Summary
We have synthesized insulins acylated by fatty acids in the ε-amino group of LysB29. Soluble preparations can be made in the usual concentration of 600 nmol/ml (100 IU/ml) at neutral pH. The time for 50% disappearance after subcutaneous injection of the corresponding TyrA14(125I)-labelled insulins in pigs correlated with the affinity for binding to albumin (r=0.97), suggesting that the mechanism of prolonged disappearance is binding to albumin in subcutis. Most protracted was LysB29-tetradecanoyl des-(B30) insulin. The time for 50% disappearance was 14.3±2.2 h, significantly longer than that of Neutral Protamine Hagedorn (NPH) insulin, 10.5±4.3 h (p<0.001), and with less inter-pig variation (p<0.001). Intravenous bolus injections of LysB29-tetradecanoyl des-(B30) human insulin showed a protracted blood glucose lowering effect compared to that of human insulin. The relative affinity of LysB29-tetradecanoyl des-(B30) insulin to the insulin receptor is 46%. In a 24-h glucose clamp study in pigs the total glucose consumptions for LysB29-tetradecanoyl des-(B30) insulin and NPH were not significantly different (p=0.88), whereas the times when 50% of the total glucose had been infused were significantly different, 7.9±1.0 h and 6.2±1.3 h, respectively (p<0.04). The glucose disposal curve caused by LysB29-tetradecanoyl des-(B30) insulin was more steady than that caused by NPH, without the pronounced peak at 3 h. Unlike the crystalline insulins, the soluble LysB29-tetradecanoyl des-(B30) insulin does not elicit invasion of macrophages at the site of injection. Thus, LysB29-tetradecanoyl des-(B30) insulin might be suitable for providing basal insulin in the treatment of diabetes mellitus.
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Abbreviations
- HI:
-
Human insulin
- HSA:
-
human serum albumin
- NN-304:
-
LysB29-tetradecanoyl des-(B30) human insulin
- NPH:
-
Neutral Protamine Hagedorn, a crystalline insulin-protamine preparation
- T50% :
-
time for 50% disappearance
- sIR:
-
soluble insulin receptor (extracellular parts)
- TBS:
-
Tris buffered saline pH 7.6
- Lys:
-
lysine
- Tyr:
-
tyrosine
- Gly:
-
glycine
- Phe:
-
phenylalanine
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Markussen, J., Havelund, S., Kurtzhals, P. et al. Soluble, fatty acid acylated insulins bind to albumin and show protracted action in pigs. Diabetologia 39, 281–288 (1996). https://doi.org/10.1007/BF00418343
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DOI: https://doi.org/10.1007/BF00418343