DE656950T1 - Tat-derivate transport polypeptide. - Google Patents

Tat-derivate transport polypeptide.

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Publication number
DE656950T1
DE656950T1 DE0656950T DE93920231T DE656950T1 DE 656950 T1 DE656950 T1 DE 656950T1 DE 0656950 T DE0656950 T DE 0656950T DE 93920231 T DE93920231 T DE 93920231T DE 656950 T1 DE656950 T1 DE 656950T1
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Prior art keywords
seq
amino acids
protein
fusion protein
cargo molecule
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Pending
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DE0656950T
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Inventor
James Barsoum
Stephen Fawell
R Pepinsky
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Biogen Inc
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Biogen Inc
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Publication of DE656950T1 publication Critical patent/DE656950T1/de
Pending legal-status Critical Current

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    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
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    • C12N2710/00011—Details
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    • C12N2740/16311—Human Immunodeficiency Virus, HIV concerning HIV regulatory proteins
    • C12N2740/16322—New viral proteins or individual genes, new structural or functional aspects of known viral proteins or genes

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Claims (34)

EP 93920231.3-2105 BIOGEN, INC. u.Z.: H 713 EP g Patentansprüche Übersetzung nach Art. 67 EPÜ
1. Fusionsprotein, bestehend aus einer carboxyterminalen Frachtmoleküleinheit und einer aminoterminalen Trans-
2Q porteinheit, wobei
(a) die Transporteinheit gekennzeichnet ist durch:
(i) die Gegenwart der Aminosäuren 49-57 des HIV-
tat-Proteins,
(ii) die Abwesenheit der Aminosäuren 22-36 des HIV-tat-Proteins und
(iii) die Abwesenheit der Aminosäuren 73-86 des HIV-
tat-Proteins; und
(b) die Frachtmoleküleinheitt nach der Transporteinheitabhängigen intrazellulären Bereitstellung eine signifikante biologische Aktivität behält.
2. Fusionsprotein nach Anspruch 1, wobei die Frachtmoleküleinheit ausgewählt ist aus therapeutischen Molekülen, prophylaktischen Molekülen und diagnostischen Molekülen.
3. Fusionsprotein, bestehend aus einer carboxyterminalen
Frachtmoleküleinheit und einer aminoterminalen Transporteinheit, wobei die Frachtmoleküleinheit aus einem menschlichen Papillomavirus E2-Repressor besteht, der
„Φ seine biologische Aktivität nach der Bereitstellung in der Zielzelle behält, und die Transporteinheit ausgewählt ist aus:
(a) Aminosäuren 47-58 des HIV-tat-Proteins
(SEQ ID NO:47),
__ (b) Aminosäuren 47-72 des HIV-tat-Proteins
(SEQ ID N0:48),
(c) Aminosäuren 38-72 des HIV-tat-Proteins
(SEQ ID NO:49), und
s (d) Aminosäuren 38-58 des HIV-tat-Proteins (SEQ ID NO:50).
4. Fusionsprotein nach Anspruch 3, wobei der Transporteinheit ein aminoterminales Methionin vorausgeht.
5. Fusionsprotein nach einem der Ansprüche 1 bis 4, wobei die Frachtmoleküleinheit aus den Aminosäuren 245-365 des menschlichen Papillomavirus E2-Proteins (SEQ ID NO:51) besteht.
6. Fusionsprotein JB106 (SEQ ID NO:38).
7. Fusionsprotein JB117 (SEQ ID NO:59). 15
8. Fusionsprotein JB118 (SEQ ID NO:60).
9. Fusionsprotein JB122 (SEQ ID NO:63).
