Supercharge Your Innovation With Domain-Expert AI Agents!

Compositions and methods for increasing serum half-life of Fc fusion proteins

A fusion protein, half-life technology, applied in chemical instruments and methods, drug combinations, fusion polypeptides, etc., can solve problems such as half-life changes

Inactive Publication Date: 2012-11-07
ACCELERON PHARMA INC
View PDF25 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Furthermore, half-life changes caused by such modifications are unpredictable

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
  • Compositions and methods for increasing serum half-life of Fc fusion proteins
  • Compositions and methods for increasing serum half-life of Fc fusion proteins
  • Compositions and methods for increasing serum half-life of Fc fusion proteins

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0163] Embodiment 1: Expression of sugar variant TNFR2-Fc fusion protein

[0164] Applicants constructed vectors for expressing various TNFR2-Fc fusion proteins, each of which contains additional N-linked glycosylation sites. A control TNFR2-Fc fusion protein (ie, the basic or "original" unmodified form of TNFR2-Fc) had the extracellular domain of human TNFR2 fused to the human IgGl Fc domain without an inserted linker. exist figure 1 The sequence of the original TNFR2-Fc fusion protein (SEQ ID NO:5) is shown in Figure 5 The encoding nucleic acid (including the leader sequence) is shown in (SEQ ID NO:8). figure 1 The fusion protein shown is the more fully labeled TNFR2-h(1) Fc. like Figure 9 As shown, the variant designated TNFR2-h(2)Fc has an alternative linker / border sequence (SEQ ID NO: 16) and is derived from Figure 10 The nucleotide sequence shown (SEQ ID NO: 17) encodes. The protein construct was cloned into the pAID4 vector for expression in mammalian cells (...

Embodiment 2

[0173] Embodiment 2: Design of sugar variant TNFR2-Fc fusion protein

[0174] The location to introduce additional N-linked glycosylation sites was chosen by examining the co-crystal structure of the TNFR2 ectodomain with its ligand TNF (crystal coordinates are publicly available), and by applying the principles described herein. Select Changes in the Figure 7 As shown in , changes in this position are predicted to preserve the TNF antagonist activity of the protein while conferring an extended half-life. Note that the amino acid numbering is based on the native unprocessed TNFR2 amino acid sequence, eg Figure 7 shown. Altered TNFR2-Fc fusion proteins were designed with an additional N-linked carbohydrate moiety at one of the following positions: 47, 48, 62, 114, 155, 202, 203, 222, and 253. Note that native TNFR2-Fc has N-linked sugar moieties at positions 171 and 193, which means that the ratio of N-linked sugar moieties to amino acids in the heterologous domain of th...

Embodiment 3

[0175] Embodiment 3: Test TNFR2-Fc sugar variant protein

[0176] Glycovariant variants of TNFR2-h(1) Fc were tested in a series of assays to evaluate the functionality of the modified molecules and the effect of the modifications on the pharmacokinetic properties of the molecules.

[0177] For the evaluation of ligand binding in cell-free biochemical assays, a Biacore® 3000 biosensor was used. Briefly, TNFR2-h(1) Fc variants were added to a flow cell and then exposed to TNF. By evaluating the kinetic parameters (k a and k d ) to calculate the dissociation constant (K D ).

[0178] To assess ligand inhibition, a cell-based assay for TNF signaling was used essentially as described by Khabar et al. Immunol. Lett. 46 (1995): 107-110. Briefly, WEHI cells (ATCC) were cultured in the presence of TNF-α (R&D Systems, Minneapolis, Minn.) and actinomycin D. TNF-α induced apoptosis of these cells, and in A 490nm Check the cleavage rate. The presence of a TNF-[alpha] antagonist ...

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
molecular weightaaaaaaaaaa
Login to View More

Abstract

Provided herein are glycovariant Fc fusion proteins having increased serum half lives. Also provided are methods for increasing the serum half life of an Fc fusion protein by introducing one or more non-endogenous glycosylation sites.

Description

[0001] related application [0002] This application claims the benefit of US Provisional Application No. 61 / 266,095, filed December 2, 2009, and US Provisional Application No. 61 / 327,582, filed April 23, 2010. The entire teachings of the aforementioned applications are incorporated herein by reference. Background of the invention [0003] Therapeutic proteins or polypeptides in their native state or when recombinantly produced are generally unstable molecules exhibiting short-term stability or a short serum half-life. Furthermore, these molecules are often extremely unstable when formulated, especially when formulated in aqueous solutions for diagnostic and therapeutic purposes. Few practical solutions exist to prolong or improve the in vivo or in vitro stability of protein therapeutics. Many therapeutic drugs, especially peptide drugs, suffer from an inappropriate serum half-life in vivo. This necessitates administration of the therapeutic at frequent and / or higher doses,...

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(China)
IPC IPC(8): C07K19/00A61K38/16C12N15/62
CPCC07K14/7151A61K38/00C07K2319/30A61P25/16A61P25/28A61P29/00A61P31/00A61P31/04A61P31/10A61P31/12A61P33/00A61P37/06C07K19/00C12N15/62A61K38/16C07K16/00C07K2317/94C07K2319/00
Inventor J.西拉J.克诺普夫R.库马
Owner ACCELERON PHARMA INC
Features
  • R&D
  • Intellectual Property
  • Life Sciences
  • Materials
  • Tech Scout
Why Patsnap Eureka
  • Unparalleled Data Quality
  • Higher Quality Content
  • 60% Fewer Hallucinations
Social media
Patsnap Eureka Blog
Learn More

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

© 2025 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com