HLM006474

Cas No.:	353519-63-8
Chemical Name:	7-((4-ethoxy-3-methylphenyl)(pyridin-2-ylamino)methyl)-2-methylquinolin-3-ol
Synonyms:	HLM006474; HLM-006474; HLM 006474;
SMILES:	CC1=C(O)C=C2C=CC(C(NC3=NC=CC=C3)C4=CC=C(OCC)C(C)=C4)=CC2=N1
Formula:	C25H25N3O2
M.Wt:	399.49
Purity:	>98%
Sotrage:	2 years -20°C Powder, 2 weeks 4°C in DMSO, 6 months -80°C in DMSO
Publication:	[1]. Ma Y, et al. A small-molecule E2F inhibitor blocks growth in a melanoma culture model. Cancer Res. 2008 Aug 1;68(15):6292-9. [2]. Kurtyka CA, et al. E2F inhibition synergizes with paclitaxel in lung cancer cell lines. PLoS One. 2014 May 15;9(5):e96357.

Description:	HLM006474 is a pan E2F inhibitor, which inhibits E2F4 DNA-binding with an IC50 of 29.8 µM in A375 cells.
Target:	IC50: 29.8 µM (E2F4 DNA-binding)[1]
In Vitro:	HLM006474 shows little activities against E2F4 DNA-binding in A375 cells at 10 and 20 µM, apparently inhibits E2F4 DNA-binding at 40 µM, and increasingly suppressses the effect at 60 and 80 µM concentrations. HLM006474 (40 µM) inhibits E2F4 activity via suppression of its DNA-binding activity and down regulation of its expression. HLM006474 (40 µM) also significantly induces apoptosis in A375 and 231 cell lines for 24 hours. HLM006474 dramaticly reduces cyclin D3 protein expression, and is a potent inducer of PARP cleavage[1]. HLM006474 reduces the viability of both SCLC and NSCLC lines with IC50 ranging from 15 to 75 µM. HLM006474 (60 µM) increases the expression of several known E2F-regulated genes after short treatments in H292 and H1299 cell lines. HLM006474 (20 µM) weakly synergizes with paclitaxel, but there is antagonism between HLM006474 and cisplatin and gemcitabine in H1299 cells[2]. HLM006474 leads to a decrease in the mRNA levels of MAD2. Furthmore, HLM006474 apparently suppresses the increase of Mad2 protein and pRb-S780 signal but not the level of Skp2 protein in human lung cancer A549 cellss[3].
Cell Assay:	The antiproliferative activity of compounds and their combinations is evaluated using a high-throughput CellTiter-Blue cell viability assay. For the assays, 1000 cells in 24 µL are plated in 384-well plates and incubated overnight at 37°C, 5% CO2. The next day, the drugs are diluted in media and 6 µL of these dilutions added to appropriate wells using an automated pipetting station. Four replicate wells are used for each drug concentration. The cells are incubated with the drug for 120 hours, at which time, 5 µL CellTiter-Blue reagent is added. Cell viability is assessed by the ability of the remaining treated cells to bioreduce resazurin to resorufin (579 nm Ex/584 nm Em). Fluorescence is read with a Synergy HT microplate reader. IC50s are determined using a sigmoidal equilibrium model regression using XLfit version 4.3.2 and is defined as the concentration of drug required for a 50% reduction in growth/viability. For 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium (MTS) cell viability assays, CellTiter 96 AQueous One Solution is added according to vendor instructions to cells for 2 hours following treatment with drug at the noted dose for 72 hours. All experiments are performed in triplicate and repeated at least three times[2].
References:	[1]. Ma Y, et al. A small-molecule E2F inhibitor blocks growth in a melanoma culture model. Cancer Res. 2008 Aug 1;68(15):6292-9. [2]. Kurtyka CA, et al. E2F inhibition synergizes with paclitaxel in lung cancer cell lines. PLoS One. 2014 May 15;9(5):e96357