10. Fusxonsprotein, bestehend aus einer carboxyterminalen
Frachtmoleküleinheit und einer aminoterminalen Transporteinheit, wobei die Frachtmoleküleinheit aus einem Rinder-Papillomavirus E2-Repressor besteht, der seine biologische Aktivität nach Bereitstellung in einer Zielzelle behält, und die Transporteinheit ausgewählt ist aus:
(a) Aminosäuren 47-62 des HIV-tat-Proteins
(SEQ ID NO:52) und
(b) Aminosäuren 38-62 des HIV-tat-Proteins (SEQ ID NO:53).
11. Fusionsprotein nach Anspruch 10, wobei der Transporteinheit ein aminoterminales Methionin vorausgeht.
;
12. Fusionsprotein nach einem der Ansprüche 1, 2, 10 oder 11, wobei die Frachtmoleküleinheit ein E2-Repressor ist, der aus den Aminosäuren 250-410 des Rinder-Papillomavi-
rus E2-Proteins (SEQ ID NO:56) besteht. 5
13. Fusionsprotein JB119 (SEQ ID NO:61).
14. Fusionsprotein JB120 (SEQ ID NO:62).
15. Kovalent verknüpftes chemisches Konjugat, bestehend aus einer Transportpolypeptid-Einheit und einer Frachtmoleküleinheit, wobei:
(a) die Transportpolypeptid-Einheit des Konjugats gekennzeichnet ist durch:
(i) die Gegenwart der Aminosäuren 49-57 des HIV-
tat-Proteins,
(ii) die Abwesenheit der Aminosäuren 22-36 des HIV-
tat-Proteins und
(iii) die Abwesenheit der Aminosäuren 73-86 des HIV-tat-Proteins, und
(b) die Frachtmoleküleinheit des Konjugats nach der Transporteinheit-abhängigen intrazellulären Bereitstellung eine signifikante biologische Aktivität behält.
16. Kovalent verknüpftes chemisches Konjugat nach Anspruch
15, wobei die Transportpolypeptid-Einheit aus den Aminosäuren 37-72 des HIV-tat-Proteins (SEQ ID NO:2) besteht.
17. Kovalent verknüpftes chemisches Konjugat nach Anspruch
16, wobei die Frachtmoleküleinheit ausgewählt ist aus:
(a) Aminosäuren 245-365 des menschlichen Papillomavirus E2-Proteins (SEQ ID-NO:51); und
(b) Aminosäuren 245-3 65 des menschlichen Papillomavirus E2-Proteins, wobei die Aminosäuren 300 und 309 zu Cystein geändert wurden (SEQ ID NO:55).
18. Kovalent verknüpftes chemisches Konjugat, bestehend aus einer Transporteinheit und einer Frachtmoleküleinheit, wobei das Transportpolypeptid aus den Aminosäuren 37-72 des HIV-tat-Proteins (SEQ ID NO:2) besteht und die Frachtmoleküleinheit ausgewählt ist aus:
(a) Aminosäuren 245-365 des menschlichen Papillomavirus E2-Proteins (SEQ ID NO:51) und
(b) Aminosäuren 245-365 des menschlichen Papillomavirus E2-Proteins, wobei die Aminosäuren 3 00 und 309 zu Cystein geändert wurden (SEQ ID NO:55).
19. Fusionsprotein, bestehend aus einer carboxyterminalen Frachtmoleküleinheit und einer aminoterminalen Transporteinheit, wobei die Frachtmoleküleinheit aus den Ami-
nosäuren 43-412 des HSV-VP16-Proteins besteht und die Transporteinheit aus den Aminosäuren 47-58 des HIV-tat-Proteins besteht.
20. Fusionsprotein nach Anspruch 19, wobei der Transporteinheit ein aminoterminales Methionin vorausgeht.
21. Kovalent verknüpftes chemisches Konjugat, bestehend aus einer Transportpolypeptid-Einheit und einer Frachtmoleküleinheit, wobei die Transportpolypeptid-Einheit aus
den Aminosäuren 37-72 des HIV-tat-Proteins (SEQ ID N0:2) besteht und die Frachtmoleküleinheit eine doppelsträngige DNA ist, ausgewählt aus:
(a) Oligonucleotid NFl (SEQ ID NO:43) angelagert an Oli-
gonucleotid NF2 (SEQ ID N0:44), und
(b) Oligonucleotid NF3 (SEQ ID NO:45) angelagert an Oligonucleotid NF4 (SEQ ID NO:46).
22. Verwendung eines Fusionsproteins nach einem der Ansprüche 1 bis 14, 19 oder 20 für die intrazelluläre Bereit-Stellung eines Frachtmoleküls.
23. Verwendung eines !covalent verknüpften chemischen Konjugats nach einem der Ansprüche 15 bis 17 oder 21 für die intrazelluläre Bereitstellung eines Frachtmoleküls.
24. Arzneimittel, umfassend eine pharmazeutisch wirksame Menge eines Fusionsproteins nach einem der Ansprüche 1 bis 14.
25. Arzneimittel, umfassend eine pharmazeutisch wirksame Menge eines Fusionsproteins nach Anspruch 19 oder 20.
26. Arzneimittel, umfassend eine pharmazeutisch wirksame Menge eines kovalent verknüpften chemischen Konjugats
nach einem der Ansprüche 15 bis 18 oder 21.
15
27. DNA-Molekül, umfassend eine Nucleotidsequenz, die ein Fusionsprotein codiert, ausgewählt aus:
(a) JB106 (SEQ ID NO:38),
(b) JB117 (SEQ ID NO:59),
(c) JB118 (SEQ ID NO:60),
(d) JB119 (SEQ ID NO:61),
(e) JB120 (SEQ ID NO:62), und
(f) JB122 (SEQ ID NO:63).
28. DNA-Molekül, umfassend eine Nucleotidsequenz, die das Fusionsprotein tat-VP16R.GF (SEQ ID NO:58) codiert.
29. DNA-Molekül nach Anspruch 27, wobei die das Fusionsprotein codierende Nucleotidsequenz funktionell mit Expres-
° sionskontrollsequenzen verknüpft ist.
30. DNA-Molekül nach Anspruch 28, wobei die das Fusionsprotein codierende Nucleotidsequenz funktionell mit Expressionskontroll Sequenzen verknüpft ist.
31. Einzelliger Wirt, transformiert mit einem DNA-Molekül nach Anspruch 29.
32. Einzelliger Wirt, transformiert mit einem DNA-Molekül nach Anspruch. 30.
33. Verfahren zur Herstellung eines Fusionsproteins, ausgewählt aus:
(a) JB106 (SEQ IDNO.-38),
(b) JB117 (SEQ ID NO:59), (C) JB118 (SEQ IDNO:60),
(d) JB119 (SEQ ID NO:61),
(e) JB120 (SEQ ID NO:62), und
(f) JB122 (SEQ IDNO:63),
wobei das Verfahren folgende Schritte umfaßt:
(a) Züchtung eines transformierten einzelligen Wirts
nach Anspruch 31 und
(b) Gewinnung des Fusionsproteins aus der Kultur.
34. Verfahren zur Herstellung eines Fusionsproteins, das aus den Aminosäuren 47-58 des HIV-tat-Proteins gefolgt von den Aminosäuren 43-412 des HSV-VP16-Proteins besteht, wobei das Verfahren folgende Schritte umfaßt:
(a) Züchtung eines transformierten einzelligen Wirts nach Anspruch 32 und
(b) Gewinnung des Fusionsproteins aus der Kultur. 25
DE0656950T 1992-08-21 1993-08-19 Tat-derivate transport polypeptide. Pending DE656950T1 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
US93437592A 1992-08-21 1992-08-21
PCT/US1993/007833 WO1994004686A1 (en) 1992-08-21 1993-08-19 Tat-derived transport polypeptides

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DE656950T1 true DE656950T1 (de) 1996-03-14

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DE69321962T Expired - Lifetime DE69321962T2 (de) 1992-08-21 1993-08-19 Von tat abgeleitete transportpolypeptide
DE0656950T Pending DE656950T1 (de) 1992-08-21 1993-08-19 Tat-derivate transport polypeptide.

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DE69321962T Expired - Lifetime DE69321962T2 (de) 1992-08-21 1993-08-19 Von tat abgeleitete transportpolypeptide

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EP (3) EP0656950B1 (de)
JP (2) JP2702285B2 (de)
KR (1) KR0153027B1 (de)
AT (1) ATE173016T1 (de)
AU (1) AU667244B2 (de)
CA (1) CA2135642C (de)
DE (2) DE69321962T2 (de)
DK (1) DK0656950T3 (de)
ES (1) ES2123062T3 (de)
FI (1) FI120495B (de)
HK (1) HK1012678A1 (de)
NO (1) NO316761B1 (de)
NZ (1) NZ255831A (de)
WO (1) WO1994004686A1 (de)

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