Abstract
Purpose. To study the oxidation of the methionine residue of antiflammin 2 (HDMNKVLDL, AF2) as a function of pH, buffer concentration, ionic strength, and temperature using different concentrations of hydrogen peroxide and to determine the accessibility of methionine residue to oxidation.
Methods. Reversed-phase high-performance liquid chromatography (RPHPLC) was used as the main analytical method in determining the oxidation rates of AF2. Calibration curves for AF2 and the oxidation product, methionine sulfoxide of AF2 (Met(O)-3-AF2), were constructed for each measurement using standard materials. Fast Atom Bombardment Mass Spectroscopy (FABMS) was used to characterize the product.
Results. Met(O)-3-AF2 was the only oxidation product detected at pH 3.0 to 8.0. The oxidation rates were independent of buffer concentrations, ionic strength, and pH from 3.0 to 7.0. However, there was an acceleration of the rates at basic pHs, and small amounts of degradation products other than Met(O)-3-AF2 were observed in this alkaline region.
Conclusions. Oxidation of methionine in AF2 does not cause the biological inactivation reported by other laboratories since this drug is relatively stable under neutral conditions in the absence of oxiding agent.
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REFERENCES
L. Miele, E. Cordella-Miele, A. Facchiano, and A. B. Mukherjee. Novel anti-inflammatory peptides from the region of highest similarity between uteroglobin and lipocortin I. Nature 335:726–730 (1988).
J. G. Vostal, A. B. Mukherjee, L. Miele, and N. R. Shulman. Novel peptides derived from a region of local homology between uteroglobin and lipocortin-1 inhibit platelet aggregation and secretion. Biochem. Biophys. Res. Comm. 165:27–36 (1989).
A. Ialenti, P. M. Doyle, G. N. Hardy, D. S. E. Simpkin, and M. D. Rosa. Anti-inflammatory effects of vasocortin and nonapeptide fragments of uteroglobin and lipocortin I (antiflammins). Agents and Actions 29:48–49 (1990).
P. H. Cartwright, E. Ilderton, J. M. Snowden, and H. J. Yardley. Inhibition of normal and psoriatic epidermal phospholipase A2 by picomolar concentrations of recombinant human lipocortin I. Brit. J. Dermatol. 121:155–160 (1989).
G. Camussi, C. Tetta, F. Bussolino, and C. Baglioni. Antiinflammatory peptides (antiflammins) inhibit synthesis of platelet-activating factor, neutrophil aggregation and chemotaxis, and intradermal inflammatory reactions. J. Exp. Med. 171:913–927 (1990).
S. Lloret and J. J. Moreno. In vitro effects of the anti-inflammatory peptides, antiflammins. Biochem. Pharmacol. 44:1437–1441 (1992).
A. Facchiano, E. Cordella-Miele, L. Miele, and A. B. Mukherjee. Inhibition of pancreatic phospholipase A2 activity by uteroglobin and antiflammin peptides: possible mechanism of action. Life Sci. 48:453–464 (1991).
C.-C. Chan, M. Ni, L. Miele, E. Cordella-Miele, M. Ferrick, A. B. Mukherjee, and R. B. Nussenblatt. Effects of antiflammins on endotoxin-induced uveitis in rats. Arch Ophthalmol. 109:278–281 (1991).
J. L. Wolfe, G. E. Lee, G. K. Potti, and J. F. Gallelli. Degradation of antiflammin 2 in aqueous solution. J. Pharm. Sci. 83:1762–1764 (1994).
J. van Binsbergen, A. J. Slotboom, A. J. Arsman, and G. H. de Haas. Synthetic peptide from lipocortin I has no phospholipase A2 inhibitory activity. FEBS 247:293–297 (1989).
F. Märki, J. Pfeilschifter, H. Rink, and I. Wiesenberg. ‘Antiflammins’: two nonapeptide fragments of uteroglobin and lipocortin I have no phospholipase A2-inhibitory and anti-inflammatory activity. FEBS 264:171–175 (1990).
W. C. Hope, B. J. Patal, and D. R. Bolin. Antiflammin-2 (HDMNKVLDL) dose not inhibit phospholipase A2 activities. Agents and Actions 34:77–80 (1991).
N. Brot and H. Weissbach. The biochemistry of methionine sulfoxide residues in proteins. Trends in Biochem. Sci. 7:137–139 (1982).
D. Bannard, L. Bateman, and J. L. Cuneen. Oxidation of organic sulfides. In N. Kharasch and C. Y. Meyers, The chemistry of organic sulfur compounds, Pregamon Press, New York, 1961.
T. H. Nguyen, J. Burnier, and W. Meng. The kinetics of relaxin oxidation by hydrogen peroxide. Pharm. Res. 10:1563–1571 (1993).
S. Li, T. H. Nguyen, C. Schöneich, and R. T. Borchardt. Aggregation and precipitation of human relaxin induced by metal-catalyzed oxidation. Biochem. 34:5762–5772 (1995).
S. Li, C. Schöneich, G. S. Wilson, and R. T. Borchardt. Chemical pathways of peptide degradation. V. Ascorbic acid promotes rather than inhibits the oxidation of methionine to methionine sulfoxide in small model peptides. Pharm. Res. 10:1572–1579 (1993).
C. Schöneich, F. Zhao, G. S. Wilson, and R. T. Borchardt. Ironthiolate induced oxidation of methionine to methionine sulfoxide in small model peptides. Intramolecular catalysis by histidine. Biochim. Biophy. Acta 1158:307–322 (1993).
P. R. Young and L.-S. Hsieh. Mechanism of buffer catalysis in the iodine oxidation of N-acetylmethionine methyl ester. J. Am. Chem. Soc. 104:1612–1616 (1982).
P. R. Young and L.-S. Hsieh. Mechanism of buffer catalysis in the iodine oxidation of N-acetylmethionine. J. Org. Chem. 47:1419–1423 (1982).
P. R. Young and L.-S. Hsieh. General base catalysis and evidence for a sulfurane intermediate in the iodine oxidation of methionine. J. Am. Chem. Soc. 100:7121–7122 (1978).
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Ye, J.M., Wolfe, J.L. Oxidative Degradation of Antiflammin 2. Pharm Res 13, 250–255 (1996). https://doi.org/10.1023/A:1016095131836
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DOI: https://doi.org/10.1023/A:1016095131836