HRP20110179T1 - Kimerni adenovirusi za uporabu u liječenju raka - Google Patents
Kimerni adenovirusi za uporabu u liječenju raka Download PDFInfo
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- HRP20110179T1 HRP20110179T1 HR20110179T HRP20110179T HRP20110179T1 HR P20110179 T1 HRP20110179 T1 HR P20110179T1 HR 20110179 T HR20110179 T HR 20110179T HR P20110179 T HRP20110179 T HR P20110179T HR P20110179 T1 HRP20110179 T1 HR P20110179T1
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- 241000701161 unidentified adenovirus Species 0.000 title claims abstract 49
- 206010028980 Neoplasm Diseases 0.000 title claims 4
- 201000011510 cancer Diseases 0.000 title claims 4
- 210000004881 tumor cell Anatomy 0.000 claims abstract 15
- 150000007523 nucleic acids Chemical group 0.000 claims abstract 9
- 108091028043 Nucleic acid sequence Proteins 0.000 claims abstract 8
- 230000000174 oncolytic effect Effects 0.000 claims abstract 4
- 231100001274 therapeutic index Toxicity 0.000 claims abstract 4
- 238000000034 method Methods 0.000 claims 9
- 108090000623 proteins and genes Proteins 0.000 claims 8
- 210000004027 cell Anatomy 0.000 claims 7
- 239000006228 supernatant Substances 0.000 claims 5
- 102000004169 proteins and genes Human genes 0.000 claims 4
- 230000010076 replication Effects 0.000 claims 4
- 208000029742 colonic neoplasm Diseases 0.000 claims 3
- 230000002950 deficient Effects 0.000 claims 3
- 238000003306 harvesting Methods 0.000 claims 3
- 239000002773 nucleotide Substances 0.000 claims 3
- 125000003729 nucleotide group Chemical group 0.000 claims 3
- 206010009944 Colon cancer Diseases 0.000 claims 2
- 101710094396 Hexon protein Proteins 0.000 claims 2
- NTIZESTWPVYFNL-UHFFFAOYSA-N Methyl isobutyl ketone Chemical compound CC(C)CC(C)=O NTIZESTWPVYFNL-UHFFFAOYSA-N 0.000 claims 2
- 206010061535 Ovarian neoplasm Diseases 0.000 claims 2
- 101710173835 Penton protein Proteins 0.000 claims 2
- 210000000481 breast Anatomy 0.000 claims 2
- 210000001072 colon Anatomy 0.000 claims 2
- 108091005899 fibrous proteins Proteins 0.000 claims 2
- 208000019691 hematopoietic and lymphoid cell neoplasm Diseases 0.000 claims 2
- 230000002401 inhibitory effect Effects 0.000 claims 2
- 210000004072 lung Anatomy 0.000 claims 2
- 239000000203 mixture Substances 0.000 claims 2
- 210000000496 pancreas Anatomy 0.000 claims 2
- 210000002307 prostate Anatomy 0.000 claims 2
- 238000000746 purification Methods 0.000 claims 2
- 230000001225 therapeutic effect Effects 0.000 claims 2
- 230000004614 tumor growth Effects 0.000 claims 2
- 206010006187 Breast cancer Diseases 0.000 claims 1
- 208000026310 Breast neoplasm Diseases 0.000 claims 1
- 102000019034 Chemokines Human genes 0.000 claims 1
- 108010012236 Chemokines Proteins 0.000 claims 1
- 102000004127 Cytokines Human genes 0.000 claims 1
- 108090000695 Cytokines Proteins 0.000 claims 1
- 108090000790 Enzymes Proteins 0.000 claims 1
- 102000004190 Enzymes Human genes 0.000 claims 1
- 206010033128 Ovarian cancer Diseases 0.000 claims 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 claims 1
- 102000006601 Thymidine Kinase Human genes 0.000 claims 1
- 108020004440 Thymidine kinase Proteins 0.000 claims 1
- 241000700605 Viruses Species 0.000 claims 1
- 238000004113 cell culture Methods 0.000 claims 1
- 230000030833 cell death Effects 0.000 claims 1
- 201000011024 colonic benign neoplasm Diseases 0.000 claims 1
- 239000012228 culture supernatant Substances 0.000 claims 1
- 238000012217 deletion Methods 0.000 claims 1
- 230000037430 deletion Effects 0.000 claims 1
- 201000010099 disease Diseases 0.000 claims 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims 1
- 230000004957 immunoregulator effect Effects 0.000 claims 1
