Topoisomerase II inhibitor 4 (compound E17) is a potent inhibitor of topoisomerase II (Topoisomerase II). Topoisomerase II inhibitor 4 Has potential anticancer/anti-tumor effects, causing G2/M phase cell cycle arrest, inhibiting cancer/tumor cell growth/proliferation and showing cell toxicity/cytotoxicity.

Physicochemical Properties

Molecular Formula	C25H25N5O4
Molecular Weight	459.497105360031
Exact Mass	459.19
CAS #	2560590-49-8
PubChem CID	162671109
Appearance	Typically exists as solid at room temperature
LogP	3.6
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	4
Heavy Atom Count	34
Complexity	731
Defined Atom Stereocenter Count	0
InChi Key	JWQPGKCTVPZVGK-UHFFFAOYSA-N
InChi Code	InChI=1S/C25H25N5O4/c1-28-10-12-29(13-11-28)15-16-2-4-17(5-3-16)26-21-8-9-22(30(33)34)24-23(21)25(32)19-14-18(31)6-7-20(19)27-24/h2-9,14,26,31H,10-13,15H2,1H3,(H,27,32)
Chemical Name	7-hydroxy-1-[4-[(4-methylpiperazin-1-yl)methyl]anilino]-4-nitro-10H-acridin-9-one
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	Topoisomerase II inhibitor 4 (compound E17) has an IC50 value of 4.55 μM for MDA-MB-231, 6.61 μM for A549), and 2.18 μM for KG1, which indicates that it inhibits cancer cells [1]. Inducing G2/M cell cycle arrest, topoisomerase II inhibitor 4 (1 μM; 0-22 hours) is effective [2]. In CRC cells, topoisomerase II inhibitor 4 (5 μM; 4 hours) blocks topoisomerase II-mediated chromosome condensation, whereas topoisomerase II-DNA is induced by (100 μM; 0.5, 1, 2 hours). The complex builds up without degrading topoisomerase II[2]. Apoptosis and DNA damage are avoided, and topoisomerase II inhibitor 4 (1 μM; 4 hours) has anticancer action in HCT116 without raising gH2AX levels [2].
Cell Assay	Cell migration assay [2] Cell Types: HCT116 and WI-38 Cell Tested Concentrations: 0.4, 0.6, 0.8, 1.0, 1.2 μM Incubation Duration: 0, 12, 24, 48 hrs (hours) Experimental Results: Migration inhibition in HCT116 and SW480 cells. Cell viability assay[2] Cell Types: HCT116 and WI-38 Cell Tested Concentrations: 0.39, 0.78, 1.56, 3.12, 6.25, 12.5, 25 μM Incubation Duration: 72 hrs (hours) Experimental Results: Anti-proliferative effect in a dose-dependent manner.
References	[1]. Li ZY, et al. Structural optimizations and bioevaluation of N-substituted acridone derivatives as strong topoisomerase II inhibitors. Bioorg Chem. 2022 Feb. 119:105543. [2]. Chen JN, et al. E17 exerts anti-tumor activity through inhibiting topo II-mediated chromosomes condensation in CRC cells. Biochem Biophys Res Commun. 2019 May 28. 513(2):313-318.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

Solubility (In Vivo)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples. Injection Formulations (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Oral Formulations Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.1763 mL	10.8814 mL	21.7628 mL
	5 mM	0.4353 mL	2.1763 mL	4.3526 mL
	10 mM	0.2176 mL	1.0881 mL	2.1763 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.