DK1234031T3 - B7-h1, et hidtil ukendt immunregulerende molekyle - Google Patents

B7-h1, et hidtil ukendt immunregulerende molekyle Download PDF

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DK1234031T3
DK1234031T3 DK00983821.0T DK00983821T DK1234031T3 DK 1234031 T3 DK1234031 T3 DK 1234031T3 DK 00983821 T DK00983821 T DK 00983821T DK 1234031 T3 DK1234031 T3 DK 1234031T3
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Lieping Chen
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Claims (26)

1. Isoleret polypeptid, hvor polypeptidet består af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1 eller SEQ ID NO: 3.
2. Polypeptid ifølge krav 1 til anvendelse som et medikament.
3. Polypeptid til anvendelse ifølge krav 2, hvor medikamentet er et medikament til administration til et pattedyr, især et pattedyr, som mistænkes for at have en immundefektsygdom, en inflammatorisk tilstand eller en autoimmunsygdom.
4. Nukleinsyre, der koder for polypeptidet ifølge krav 1, hvor nukleinsyren er til anvendelse som et medikament.
5. Rekombinant celle, som er en dattercelle af en celle opnået fra et pattedyr, der er blevet transficeret eller transformeret ex vivo med en nukleinsyre, der koder for polypeptidet ifølge krav 1, således at cellen udtrykker polypeptidet, til anvendelse som et medikament, hvor cellen opnået fra pattedyret ikke er en human embryocelle, og hvor transfektionen eller transformationen med nukleinsyren ikke er en proces til modificering af den genetiske identitet af kimlinjen hos et menneske.
6. Rekombinant celle, som er en dattercelle af en celle opnået fra et pattedyr, der er blevet transficeret eller transformeret ex vivo med et DNA, der omfatter en polypeptidkodende nukleinsyre, hvor nukleinsyren under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3, hvor cellen udtrykker polypeptidet, der kodes af nukleinsyren, og hvor den rekombinante celle er til anvendelse som et medikament til behandling af en immundefektsygdom, en inflammatorisk tilstand eller en autoimmunsygdom hos pattedyret, hvor cellen opnået fra pattedyret ikke er en human embryocelle, og hvor transfektionen eller transformationen med nukleinsyren ikke er en proces til modificering af den genetiske identitet af kimlinjen hos et menneske.
7. Rekombinant celle til anvendelse ifølge krav 5 eller 6, hvor cellen er en antigenpræsenterende celle (APC), og cellen udtrykker polypeptidet på dens overflade.
8. Rekombinant celle til anvendelse ifølge krav 7, hvor APC'en er en APC, der er pulseret med et antigen eller et antigent peptid.
9. Isoleret polypeptid, der kodes af et DNA, der omfatter: en polypeptidkodende nukleinsyre, hvor nukleinsyren under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ NO: 1 eller SEQ NO: 3, hvor det isolerede polypeptid er til anvendelse som et medikament til administration til et pattedyr, som mistænkes for at have en inflammatorisk tilstand.
10. In vitro-fremgangsmåde til etablering af kontakt med en T-celle, hvilken fremgangsmåde omfatter dyrkning af T-cellen in vitro med polypeptidet ifølge krav 1.
11. Fremgangsmåde til identificering af en forbindelse, der hæmmer et immunrespons, hvilken fremgangsmåde omfatter: (a) tilvejebringelse af en testforbindelse; (b) dyrkning af forbindelsen sammen med et isoleret polypeptid, en T-celle og et molekyle, der leverer et aktiveringssignal til en T-celle; og (c) bestemmelse af, om testforbindelsen hæmmer T-cellens respons på molekylet som indikation af, at testforbindelsen hæmmer et immunrespons, hvor det isolerede polypeptid kodes af et DNA, der omfatter: en polypeptidkodende nukleinsyre, hvilken nukleinsyren under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ NO: 1 eller SEQ NO: 3.
12. Fremgangsmåde ifølge krav 11, hvor molekylet er et antistof, der binder til en T-cellereceptor eller et CD3-polypeptid, eller hvor molekylet er et alloantigen eller et antigent peptid, der er bundet til et MHC (major histocompatibility complex)-molekyle på overfladen af en antigenpræsenterende celle (APC) , især en APC, der er transficeret eller transformeret med en nukleinsyre, der koder for polypeptidet, og polypeptidet udtrykkes på overfladen af APC'en.
13. Fremgangsmåde til identificering af en forbindelse, der forstærker et immunrespons, hvilken fremgangsmåde omfatter: (a) tilvejebringelse af en testforbindelse; (b) dyrkning af forbindelsen sammen med et isoleret polypeptid, en T-celle og et molekyle, der leverer et aktiveringssignal til en T-celle; og (c) bestemmelse af, om testforbindelsen forstærker T-cellens respons på molekyle som indikation af, at testforbindelsen forstærker et immunrespons, hvor det isolerede polypeptid kodes af et DNA, der omfatter: en polypeptidkodende nukleinsyre, hvilken nukleinsyre under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ NO: 1 eller SEQ NO: 3.
