N-tert-Butyl-α-phenylnitrone is a nitrone-based free radical scavenger that forms nitric oxide spin adducts. N-tert-Butyl-α-phenylnitrone inhibits the catalytic activity of COX2. N-tert-Butyl-α-phenylnitrone has potent ROS scavenging, anti-inflammatory, neuro-protection, anti-aging and anti-diabetic effects, and can penetrate the BBB (blood-brain barrier).

Physicochemical Properties

Molecular Formula	C11H15NO
Molecular Weight	177.24
Exact Mass	177.115
CAS #	3376-24-7
PubChem CID	638877
Appearance	White to off-white solid powder
Density	1.0±0.1 g/cm3
Boiling Point	283.3±23.0 °C at 760 mmHg
Melting Point	71-75ºC
Flash Point	118.5±15.4 °C
Vapour Pressure	0.0±0.6 mmHg at 25°C
Index of Refraction	1.552
LogP	1.25
Hydrogen Bond Donor Count	0
Hydrogen Bond Acceptor Count	1
Rotatable Bond Count	2
Heavy Atom Count	13
Complexity	185
Defined Atom Stereocenter Count	0
SMILES	CC(C)(C)/[N+](=C/C1=CC=CC=C1)/[O-]
InChi Key	IYSYLWYGCWTJSG-XFXZXTDPSA-N
InChi Code	InChI=1S/C11H15NO/c1-11(2,3)12(13)9-10-7-5-4-6-8-10/h4-9H,1-3H3/b12-9-
Chemical Name	N-tert-butyl-1-phenylmethanimine oxide
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

Targets	COX-2 Reactive oxygen species (ROS)
ln Vitro	The formation of intracellular ROS caused by 2,2'-azobis (2-amidinopropane) dihydrochloride (AAPH) is significantly reduced when treated with N-tert-Butyl-α-phenylnitrone (PBN) at concentrations ranging from 25 to 100 µM. Additionally, AAPH-induced cytotoxicity, matrix disintegration, and apoptosis are attenuated by N-tert-Butyl-α-phenylnitrone. N-tert-Butyl-α-phenylnitrone inhibits the activation of the ERK/MAPK pathway caused by AAPH. There is potential for studying intervertebral disc degeneration (IDD) with N-tert-Butyl-α-phenylnitrone[1].
ln Vivo	Treatment with N-tert-Butyl-α-phenylnitrone (PBN; 100 mg/kg; intraperitoneal injection; twice daily; C57Bl/6 mice) eliminates the lipid peroxidation, nitrotyrosine residue levels, and GSH depletion caused by LPS and reduces the occurrence of external malformations[2].
Animal Protocol	Animal/Disease Models: C57Bl/6 mice induced by lipopolysaccharide (LPS)[2] Doses: 100 mg/kg Route of Administration: intraperitoneal (ip) injection; twice a day (on gestational day 8) Experimental Results: Abolished LPS -induced lipid peroxidation, nitrotyrosine residues, and GSH depletion.
References	[1]. PBN Protects NP Cells From AAPH-induced Degenerative Changes by Inhibiting the ERK1/2 Pathway. Connect Tissue Res. 2020 Mar 30;1-10. [2]. Reactive Oxygen Species Contribute to Lipopolysaccharide-Induced Teratogenesis in Mice. Toxicol Sci. 2008 May;103(1):149-57. [3]. Inhibition of NF-kappaB, iNOS mRNA, COX2 mRNA, and COX Catalytic Activity by phenyl-N-tert-butylnitrone (PBN). Biochim Biophys Acta. 1998 Nov 19;1448(1):77-84. [4]. R A Floyd. Antioxidants, Oxidative Stress, and Degenerative Neurological Disorders. Proc Soc Exp Biol Med. 1999 Dec;222(3):236-45.

Solubility Data

Solubility (In Vitro)

DMSO: 100 mg/mL (564.21 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (14.11 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 25.0 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 2: ≥ 2.5 mg/mL (14.11 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 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 3: ≥ 2.5 mg/mL (14.11 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 25.0 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	5.6421 mL	28.2103 mL	56.4207 mL
	5 mM	1.1284 mL	5.6421 mL	11.2841 mL
	10 mM	0.5642 mL	2.8210 mL	5.6421 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.