Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Stabilized insulin-like growth factor polypeptides

a growth factor and polypeptide technology, applied in the direction of peptide/protein ingredients, extracellular fluid disorder, metabolism disorder, etc., can solve the problems that igf-1 and igf-2 appear to be poor drug candidates, and achieve the effect of avoiding or reducing this cleavag

Inactive Publication Date: 2010-07-08
NOVARTIS AG
View PDF10 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

The invention is based on the discovery that a precursor protein called IGF-1 can be modified to prevent the cleavage of a specific peptide called E-peptide, which is important for the stability and function of the protein. By mutating or deleting certain amino acids in the precursor protein, the E-peptide can be reduced or eliminated, resulting in a more stable and effective pharmaceutical. The modified precursor protein can also have increased or decreased affinity for certain proteins, making it more effective for treating certain diseases. The invention also includes a method for treating muscle-related diseases, diabetes, and neural cell death by administering the modified polypeptide. The invention also includes veterinary methods for using the modified polypeptide for enhancing growth, efficiency of feed conversion, and improving neonatal health in animals.

Problems solved by technology

Both IGF-1 and IGF-2 appear to be poor drug candidates, since these proteins are quickly degraded by endogenous proteases in the serum of patients.

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
  • Stabilized insulin-like growth factor polypeptides
  • Stabilized insulin-like growth factor polypeptides
  • Stabilized insulin-like growth factor polypeptides

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0111]A DNA expression vector encoding the hIGF-1-Ea precursor polypeptide containing the following modifications was constructed: deletion of G1, deletion of P2, and deletion of E3; mutation of R37 to A; and deletion of R71 and deletion of S72. These mutations are sometimes referred to as “3mut” throughout the present disclosure. This results in the following secreted protein sequence:

(SEQ ID NO: 8)tlcgaelvdalqfvcgdrgfyfnkptgygsssraapqtgivdeccfrscdlrrlemycaplkpaksavraqrhtdmpktqkevhlknasrgsagnknyrm

[0112]Cos7 cells (available from ATCC) were maintained in DMEM containing 10% fetal bovine serum, 100 U / ml penicillin, and 100 μg / ml streptomycin and plated at a density of 1×106 cells per 10-cm plate. These cell cultures were transfected with 8 μg of expression plasmid using Fugene (Roche) according to manufacturer's instructions. Twenty-four hours post-transfection, cells were washed once and cultured in serum-free medium for 48 hours. Supernatants were collected and stored at −80° C.

[01...

example 2

[0116]A DNA expression vector encoding the hIGF-1-Eb precursor polypeptide containing the following mutations was constructed: deletion of G1, deletion of P2, and deletion of E3; mutation of R37 to A; and deletion of R71 and deletion of S72 (i.e., the “3mut”). This results in the following secreted protein sequence:

(SEQ ID NO: 9)tlcgaelvdalqfvcgdrgfyfnkptgygsssarapqtgivdeccfrscdlrrlemycaplkpaksavraqrhtdmpktqkyqppstnkntksqrrkgwpkthpggeqkegteaslqirgkkkeqrreigsrnaecrgkkgk

[0117]The polypeptide was assayed in accordance with the procedures described in Example 1 above. FIG. 1B and use of densitometry indicated that the ratio of uncleaved to cleaved IGF-1 was about 1:9, while the ratio for hIGF-1Eb3mut was about 1:1, showing that these modifications result in a stabilized polypeptide. FIG. 2B indicates that the hIGF-1Eb3mut was able to activate the IGF-1R cellular pathway to a similar extent as the long-R3-IGF-1 positive control reagent and the recombinant IGF-1. In addition, the data in ...

example 3

[0118]A DNA expression vector encoding the hIGF-1-Ec precursor polypeptide containing the following mutations was constructed: deletion of G1, deletion of P2, and deletion of E3; mutation of R37 to A; and deletion of R71 and deletion of S72 (i.e., the “3mut”). This results in the following secreted protein sequence:

(SEQ ID NO: 10)tlcgaelvdalqfvcgdrgfyfnkptgygsssrapqtgivdeccfrscdlrrlemycaplkpaksavraqrhtdmpktqkyqppstnkntksqrrkgstfeerk

[0119]The polypeptide was assayed in accordance with the procedures described in Example 1 above. FIG. 1C and use of densitometry indicated that the ratio of uncleaved to cleaved IGF-1 was about 1:5, while the ratio for hIGF-1Ec3mut was about 1:0.96, showing that these modifications result in a stabilized polypeptide. FIG. 2D indicates that the hIGF-1Ec3mut was able to activate the IGF-1R cellular pathway to a similar extent as the long-R3-IGF-1 positive control reagent and the recombinant IGF-1. In addition, the data in FIG. 5 directly shows that hIGF-1E...

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

PropertyMeasurementUnit
body weightaaaaaaaaaa
densityaaaaaaaaaa
Eaaaaaaaaaaa
Login to View More

Abstract

The invention relates to stabilized polypeptides having an IGF-1 or IGF-2 sequence and an E-peptide sequence, where the natural physiological cleavage of the E-peptide from the IGF is prevented.

Description

BACKGROUND OF THE INVENTION[0001]Insulin-like growth factors (IGFs) are part of a complex system that cells use to communicate with their physiologic environment. This complex system (often referred to as the insulin-like growth factor axis) consists of two cell-surface receptors (IGF-1R and IGF-2R), two ligands (IGF-1 and IGF-2), a family of six high-affinity IGF-binding proteins (IGFBP 1-6), and associated IGFBP degrading enzymes (proteases). This system is important not only for the regulation of normal physiology but also for a number of pathological states (Glass, Nat Cell Biol 5:87-90, 2003).[0002]The IGF axis has been shown to play roles in the promotion of cell proliferation and the inhibition of cell death (apoptosis). IGF-1 is mainly secreted by the liver as a result of stimulation by human growth hormone (hGH). Almost every cell in the human body is affected by IGF-1, especially cells in muscles, cartilage, bones, liver, kidney, nerves, skin and lungs. In addition to the ...

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): A61K38/16C07K14/00A61P43/00
CPCA61K38/30C07K14/475C07K14/65A61P11/00A61P17/02A61P19/00A61P19/08A61P19/10A61P21/00A61P25/00A61P25/28A61P27/02A61P3/10A61P43/00A61P5/48A61P7/06
Inventor GLASS, DAVID JONATHAN
Owner NOVARTIS AG
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com