PULMONARY BIOTRANSFORMATION OF INSULIN IN RAT AND RABBIT

In vitro biodegradation of insulin in rabbit and rat lung homogenates was investigated. Insulin can be sequentially metabolized into two primary fragments in rabbit lung homogenate by an aminopeptidase. The amino acid sequences of the fragments were found to be the des-Phe-Insulin(B1) (Metabolite I) and des-Phe-Val-Insulin(B1-2) (Metabolite II). However, only the former metabolite (Metabolite I) was identified in the rat lung homogenate. The k(m) and V(m) values associated with rabbit lung homogenate were 0.29 +/- 0.14 mM and 16.4 +/- 6.9 muM/hr/mg protein, respectively, whereas those for a rabbit lung preparation containing both microsomes and cytosol were 0.22 +/- 0.07 mM and 17.9 +/- 5.4 muM/hr/mg protein, respectively. The k(m) and V(m) associated with the cytosolic fraction of rabbit lung were 0.32 +/- 0.16 and 20.6 +/- 6.1 muM/hr/mg protein, respectively. The results indicate that the lung aminopeptidase may be a cytosolic enzyme. The degradation of dimeric insulin in the lung homogenate was faster than that of hexameric insulin due to the difference in collision frequency between the enzyme and insulin aggregates. The major metabolites in the lungs reportedly retain almost the same bioactivity of insulin, suggesting that the pulmonary route of insulin delivery will not adversely affect its hypoglycemic activity.