DK1685251T3 - Nukleinsyrekonstruktioner - Google Patents

Nukleinsyrekonstruktioner Download PDF

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DK1685251T3
DK1685251T3 DK04768811.4T DK04768811T DK1685251T3 DK 1685251 T3 DK1685251 T3 DK 1685251T3 DK 04768811 T DK04768811 T DK 04768811T DK 1685251 T3 DK1685251 T3 DK 1685251T3
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sequence
nucleic acid
seq
intron
acid construct
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James Fuller
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Powderject Vaccines Inc
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Claims (29)

1. Nukleinsyrekonstruktion omfattende: (i) en kimær promotorsekvens, som omfatter: (a) en umiddelbar tidlig promotorsekvens (immediate early promoter sequence) fra hCMV; (b) exon 1 og i det mindste en del af exon 2 fra det store umiddelbare tidlige hCMV-gen, idet exon 1 og i det mindste en del af exon 2 er sammenhængende; og (c) et heterologt intron, som helt eller delvis erstatter det native intron A i det store umiddelbare tidlige hCMV-gen; (ii) en kodende sekvens, der er funktionelt koblet til promotorsekvensen (i), og; (iii) en forstærkersekvens 3’ for og funktionelt koblet til den kodende sekvens (ii); hvor forstærkersekvensen er afledt fra en 3’UTR i en HBsAg-sekvens, og den kodende sekvens (ii) er heterolog til forstærkersekvensen.
2. Nukleinsyrekonstruktion ifølge krav 1, som endvidere omfatter en ikke-translateret ledersekvens afledt af HBVpreS2-antigensekvensen, HBV-e-antigensekvensen og HSV-type-2gD-antigensekvensen, idet ledersekvensen er funktionelt koblet til promotoren og den kodende sekvens, og den kodende sekvens er heterolog til den utranslaterede ledersekvens.
3. Nukleinsyrekonstruktion omfattende: (i) en kimær promotorsekvens, som omfatter: (a) en umiddelbar tidlig promotorsekvens (immediate early promoter sequence) fra hCMV; (b) exon 1 og i det mindste en del af exon 2 fra det store umiddelbare tidlige hCMV-gen, idet exon 1 og i det mindste en del af exon 2 er sammenhængende; og (c) et heterologt intron, som helt eller delvis erstatter det native intron A i det store umiddelbare tidlige hCMV-gen; (ii) en kodende sekvens i funktionel kobling med den kimære promotor; (iii) en utranslateret ledersekvens, som er valgt blandt HBVpreS2-antigensekvensen, HBV-e-antigensekvensen og HSV-type-2D-antigensekvensen, og som er i funktionel kobling med den kimære promotor; og (iv) en forstærkersekvens, som er afledt af en 3’-utranslateret region (UTR) i en HBsAg-sekvens, som er i funktionel kobling med den kimære promotor, og som findes nedstrøms for den kodende sekvens.
4. Nukleinkonstruktion ifølge et hvilket som helst af de foregående krav, hvori (i) den umiddelbare tidlige promotorsekvens (a) fra hCMV omfatter SEQ ID NO: 1, en funktion variant deraf med mindst 80% identitet eller et funktionelt fragment af en af disse; og/eller (ii) exonsekvens (b) omfatter SEQ ID NO: 2, en funktionel variant deraf med mindst 80% identitet eller et funktionelt fragment af en af disse.
5. Nukleinsyrekonstruktion ifølge et hvilket som helst af de foregående krav, hvori det heterologe intron (c) omfatter en sekvens valgt blandt en rotteinsulingen-intron-A-sekvens, en kyllingekeratingen-intron-A-sekvens, en kyllingehjerteactingen-intron-A-sekvens, en funktionel variant med mindst 80% identitet med intron-A-sekvensen og et funktionelt fragment af intron-A-sekvensen eller af den funktionelle variant deraf.
6. Nukleinsyrekonstruktion ifølge krav 5, hvori rotteinsulingen-intron-A-sekvensen omfatter SEQ ID NO: 3, en funktionel variant deraf med mindst 80% identitet eller et funktionelt fragment af en af disse.
7. Nukleinsyrekonstruktion ifølge et hvilket som helst af de foregående krav, hvori (i) den utranslaterede ledersekvens er valgt blandt SEQ ID NO: 5, SEQ ID NO: 6, SEQ ID NO: 7, en funktionel variant med mindst 80% identitet med en hvilken som helst af SEQ ID NOS: 5 til 7 eller et funktionelt fragment af en hvilken som helst af SEQ ID NOS: 5 til 7 eller af varianten deraf; og/eller (ii) forstærkersekvensen er valgt blandt SEQ ID NO: 8, en funktionel variant med mindst 80% identitet med SEQ ID NO: 8 eller et funktionelt fragment af SEQ ID NO: 8 eller af varianten deraf.
8. Nukleinsyrekonstruktion ifølge et hvilket som helst af de foregående krav, som omfatter: (i) en polyadenyleringssekvens og/eller (ii) en sekvens, der koder for et signalpeptid.
