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Oral Delivery of Glucagon Like Peptide-1 by a Recombinant Lactococcus lactis

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Abstract

Purpose

To develop a live oral delivery system of Glucagon like peptide-1 (GLP-1), for the treatment of Type-2 Diabetes.

Methods

LL-pUBGLP-1, a recombinant Lactococcus lactis (L. lactis)) transformed with a plasmid vector encoding GLP-1 cDNA was constructed and was used as a delivery system. Secretion of rGLP-1 from LL-pUBGLP-1 was characterized by ELISA. The bioactivity of the rGLP-1 was examined for its insulinotropic activity on HIT-T15 cells. Transport of rGLP-1 across MDCK cell monolayer when delivered by LL-pUBGLP-1 was studied. The therapeutic effect of LL-pUBGLP-1 after oral administration was investigated in ZDF rats.

Results

DNA sequencing and ELISA confirmed the successful construction of the LL-pUBGLP-1 and secretion of the active form of rGLP-1. In vitro insulinotropic studies demonstrated that LL-pUBGLP-1 could significantly (p < 0.05) stimulate HIT-T15 cells to secrete insulin as compared to the controls. When delivered by LL-pUBGLP-1, the GLP-1 transport rate across the MDCK cell monolayer was increased by eight times (p < 0.01) as compared to the free solution form. Oral administration of LL-pUBGLP-1 in ZDF rats resulted in a significant decrease (10–20%, p < 0.05) in blood glucose levels during 2–11 h post dosing and a significant increase in insulin AUC0-11h (2.5 times, p < 0.01) as compared to the free solution.

Conclusion

The present study demonstrates that L. lactis when genetically modified with a recombinant plasmid can be used for the oral delivery of GLP-1.

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Abbreviations

GLP-1:

Glucagon like peptide-1

L. lactis :

Lactococcus lactis sub sp. lactis

LL-pUB1000:

Lactococcus lactis transformed with pUB1000 plasmid

LL-pUBGLP-1:

Lactococcus lactis transformed with pUBGLP-1 plasmid

M17G:

M17 growth media supplemented with 5% glucose

pUB1000:

Expression host plasmid vector

pUBGLP-1:

Recombinant plasmid with GLP-1 c-DNA

rGLP-1:

recombinant GLP-1 secreted extracellularly by LL-pUBGLP-1

T2DM:

Type 2 Diabetes mellitus

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ACKNOWLEDGMENTS AND DISCLOSURES

We would like to thank Dr. H. F. Jenkinson and Dr. J. Brittan (Department of Oral and Dental Science, University of Bristol, Bristol, UK) for providing us with the L. lactis strain with pUB1000 plasmid and also for their valuable assistance. The animal work was supported by the Seed grant/Venture Capital Fund from St. John’s University, NY.

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Authors and Affiliations

  1. School of Pharmacy, Notre Dame of Maryland University, Baltimore, Maryland, 21210, USA

    Payal Agarwal

  2. Department of Pharmaceutical Sciences, College of Pharmacy and Health Sciences, St. John’s University, 8000 Utopia Parkway, Queens, New York, 11439, USA

    Pulkit Khatri, Blasé Billack, Woon-Kai Low & Jun Shao

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  1. Payal Agarwal
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  2. Pulkit Khatri
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  4. Woon-Kai Low
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  5. Jun Shao
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Correspondence to Jun Shao.

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Agarwal, P., Khatri, P., Billack, B. et al. Oral Delivery of Glucagon Like Peptide-1 by a Recombinant Lactococcus lactis . Pharm Res 31, 3404–3414 (2014). https://doi.org/10.1007/s11095-014-1430-3

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