Molecular Weight (MW)

394.12

Formula

C13H20Cl4N2O3

CAS No.

685898-44-6

Storage

-20℃ for 3 years in powder form

-80℃ for 2 years in solvent

Solubility (In vitro)

DMSO: 78 mg/mL (197.9 mM)

Water: 78 mg/mL (197.9 mM)

Ethanol: N/A

Solubility (In vivo)

Chemical Name: (S)-4-(2-amino-2-carboxyethyl)-N,N-bis(2-chloroethyl)aniline oxide dihydrochloride

InChi Key: GIGCDIVNDFQKRA-LTCKWSDVSA-N

InChi Code: InChI=1S/C13H18Cl2N2O3.2ClH/c14-5-7-17(20,8-6-15)11-3-1-10(2-4-11)9-12(16)13(18)19;;/h1-4,12H,5-9,16H2,(H,18,19);2*1H/t12-;;/m0../s1

SMILES Code: ClCC[N+](CCCl)([O-])C1=CC=C(C[C@H](N)C(O)=O)C=C1.[H]Cl.[H]Cl

Synonyms

In Vitro

In vitro activity: PC3 and DU 145 cells express HIF-1α protein are treated with PX-478 for 20 hr under normoxia. PC3 cells are more sensitive to PX-478 as compared with DU 145 cells. Densitometric analysis shows that the IC50 for HIF-1α inhibition for PC3 cells under normoxic condition is 20-25 μM, whereas the IC50 for HIF-1α inhibition for the DU 145 cells is 40-50 μM. PC3 and DU 145 cells are treated with different concentrations of PX-478 (10, 20, 30, 40, 50, and 60 μM) for 18-20 hr under normoxia or hypoxia. Under normoxia, PC3 cells are more sensitive to PX-478 than DU 145 cells. IC50 for clonogenic survival (n=3) is 17 μM for PC3 cells and 35 μM for DU 145 cells. When cells are treated with the drug under hypoxic condition for 18 hr, the IC50 is 16 μM for PC3 cells and 22 μM for DU 145 cells. Thus DU 145 cells are more sensitive to PX-478 under hypoxic condition

Cell Assay: To determine the effect of PX-478 in combination with radiation, cells are treated with PX-478 for 24 hr under normoxic condition, irradiated and plated after 1 hr. Colonies are stained with crystal violet after 12 days and the colonies of>50 cells are counted. For combination treatments, net survival is calculated by correcting the toxicity of PX-478 alone. Enhancement factor (EF) is calculated by dividing the dose of radiation required to reduce plating efficiency to 10% when cells are treated with radiation alone by the dose of radiation required to reduce plating efficiency to 10% when cells are treated with PX-478 and radiation

In Vivo

In HT-29 human colon cancer xenografts, PX-478 suppresses HIF-1alpha levels and inhibits the expression of HIF-1 target genes including vascular endothelial growth factor and the glucose transporter-1. In addition, PX-478 (100 or 120 mg/kg i.p.) also shows antitumor activity including cures against several established human tumor xenografts that is related to the levels of HIF-1α. In high-fat-diet mice, PX-478 causes reduced fibrosis and fewer inflammatory infiltrates in their adipose tissues

Animal model

Mice bearing MCF-7 human breast cancer, HT-29 colon cancer, PC-3 prostate cancer, DU-145 prostate cancer, OvCar-3 ovarian cancer, A-549 non-small cell lung cancer, SHP-77 small cell lung cancer, and Caki-1 renal cancer, Panc-1, MiaPaCa, or BxPC-3 pancreatic cancer xenografts.

Formulation & Dosage

Formulation 1: Dissolved in 0.9% NaCl; 60, 80, 100, 120, 200 mg/kg; i.p. injection; or 25mg/kg, p.o. [Mol Cancer Ther. 2004 Mar;3(3):233-44];

Formulation 2: Dissolved in PBS solution; 100 mg/kg; i.p. injection [Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):E338-47.].

References

Mol Cancer Ther. 2004 Mar;3(3):233-44; Mol Cancer Ther. 2008 Jan;7(1):90-100; Int J Cancer. 2008 Nov 15;123(10):2430-7; Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):E338-47..

Western blot analysis of HIF-1α in PC3 and DU 145 cells. Int J Cancer. 2008 Nov 15;123(10):2430-7.

Clonogenic cell survival of PC3 and DU 145 cells treated with PX-478 for 20 hr under normoxic (N) and 18 hr under hypoxic (H) condition. Int J Cancer.2008 Nov 15;123(10):2430-7.

Analysis of γH2AX following treatment with PX-478 and radiation. Int J Cancer. 2008 Nov 15;123(10):2430-7.