Pharmaceutical Research

, Volume 31, Issue 12, pp 3404–3414 | Cite as

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

  • Payal Agarwal
  • Pulkit Khatri
  • Blasé Billack
  • Woon-Kai Low
  • Jun Shao
Research Paper

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.

KEY WORDS

GLP-1 L. lactis normal flora oral protein delivery type-2 diabetes 

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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Payal Agarwal
    • 1
  • Pulkit Khatri
    • 2
  • Blasé Billack
    • 2
  • Woon-Kai Low
    • 2
  • Jun Shao
    • 2
  1. 1.School of PharmacyNotre Dame of Maryland UniversityBaltimoreUSA
  2. 2.Department of Pharmaceutical Sciences, College of Pharmacy and Health SciencesSt. John’s UniversityQueensUSA

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