PF-06424439

Cell lines

MCF7 cells

Preparation Method

Cell viability was determined using PrestoBlue Reagent 10×. MCF7 cells were seeded at 4.7×10³ cells/well in 96-well black polystyrene microplates and incubated overnight at 37°C to allow for cell attachment. The next day, cells were treated with 1, 10, 50, 100, 200μM of PF-06424439 for 24, 48, 72 and 96h. At each time point, cells were washed with HBSS and incubated in the dark at 37°C with 10μL of Presto Blue Cell Viability Reagent diluted in 90μL of fresh complete medium for 90min. Cell viability was measured at 560nm and 590nm using a CLARIOstar® microplate reader. Anti-proliferative activity of PF-06424439 was calculated as 50% inhibitory concentration (IC₅₀) at 72h and curve-fitting was performed by non-linear regression analysis by using GraphPad Prism 9 software.

Reaction Conditions

1, 10, 50, 100, 200μM; 24, 48, 72 , 96h

Applications

PF-06424439 can reduce the expression of key enzymes involved in lipid metabolism, namely 3-hydroxy-3-methylglutaryl-Coenzyme A reductase (HMGCR), fatty acid synthase (FASN), and DGAT2, and inhibit lipid droplet formation.

Animal models

Male Sprague–Dawley rats

Preparation Method

Male Sprague–Dawley rats were fed a high-sucrose, low-fat diet 2 days prior to the study. On the day of the study, animals were fasted for 4h prior to treatment. Immediately before dosing, blood was drawn from the lateral tail vein into a dipotassium ethylenediaminetetraacetic acid-containing tube. Following centrifugation, the plasma was transferred to a fresh tube for the determination of TG levels using a Roche Hitachi Chemistry analyzer. Animals were dosed with 0.01, 0.03, 0.1, 0.3, 1, 3, 10mg/kg PF-06424439 as a solution in 0.5% w/v methylcellulose (10mL/kg dosing volume). Vehicle-treated animals received 0.5% methylcellulose alone. After a further 2h, blood was drawn and TG determined as described above.

Dosage form

0.01, 0.03, 0.1, 0.3, 1, 3, 10mg/kg; p.o.

Applications

PF-06424439 reduces the plasma TG level in a dose-dependent manner and significantly inhibits TG synthesis.

References:
[1]Nisticò C, Pagliari F, Chiarella E, Fernandes Guerreiro J, Marafioti MG, Aversa I, Genard G, Hanley R, Garcia-Calderón D, Bond HM, Mesuraca M, Tirinato L, Spadea MF, Seco JC. Lipid Droplet Biosynthesis Impairment through DGAT2 Inhibition Sensitizes MCF7 Breast Cancer Cells to Radiation. Int J Mol Sci. 2021 Sep 18;22(18):10102.
[2]Futatsugi K, Kung DW, Orr ST, Cabral S, Hepworth D, Aspnes G, Bader S, Bian J, Boehm M, Carpino PA, Coffey SB, Dowling MS, Herr M, Jiao W, Lavergne SY, Li Q, Clark RW, Erion DM, Kou K, Lee K, Pabst BA, Perez SM, Purkal J, Jorgensen CC, Goosen TC, Gosset JR, Niosi M, Pettersen JC, Pfefferkorn JA, Ahn K, Goodwin B. Discovery and Optimization of Imidazopyridine-Based Inhibitors of Diacylglycerol Acyltransferase 2 (DGAT2). J Med Chem. 2015 Sep 24;58(18):7173-85.