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Figure 3. Metabolic activity of insulin glargine and its metabolites in rat adipocytes.

To compare the metabolic activity of glargine and its metabolites insulin-stimulated lipid synthesis in isolated primary rat adipocytes was analyzed by incorporation of [3- 3 H]glucose into toluene-extractable lipids and subsequent measurement of radioactivity by liquid scintillation counting. The adipocytes were treated with increasing concentrations of insulin, analogs, metabolites or IGF-1 for 90 min at 37°C in the presence of [3- 3 H]glucose. Each point represents the mean ± SEM of five different adipocyte preparations with activity measurements done in duplicate.

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The relative metabolic activities of human insulin > insulin glargine  =  glargine metabolites correlate well with their binding affinities and autophosphorylation of IR-A and IR-B. This is compatible with previous studies demonstrating that ligand affinity and activation (i.e. autophosphorylation) of the IR are the key events for initiation of downstream signaling to the metabolic effector systems of insulin and its analogs Saint Laurent Pony Hair amp; Suede Court Classic Sneakers in Animal Print ORpywdsc
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The maintenance of binding, autophosphorylation and metabolic activities of insulin analogs with modified carboxy-terminus confirm the previous findings that amino acid modifications at the insulin B-chain beyond position B25, in particular the B26-B30 region, do not play a critical role in its recognition by both isoforms of the IR YUUL YIE Womens Medallion Slingback Pump H8EcY6
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and thus in metabolic signaling in insulin target cells [6] , [43] . The slightly lower binding, autophosphorylation and metabolic activities of M1 relative to those of human insulin are in agreement with the previous suggestion that the minor structural change caused by loss of the salt bridge between Asn A21 and Arg B22 leads to a (small) reduction in IR affinity Alexander McQueen McQueen Leather Runner Sneakers D2ObcW9SG
. In consequence, the slight reduction in metabolic activity of insulin glargine vs. human insulin derives predominantly from altered structural features at the carboxy-terminus of the B-chain and rather than from the amino acid substitution at A21.

Taken together, the present data clearly demonstrate that the major metabolites of insulin glargine detected in rats, dogs, and humans [9] , [10] are metabolically active. Since a ratio of 50∶50 for parent compound and M1/M2 was reported in tissue samples derived from the site of injection in humans [10] and in vitro metabolism of insulin glargine resulted into 46% to 98% appearance of M1 [11] , it is reasonable to assume that the metabolites of insulin glargine will contribute to the blood glucose lowering profile observed after administration of insulin glargine, dependent on the extent of their generation at the subcutaneous injection site and in plasma in the course of carboxypeptidase action [11] .

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