NS-398 [also called NS 398, N-(2-cyclohexyloxy-4-nitrophenyl)methane sulfonamide] is a novel, highly potent and selective inhibitor of cyclooxygenase-2 (COX-2) that is used as a non-steroidal an-inflammatory agent with analgesic and antipyretic effects. The IC50 values for human recombinant COX-1 and -2 are 75 and 1.77 μM, respectively, which means the COX-1 activity is completely unaffected by 75 μM NS-398, whereas the COX-2 activity was concentration-dependently inhibited with the IC50 value being 1.77 μM. NS-398 inhibits proliferation and induces apoptosis in human osteosarcoma cells via downregulation of the survivin pathway. NS-398-enhanced apoptosis of esophageal carcinoma cell EC9706 by adjusting expression of survivin and caspase-3.

Physicochemical Properties

Molecular Formula	C13H18N2O5S
Molecular Weight	314.36
Exact Mass	314.093
CAS #	123653-11-2
Related CAS #	123653-11-2
PubChem CID	4553
Appearance	Light yellow to yellow solid powder
Density	1.4±0.1 g/cm3
Boiling Point	490.6±55.0 °C at 760 mmHg
Melting Point	139 °C
Flash Point	250.5±31.5 °C
Vapour Pressure	0.0±1.2 mmHg at 25°C
Index of Refraction	1.593
LogP	3.67
Hydrogen Bond Donor Count	1
Hydrogen Bond Acceptor Count	6
Rotatable Bond Count	4
Heavy Atom Count	21
Complexity	450
Defined Atom Stereocenter Count	0
SMILES	0
InChi Key	KTDZCOWXCWUPEO-UHFFFAOYSA-N
InChi Code	InChI=1S/C13H18N2O5S/c1-21(18,19)14-12-8-7-10(15(16)17)9-13(12)20-11-5-3-2-4-6-11/h7-9,11,14H,2-6H2,1H3
Chemical Name	N-(2-cyclohexyloxy-4-nitrophenyl)methanesulfonamide
Synonyms	NS-398; NS 398; NS398; N-(2-cyclohexyloxy-4-nitrophenyl)methane sulfonamide
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	With an IC50 of 3.8 μM and no effect on COX-1 at 100 μM, NS-398 is a nonsteroidal anti-inflammatory drug that specifically inhibits prostaglandin G/H synthase/cyclooxygenase (COX-2) activity. [1]. In sheep seminal vesicle microsomes, NS-398 weakly inhibits PG endoperoxide synthase activity (IC50: 11 μM)[2].
ln Vivo	Rats with an ED30 of 1.14 mg/kg showed dose-dependent inhibition of paw edema when treated with NS-398 (0.5-10 mg/kg, po), and this treatment was effective in managing adjuvant arthritis (ED30, 4.69 mg/kg). In rats, it has an antipyretic effect (ED50, 1.84 mg/kg) and a dose-dependent analgesic effect (ED50, 1.65 mg/kg). With an ED50 of 8.2 mg/kg, NS-398 suppresses the writhing response brought on by acetic acid in mice[2].
Animal Protocol	Dissolved in saline; 20 mg/kg; i.p. CD1 mice
References	[1]. NS-398, a new anti-inflammatory agent, selectively inhibits prostaglandin G/H synthase/cyclooxygenase (COX-2) activity in vitro. Prostaglandins. 1994 Jan;47(1):55-9. [2]. NS-398, a novel non-steroidal anti-inflammatory drug with potent analgesic and antipyretic effects, which causes minimal stomach lesions. Gen Pharmacol. 1993 Jan;24(1):105-10.
Additional Infomation	NS-398 is a C-nitro compound that is N-methylsulfonyl-4-nitroaniline bearing an additional cyclohexyloxy substituent at position 2. It has a role as a cyclooxygenase 2 inhibitor and an antineoplastic agent. It is a sulfonamide, an aromatic ether and a C-nitro compound. It is functionally related to a 4-nitroaniline. NS-398 is a COX-2 inhibitor. It was developed as part of the mechanistic study of the cyclooxygenases. N-(2-Cyclohexyloxy-4-nitrophenyl)methanesulfonamide has been reported in Plantago major with data available.

Solubility Data

Solubility (In Vitro)

DMSO:62 mg/mL (197.2 mM) Water:<1 mg/mL Ethanol:<1 mg/mL

Solubility (In Vivo)

Solubility in Formulation 1: 2.5 mg/mL (7.95 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 sonication. 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 2: ≥ 1.67 mg/mL (5.31 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 16.7 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 3: ≥ 1.67 mg/mL (5.31 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 16.7 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	3.1811 mL	15.9053 mL	31.8107 mL
	5 mM	0.6362 mL	3.1811 mL	6.3621 mL
	10 mM	0.3181 mL	1.5905 mL	3.1811 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.