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Diabetologia

, Volume 39, Issue 3, pp 281–288 | Cite as

Soluble, fatty acid acylated insulins bind to albumin and show protracted action in pigs

  • J. Markussen
  • S. Havelund
  • P. Kurtzhals
  • A. S. Andersen
  • J. Halstrøm
  • E. Hasselager
  • U. D. Larsen
  • U. Ribel
  • L. Schäffer
  • K. Vad
  • I. Jonassen
Originals

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.

Keywords

Insulin analogues albumin binding prolonged action basal insulin fatty acids tetradecanoic acid myristic acid lysineB29 acylation receptor affinity 

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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Copyright information

© Springer-Verlag 1996

Authors and Affiliations

  • J. Markussen
    • 1
  • S. Havelund
    • 1
  • P. Kurtzhals
    • 1
  • A. S. Andersen
    • 1
  • J. Halstrøm
    • 1
  • E. Hasselager
    • 1
  • U. D. Larsen
    • 1
  • U. Ribel
    • 1
  • L. Schäffer
    • 1
  • K. Vad
    • 1
  • I. Jonassen
    • 1
  1. 1.Department of Insulin ResearchNovo Nordisk A/SBagsvaerdDenmark

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