Chemical and alpha-chymotrypsin-mediated proteolytic degradation of insulin in bile salt-unsaturated fatty acid mixed micellar systems.
The proteolytic degradation of porcine zinc insulin by alpha-chymotrypsin was previously found to depend markedly on the state of insulin aggregation (Pharm. Res. 9:864-869, 1992). In this study, the effect of bile salt-unsaturated fatty acid mixed micelles on alpha-chymotryptic degradation of insulin was further characterized. The incorporation of linoleic acid has greatly accelerated insulin degradation with the apparent first order rate constant being linearly related to the concentration of linoleic acid. At a 10 mM linoleic acid concentration solubilized in 10 mM sodium glycocholate, the proteolytic degradation rate constant increased by 16 times, which could not be explained solely by the mechanism of insulin oligomer dissociation. Further, this effect is significantly reduced when the free carboxylic group of linoleic acid is methylated. The catalytic role of mixed micelles on chemical degradation of insulin was found to depend on the concentration of linoleic acid incorporated. When solubilized in the form of mixed micelles, linoleic acid chemically catalyzes peptide bond cleavage in a concentration-dependent manner.
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Department of Industrial and Physical Pharmacy, School of Pharmacy and Pharmacal Sciences, Purdue University, West Lafayette, Indiana 47907-1336.
Shao
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Mitra
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D000818:Animals; D001647:Bile Acids and Salts / Q000737:chemistry*; D001673:Biodegradation, Environmental / Q000187:drug effects; D002918:Chymotrypsin / Q000494:pharmacology*; D004355:Drug Stability; D007328:Insulin / Q000378:metabolism*; D019787:Linoleic Acid; D008041:Linoleic Acids / Q000737:chemistry*; D008823:Micelles; D013552:Swine
D001647:Bile Acids and Salts; D007328:Insulin; D008041:Linoleic Acids; D008823:Micelles; D019787:Linoleic Acid; D002918:Chymotrypsin