Evidence for a novel pentyl radical adduct of the cyclic nitrone spin trap MDL 101,002.
3,4-Dihydro-3,3-dimethyl-isoquinoline-2-oxide (MDL 101,002) is a conformationally constrained cyclic analog of the known spin trap alpha-phenyl N-tert-butyl nitrone (PBN). Because of PBN's ability to scavenge free radicals, MDL 101,002 is currently being evaluated in stroke models as a means to ameliorate the oxidative insult associated with reperfusion injury. To augment our understanding of the radical scavenging mechanism of this potential drug, MDL 101,002 was incubated with soybean lipoxygenase in the presence of linoleic acid to study the interaction between MDL 101,002 and free radicals formed during lipid peroxidation. Analysis of the reaction mixture was performed by high performance liquid chromatography using normal phase conditions with detection by atmospheric pressure chemical ionization mass spectrometry (APCI-MS). Similar to the work by Iwahashi et al. [Arch. Biochem. Biophys., 1991, 285, 172], who studied the spin trap alpha-(4-pyridyl-1-oxide)-N-tert-butyl nitrone (4-POBN), an adduct that suggested the trapping of pentyl radicals by MDL 101,002 was observed. However, the apparent molecular ion for this adduct (246 Da) was 1 Da lower than would be predicted if a pentyl radical had simply added to MDL 101,002. In addition, the adduct exhibited significant absorbance at 304 nm, consistent with the unsaturated nitrone structure of MDL 101,002. To account for these observations, it is postulated that, after the initial capture of a pentyl radical, subsequent abstraction of a hydrogen atom by a neighboring radical occurs to regenerate a nitrone (1-pentyl analog of MDL 101,002). We present evidence for this adduct and offer a mechanism for its formation. These findings indicate that mass spectroscopic analysis of stable nitrone radical adducts may be useful in the identification of radical-dependent damage in vivo and possibly in clinical development of MDL 101,002 as an antioxidant pharmaceutical.
Dage
J L
JL
Hoechst Marion Roussel, Inc., Cincinnati, OH USA.
Ackermann
B L
BL
Barbuch
R J
RJ
Bernotas
R C
RC
Ohlweiler
D F
DF
Haegele
K D
KD
Thomas
C E
CE
D000975:Antioxidants / Q000737:chemistry; D002851:Chromatography, High Pressure Liquid; D016166:Free Radical Scavengers / Q000737:chemistry; D005609:Free Radicals / Q000737:chemistry; D007546:Isoquinolines / Q000737:chemistry*; D015227:Lipid Peroxidation; D008084:Lipoxygenase / Q000378:metabolism; D013058:Mass Spectrometry; D009589:Nitrogen Oxides / Q000737:chemistry*; D013025:Glycine max / Q000201:enzymology; D013113:Spin Labels
C091566:3,4-dihydro-3,3-dimethylisoquinoline-N-oxide; D000975:Antioxidants; D016166:Free Radical Scavengers; D005609:Free Radicals; D007546:Isoquinolines; D009589:Nitrogen Oxides; D013113:Spin Labels; D008084:Lipoxygenase