GSK2879552

LSD1 enzyme assay

LSD1 activity was measured using a horseradish peroxidase (HRP) coupled assay with amplex red as an electron donor. The formation of product over time was measured using fluorescence intensity, Ex 531 nm and Em 595 nm, in a PerkinElmer EnVision plate reader. Final assay conditions were: 5 nM LSD1, 2.5 μM H3K4me2 peptide, 50 mM HEPES pH 7, 1 U/mL of HRP, 1 mM CHAPS, 0.03% dBSA and 10 μM amplex red.

Cell lines

SCLC cell lines

Preparation method

The solubility of this compound in DMSO is > 12.8 mg/mL. General tips for obtaining a higher concentration: Please warm the tube at 37 °C for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below - 20 °C for several months.

Reacting condition

0 ~ 5000 nM; 10 days

Applications

At the 4th day after treatment, GSK2879552 started to significantly inhibit the growth of NCI-H1417 cells in a dose-dependent manner. In a 6-day proliferation assay, the maximal growth inhibition on NCI-H1417 cells was largely below 100%, which implied the effect was predominantly cytostatic. Moreover, GSK2879552 did not cause any cytotoxic response to SCLC cell lines.

Animal models

Mice bearing NCI-H526 or NCI-H1417 xenografts

Dosage form

1.5 mg/kg; p.o.

Applications

In mice bearing NCI-H526 or NCI-H1417 xenografts, GSK2879552 significantly inhibited tumor growth with the TGI values of 57% and 83%, respectively. Meanwhile, GSK2879552 did not cause thrombocytopenia at the indicated dose. On the other hand, the results of immunohistochemistry showed that there were 98 percent of SCLC tumors with a very high expression of LSD1, implying the inhibition effect of GSK2879552 on tumor growth might be exerted by targeting LSD1.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1]. Mohammad HP, Smitheman KN, Kamat CD, Soong D, Federowicz KE, Van Aller GS, Schneck JL, Carson JD, Liu Y, Butticello M, Bonnette WG, Gorman SA, Degenhardt Y, Bai Y, McCabe MT, Pappalardi MB, Kasparec J, Tian X, McNulty KC, Rouse M, McDevitt P, Ho T, Crouthamel M, Hart TK, Concha NO, McHugh CF, Miller WH, Dhanak D, Tummino PJ, Carpenter CL, Johnson NW, Hann CL, Kruger RG. A DNA Hypomethylation Signature Predicts Antitumor Activity of LSD1 Inhibitors in SCLC. Cancer Cell. 2015 Jul 13;28(1):57-69.