GN25 is a specific p53-Snail binding inhibitor (antagonist) with anti-tumor effects.

Physicochemical Properties

Molecular Formula	C15H14O6S
Molecular Weight	322.33
Exact Mass	322.051
CAS #	1227401-27-5
PubChem CID	46218884
Appearance	Light yellow to orange solid powder
Density	1.4±0.1 g/cm3
Boiling Point	608.4±55.0 °C at 760 mmHg
Flash Point	321.7±31.5 °C
Vapour Pressure	0.0±1.8 mmHg at 25°C
Index of Refraction	1.622
LogP	2.19
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	7
Rotatable Bond Count	6
Heavy Atom Count	22
Complexity	500
Defined Atom Stereocenter Count	0
InChi Key	DIIRLGMLDOPLPV-UHFFFAOYSA-N
InChi Code	InChI=1S/C15H14O6S/c1-20-9-3-4-10(21-2)14-13(9)8(16)7-11(15(14)19)22-6-5-12(17)18/h3-4,7H,5-6H2,1-2H3,(H,17,18)
Chemical Name	3-(5,8-dimethoxy-1,4-dioxonaphthalen-2-yl)sulfanylpropanoic acid
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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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	GN25 (10 and 20 μM; 24 h) decreases the viability of MEF cells with a K-Ras mutation[1]. In a K-Ras-dependent manner, p53 is activated by GN25 (5 μM; 4 h)[1]. In K-Ras-mutant A549 and HCT116 cell lines, GN25 (1 -10 μM; 1-6 h) activates p53 and p21 in a dose- and time-dependent manner[1]. In p53WT/MT cells, GN25 activates wild-type p53[1]. The masking effect of p53 on CK1/GSK3b-mediated Snail phosphorylation is recovered by GN25 (0.25 μM)[1].
ln Vivo	In mice, GN25 (10 and 20 mg/kg; i.p. once weekly for 10 weeks) inhibits the growth of tumors and causes tumor regression[1].
Cell Assay	Cell Viability Assay[1] Cell Types: HCT116, A459 and MKN45 cell lines Tested Concentrations: 10 μM Incubation Duration: 24 h Experimental Results: Obviously suppressed the cell viability of HCT116 and A459 cell lines. Cell Proliferation Assay[1] Cell Types: K-Ras-transformed and N-Ras/Myc-transformed MEF cells Tested Concentrations: 10 μM and 20 μM Incubation Duration: 24 h Experimental Results: Inhibited the viability of K-Ras-transformed but not N-Ras/Myc-transformed MEF cells. Western Blot Analysis[1] Cell Types: K-Ras-MEF, N-Ras/Myc-MEF and Nontransformed MEF cells Tested Concentrations: 5 μM Incubation Duration: 4 h Experimental Results: Only induced p53 and p21 in K-Ras-MEF cells. Western Blot Analysis[1] Cell Types: K-Ras-mutated A549, HCT116 cell lines and MKN45 Tested Concentrations: 1, 5, and 10 μM Incubation Duration: 1, 3, and 6 h Experimental Results: Induced p53 and p21 in a dose- and time-dependent manner in A549 and HCT116 cells.
Animal Protocol	Animal/Disease Models: Athymic mice with A549 xenografts[1] Doses: 10 and 20 mg/kg Route of Administration: intraperitoneal (ip)injection; once a week for 10 weeks Experimental Results: Obviously regressed tumors in mice. demonstrated no significant toxicity in liver, pancreas and kidney.
References	[1]. Lee SH, Shen GN, Jung YS, Lee SJ, Chung JY, Kim HS, Xu Y, Choi Y, Lee JW, Ha NC, Song GY, Park BJ. Antitumor effect of novel small chemical inhibitors of Snail-p53 binding in K-Ras-mutated cancer cells. Oncogene. 2010 Aug 12;29(32):4576-87.

Solubility Data

Solubility (In Vitro)

DMSO : 100 mg/mL (310.24 mM)

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	3.1024 mL	15.5121 mL	31.0241 mL
	5 mM	0.6205 mL	3.1024 mL	6.2048 mL
	10 mM	0.3102 mL	1.5512 mL	3.1024 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.