Summary
This paper presents a survey of the biological potencies of a variety of naturally-occurring and semi-synthetic insulin analogues and a study of the joint biological action of some of these materials with native insulin. Biological activity was tested on isolated rat fat cells using lipogenesis from glucose as the metabolic index. A brief comparison using inhibition of fat cell lipolysis was included. The results indicated: 1. Analogue potencies varied considerably (0.4–100% insulin activity). Values obtained were mainly confirmatory but included two further B1-modified materials and a tricarbamylated insulin. The results supported previous indications on the relative roles of the A1, B1, and B29 residues of insulin for hormone activity. 2. Analogue bioactivities, whether assessed by stimulation of lipogenesis or inhibition of lipolysis, were similar for the four materials tested in both systems. The response of fat cells with respect to both metabolic indices occurred over a comparable range of insulin concentrations, with half maximal effects at 30–35 pmol 1−1 insulin. 3. The presence of modified insulins appeared to alter the biological action of native insulinin vitro. Small effects of both potentiation and antagonism were identified.
Article PDF
Similar content being viewed by others
References
Brandenburg, D.: Preparation of NαAl, NɛB29 adipoyl insulin. An intramolecular cross-linked derivative of beef insulin. Hoppe Seylers Z. Physiol. Chem.353, 869–873 (1972)
Lindsay, D. G., Shall, S.: The acetylation of insulin. Biochem. J.121, 737–745 (1971)
Lindsay, D. G.: Intramolecular cross-linked insulin. FEBS Lett.21, 105–108 (1972)
Brandenburg, D., Busse, W.-D., Gattner, H-G., Zahn, H., Wollmer, A., Gliemann, J., Puls, W.: Structure-function studies with chemically modified insulins. Proceedings of the Twelfth European Peptide Symposium, p. 270–284. Amsterdam: North Holland Publishing Co. 1973
Gliemann, J., Gammeltoft, S.: The biological activity and binding affinity of modified insulins determined on isolated rat fat cells. Diabetologia10, 105–113 (1974)
Freychet, P., Roth, J., Neville, D. M.: Insulin receptors in the liver: specific binding of125I insulin to the plasma membrane and its relation to insulin bioactivity. Proc. Natl. Acad. Sci. USA68, 1833–1837 (1971)
Blundell, T. L., Dodson, C. G., Hodgkin, D. C., Mercola, D.: Insulin: The structure in the crystal and its reflection in chemical and biological properties. Adv. Protein Chem.26, 279–402 (1972)
Pullen, R. A., Lindsay, D. G., Wood, S. P., Tickle, I. J., Blundell, T. L., Wollmer, A., Krail, G., Brandenburg, D., Zahn, H., Gliemann, J., Gammeltoft, S.: Receptor-binding region of insulin. Nature259, 369–373 (1976)
Jones, R. H., Dron, D. I., Ellis, M. J., Sönksen, P. H., Brandenburg, D.: Biological properties of chemically modified insulins. 1. Biological activity of proinsulin and insulin modified at A1-glycine and B29-lysine. Diabetologia12, 601–608 (1976)
Perry, W. F., Bowen, H. P.: Factors affecting thein vitro production of non-esterified fatty acid from adipose tissue. Canad. J. Biochem. Physiol.40, 749–755 (1962)
Jungas, R. L., Ball, E. G.: The effects of insulin and epinephrine on free fatty acids and glycerol production in the presence and absence of glucose. Biochemistry2, 383–388 (1963)
Fain, J. N., Kovacev, V. P., Scow, R. D.: Anti-lipolytic effect of insulin in isolated fat cells of the rat. Endocrinology78, 773–778 (1966)
Renner, R., Hepp, K. D.: Evidence for different membrane receptors at the fat cell mediating glucose transport and antilipolysis. Diabetologia10, 384 (1975)
Koelle, G. B.: The pharmacological basis of therapeutics, 4th ed., pp. 442–465. New York: Macmillan Company 1970
Brandenburg, D., Gattner, H.-G., Wollmer, A.: Darstellung und Eigenschaften von Acetylderivaten des Rinderinsulins. I. Hoppe Seylers Z. Physiol. Chem.353, 599–617 (1972)