- 238000000338 in vitro Methods 0.000 claims 1
- 238000002955 isolation Methods 0.000 claims 1
- 208000020816 lung neoplasm Diseases 0.000 claims 1
- 208000037841 lung tumor Diseases 0.000 claims 1
- 230000001613 neoplastic effect Effects 0.000 claims 1
- 108020004707 nucleic acids Proteins 0.000 claims 1
- 102000039446 nucleic acids Human genes 0.000 claims 1
- 210000001672 ovary Anatomy 0.000 claims 1
- 230000002028 premature Effects 0.000 claims 1
- 229940002612 prodrug Drugs 0.000 claims 1
- 239000000651 prodrug Substances 0.000 claims 1
- 208000023958 prostate neoplasm Diseases 0.000 claims 1
- 230000006798 recombination Effects 0.000 claims 1
- 238000005215 recombination Methods 0.000 claims 1
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Abstract
Rekombinantni kimerni adenovirus koji ima genom koji sadrži E2B područje, naznačen time da navedeno E2B područje sadrži sekvencu nukleinske kiseline koja proizlazi iz prvog adenovirusnog serotipa i sekvencu nukleinske kiseline koja proizlazi iz drugog adenovirusnog serotipa; time da su navedeni prvi i drugi serotipovi svaki odabran iz adenovirusnih podskupina B, C, D, E, ili F i koji su udaljeni jedan od drugoga; itime da je navedeni kimerni adenovirus onkolitički i pokazuje poboljšani terapijski indeks za stanicu tumora. Patent sadrži još 24 patentna zahtjeva.
Claims (25)
1. Rekombinantni kimerni adenovirus koji ima genom koji sadrži E2B područje, naznačen time da navedeno E2B područje sadrži sekvencu nukleinske kiseline koja proizlazi iz prvog adenovirusnog serotipa i sekvencu nukleinske kiseline koja proizlazi iz drugog adenovirusnog serotipa;
time da su navedeni prvi i drugi serotipovi svaki odabran iz adenovirusnih podskupina B, C, D, E, ili F i koji su udaljeni jedan od drugoga; i
time da je navedeni kimerni adenovirus onkolitički i pokazuje poboljšani terapijski indeks za stanicu tumora.
2. Adenovirus prema zahtjevu 1, koji nadalje sadrži područja koja enkodiraju vlaknaste, heksonske i pentonske proteine, naznačen time da nukleinska kiselina koja enkodira vlaknaste, heksonske i pentonske proteine navedenog adenovirusa je iz istog adenovirusnog serotipa.
3. Adenovirus prema zahtjevu 1 ili 2, naznačen time da nadalje sadrži izmijenjeno E3 područje ili nadalje sadrži izmijenjeno E4 područje.
4. Adenovirus prema bilo kojem od zahtjeva 1 do 3, naznačen time da navedena stanica tumora je stanica raka debelog crijeva, dojke, gušterače, pluća, prostate, jajnika, ili hematopoetska stanica tumora.
5. Adenovirus prema zahtjevu 4, naznačen time da navedena stanica tumora je stanica raka debelog crijeva.
6. Adenovirus prema bilo kojem od zahtjeva 1 do 5, naznačen time da nukleotidna sekvenca E2B područja navedenog adenovirusa sadrži SEQ ID NO: 3.
7. Adenovirus prema bilo kojem od zahtjeva 1 do 5, naznačen time da nukleotidna sekvenca navedenog adenovirusa sadrži SEQ ID NO: 1.
8. Rekombinantni kimerni adenovirus koji ima genom koji sadrži E2B područje, naznačen time da navedeno E2B područje sadrži sekvencu nukleinske kiseline koja proizlazi iz prvog adenovirusnog serotipa i sekvencu nukleinske kiseline koja proizlazi iz drugog adenovirusnog serotipa, i
time da navedeni prvi i drugi adenovirusni serotipovi su svaki odabran iz adenovirusnih podskupina B, C, D, E, ili F i koji su udaljeni jedan od drugoga, i
time da navedeni kimerni adenovirus je onkolitički i pokazuje poboljšani terapijski indeks za stanicu tumora, i
time da navedeni kimerni adenovirus je napravljen kao adenovirus s nedostatkom replikacije putem brisanja jednog ili više adenovirusnih područja koja enkodiraju proteine uključene u adenovirusnu replikaciju odabranih iz skupine koja se sastoji od E1, E2, E3 ili E4.
