HRP20140327T1 - Lentiviralni vektori pseudotipizirani sa glikoproteinom ovojnice virusa sindbis - Google Patents
Lentiviralni vektori pseudotipizirani sa glikoproteinom ovojnice virusa sindbis Download PDFInfo
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- HRP20140327T1 HRP20140327T1 HRP20140327AT HRP20140327T HRP20140327T1 HR P20140327 T1 HRP20140327 T1 HR P20140327T1 HR P20140327A T HRP20140327A T HR P20140327AT HR P20140327 T HRP20140327 T HR P20140327T HR P20140327 T1 HRP20140327 T1 HR P20140327T1
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Claims (23)
1. Čestica lentiviralnog vektora naznačena time da sadrži:
(a) ovojnicu koja sadrži
(i) glikoprotein E2 virusa Sindbis sa sekvencom SEQ ID NO:1 u kojoj je 160X odsutan ili je amino kiselina koja nije glutaminska kiselina, ili varijanta SEQ ID NO:1 koja ima barem 80% identičnosti sa SEQ ID NO:1, te pri čemu je 160X odsutan ili je amino kiselina koja nije glutaminska kiselina, koja može inficirati dendritske stanice; pri čemu E2 nije dio kondenziranog proteina sa Sindbis virusom E3; te
(b) genom lentiviralnog vektora koji sadrži jednu ili više sekvenci od interesa.
2. Čestica lentiviralnog vektora prema zahtjevu 1, naznačena time da 160X je odsutan ili glicin, alanin, valin, leucin ili izoleucin, kao što je na primjer izabran od glicina, valina, leucina ili izoleucina.
3. Čestica lentiviralnog vektora prema zahtjevu 1 ili 2, naznačena time da varijanta glikoproteina E2 ima barem jednu izmijenjenu amino kiselinu, da bi se smanjio njegov neto pozitivni naboj.
4. Čestica lentiviralnog vektora prema zahtjevu 3, naznačena time da je barem jedna amino kiselina koja je promijenjena da bi se smanjio njegov neto pozitivni naboj odabrana od lizina 70, lizina 76 ili lizina 159 iz SEQ ID NO:1, te poželjno gdje su supstitucije neovisno odabrane od glutaminske kiseline ili asparaginske kiseline.
5. Čestica lentiviralnog vektora prema zahtjevu 1, naznačena time da varijanta E2 sekvence je ostatak amino kiselina 66 do 488 iz SEQ ID NO:3, ostatci amino kiselina 66 do 488 iz SEQ ID NO: 4, ili ostatci amino kiselina 66 do 486 iz SEQ ID NO:5; (varijante 1, 2, i 3).
6. Čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-5, naznačena time da je varijanta u kojoj je sekvenca sa ostatcima 71-75 iz SEQ ID NO:1 nepromijenjena ili ima jednu ili dvije supstitucije koje ne utječu na sposobnost varijante da inficira DC-ove, ali ne mijenja broj amino kiselina u toj regiji.
7. Čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-6, naznačena time da sekvenca od interesa eksprimira produkt koji je antigen agensa koji uzrokuje bolest ili oboljela stanica.
8. Čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-6 naznačena time da sekvenca od interesa kodira tumor-specifični antigen, proizvoljno NY-ESO-1, MAGE, MART-1/Melan-A, BAGE, RAGE, antigen linije melanocit-melanom, kao gp100, gp75, mda-7, tirozinaza ili sa tirozinazom povezan protein; antigen karcinoma renalne stanice, 5T4, SM22-alfa, karbonska anhidraza I, karbonska anhidraza IX (također poznata kao G250), faktori, koji induciraju hipoksiju (proizvoljno, HIF-1 alfa ili HIF-2alfa), VEGF ili membranski antigen specifičan za prostatu (PSMA), prostata specifični antigen (PSA), prostatičnu kiselu fosfatazu, šest-transmembranski epoitelialni antigen prostate (STEAP), NKX3.1, epitop proteini/peptidi dobiveni iz gena koji su mutirali u stanicama tumora in ili genima transkribiranim na različitim razinama u tumoru u usporedbi sa normalnim stanicama, kao što su enzim telomeraze, survivin, mezotelin, mutirani ras, preuređeni bcr/abl, Her2/neu, mutiran ili divljeg soja p53, citokrom P450 1B1, abnormalno eksprimirane intronske sekvence kao što su N-acetilglukozaminiltransferaza-V; klonalno preuređene imunoglobulinske gene koji generiraju jedinstvene idiotipove u mijelomu ili u limfomima B-stanica; epitop proteini/peptidi dobiveni iz onkoviralnih procesa, kao što su proteini E6 ili E7humanog papiloma virusa; ili ne-mutirani onkofetalni proteini sa tumorskom-selektivnom ekspresijom, kao što su karcinoembrionski antigen ili alfa-fetoprotein.
9. Čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-7, naznačena time da sekvenca od interesa kodira antigen dobiven iz virusa, kao HIV ili SIV antigen, polipeptid adenovirusa, polipeptid alfavirusa, polipeptid kalicivirusa, npr., antigen kalicivirus kapsida, polipeptid koronavirusa, polipeptid distemper virusa, polipeptid Ebola virusa, polipeptid enterovirusa, polipeptid flavivirusa, polipeptid virusa hepatitisa (AE), npr., jezgru ili površinski antigen hepatitisa B, ili glikoproteine E1 ili E2virusa hepatitisa C, jezgru, ili ne-strukturne proteine, polipeptid virusa herpesa, npr., virus herpes simplex ili glikoprotein virusa varicela zoster, polipeptid virusa imunodeficijencije, npr., ovojnicu ili proteazu humanog virusa imunodeficijencije, polipeptid virusa infekcijskog peritonitisa, polipeptid virusa influence, npr., hemaglutinin influence A, neuraminidaza; ili nukleoprotein, polipeptid virusa leukemije, polipeptid virusa Marburg, polipeptid ortomikso virusa, polipeptid papiloma virusa, polipeptide parainfluenca virusa, npr., hemaglutinin/neuraminidaza, polipeptid paramiksovirusa, polipeptid parvovirusa, polipeptid pestivirusa, polipeptid picorna virusa, npr., polipeptid poliovirusa kapsid, polipeptid virus boginja, npr., polipeptid vaccinia virusa, polipeptid virusa bjesnoće, npr., glikoprotein G virusa bjesnoće, polipeptid reovirusa, polipeptid retrovirusa, ili polipeptid rotavirusa.
10. Pripravak koji sadrži česticu lentiviralnog vektora prema bilo kojem zahtjevu od 1-9, naznačen time da pripravak ima IU od barem 105/mL.
11. Sustav pakiranja lentiviralnog vektora za proizvodnju pseudotipizirane čestice lentiviralnog vektora prema bilo kojem zahtjevu od 1-9, ili pripravak prema zahtjevu 10, naznačen time da sadrži:
(i) prvu molekulu nukleinske kiseline koja kodira glikoprotein E2 virusa Sindbis prema zahtjevu 1 (a)(i), te;
(ii) drugu molekulu nukleinske kiseline koja kodira gag i pol proteine,
(iii) treću molekulu nukleinske kiseline koja kodira rev;
(iv) genom lentiviralnog vektora koji sadrži sekvencu od interesa; te
(v) pakirajuću stanicu.
12. Sustav pakiranja lentiviralnog vektora prema zahtjevu 11, naznačen time da:
(a) pol protein ima ne-funkcionalnu integrazu; i/ili
(b) pol protein ima ne-funkcionalnu integrazu sa D64V mutacijom; i/ili
(c) genom lentiviralnog vektora je ne-integrirajući lentiviralni genom; i/ili
(d) pakirajuća stanica je stabilno pretvorena sa (ii) i (iii); i/ili
(e) pakirajuća stanica je transfektirana sa (i) i (iv).