9. Nukleinsyrekonstruktion ifølge krav 8, hvori: (i) polyadenyleringssekvensen er en polyadenyleringssekvens af et gen valgt blandt kanin-3-globingenet, det tidlige eller det sene gen fra humant papillom-virus (HPV), HSV-2gB-genet, et umiddelbart tidligt abe-CMV-gen og et sent HSVgD-gen, en funktionel variant med mindst 80% identitet med en hvilken som helst af de nævnte sekvenser eller et funktionelt fragment af en hvilken hvilken som helst af polyadenyleringssekvenserne eller de funktionelle varianter deraf; og/eller (ii) signalpeptidet er valgt blandt humant vævsplasminogenaktivator-signalpeptid (hTPAsp), aprotinin-signalpeptid, tobak-extensin-signalpeptid, kyllingelysozym-signalpeptid, en funktionel variant med mindst 80% identitet med en hvilken som helst af sekvenserne eller et funktionelt fragment af en hvilken som helst af sekvenserne, som koder for signalpeptidet eller den funktionelle variant deraf.
10. Nukleinsyrekonstruktion ifølge krav 9, hvori polyadenyleringssekvensen er valgt blandt SEQ ID NO: 10, SEQ ID NO: 11, SEQ ID NO: 12, SEQ ID NO: 13, en funktionel variant med mindst 80% identitet med en vilkårlig af SEQ ID NOS: 10 til 13 eller et funktionelt fragment afen vilkårlig af SEQ ID NOS: 10 til 13 eller af varianten deraf.
11. Nukleinsyrekonstruktion ifølge et hvilket som helst af de foregående krav, hvori den kodende sekvens koder for et antigen.
12. Nukleinsyrekonstruktion ifølge krav 11, hvori antigenet er et antigen til et viralt, bakterielt, parasitisk eller fungalt pathogen, et allergiantigen eller et cancerantigen.
13. Nukleinsyrekonstruktion ifølge krav 12, hvori det virale antigen er et HPV-, HIV-, HSV1-, HSV2-, influenzavirus-, Hepatitis-A-virus- eller Hepatitis-B-virusantigen.
14. Nukleinsyrekonstruktion ifølge krav 13, hvori antigenet er HBsAg.
15. Nukleinsyrekonstruktion ifølge et hvilket som helst af kravene 1 til 10, hvori den kodende sekvens koder for en ADP-ribosylerende bakteriel underenhed A og/eller B-underenhed, en aktiv homolog til én eller begge de underenheder, idet hver homolog har mindst 80% identitet med den pågældende underenhed eller et aktivt fragment af underenheden eller af en aktiv homolog dertil.
16. Nukleinsyrekonstruktion ifølge krav 15, hvori den bakterielle underenhed er valgt blandt choleratoxin-underenhed A, choleratoxin-underenhed B, varmelabil toxin-underenhed A fra E.coli, varmelabil toxin-underenhed B fra E.coli, en aktiv homolog med mindst 80% identitet med en vilkårlig af de nævnte underenheder eller et aktivt fragment af en vilkårlig af de nævnte underenheder eller de aktive homologe.
17. Nukleinsyrekonstruktion ifølge et hvilket som helst af de foregående krav, som er et DNA-plasmid.
18. Belagte partikler, der er egnede til afgivelse fra en partikelformidlet afgivelsesindretning, idet partiklerne omfatter bærepartikler, der er belagt med en nukleinsyrekonstruktion ifølge et hvilket som helst af de foregående krav.
19. Belagte partikler ifølge krav 18, hvori bærepartiklerne er guld eller wolfram.
20. Doseringsbeholder til en partikelformidlet afgivelsesindretning omfattende belagte partikler ifølge krav 18 eller 19.
21. Partikelformidlet afgivelsesindretning fyldt med belagte partikler ifølge krav 18 eller 19.
22. Partikelformidlet afgivelsesindretning ifølge krav 21, som er en nåleløs sprøjte.
23. Farmaceutisk præparat omfattende en nukleinsyrekonstruktion ifølge et hvilket som helst af kravene 1 til 17 og et farmaceutisk acceptabelt excipiens.
24. Vaccine omfattende en nukleinsyrekonstruktion ifølge et hvilket som helst af kravene 11 til 14.
25. Vaccine ifølge krav 24, som endvidere omfatter en adjuvanskonstruktion ifølge krav 15 eller 16.
26. Fremgangsmåde in vitro til opnåelse af ekspression af et polypeptid af interesse i pattedyrsceller, hvilken fremgangsmåde omfatter overførsel til cellerne af en nukleinsyrekonstruktion ifølge et hvilket som helst af kravene 1 til 17 eller belagte partikler ifølge krav 18 eller 19.
27. Nukleinsyrekonstruktion ifølge et hvilket som helst af kravene 11 til 14 til anvendelse ved immunisering.
28. Nukleinsyrekonstruktion ifølge krav 27, hvor immuniseringen er mod infektion med et pathogen, allergi eller cancer.
29. Nukleinsyrekonstruktion ifølge krav 27 eller 28, hvor afgivelsen skal ske ved injektion, transdermal partikelafgivelse, inhalation, topisk, oralt, intranasalt eller transmucosalt.
DK04768811.4T 2003-10-10 2004-10-11 Nukleinsyrekonstruktioner DK1685251T3 (da)

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PCT/GB2004/004279 WO2005035771A2 (en) 2003-10-10 2004-10-11 Nucleic acid constructs

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PL1685251T3 (pl) 2014-07-31
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