Peptides acting as both GLP-1 receptor agonists and glucagon receptor antagonists and their pharmacological methods of use

a technology of glucagon receptor and glp-1 receptor, which is applied in the field of newly identified polypeptides, can solve the problems of limiting their effectiveness, affecting the effect of glucagon receptor, and limiting insulin secretion, so as to improve the effect of fasting glucose, mild alteration of insulin secretion, and reduced endogenous insulin secretion

Inactive Publication Date: 2006-01-05
BAYER HEALTHCARE LLC
View PDF3 Cites 80 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, gastrointestinal side effects have been reported in the literature and limit their effectiveness.
The insulin secretagogues have limitations that include a potential for inducing hypoglycemia, weight gain, and high primary and secondary failure rates.
However, GLUCOPHAGE™ has a number of side effects, including gastrointestinal disturbances and lactic acidosis.
This treatment has several drawbacks: it is an injectable, it can produce hypoglycemia, and it can cause weight gain.
The possibility of inducing hypoglycemia with insulin limits the extent that hypoglycemia can be controlled.
Problems with current treatments necessitate new therapies to treat type 2 diabetes.
However, GLP-1 is rapidly degraded and has a very short half-life in the body.
Furthermore, GLP-1 causes gut motility side effects at or near its therapeutic doses.
Therefore, GLP-1 itself has significant limitations as a therapeutic agent, and modified versions of the peptide with enhanced stability are being pursued.
However, no peptide glucagon antagonist has moved beyond preclincal development.
However, attempts to identify small molecule inhibitors of the glucagon receptor have met with limited success in vivo.
This could be a major obstacle to the development of such a therapeutic.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Peptides acting as both GLP-1 receptor agonists and glucagon receptor antagonists and their pharmacological methods of use

Examples

Experimental program
Comparison scheme
Effect test

example 1

Peptide Synthesis Methodology

[0098] The following general procedure was followed to synthesize some of polypeptides of the invention. Peptide synthesis was carried out by the FMOC / t-Butyl strategy (Peptide Synthesis Protocols (1994), Volume 35 by Michael W. Pennington & Ben M. Dunn) under continuous flow conditions using Rapp-Polymere PEG-Polystyrene resins (Rapp-Polymere, Tubingen, Germany). At the completion of synthesis, peptides were cleaved from the resin and de-protected using TFA / DTT / H2O / Triisopropyl silane (88 / 5 / 5 / 2). Peptides were precipitated from the cleavage cocktail using cold diethyl ether. The precipitate was washed three times with the cold ether and then dissolved in 5% acetic acid prior to lyophilization. Peptide identity was confirmed by reversed-phase chromatography on a YMC-Pack ODS-AQ column (YMC, Inc., Wilmington, N.C.) on a Waters ALLIANCE® system (Waters Corporation, Milford, Mass.) using water / acetonitrile with 3% TFA as a gradient from 0% to 100% acetoni...

example 2

Peptide Cloning

[0099] To establish a robust method for expressing polypeptides of the invention and mutants thereof, nucleotide sequences encoding polypeptides were cloned C-terminal to GST with a single Factor Xa recognition site separating the monomeric peptide and GST. The gene encoding the Factor Xa recognition site fused to the DNA sequence of the peptide to be produced has been synthesized by hybridizing two overlapping single-stranded DNA fragments (70-90mers) containing a BamHI or EcoRI restriction enzyme site immediately 5′ to the DNA sequence of the polynucleotide to be cloned, followed by DNA synthesis of the opposite strands via the large fragment of DNA polymerase I (Life Technologies, Inc., Gaithersburg, Md.).

[0100] The DNA sequence chosen for each polynucleotide was based on the reverse translation of the designed amino acid sequence of each peptide. In some cases, the polynucleotide encoding the peptide is generated by PCR mutagenesis (Picard et al., Nucleic Acids...

example 3

Peptide Recombinant Expression and Purification

[0103] BL21 (DE3) cells (Stratagene) transformed with a GST-peptide fusion containing plasmids were grown at 37° C. until OD600 reached 0.6 to 1.0 and induced by 1 mM IPTG (Life Technologies) for 2 hours at 37° C. Two liters of cells were spun at 7,700×g for 15 minutes, and stored at −20° C. The cell pellet was resuspended in 80 ml B-PER bacterial protein extraction reagent (Cat No. 78248, Pierce) and 1× Complete Protease Inhibitor (Roche) until the cell suspension was homogenous. The homogenous mixture was gently shaken at room temperature for 10 minutes. Lysosome and Dnase I were added to a final concentration of 200 μg / ml and 10 μg / ml to further solubilize proteins and to reduce viscosity, respectively. The mixture was incubated for an additional 5 minutes. Cellular debris was spun down at 27,000 g for 20 minutes. The supernatant was mixed with 2 mL of pre-washed Glutathione Sepharose 4B resin (Pharmacia) on a shaker overnight at 4...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention provides polypeptides that act both as an agonist of the GLP-1 receptor and an antagonist of the glucagon receptor. Such polypeptides are useful for treating individuals with type 2 diabetes or other metabolic disorders.

Description

[0001] This application is a continuation-in-part of U.S. Ser. No. 10 / 265,345, filed Oct. 3, 2002.FIELD OF THE INVENTION [0002] This invention relates to newly identified polypeptides that act both as an agonist of the GLP-1 receptor and an antagonist of the glucagon receptor and the use of such polypeptides for therapeutic purposes. More particularly, polypeptides of the present invention are useful in stimulating the release of insulin from pancreatic beta cells in a glucose-dependent manner and reducing glucagon-mediated secretion of glucose from the liver, thereby providing a treatment option for those individuals afflicted with a metabolic disorder such as diabetes, hyperglycemia, impaired fasting glucose, impaired glucose tolerance, prediabetic states, and obesity. BACKGROUND OF THE RELATED ART [0003] Diabetes is characterized by impaired insulin secretion manifesting itself among other things by an elevated blood glucose level in the diabetic patient. Underlying defects lead ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(United States)
IPC IPC(8): A61K38/17A61K38/00C07K14/605
CPCA61K38/1709C07K14/605A61K38/26
Inventor PAN, CLARKWHELAN, JAMESCLAIRMONT, KEVIN
Owner BAYER HEALTHCARE LLC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap