FI122725B - Derivat av zinkfingerprotein och förfaranden därav - Google Patents

Derivat av zinkfingerprotein och förfaranden därav Download PDF

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FI122725B
FI122725B FI20105336A FI20105336A FI122725B FI 122725 B FI122725 B FI 122725B FI 20105336 A FI20105336 A FI 20105336A FI 20105336 A FI20105336 A FI 20105336A FI 122725 B FI122725 B FI 122725B
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nucleotide sequence
zinc finger
sequence
variant
dna
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FI20105336A
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FI20105336A (sv
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Iii Carlos F Barbas
Joel M Gottesfeld
Peter E Wright
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Scripps Research Inst
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    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
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    • A—HUMAN NECESSITIES
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    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • A—HUMAN NECESSITIES
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    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
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    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4702—Regulators; Modulating activity
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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    • G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/502—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing non-proliferative effects
    • G—PHYSICS
    • G01—MEASURING; TESTING
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    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/569—Immunoassay; Biospecific binding assay; Materials therefor for microorganisms, e.g. protozoa, bacteria, viruses
    • G01N33/56983—Viruses
    • G01N33/56988—HIV or HTLV
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K48/00—Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/02—Fusion polypeptide containing a localisation/targetting motif containing a signal sequence
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/01—Fusion polypeptide containing a localisation/targetting motif
    • C07K2319/09—Fusion polypeptide containing a localisation/targetting motif containing a nuclear localisation signal
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/24—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a MBP (maltose binding protein)-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/40—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation
    • C07K2319/42—Fusion polypeptide containing a tag for immunodetection, or an epitope for immunisation containing a HA(hemagglutinin)-tag
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/71—Fusion polypeptide containing domain for protein-protein interaction containing domain for transcriptional activaation, e.g. VP16
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/73—Fusion polypeptide containing domain for protein-protein interaction containing coiled-coiled motif (leucine zippers)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/70—Fusion polypeptide containing domain for protein-protein interaction
    • C07K2319/735—Fusion polypeptide containing domain for protein-protein interaction containing a domain for self-assembly, e.g. a viral coat protein (includes phage display)
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/80—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor
    • C07K2319/81—Fusion polypeptide containing a DNA binding domain, e.g. Lacl or Tet-repressor containing a Zn-finger domain for DNA binding

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Claims (34)

