HLM006474 is a broad spectrum (a wide range) E2F inhibitor that can suppress E2F4 DAN binding in A375 cells with IC50 of 29.8 µM.

Physicochemical Properties

Molecular Formula	C25H25N3O2
Molecular Weight	399.484905958176
Exact Mass	399.195
CAS #	353519-63-8
PubChem CID	2895500
Appearance	White to off-white solid powder
LogP	5.625
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	30
Complexity	533
Defined Atom Stereocenter Count	0
SMILES	OC(C1=NC(C)=CC=C1C=C2)=C2C(NC3=NC=CC=C3)C(C=C4C)=CC=C4OCC
InChi Key	CYNZBLNMIJNBSF-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H25N3O2/c1-4-30-21-13-11-19(15-16(21)2)23(28-22-7-5-6-14-26-22)20-12-10-18-9-8-17(3)27-24(18)25(20)29/h5-15,23,29H,4H2,1-3H3,(H,26,28)
Chemical Name	7-[(4-ethoxy-3-methylphenyl)-(pyridin-2-ylamino)methyl]-2-methylquinolin-8-ol
HS Tariff Code	2934.99.9001
Storage	Powder-20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Shipping Condition	Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Biological Activity

ln Vitro

At 10 and 20 µM, HLM006474 has little effect on E2F4 DNA binding in A375 cells. At 40 µM, it significantly inhibits E2F4 DNA binding, and at 60 and 80 µM, it gradually reduces this effect. By inhibiting its DNA-binding activity and downregulating its expression, HLM006474 (40 µM) suppresses E2F4 activity. In A375 and 231 cell lines, HLM006474 (40 µM) also markedly induced apoptosis over the course of 24 hours. HLM006474 is a strong inducer of PARP cleavage and significantly lowers the expression of the cyclin D3 protein [1]. HLM006474, whose IC50 ranges from 15 to 75 µM, decreases the viability of SCLC and NSCLC cell lines. In H292 and H1299 cell lines, HLM006474 (60 µM) temporarily increases the expression of several known E2F-regulated genes. In H1299 cells, HLM006474 (20 µM) exhibits weak synergistic effects with paclitaxel, but antagonistic effects with cisplatin and gemcitabine [2]. HLM006474 causes the levels of MAD2 mRNA to drop. Furthermore, in human lung cancer A549 cells, HLM006474 significantly reduced the rise in Mad2 protein and pRb-S780 signal, but not the level of Skp2 protein [3].

References

[1]. A small-molecule E2F inhibitor blocks growth in a melanoma culture model. Cancer Res. 2008 Aug 1;68(15):6292-9. [2]. E2F inhibition synergizes with paclitaxel in lung cancer cell lines. PLoS One. 2014 May 15;9(5):e96357.

Solubility Data

Solubility (In Vitro)

DMSO : ~16.67 mg/mL (~41.73 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.26 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: 2.5 mg/mL (6.26 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. Solubility in Formulation 3: ≥ 2.5 mg/mL (6.26 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.5033 mL	12.5163 mL	25.0325 mL
	5 mM	0.5007 mL	2.5033 mL	5.0065 mL
	10 mM	0.2503 mL	1.2516 mL	2.5033 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.