Lindsay, D. G., Loge, P., Losert, W., Shall, S.: Carbamyl and methylthiocarbamyl insulins. Biochim. Biophys. Acta263, 658–665 (1972)
Geiger, R., Geisen, K., Summ, H.-D., Langer, D.: [A 1-D-Alanin] insulin. Hoppe Seylers Z. Physiol. Chem.356, 1635–1649 (1975)
Teetz, V., Eckert, H. G., Geiger, R.: [Bl-3,5-diiodo-L-tyrosine] insulin. Proc. 14th European Peptide Symposium Belgium, 1976. Editions de l'Universite de Bruxelles (in press)
Sodeyez, J. C., Sodoyez-Goffaux, F., Goff, M. M., Zimmerman, A. E., Arquilla, E. R.: [127I] or carrier-free [125I] monoiodoinsulin: Preparation, physical, immunological and biological properties and susceptibility to “insulinase” degradation. J. Biol. Chem.250, 4268–4277 (1975)
Emdin, S. O., Gammeltoft, S., Gliemann, J.: The biological activity on rat fat cells of insulin from the Atlantic hagfish. Diabetologia11, 340 (1975)
Gliemann, J.: Assay of insulin-like activity by the isolated fat cell method. I. Factors affecting the crystalline response to insulin. Diabetologia3, 382–388 (1967)
Cohen, P. P.: Monometric techniques, 3rd ed., p. 149. Minneapolis: Burgess Publ. Co. 1957
Moody, A. J., Stan, M. A., Stan, M., Gliemann, J.: A simple free fat cell bioassay for insulin-like activity. Horm. Metab. Res.6, 12–16 (1975)
Eggstein, M., Kreutz, F. H.: Eine neue Bestimmung der Neutralsette in Blutserum und Gewebe. Klin. Wochenschr.44, 262–267 (1966)
Finney, D. J.: Statistical methods in biological assay. 2nd ed. London: Charles Griffin & Co. Ltd. 1964
McArthur, J. W., Ulfelder, H., Finney, D. J.: A flexible computer programme for the composite analysis of symmetrical and asymmetrical biologic assays of parallel-line type. J. Pharmacol. Exp. Ther.153, 573–580 (1966)
Darby, S. C., Ellis, M. J.: A test for synergism between two drugs. Appl. Statist.25, 296–299 (1976)
Loewe, S.: Antagonisms and antagonists. Pharmacol. Rev.9, 237–242 (1957)
Ellis, M. J., Jones, R. H., Thomas, J. H., Geiger, R., Teetz, V., Sönksen, P. H.: B1-3,5-diiodotyrosine insulin: A valid tracer for insulin. Diabetologia13, 257–261 (1977)
Henrichs, H. R., Buche, Th., Hofbauer, Kerp, L.: In vitro studies on receptor binding and biological activity of porcine insulin with chemical modifications in position A1. Diabetologia12, 397 (1976)
Gliemann, J., Sørensen, H. H.: Assay of insulin-like activity by the isolated fat cell method: IV. The biological activity of proinsulin. Diabetologia6, 499–504 (1970)
Zimmerman, A. E., Kells, D. I. C., Yip, C. C.: Physical and biological properties of guinea-pig insulin. Biochem. Biophys. Res. Commun.46, 2127–2133 (1972)
Stoll, R. W., Ensinck, J. W., Williams, R. H.: Immunologic and biologic activities of the heterogeneous components of insulin. Diabetes18, 392–396 (1969)
Weitzel, G. Stratling, W.-M., Hahn, J., Martini, O.: Insulin vom Schleimfisch. Hoppe Seylers Z. Physiol. Chem.348, 525–532 (1967)
Hollenberg, M. D.: Action of insulin analogues on cultured human fibroblasts reflects biological potency. Life Sci.18, 521–528 (1976)
Lavis, V. R., Williams, R. H.: Comparison of the lipolytic effects of insulin and proinsulin on isolated fat cells. Diabetes24, 238–239 (1975)
Sönksen, P. H., Tompkins, C. V. T., Srivastava, M. C., Nabarro, J. D. N.: A comparative study on the metabolism of human insulin and porcine proinsulin in man. Clin. Sci. Mol. Med.45, 633–654 (1973)
Rudman, D., Garcia, A. L., Del Rio, A., Akgun, S.: Effects on mammalian adipose tissue of fragments of bovine insulin and of certain synthetic peptides. Biochemistry7, 1864–1874 (1968)
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Ellis, M.J., Darby, S.C., Jones, R.H. et al. In vitro bioactivity of insulin analogues: Lipogenic and anti-lipolytic potency and their interaction with the effect of native insulin. Diabetologia 15, 403–410 (1978). https://doi.org/10.1007/BF01219650
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF01219650