14. Fremgangsmåde ifølge krav 13, hvor molekylet er et antistof, der binder til en T-cellereceptor eller et CD3-polypeptid, eller hvor molekylet er et alloantigen eller et antigent peptid, der er bundet til et MHC-molekyle på overfladen af en APC, især en APC, der er transficeret eller transformeret med en nukleinsyre, der koder for polypeptidet, og polypeptidet udtrykkes på overfladen af APC'en.
15. Antistof, der binder til et polypeptid, der består af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1 eller SEQ ID NO: 3.
16. B7-Hl-molekyle til anvendelse til behandling af en inflammatorisk tilstand, rheumatoid arthritis (RA), multipel sklerose (MS) eller insulinafhængig diabetes mellitus (IDDM), hvor B7-Hl-molekylerne er polypeptider, der kodes af et DNA, der indbefatter en nukleinsyre, der (i) koder for et polypeptid med evnen til at co-stimulere en T-celle og (11) under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3, eller polypeptider, der omfatter aminosyrer med sekvensen af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1 eller SEQ ID NO: 3, eller polypeptider, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3 eller den ene eller den anden af disse aminosyrer, som kun afviger ved en eller flere konservative substitutioner, eller funktionelle fragmenter af et hvilket som helst af disse polypeptider, hvor et funktionelt fragment af et polypeptid er et fragment af polypeptidet, der er kortere end polypeptidets fulde længde og har evnen til at co-stimulere en T-celle, eller fusionsproteiner, der indeholder et første domæne og mindst ét yderligere domæne, hvor det første domæne er et hvilket som helst af polypeptiderne eller de funktionelle fragmenter, eller et hvilket som helst af polypeptiderne og de funktionelle fragmenter, der er modificeret ved tilføjelse i den amino-og/eller carboxylterminale ende af et blokeringsmiddel til fremme af overlevelse af det relevante polypeptid in vivo, eller et hvilket som helst af polypeptiderne og de funktionelle fragmenter, der er kovalent eller ikke-kovalent bundet til farmaceutisk acceptable bæreproteiner.
17. B7-Hl-molekyler til anvendelse ifølge krav 16, hvor den inflammatoriske tilstand er en inflammatorisk tilstand, der er induceret af et smitstof.
18. Fremgangsmåde til co-stimulering af en T-celle, der omfatter etablering af kontakt af en kombination, der omfatter et B7-Hl-polypeptid og (a) et antistof, der binder til en T-cellereceptor (TCR), eller (b) et polypeptid af CD3- komplekset, der er fysisk forbundet med TCR'en, med en CD4+-T-celle eller en CD8+-T-celle in vitro, eller etablering af kontakt af en kombination, der omfatter et B7-Hl-polypeptid og et MHC-alloantigen, med en CD8+-T-celle in vitro, hvor B7-H1-polypeptidet er et polypeptid, der kodes af et DNA, der omfatter en nukleinsyre med en sekvens, der (i) koder for et polypeptid med evnen til at co-stimulere en T-celle og (11) under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3, eller et polypeptid, der omfatter aminosyrer med sekvensen af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1 eller SEQ ID NO: 3, eller et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3 eller den ene eller den anden af disse aminosyrer, som kun afviger ved en eller flere konservative substitutioner, eller et funktionelt fragment af et hvilket som helst af disse polypeptider, hvor et funktionelt fragment af et polypeptid er et fragment af polypeptidet, der er kortere end polypeptidets fulde længde og har evnen til at co-stimulere en T-celle, eller et fusionsprotein, der indeholder et første domæne og mindst ét yderligere domæne, hvor det første domæne er et hvilket som helst af polypeptiderne eller det funktionelle fragment, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er modificeret ved tilføjelse i den amino-og/eller carboxylterminale ende af et blokeringsmiddel til fremme af overlevelse af det relevante polypeptid in vivo, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er kovalent eller ikke-kovalent bundet til et farmaceutisk acceptabelt bæreprotein.
19. Fremgangsmåde ifølge krav 18, hvor antistoffet binder til et polypeptid af CD3-komplekset, der er fysisk forbundet TCR'en.
