DK2004683T3 - Pc5 som et faktor ix-propeptid-forarbejdende enzym - Google Patents

Pc5 som et faktor ix-propeptid-forarbejdende enzym Download PDF

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Publication number
DK2004683T3
DK2004683T3 DK07753848.6T DK07753848T DK2004683T3 DK 2004683 T3 DK2004683 T3 DK 2004683T3 DK 07753848 T DK07753848 T DK 07753848T DK 2004683 T3 DK2004683 T3 DK 2004683T3
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profix
fix
fusion protein
gly
ser
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DK07753848.6T
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Robert T Peters
Alan Bitonti
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Biogen Hemophilia Inc
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/644—Coagulation factor IXa (3.4.21.22)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/64—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue
    • C12N9/6421—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from animal tissue from mammals
    • C12N9/6424—Serine endopeptidases (3.4.21)
    • C12N9/6454—Dibasic site splicing serine proteases, e.g. kexin (3.4.21.61); furin (3.4.21.75) and other proprotein convertases
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • C12Y304/21—Serine endopeptidases (3.4.21)
    • C12Y304/21022—Coagulation factor IXa (3.4.21.22)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/30—Non-immunoglobulin-derived peptide or protein having an immunoglobulin constant or Fc region, or a fragment thereof, attached thereto

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Engineering & Computer Science (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Molecular Biology (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Claims (22)

1. Eukaryot celle omfattende et første polynukleotid, der koder for et proprotein af Faktor IX (proFIX), eller et fusionsprotein deraf, og et andet polynukleotid, der koder for en funktionel pattedyr-subtilisin/Kex2p-lignende proproteinkonvertase PC5.
2. Eukaryot celle ifølge krav 1, hvor den funktionelle PC5 omfatter en aminosyresekvens fra SEQ ID NO: 1.
3. Eukaryot celle ifølge krav 1, hvor det første polynukleotid koder for proFIX.
4. Eukaryot celle ifølge krav 1, hvor det første polynukleotid koder for et fusionsprotein af proFIX.
5. Eukaryot celle ifølge krav 4, hvor fusionsproteinet er et proFIX-FcRn- bindingspartnerfusionsprotein.
6. Eukaryot celle ifølge krav 5, hvor proFIX-FcRn-bindingspartnerfusionsproteinet er et proFIX-Fc-fusionsprotein.
7. Eukaryot celle ifølge krav 6, hvor proFIX-Fc-fusionsproteinet omfatter et human Fc- gamma.
8. Eukaryot celle ifølge krav 6, hvor cellen producerer en proFIX-Fc-homodimer.
9. Eukaryot celle ifølge krav 6, hvor cellen producerer en proFIX-Fc-monomer-dimer hybrid.
10. Eukaryot celle ifølge krav 4, hvor fusionsproteinet er et proFIX-albumin- fusionsprotein.
11. Eukaryot celle ifølge krav 4, hvor fusionsproteinet er et proFIX-transferrin-fusionsprotein.
12. Eukaryot celle ifølge krav 4, hvor fusionsproteinet omfatter en linker.
13. Eukaryot celle ifølge krav 12, hvor linkereb er GGGGSGGGGSGGGGSGGGGSGGGGSGGGGS (SEQ ID NO: 16).
14. Eukaryot celle ifølge krav 1, hvor den eukaryote celle er en pattedyrecelle.
15. Eukaryot celle ifølge krav 1, hvor den eukaryote celle er en HEK 293-celle.
16. Eukaryot celle ifølge krav 1, hvor den eukaryote celle er en CHO-celle.
17. Eukaryot celle ifølge krav 1, hvor det første polynukleotid, der koder for proFIX eller fusionsproteinet deraf, og det andet polynukleotid, der koder for den funktionelle PC5, er i en enkelt ekspressionsvektor.
18. Ekspressionsvektor omfattende en første polynukleotidsekvens, der koder for et proprotein af Faktor IX (proFIX), eller et fusionsprotein deraf, der er operativt bundet til en ekspressionskontrolsekvens, der tillader ekspression af proFIX eller fusionsproteinet deraf, og en anden polynukleotidsekvens, der koder for en funktionel pattedyr-subtilisin/Kex2p-lignende proproteinkonvertase PC5, der er operativt bundet til en ekspressionskontrolsekvens, der tillader ekspression af den funktionelle PC5.
19. Fremgangsmåde til fremstilling af et maturt Faktor IX-holdigt polypeptid ud af et proprotein af Faktor IX (proFIX), eller et fusionsprotein deraf, hvilken fremgangsmåde omfatter dyrkning af eukaryot celle ifølge krav 1 under forhold, der tillader: ekspression af både proFIX eller fusionsproteinet deraf og den funktionelle PC5 og forarbejdning af proFIX eller fusionsproteinet deraf ved den funktionelle PC5.
20. Fremgangsmåde til at øge udbyttet af et maturt Faktor IX-holdigt polypeptid fra et proprotein af Faktor IX (proFIX), eller et fusionsprotein deraf, hvilken fremgangsmåde omfatter dyrkning af den eukaryote celle ifølge krav 1 under forhold, der tillader: ekspression af både proFIX eller fusionsproteinet deraf og den funktionelle PC5, og forarbejdning af proFIX eller fusionsproteinet deraf ved den funktionelle PC5, hvor udbyttet af maturt Faktor IX-holdigt polypeptid er øget sammenlignet med udbyttet af maturt Faktor IX-holdigt polypeptid, der er fremstillet under tilsvarende forhold uden forarbejdning ved den funktionelle PC5.
21. Fremgangsmåde in vitro til fremstilling af et maturt Faktor IX-holdigt polypeptid ud fra et proprotein af Faktor IX (proFIX), et fusionsprotein deraf eller et konjugat deraf, hvilken fremgangsmåde omfatter etablering af kontakt mellem proFIX eller konjugatet deraf med en effektiv mængde af en funktionel pattedyr-subtilisin/Kex2p-lignende proproteinkonvertase PC5.
22. Fremgangsmåde ifølge 21, hvor proFIX eller konjugatet deraf er et PEGyleret proFIX.
DK07753848.6T 2006-03-24 2007-03-23 Pc5 som et faktor ix-propeptid-forarbejdende enzym DK2004683T3 (da)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US78542106P 2006-03-24 2006-03-24
PCT/US2007/007252 WO2007112005A2 (en) 2006-03-24 2007-03-23 Pc5 as a factor ix propeptide processing enzyme

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DK2004683T3 true DK2004683T3 (da) 2016-08-22

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US (3) US7566565B2 (da)
EP (2) EP2004683B1 (da)
DK (1) DK2004683T3 (da)
ES (1) ES2987558T3 (da)
HR (1) HRP20160990T1 (da)
HU (2) HUE030567T2 (da)
LT (1) LT2004683T (da)
PL (1) PL2650305T3 (da)
SI (1) SI2004683T1 (da)
TW (1) TWI428448B (da)
WO (1) WO2007112005A2 (da)

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