A co-formulation of supramolecularly stabilized insulin and pramlintide enhances mealtime glucagon suppression in diabetic pigs

Abstract

Treatment of patients with diabetes with insulin and pramlintide (an amylin analogue) is more effective than treatment with insulin only. However, because mixtures of insulin and pramlintide are unstable and have to be injected separately, amylin analogues are only used by 1.5% of people with diabetes needing rapid-acting insulin. Here, we show that the supramolecular modification of insulin and pramlintide with cucurbit[7]uril-conjugated polyethylene glycol improves the pharmacokinetics of the dual-hormone therapy and enhances postprandial glucagon suppression in diabetic pigs. The co-formulation is stable for over 100 h at 37 °C under continuous agitation, whereas commercial formulations of insulin analogues aggregate after 10 h under similar conditions. In diabetic rats, the administration of the stabilized co-formulation increased the area-of-overlap ratio of the pharmacokinetic curves of pramlintide and insulin from 0.4 ± 0.2 to 0.7 ± 0.1 (mean ± s.d.) for the separate administration of the hormones. The co-administration of supramolecularly stabilized insulin and pramlintide better mimics the endogenous kinetics of co-secreted insulin and amylin, and holds promise as a dual-hormone replacement therapy.

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Fig. 1: CB[7]–PEG binds to insulin and pramlintide and alters diffusion rates in formulation.
Fig. 2: Formulation with CB[7]–PEG stabilizes a co-formulation of pramlintide and Novolog or Humalog at physiological pH.
Fig. 3: Aspart and pramlintide pharmacokinetics following different administration routes in diabetic rats.
Fig. 4: Administration of aspart and pramlintide as a co-formulation in diabetic rats enhances the pharmacokinetic overlap.
Fig. 5: Lispro and pramlintide pharmacokinetics following different administration routes in diabetic pigs.
Fig. 6: Overlap between the pharmacokinetic curves of lispro and pramlintide, and glucagon suppression following the treatment of diabetic pigs with different formulations.

Data availability

All data supporting the results in this study are available within the article and its Supplementary information. The broad range of raw datasets acquired and analysed (or any subsets thereof), which would require contextual metadata for reuse, are available from the corresponding author on reasonable request.

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Acknowledgements

This work was funded in part by a NIDDK R01 (the National Institutes of Health grant no. R01DK119254), a Pilot and Feasibility funding from the Stanford Diabetes Research Center (NIH grant no. P30DK116074) and the Stanford Child Health Research Institute, as well as a Research Starter Grant from the PhRMA Foundation. C.L.M. was supported by the NSERC Postgraduate Scholarship and the Stanford Bio-X Bowes Graduate Student Fellowship. A.A.A.S. was funded by grant no. NNF18OC0030896 from the Novo Nordisk Foundation and the Stanford Bio-X Program, as well as by the Danish Council of Independent Research (grant no. DFF5054-00215). The authors thank the Stanford Animal Diagnostic Lab and the Veterinary Service Centre staff for their technical assistance.

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C.L.M., A.A.A.S. and E.A.A. designed experiments and wrote the manuscript. C.L.M., A.A.A.S., L.Z., G.A.R., L.M.S., E.C.G., A.C.Y., J.L.M., S.C., A.K.G., C.M.M., D.C. and C.S.L. performed the experiments. S.W.B. performed the pig surgeries and provided scientific input. C.L.M., A.A.A.S., G.A.R. and M.T. analysed data. D.M.M., B.A.B. and M.J.W. provided scientific input. All authors provided feedback and contributed to writing.

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Correspondence to Eric A. Appel.

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E.A.A., B.A.B., D.M.M., C.L.M. and G.A.R. are inventors on a patent filing (provisional application no. 62/804,357) describing the work reported in this manuscript.

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Maikawa, C.L., Smith, A.A.A., Zou, L. et al. A co-formulation of supramolecularly stabilized insulin and pramlintide enhances mealtime glucagon suppression in diabetic pigs. Nat Biomed Eng 4, 507–517 (2020). https://doi.org/10.1038/s41551-020-0555-4

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