20. Fremgangsmåde til co-stimulering af en T-celle, der omfatter etablering af kontakt af en kombination, der omfatter et B7-Hl-polypeptid og (a) et MHC-alloantigen eller (b) et antistof, der binder til en T-cellereceptor (TCR), eller (c) et polypeptid af CD3-komplekset, der er fysisk forbundet med TCR'en, med T-cellen in vitro, hvor B7-Hl-polypeptidet er bundet til bunden af en dyrkningsbeholder, især en brønd på en plastikmikrotiterplade, og hvor B7-Hl-polypeptidet er et polypeptid, der kodes af et DNA, der omfatter en nukleinsyre med en sekvens, der (i) koder for et polypeptid med evnen til at co-stimulere en T-celle og (ii) under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3, eller et polypeptid, der omfatter aminosyrer med sekvensen af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1 eller SEQ ID NO: 3, eller et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3 eller den ene eller den anden af disse aminosyrer, som kun afviger ved en eller flere konservative substitutioner, eller et funktionelt fragment af et hvilket som helst af disse polypeptider, hvor et funktionelt fragment af et polypeptid er et fragment af polypeptidet, der er kortere end polypeptidets fulde længde og har evnen til at co-stimulere en T-celle, eller et fusionsprotein, der indeholder et første domæne og mindst ét yderligere domæne, hvor det første domæne er et hvilket som helst af polypeptiderne eller det funktionelle fragment, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er modificeret ved tilføjelse i den amino-og/eller carboxylterminale ende af et blokeringsmiddel til fremme af overlevelse af det relevante polypeptid in vivo, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er kovalent eller ikke-kovalent bundet til et farmaceutisk acceptabelt bæreprotein.
21. Fremgangsmåde ifølge krav 20, hvor antistoffet binder til et polypeptid af CD3-komplekset, der er fysisk forbundet med TOR'en.
22. Anvendelse af en opløsning eller en fast understøtning, der omfatter et B7-Hl-polypeptid eller -polypeptidfragment og et molekyle, der leverer et aktiveringssignal til en T-celle, til ex vivo-behandling af mononukleære celler fra perifert blod (PBMC), hvor B7-Hl-polypeptidet er et polypeptid, der kodes af et DNA, der omfatter en nukleinsyre med en sekvens, der (i) koder for et polypeptid med evnen til at co-stimulere en T-celle og (ii) under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3, eller et polypeptid, der omfatter aminosyrer med sekvensen af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1 eller SEQ ID NO: 3, eller et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller SEQ ID NO: 3 eller den ene eller den anden af disse aminosyrer, som kun afviger ved en eller flere konservative substitutioner, eller et funktionelt fragment af et hvilket som helst af disse polypeptider, hvor et funktionelt fragment af et polypeptid er et fragment af polypeptidet, der er kortere end polypeptidets fulde længde og har evnen til at co-stimulere en T-celle, eller et fusionsprotein, der indeholder et første domæne og mindst ét yderligere domæne, hvor det første domæne er et hvilket som helst af polypeptiderne eller det funktionelle fragment, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er modificeret ved tilføjelse i den amino-og/eller carboxylterminale ende af et blokeringsmiddel til fremme af overlevelse af det relevante polypeptid in vivo, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er kovalent eller ikke-kovalent bundet til et farmaceutisk acceptabelt bæreprotein.
23. Anvendelse ifølge krav 22, hvor aktiveringssignalet leveres til T-cellen via den antigenspecifikke T-cellereceptor (TCR).
24. Anvendelse ifølge krav 22 eller 23, hvor molekylet er et antistof, der binder til TCR'en eller til et polypeptid af CD3-komplekset, der er fysisk forbundet med TCR'en på Τ'-celleoverfladen, et alloantigen eller et antigent peptid, der er bundet til et MHC-molekyle.
25. Fusionsprotein, der omfatter det ekstracellulære domæne af human B7-H1, som er fusioneret med Fc-delen af muse-IgG2a (hB7-HHg) , hvor B7-H1 er et polypeptid, der kodes af et DNA, der omfatter en nukleinsyre med en sekvens, der (i) koder for et polypeptid med evnen til at co-stimulere en T-celle og (i i) under stringente betingelser hybridiserer til komplementet af en nukleinsyre, der koder for et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1, eller et polypeptid, der omfatter aminosyrer med sekvensen af aminosyrerest 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31 eller 32 til aminosyrerest 290 af SEQ ID NO: 1, eller et polypeptid, der omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1 eller omfatter aminosyrer med sekvensen ifølge SEQ ID NO: 1, som kun afviger ved en eller flere konservative substitutioner, eller et funktionelt fragment af et hvilket som helst af disse polypeptider, hvor et funktionelt fragment af et polypeptid er et fragment af polypeptidet, der er kortere end polypeptidets fulde længde og har evnen til at co-stimulere en T-celle, eller et fusionsprotein, der indeholder et første domæne og mindst ét yderligere domæne, hvor det første domæne er et hvilket som helst af polypeptiderne eller det funktionelle fragment, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er modificeret ved tilføjelse i den amino-og/eller carboxylterminale ende af et blokeringsmiddel til fremme af overlevelse af det relevante polypeptid in vivo, eller et hvilket som helst af polypeptiderne eller det funktionelle fragment, der er kovalent eller ikke-kovalent bundet til et farmaceutisk acceptabelt bæreprotein.
26. Fremgangsmåde til hæmning af en co-stimulerende virkning af human B7-H1 på T-celler, hvor T-cellerne bringes i kontakt med antistoffer, der binder specifikt til et polypeptid med sekvensen ifølge SEQ ID NO: 1 in vitro, hvor den co-stimulerende virkning af B7-H1 på T-cellerne omfatter forstærkning af et effektorrespons, et hjælperespons eller et suppressivt respons af T-cellerne.
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