1. Zinkfingerpolypeptidvariant som binds vid nukleotider, kanne-t e c k n a d av att den omfattar minst tvä zinkfingermoduler av den kända 5 zinkfingerpolypeptiden som binds vid nukleotider, vilken polypeptid binds vid en cellular nukleotidsekvens och som modulerar den cellulära nukleotid-sekvensens funktion, varvid (a) en modul av nämnda variant uppvisar minst en aminosyra-sekvensmodifikation; 10 (b) den cellulära nukleotidsekvensen är en andra cellular nukleo tidsekvens, som är olik än den första cellulära nukleotidsekvensen, vid vilken zinkfingerpolypeptiden som binds vid nukleotider är bunden fore modifikation av modulen; och (c) var och en aminosyrasekvens av modulen som binder den andra 15 cellulära nukleotidsekvensen omfattar tvä cystein och tvä histidin, varvid bäda cysteinerna är aminoproximala i förhällande till bäda histidinerna; och (d) peptidkopplaren sammanfogar minst tvä av modulerna.
2. Variant enligt patentkrav 1, kännetecknad av att zinkfingerpolypeptiden som binds vid nukleotider effektiverar transkriptionen av en 20 gen som ärfunktionellt kopplad till den cellulära nukleotidsekvensen.
3. Variant enligt patentkrav 1, kännetecknad av att zinkfingerpolypeptiden som binds vid nukleotider försvagar transkriptionen av ge-nen som ärfunktionellt kopplad till den cellulära nukleotidsekvensen.
4. Variant enligt nägot av patentkraven 1-3, kännetecknad av 25 att den är härledd frän zinkfingerpolypeptiden som binds vid nukleotider och som väljs frän en grupp, som bestär av zif268 och TFIIIA.
5. Variant enligt nägot av patentkraven 1-4, kännetecknad av CM ^ att den cellulära nukleotidsekvensen är DNA.
° 6. Variant enligt nägot av patentkraven 1-4, kännetecknad av att ^ 30 den cellulära nukleotidsekvensen är RNA.
7. Variant enligt nägot av patentkraven 1-6, kännetecknad av to att polypeptiden innehäller mellan zinkfingrarna ett kopplaromräde, som vä- 00 £3 sentligen bestär av aminosyrasekvensen TGEKP.
° 8. Variant enligt nägot av patentkraven 1-7, kännetecknad av ^ 35 att den cellulära nukleotidsekvensen är en strukturgens nukleotidsekvens. 96
9. Variant enligt nägot av patentkraven 1-5 eller 7, känneteck-n a d av att den cellulära nukleotidsekvensen är en promotors nukleo-tidsekvens.
10. Variant enligt patentkrav 9, k ä n n e t e c k n a d av att promo-5 torn är en onkopromotor.
11. Variant enligt patentkrav 10, k ä n n e t e c k n a d av att promo-torn är ett virus promotor.
12. Variant enligt nägot av patentkraven 1-11, kännetecknad av att den cellulära nukleotidsekvensen är ett retrovirus nukleotidsekvens.
13. Variant enligt patentkrav 12, k ä n n e t e c k n a d av att retro- viruset är ett humant T-lymfotropiskt virus (HTLV).
14. Variant enligt patentkrav 13, kännetecknad av att retro-viruset är HTLV-1 eller HTLV-2.
15. Variant enligt patentkrav 12, k ä n n e t e c k n a d av att retro- 15 viruset är ett humant immunbristvirus (HIV).
16. Variant enligt patentkrav 15, k ä n n e t e c k n a d av att retro-viruset är HIV-1 eller HIV-2.
17. Variant enligt nägot av patentkraven 1-8, kännetecknad av att den cellulära nukleotidsekvensen är en onkogens nukleotidsekvens.
18. Variant enligt nägot av patentkraven 1-8, kännetecknad av att den cellulära nukleotidsekvensen är en växts cellulära nukleotidsekvens.
19. Nukleotidsekvens, kännetecknad av att den kodar för en zinkfingerpolypeptidvariant som binds vid nukleotider enligt nägot av patentkraven 1-18.
20. Rekombinant expressionsvektor, kännetecknad av att den innehäller en nukleotidsekvens enligt patentkrav 19. o
21. Farmaceutisk komposition, kännetecknad av att den om- <M ^ fattar en terapeutiskt effektiv mängd av en zinkfingerpolypeptidvariant som ^ binds vid nukleotider enligt nägot av patentkraven 1-18 eller en terapeutiskt ™ 30 effektiv mängd av en nukleotidsekvens enligt patentkrav 19 tillsammans med | en farmaceutiskt godtagbar bärare. cd
22. Farmaceutisk komposition enligt patentkrav 21, känne- 00 tecknadavatt moduleringen är effektivering av transkriptionen av en gen £ som är funktionellt kopplad tili den cellulära nukleotidsekvensen. C\l 97
23. Farmaceutisk komposition enligt patentkrav 21, känne-t e c k n a d av att moduleringen är försvagning av transkriptionen av en gen som ärfunktionellt kopplad tili den cellulära nukleotidsekvensen.
24. Farmaceutisk komposition enligt nägot av patentkraven 21-23, 5 kännetecknad av att zinkfingerpolypeptidvarianten som binds vid nuk- leotider är en inkortad zinkerfingerdomän som binds vid nukleotider av vildtyp.
25. In vitro-förfarande för modifiering av funktionen av en cellular nukleotidsekvens som omfattar ett zinkfingermotiv som binds vid nukleotider, kännetecknat av att motivet bringas i kontakt med en effektiv mängd av 10 en zinkfingerpolypeptidvariant som binds vid nukleotider enligt nägot av patentkrav 1-18.
26. Zinkfingerpolypeptidvariant som binds vid nukleotider enligt nägot av patentkraven 1-18 eller nukleotidsekvens enligt patentkrav 19 för att användas för framställning av en farmaceutisk komposition för behandling av 15 sjukdom som kan behandlas genom att bringa en cellulär nukleotidsekvens som omfattar ett zinkfingermotiv som binds vid nukleotider i kontakt med en effektiv mängd av zinkfingerpolypeptidvarianten som binds vid nukleotider.
27. Variant eller nukleotidsekvens enligt patentkrav 26, kännetecknad av att sjukdomen är en sjukdom, som väljs frän en grupp, som 20 bestär av adenocarcinom, tjocktarmscancer, njurcellscarcinom, icke-smäcells-lungcarcinom, tunntarmscancer, matstrupscancer, psoriasis, vanligt bläsutslag, Behcets syndrom och lipidhistiocytos.
28. Förfarande enligt patentkrav 25 eller variant eller nukleotidsekvens enligt patentkrav 27, k ä n n e t e c k n a t av att zinkfingerpolypeptid- 25 varianten som binds vid nukleotider är ett avkortat zinkfingerprotein.
29. Förfarande enligt patentkrav 25 eller variant eller nukleotidit sekvens enligt nägot av patentkraven 26-28, kännetecknat av att den (M ^ cellulära nukleotidsekvensen är DNA.
° 30. Förfarande enligt patentkrav 25 eller variant eller nukleotid- ^ 30 sekvens enligt nägot av patentkraven 26-28, kännetecknat av att den | cellulära nukleotidsekvensen är RNA. cd
31. Förfarande enligt patentkrav 25 eller variant eller nukleotid- 00 £3 sekvens enligt nägot av patentkraven 26-30, kännetecknat av att den ^ cellulära nukleotidsekvensen är en strukturgens nukleotidsekvens. (M 98
32. Förfarande enligt patentkrav 25 eller variant eller nukleotid-sekvens enligt nägot av patentkraven 26-29, kännetecknat avatt den cellulära nukleotidsekvensen är en promotors nukleotidsekvens.
33. Förfarande enligt patentkrav 25 eller variant eller nukleotid-5 sekvens enligt nägot av patentkraven 26-31, kännetecknat avatt den cellulära nukleotidsekvensen är en onkogens nukleotidsekvens.
34. Förfarande enligt patentkrav 25 eller variant eller nukleotidsekvens enligt nägot av patentkraven 26-32, kännetecknat avatt den cellulära nukleotidsekvensen är en växts cellulära nukleotidsekvens. (M δ (M i CO o CO (M X en CL CD CO CO in o δ (M
FI20105336A 1994-01-18 2010-03-31 Derivat av zinkfingerprotein och förfaranden därav FI122725B (sv)

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US18311994A 1994-01-18 1994-01-18
US18311994 1994-01-18
US31260494A 1994-09-28 1994-09-28
US31260494 1994-09-28
US9500829 1995-01-18
PCT/US1995/000829 WO1995019431A1 (en) 1994-01-18 1995-01-18 Zinc finger protein derivatives and methods therefor

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