53 and suppress tumor growth.Cancer Biol Ther. Cancer Biol Ther. 2012 Aug;13(10):915-24.

  [3]. Supiot S, et al. Nutlin-3 radiosensitizes hypoxic prostate cancer cells independent of p53. Mol Cancer Ther. 2008 Apr;7(4):993-9.

581.49

C₃₀H₃₀Cl₂N₄O₄

548472-68-0

O=C(N1C(C2=C(C=C(C=C2)OC)OC(C)C)=N[C@H](C3=CC=C(C=C3)Cl)[C@@H]1C4=CC=C(C=C4)Cl)N5CC(NCC5)=O.[relative stereochemistry]

Room temperature in continental US; may vary elsewhere