NSC-658497 is a potent SOS1 inhibitor. NSC-658497 binds to SOS1, competitively inhibits SOS1-Ras interaction, and inhibits SOS1-GEF activity in a dose-dependent fashion. NSC-658497 demonstrated dose-dependent efficacy in inhibiting Ras, downstream signaling activities, and associated cell growth/proliferation.

Physicochemical Properties

Molecular Formula	C20H10N2O6S2
Molecular Weight	438.42
Exact Mass	437.998
CAS #	909197-38-2
PubChem CID	54608884
Appearance	Typically exists as solid at room temperature
LogP	3.942
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	8
Rotatable Bond Count	2
Heavy Atom Count	30
Complexity	869
Defined Atom Stereocenter Count	0
SMILES	S1C(N(C(C1=CC1C=CC(=CC=1)[N+](=O)[O-])=O)C=C1C(=O)OC2C=CC=CC=2C1=O)=S
InChi Key	DJFUFNNDOAUACG-FUVGAYRCSA-N
InChi Code	InChI=1S/C20H10N2O6S2/c23-17-13-3-1-2-4-15(13)28-19(25)14(17)10-21-18(24)16(30-20(21)29)9-11-5-7-12(8-6-11)22(26)27/h1-10H/b14-10-,16-9-
Chemical Name	(3Z)-3-[[(5Z)-5-[(4-nitrophenyl)methylidene]-4-oxo-2-sulfanylidene-1,3-thiazolidin-3-yl]methylidene]chromene-2,4-dione
Synonyms	NSC658497; NSC 658497; NSC-658497
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	NSC-658497 (0–20 μM; 2 hours; NIH/3T3 cells) suppresses EGFR activation, but not EGF (50 ng/mL)–stimulated Ras in a dose-dependent manner [1]. NSC-658497 (0-100 μM; 2 hours; NIH/3T3 cells) concurrently suppresses EGF activation, Ras downstream targets ERK1/2, and AKT in a dose-dependent manner[1]. These findings are supported by the fact that NSC-658497 dose-dependently suppresses Ras signaling that is mediated by the overexpression of an active SOS1 mutant (W729L), which was first discovered in human embryonic kidney cells to cause Noonan syndrome [1]. NSC-658497 dose-dependently inhibits 50 nM SOS1-cat-mediated GDP/GTP nucleotide exchange at 2 μM H-Ras (aa. 1-166) (IC50=15.4 μM) in the BODIPYFL-GDP dissociation assay[1]. The proliferation of PC-3 and DU-145 cells is inhibited by NSC-658497 (0-60 μM; 3 days) [1]. Ras-GTP activity and downstream p-ERK1/2 and p-Akt activities are dose-dependently inhibited by NSC-658497 (0-60 μM; 2 hours; PC-3 and DU-145 cells) [1].
Cell Assay	Cell proliferation assay [1] Cell Types: PC-3 and DU-145 Cell Tested Concentrations: 0-60 μM Incubation Duration: 3 days Experimental Results: Dose-dependent inhibition of PC-3 and DU-145 cell proliferation. Western Blot Analysis[1] Cell Types: PC-3 and DU-145 Cell Tested Concentrations: 0-60 μM Incubation Duration: 2 hrs (hours) Experimental Results: Dose-dependent inhibition of Ras-GTP activity and downstream p-ERK1/2 and p-Akt activities .
References	[1]. Rational design of small molecule inhibitors targeting the Ras GEF, SOS1. Chem Biol. 2014;21(12):1618-1628.

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	2.2809 mL	11.4046 mL	22.8092 mL
	5 mM	0.4562 mL	2.2809 mL	4.5618 mL
	10 mM	0.2281 mL	1.1405 mL	2.2809 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.