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Synthesis, Characterization and In Vivo Efficacy of PEGylated Insulin for Oral Delivery with Complexation Hydrogels

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Abstract

Purpose

This work evaluated the feasibility of combining insulin PEGylation with pH responsive hydrogels for oral insulin delivery.

Methods

A mono-substituted PEG–insulin conjugate was synthesized and purified. The site of conjugation was determined by MALDI-TOF MS. Uptake and release of PEGylated insulin was performed in complexation hydrogels to simulate oral dosing. The bioactivity of the conjugate and PK/PD profile was measured in vivo in rats.

Results

PEGylation was confirmed to be specifically located at the amino terminus of the B-chain of insulin. Higher loading efficiency was achieved with PEGylated insulin than regular human insulin in pH responsive hydrogels. The release of PEGylated insulin was lower than that of human insulin at all pH levels considered. Full retention of bioactivity of the PEG–insulin conjugate was confirmed by intravenous dosing while subcutaneous dosing exhibited a relative hypoglycemic effect 127.8% that of human insulin.

Conclusions

Polyethylene glycol conjugated specifically to the amino terminus of the B-chain of insulin maintained the bioactivity of the protein and significantly extended the duration of the hypoglycemic effect. Used in combination with pH responsive hydrogels, PEGylated insulin has significant potential for oral delivery.

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Abbreviations

ELISA:

enzyme-linked immunosorbent assay

FPLC:

fast protein liquid chromatography

IEC:

ion exchange chromatography

MAA:

methacrylic acid

MALDI-TOF MS:

matrix-assisted laser desorption ionization time of flight mass spectrometry

PEG:

polyethylene glycol

PEGDMA:

polyethylene glycol dimethacrylate

PEGMA:

polyethylene glycol monomethacrylate monomethyl ether

P(MAA-g-EG):

poly(methacrylic acid-graft-ethylene glycol)

RP-HPLC:

reverse-phase high performance liquid chromatography

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ACKNOWLEDGMENTS

This work was supported by funding from NIH Grant # DGE-0221664. Additional funding was provided by NSF grant number R01-EB000246.

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Authors and Affiliations

  1. Department of Chemical and Biological Engineering, Drexel University, 3141 Chestnut Street, Philadelphia, Pennsylvania, 19104, USA

    Anthony D. Tuesca & Anthony M. Lowman

  2. The Artificial Pancreas Center, Department of Anesthesiology, Thomas Jefferson University, Philadelphia, Pennsylvania, 19107, USA

    Collin Reiff & Jeffrey I. Joseph

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  1. Anthony D. Tuesca
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  2. Collin Reiff
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  3. Jeffrey I. Joseph
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  4. Anthony M. Lowman
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Correspondence to Anthony M. Lowman.

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Tuesca, A.D., Reiff, C., Joseph, J.I. et al. Synthesis, Characterization and In Vivo Efficacy of PEGylated Insulin for Oral Delivery with Complexation Hydrogels. Pharm Res 26, 727–739 (2009). https://doi.org/10.1007/s11095-008-9816-8

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