Topoisomerase I inhibitor 2 (ZML-8) is a selective inhibitor of DNA topoisomerase I (Top1). Topoisomerase I inhibitor 2 inhibits Top1 activity, causing DNA damage. Topoisomerase I inhibitor 2 can also block the G2/M phase, cause apoptosis, and Has potential anticancer/anti-tumor effects.

Physicochemical Properties

Molecular Weight	293.316604852676
Exact Mass	293.105
CAS #	2588211-44-1
PubChem CID	164886579
Appearance	Typically exists as solid at room temperature
LogP	3.3
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	4
Rotatable Bond Count	2
Heavy Atom Count	22
Complexity	391
Defined Atom Stereocenter Count	0
InChi Key	UUDRXJXFLKSENM-UHFFFAOYSA-N
InChi Code	InChI=1S/C18H15NO3/c1-20-17-8-11-7-13-12-5-3-4-6-16(12)22-10-15(13)19-14(11)9-18(17)21-2/h3-9H,10H2,1-2H3
Chemical Name	9,10-dimethoxy-6H-chromeno[3,4-b]quinoline
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 I inhibitor 2 (ZML-8) (24 hours) selectively inhibits HepG2 compared to the normal human liver cell line L-02, with an IC50 value of 0.58 μM. The selectivity index (SI) value is 55.70%[1]. Reduced anti-apoptotic expression of Bcl-2 and enhanced caspase-dependent apoptosis are two ways that topoisomerase I inhibitor 2 (1.25, 2.5 μM, 48 hours) reduces the growth of tumor cells [1]. DNA damage occurs due to DNA supercoiling caused by Topoisomerase I inhibitor 2 (1.25, 2.5 μM, 48 hours) which also lowers Top1 specific activity [1].
Cell Assay	Cell cycle analysis[1] Cell Types: HepG2 Cell Tested Concentrations: 2.5 μM Incubation Duration: 24 hrs (hours) Experimental Results: Arrests the cell cycle in the G2/M phase in a dose-dependent manner. Cell proliferation experiment [1] Cell Types: HepG2 Cell Tested Concentrations: 0.625, 1.25, 2.5 μM Incubation Duration: 48 hrs (hours) Experimental Results: Induced cell apoptosis in a dose-dependent manner, inhibited tumor cell proliferation, and the apoptosis rates were 65.0% and 77.6 respectively % , respectively. Western Blot Analysis [1] Cell Types: HepG2 cells Tested Concentrations: 0.625, 1.25, 2.5 μM Incubation Duration: 48 hrs (hours) Experimental Results: At 2.5 μM, the expression of Bcl-2 anti-apoptotic protein Dramatically diminished, and the expression of pro-apoptotic protein Bax increased. No, there's still p53. and also Dramatically enhanced caspase-dependent apoptosis and activated Cleaved caspase-3 in a dose-dependent manner.
References	[1]. Design and synthesis of Aza-boeravinone derivatives as potential novel topoisomerase I inhibitors. Bioorg Chem. 2022 May. 122:105747.

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	3.4092 mL	17.0462 mL	34.0925 mL
	5 mM	0.6818 mL	3.4092 mL	6.8185 mL
	10 mM	0.3409 mL	1.7046 mL	3.4092 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.