13. Izolirana molekula nukleinske kiseline naznačena time da sadrži nukleotidnu sekvencu koja kodira glikoprotein E2 ili varijantu prema zahtjevu 1 (a)(i), ili prema bilo kojem od zahtjeva 2-9.
14. Molekula nukleinske kiseline prema zahtjevu 13 naznačena time da protein je poliprotein Sindbis E3/E2/6K/E1 koji je obrađen tako da protein E2 nije dio spojenog proteina sa E3 kada je ugrađen u ovojnicu virusa.
15. Molekula nukleinske kiseline prema zahtjevu 14, naznačena time da E3 sekvenca odgovara (a) ostatcima 1-65 iz SEQ ID NO:20, ili (b) njihovoj varijanti koja ima barem 80% identičnosti sekvence sa ostatcima 1-65 iz SEQ ID NO:20,
pri čemu ostatci 62-65 su RSKR (SEQ ID NO: 27), a varijanta se može ugraditi u pseudotipiziranu ovojnicu virusa.
16. Postupak dobivanja čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-9, ili pripravak prema zahtjevu 10, naznačen time da sadrži kultiviranje stanica za pakiranje prema zahtjevu 11.
17. Lentiviralna čestica prema bilo kojem zahtjevu od 1-9, ili pripravak prema zahtjevu 10, naznačena time da genom lentiviralnog vektora
(a) ima deaktivirano ili samostalno-deaktivirano 3' dugo terminalno ponavljanje (LTR); i/ili
(b) sadrži U3 element kojem nedostaju barem jedan od:
(i) sekvenca pojačivača;
(ii) TATA box (iii) mjesto Sp1 (iv) mjesto NK-kapa B i (v) polipurinski trakt (PPT); i/ili
(c) sadrži nukleotidnu sekvencu iz SEQ ID NO: 21, 22 ili 23; i/ili,
(d) sadrži nukleotidnu sekvencu koja kodira faktor starenja/stimulacije dendritske stanice.
18. Čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-9, ili pripravak prema zahtjevu 10, naznačeni time da su za uporabu u postupku liječenja subjekta koji je čovjek ili životinja,te proizvoljno sadrži dostavu ex vivo čestica lentiviralnog vektora u dendritske stanice.
19. Čestica lentiviralnog vektora prema bilo kojem zahtjevu od 1-9, ili pripravak prema zahtjevu 10, naznačeni time da su za uporabu kod induciranja antigen-specifičnog imunološkog odgovora, proizvoljno humoralnog imunološkog odgovora i/ili celularnog imunološkog odgovora.
20. Cjepivo za terapiju ili profilaksu naznačeno time da sadrži čestice lentiviralnih vektora prema bilo kojem zahtjevu od 1-9, ili pripravak prema zahtjevu 10, te farmaceutski prihvatljivu pomoćnu tvar.
21. Cjepivo za terapiju ili profilaksu prema zahtjevu 20 naznačeno time da dodatno sadrži adjuvant.
22. Cjepivo za terapiju ili profilaksu prema zahtjevu 21, naznačeno time da adjuvant je adjuvant sa formulom (I):
[image]
pri čemu A1 i A2 su neovisno odabrani iz skupine koja sadrži vodik, fosfat, te soli fosfata, a dijelovi R1, R2, R3, R4, R5 i R6 su neovisno odabrani iz skupine koja sadrži hidrokarbil koji ima 3 do 23 atoma ugljika, koji predstavlja C3-C23.
23. Cjepivo za terapiju ili profilaksu prema zahtjevu 22, naznačeno time da
(a) A1 je fosfat ili fosfatna sol i A2 je vodik, te
(b)
(i) R1, R3, R5 i R6 su C11-C20 alkil, i R2 i R4 su C12-C20 hidrokarbil; ili
(ii) R1, R3, R5 i R6 su C11 alkil, i R2 i R4 su C13 hidrokarbil; ili
(iii) R1, R3, R5 i R6 su undecil, i R2 i R4 su tridecil.
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