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

Clotting factor-Fc chimeric proteins to treat hemophilia

Inactive Publication Date: 2005-07-07
BIOVERATIV THERAPEUTICS INC
View PDF70 Cites 139 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0014] The invention relates to an improved chimeric protein for treating hemostatic disorders. The invention provides a chimeric protein to treat a hemostatic disorder that can be administered parenterally or

Problems solved by technology

Traditional therapies, however, have many associated problems.
Currently clotting factors must be administered parenterally in order to attain effective doses because to date non-invasive methods have not been successful in attaining therapeutic levels.
Problems associated with parenteral administration include injection site occlusion, pain and infection.
Use of an in-line catheter increases the risk of all of these events.
Specific problems associated with the parenteral administration of clotting factors to infants and small children include central venous access.
Additionally, parenteral administration of clotting factors runs the risk of precipitating a bleeding episode.
These problems are particularly relevant when the patient is undergoing regular prophylactic administration of a clotting factor.

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
  • Clotting factor-Fc chimeric proteins to treat hemophilia
  • Clotting factor-Fc chimeric proteins to treat hemophilia
  • Clotting factor-Fc chimeric proteins to treat hemophilia

Examples

Experimental program
Comparison scheme
Effect test

example 1

Cloning of pCDNA 3.1-Flag-Fc

[0129] The sequence for the FLAG peptide (Asp-Tyr-Lys-Asp-Asp-Asp-Asp-Lys)(SEQ ID NO: 23), a common affinity tag used to identify or purify proteins, was cloned into the pcDNA 3.1-Fc plasmid, which contains the mouse IgK signal sequence followed by the Fc fragment of human IgG1 (amino acids 221-447, EU numbering). The construct was created by overlapping PCR using the following primers:

FlagFc-F1:5′- GCTGGCTAGCCACCATGGA -3′(SEQ ID NO: 10)FlagFc-R1:5′- CTTGTCATCGTCGTCCTTGTAGTCGTCA(SEQ ID NO: 11)CCAGTGGAACCTGGAAC -3′FlagFc-F2:5′- GACTACAAGG ACGACGATGA(SEQ ID NO: 12)CAAGGACAAAACTCACACAT GCCCACCGTG CCCAGCTCCGGAACTCC -3′FlagFc-R2:5′- TAGTGGATCCTCATTTACCCG -3′(SEQ ID NO: 13)

[0130] The pcDNA 3.1-Fc template was then added to two separate PCR reactions containing 50 pmol each of the primer pairs FlagFc-F1 / R1 or FlagFc-F2 / R2 in a 50 μl reaction using Pfu Ultra DNA polymerase (Stratagene, Calif.) according to manufacturer's standard protocol in a MJ Thermocycler ...

example 2

Cloning of PACE Construct

[0131] The coding sequence for human PACE (paired basic amino acid cleaving enzyme), an endoprotease, was obtained by RT-PCR. The following primers were used:

(SEQ ID NO: 15)PACE-F1:5′- GGTAAGCTTGCCATGGAGCTGAGGCCCTGGTTGC -3′(SEQ ID NO: 16)PACE-R1:5′- GTTTTCAATCTCTAGGACCCACTCGCC -3′(SEQ ID NO: 17)PACE-F2:5′- GCCAGGCCACATGACTACTCCGC -3′(SEQ ID NO: 18)PACE-R2:5′- GGTGAATTCTCACTCAGGCAGGTGTGAGGGCAGC -3′

[0132] The PACE-F1 primer adds a HindIII site to the 5′ end of the PACE sequence beginning with 3 nucleotides before the start codon, while the PACE-R2 primer adds a stop codon after amino acid 715, which occurs at the end of the extracellular domain of PACE, as well as adding an EcoRI site to the 3′ end of the stop codon. The PACE-R1 and -F2 primers anneal on the 3′ and 5′ sides of an internal BamHI site, respectively. Two RT-PCR reactions were then set up using 25 pmol each of the primer pairs of PACE-F1 / R1 or PACE-F2 / R2 with 20 ng of adult human liver RNA (Clo...

example 3

Cloning of Fc-Factor VII construct

[0133] The coding sequence for Factor VII, was obtained by RT-PCR from human fetal liver RNA (Clontech, Palo Alto, Calif.). The cloned region is comprised of the cDNA sequence from bp 36 to bp 1430 terminating just before the stop codon. A SbfI site was introduced on the N-terminus. A BspEI site was introduced on the C-terminus. The construct was cloned by PCR using the primers:

(SEQ ID NO: 19)Down-5′ GCTACCTGCAGGCCACCATGGTCTCCCAGGCCCTCAGGstream:3′(SEQ ID NO: 20)Upstream:5′ CAGTTCCGGAGCTGGGCACGGCGGGCACGTGTGAGTTTTGTCGGGAAAT GG 3′

and the following conditions: 95° C. for 5 minutes followed by 30 cycles of 95° C. for 30 seconds, 55° C. for 30 seconds, 72° C. for 1 minute and 45 seconds, and a final extension cycle of 72° C. for 10 minutes.

[0134] The fragment was digested SbfI-BspE I and inserted into pED.dC-Fc a plasmid encoding for the Fc fragment of an IgG1.

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
Fractionaaaaaaaaaa
Dimensionless propertyaaaaaaaaaa
Disorderaaaaaaaaaa
Login to View More

Abstract

The invention relates to a chimeric protein comprising at least one clotting factor and at least a portion of an immunoglobulin constant region. The invention relates to a method of treating a hemostatic disorder comprising administering a therapeutically effective amount of a chimeric protein wherein the chimeric protein comprises at least one clotting factor and at least a portion of an immunoglobulin constant region.

Description

[0001] This application claims priority to U.S. Provisional Appln. No. 60 / 468,837, filed on May 6, 2003, which is incorporated by reference.FIELD OF THE INVENTION [0002] The invention relates generally to the field of therapeutics for hemostatic disorders. More specifically, the invention relates to a chimeric protein for the treatment of a hemostatic disorder. BACKGROUND OF THE INVENTION [0003] Hemostatic disorders are characterized by uncontrolled bleeding resulting from the inability or reduced ability to form fibrin clots. Hemostatic disorders can result from a genetic defect or can be acquired as a result of an unrelated medical condition (e.g., cancer and related chemotherapy, autoimmune disease) (Kasper 2000, Hemophilia 6(Supp):13; Cohen et al. 1996, Bailiere's Clinical Hematology 9(2):331). Typically, hemostatic disorders result from the deficiency of a specific blood clotting factor. Classic examples of hemostatic disorders include hemophilia A, which results from a deficie...

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
IPC IPC(8): A61K39/395A61K47/48C07K16/46C12NC12N9/64C12P21/08
CPCA61K47/4843A61K47/48507C07K2319/30C12N9/6437C12N9/644C12N9/6454A61K45/06C12Y304/21022C07K16/18A61K38/36A61K2039/60C07K14/745A61K38/4846C12Y304/21021A61K47/6815A61K47/6835A61P7/04
Inventor RIVERA, DANIELPETERS, ROBERTBITONTI, ALAN
Owner BIOVERATIV THERAPEUTICS INC
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