9. Adenovirus s nedostatkom replikacije prema zahtjevu 8, naznačen time da E1 i E3 područja su izbrisana.
10. Adenovirus s nedostatkom replikacije prema zahtjevu 9, naznačen time da nadalje sadrži brisanje E4 područja.
11. Adenovirus prema bilo kojem od prethodnih zahtjeva, koji nadalje sadrži heterologni gen, naznačen time da je navedeni heterologni gen eksprimiran unutar stanice inficirane sa navedenim adenovirusom.
12. Adenovirus prema zahtjevu 11, naznačen time da je navedeni heterologni gen timidin kinaza.
13. Adenovirus prema zahtjevu 11, naznačen time da navedeni heterologni gen enkodira terapeutski protein odabran iz skupine koja se sastoji od citokina i kemokina, antitijela, pro-lijekova koji pretvaraju enzime i imunoregulacijskih proteina.
14. Adenovirus prema bilo kojem od zahtjeva 1 do 13 naznačen time da je za uporabu kod liječenja.
15. Adenovirus prema bilo kojem od zahtjeva 1 do 13 naznačen time da je za uporabu kod liječenja neoplastičnih bolesti ili raka, ili za inhibiciju rasta tumorskih stanica.
16. Postupak za inhibiciju rasta tumorskih stanica, naznačen time da sadrži inficiranje navedene stanice raka in vitro sa adenovirusom prema bilo kojem od zahtjeva 1 do 13.
17. Adenovirus prema zahtjevu 15 ili postupak prema zahtjevu 16, naznačen time da navedena stanica tumora je stanica raka debelog crijeva.
18. Adenovirus ili postupak prema zahtjevu 17, naznačen time da nukleotidna sekvenca navedenog adenovirusa sadrži SEQ ID NO: 1.
19. Adenovirus prema bilo kojem od zahtjeva 1 do 13, naznačen time da je za uporabu za dostavu terapeutskog proteina stanici.
20. Postupak izolacije adenovirusa prema bilo kojem od zahtjeva 1 do 13, naznačen time da navedeni postupak sadrži
a) ujedinjavanje adenovirusnih serotipova koji predstavljaju adenovirusne podskupine B-F, i time stvaraju adenovirusnu mješavinu;
b) prevođenja ujedinjene adenovirusne mješavine iz koraka (a) na aktivno rastuću kulturu tumorskih stanica u dovoljno visokom omjeru čestica po stanici za poticanje rekombinacije između serotipova, ali ne toliko visokim za uzrokovanje prerane smrti stanica;
c) žetve supernatanta iz koraka (b);
d) inficiranja neaktivne kulture tumorskih stanica sa supernatantom dobivenim u koraku (c);
e) žetve stanične kulture supernatanta iz koraka (d) prije bilo kojeg znaka CPE;
f) inficiranja neaktivne kulture tumorskih stanica sa supernatantom dobivenim u koraku (e); i
g) izolacije virusa prema bilo kojem od zahtjeva 1 do 13 iz supernatanta dobivenog u koraku (f) pomoću pročišćavanja plaka.
21. Postupak prema zahtjevu 20, naznačen time da se korak (b) provodi dva puta prije žetve supernatanta u koraku (c).
22. Postupak prema zahtjevu 20, naznačen time da se koraci (e) i (f) ponavljaju do 20 puta prije koraka (g).
23. Postupak prema zahtjevu 20, naznačen time da se drugi krug pročišćavanja plaka provodi nakon koraka (g).
24. Postupak prema zahtjevu 20, naznačen time da stanica tumora je stanica raka debelog crijeva, dojke, gušterače, pluća, prostate, jajnika, ili hematopoetska stanica tumora.
25. Rekombinantni kimerni adenovirus koji ima genom koji sadrži E2B područje, naznačen time da navedeno E2B područje sadrži sekvencu nukleinske kiseline koja proizlazi iz prvog adenovirusnog serotipa i sekvencu nukleinske kiseline koja proizlazi iz drugog adenovirusnog serotipa; i
time da navedeni prvi i drugi adenovirusni serotipovi su svaki odabrani iz adenovirusnih podskupina B, C, D, E, ili F i koji su udaljeni jedan od drugoga; i
time da je navedeni kimerni adenovirus onkolitički i pokazuje poboljšani terapijski indeks za stanicu tumora;
se dobiva postupkom prema bilo kojem od zahtjeva 20 do 24.
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