HRP20201906T1 - Označene kimerne efektorske molekule i njihovi receptori - Google Patents

Označene kimerne efektorske molekule i njihovi receptori Download PDF

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HRP20201906T1
HRP20201906T1 HRP20201906TT HRP20201906T HRP20201906T1 HR P20201906 T1 HRP20201906 T1 HR P20201906T1 HR P20201906T T HRP20201906T T HR P20201906TT HR P20201906 T HRP20201906 T HR P20201906T HR P20201906 T1 HRP20201906 T1 HR P20201906T1
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Croatia
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signal
fusion protein
binding
domain
cassette
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HRP20201906TT
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Stanley R. Riddell
Lingfeng LIU
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Fred Hutchinson Cancer Research Center
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Publication of HRP20201906T1 publication Critical patent/HRP20201906T1/hr

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Claims (25)

1. Fuzijski protein s jednim lancem, koji sadrži hidrofobni dio postavljen između izvanstanične komponente i unutarstanične komponente, pri čemu izvanstanična komponenta sadrži množinu znakovnih kazeta i spojnu regiju koja sadrži zglob, pri čemu unutarstanična komponenta sadrži efektorsku domenu, te pri čemu svaka od množine znakovnih kazeta sadrži Strep oznaku, pri čemu svaka Strep oznaka ima streptavidin, streptaktin, ili oba kao srodni vezni partner.
2. Fuzijski protein s jednim lancem prema zahtjevu 1, pri čemu spojna regija nadalje sadrži: (a) vezni modul; i/ili (b) vezni modul, pri čemu vezni modul sadrži jedan ili više GlyxSery, pri čemu x i y su nezavisno cijeli broj od 0 do 10 uz uvjet da x i y nisu oboje 0; i/ili (c) vezni modul, pri čemu vezni modul je (GlyxSer)n, pri čemu x je cijeli broj od 1 do 5 te n je cijeli broj od 1 do 10; i/ili (d) vezni modul, pri čemu vezni modul je imunoglobulin CH2CH3 ili imunoglobulin CH3.
3. Fuzijski protein s jednim lancem prema zahtjevu 1 ili 2, pri čemu spojna regija sadrži: (a) od jedne do pet množina znakovnih kazeta; i/ili (b) od jedne do pet množina znakovnih kazeta, pri čemu je svaka znakovna kazeta vezana za jedan ili dva vezna modula koji sadrže (GlyxSer)n, pri čemu je n cijeli broj od 1 do 10, te su x i y nezavisno cijeli broj od 0 do 10 uz uvjet da x i y nisu oboje 0; i/ili (c) od jedne do pet množina znakovnih kazeta, pri čemu je svaka znakovna kazeta vezana za jedan ili dva vezna modula koji imaju sekvencu amino kiselina Gly-Gly-Gly-Gly-Ser (SEQ ID NO.: 10), (Gly-Gly-Gly-Gly-Ser)2 (SEQ ID NO.: 11), (Gly-Gly-Gly-Ser)2-Gly-Gly-Ser (SEQ ID NO.: 12), ili bilo koju njihovu kombinaciju.
4. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1-3, pri čemu fuzijski protein s jednim lancem dalje sadrži His oznaku, Flag oznaku, Xpress oznaku, Avi oznaku, Calmodulin oznaku, Poliglutamat oznaku, HA oznaku, Myc oznaku, Nus oznaku, S oznaku, SBP oznaku, Softag oznaku, V5 oznaku, CBP, GST, MBP, GFP, Tioredoksin oznaku, ili bilo koju njihovu kombinaciju.
5. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1-4, pri čemu Strep oznaka sadrži sekvencu amino kiselina Trp-Ser-His-Pro-Gln-Phe-Glu-Lys (SEQ ID NO.: 1) ili Trp-Arg-His-Pro-Gln-Phe-Gly-Gly (SEQ ID NO.: 2).
6. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1 do 5, pri čemu je hidrofobni dio transmembranska domena iz CD4, CD8, CD28 ili CD27.
7. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1-6, pri čemu: (a) efektorska domena je iz CD3ε, CD3δ, CD3ζ, CD27, CD28, CD79A, CD79B, CD137, CARD11, DAP10, FcRα, FcRβ, FcRγ, Fyn, HVEM, ICOS, Lck, LAG3, LAT, LRP, NKG2D, NOTCH1, NOTCH2, NOTCH3, NOTCH4, ROR2, Ryk, SLAMF1, Slp76, pTα, TCRα, TCRβ, TRIM, Zap70, PTCH2, ili bilo koja njihova kombinacija, pri čemu, izborno, efektorska domena je iz LRP, NOTCH1, NOTCH2, NOTCH3, NOTCH4, ROR2, ili Ryk; ili (b) efektorska domena je iz (i) CD3ζ i (ii) jedna ili više od 4-1BB (CD137), CD27, CD28, i OX40 (CD134).
8. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1-7, pri čemu fuzijski protein sadrži od amino-terminusa do karboksi-terminusa: (a) znakovnu kazetu, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (b) prvu spojnu regiju, znakovnu kazetu, drugu spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (c) prvu znakovnu kazetu, prvu spojnu regiju, drugu znakovnu kazetu, drugu spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (d) prvu znakovnu kazetu, prvu spojnu regiju, drugu znakovnu kazetu, drugu spojnu regiju, treću znakovnu kazetu, treću spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (e) dvije do pet znakovnih kazeta, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (f) znakovnu kazetu, spojnu regiju koja sadrži IgG zglob, transmembransku domenu, te unutarstaničnu komponentu koja sadrži efektorsku domenu iz 4-1BB i CD3ζ; CD27 i CD3ζ; CD28 i CD3ζ; ili CD28,4-1BB i CD3ζ; ili (g) znakovnu kazetu, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu iz LRP, NOTCH1, NOTCH2, NOTCH3, NOTCH4, ROR2, ili Ryk.
9. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1-8, pri čemu izvanstanična komponenta dalje sadrži veznu domenu koja se specifično veže za metu.
10. Fuzijski protein s jednim lancem prema zahtjevu 9, pri čemu je vezna domena scFv, scTCR, receptorska ekto-domena, ili ligand.
11. Fuzijski protein s jednim lancem ili receptor prema zahtjevu 9 ili 10, pri čemu fuzijski protein sadrži od amino-terminusa do karboksi-terminusa: (a) izvanstaničnu veznu domenu, znakovnu kazetu, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (b) izvanstaničnu veznu domenu, prvu spojnu regiju, znakovnu kazetu, drugu spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (c) izvanstaničnu veznu domenu, prvu znakovnu kazetu, prvu spojnu regiju, drugu znakovnu kazetu, drugu spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (d) izvanstaničnu veznu domenu, prvu znakovnu kazetu, prvu spojnu regiju, drugu znakovnu kazetu, drugu spojnu regiju, treću znakovnu kazetu, treću spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (e) znakovnu kazetu, izvanstaničnu veznu domenu, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (f) izvanstaničnu scFv ili scTCR veznu domenu koja sadrži varijabilni veznik regija koji sadrži znakovnu kazetu postavljenu između varijabilnih regija, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu; ili (g) izvanstaničnu scFv ili scTCR veznu domenu, znakovnu kazetu, spojnu regiju koja sadrži IgG zglob, transmembransku domenu, te unutarstaničnu komponentu koja sadrži efektorsku domenu, pri čemu je efektorska domena iz: (A) 4-1BB i CD3ζ, (B) CD27 i CD3ζ, (C) CD28 i CD3ζ, (D) OX40 i CD3ζ, (E) CD28,4-1BB i CD3ζ, (F) OX40, 4-1BB i CD3ζ, ili (G) CD28, OX40 i CD3ζ; ili (h) izvanstaničnu veznu domenu koja sadrži receptorsku ektodomenu, znakovnu kazetu, spojnu regiju koja sadrži zglob, hidrofobni dio, te unutarstaničnu komponentu koja sadrži efektorsku domenu, pri čemu je efektorska domena iz 4-1BB, CD27, CD28, ili OX40.
12. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 1-8, pri čemu fuzijski protein dalje sadrži: (a) ne-kovalentno povezanu veznu domenu; i/ili (b) ne-kovalentno povezanu veznu domenu, pri čemu se ne-kovalentno povezana vezna domena veže sa znakovnom kazetom; i/ili (c) ne-kovalentno povezanu veznu domenu, pri čemu ne-kovalentno povezana vezna domena je scFv, scTCR, receptorska ektodomena, ili ligand; i/ili (d) ne-kovalentno povezanu bi-specifičnu veznu domenu, pri čemu je prvi vezni kraj specifičan za znakovnu kazetu te je drugi vezni kraj specifičan za metu koja nije znakovna kazeta; i/ili (e) ne-kovalentno povezanu bi-specifičnu veznu domenu, pri čemu su prvi i drugi vezni kraj specifični za znakovnu kazetu; i/ili (f) ne-kovalentno povezanu multi-specifičnu veznu domenu, pri čemu se prvi kraj veže za znakovnu kazetu te je drugi vezni kraj specifičan za jednu ili više meta koje nisu znakovne kazete.
13. Fuzijski protein s jednim lancem prema bilo kojem od zahtjeva 9-12, pri čemu meta sadrži CD3, CEACAM6, c-Met, EGFR, EGFRvIII, ErbB2, ErbB3, ErbB4, EphA2, IGF1R, GD2, O-acetil GD2, O-acetil GD3, GHRHR, GHR, FLT1, KDR, FLT4, CD44v6, CD151, CA125, CEA, CTLA-4, GITR, BTLA, TGFBR2, TGFBR1, IL6R, gp130, Lewis A, Lewis Y, TNFR1, TNFR2, PD1, PD-L1, PD-L2, HVEM, MAGE-A, mezotelin, NY-ESO-1, PSMA, RANK, ROR1, TNFRSF4, CD40, CD137, TWEAK-R, HLA, tumor ili peptid povezan s patogenom vezan za HLA, hTERT peptid vezan za HLA, tirozinazni peptid vezan za HLA, WT-1 peptid vezan za HLA, LTβR, LIFRβ, LRP5, MUC1, OSMRβ, TCRα, TCRβ, CD19, CD20, CD22, CD25, CD28, CD30, CD33, CD52, CD56, CD80, CD81, CD86, CD123, CD171, CD276, B7H4, TLR7, TLR9, PTCH1, WT-1, Robo1, α-fetoprotein (AFP), Frizzled, OX40, ili CD79b.
14. Molekula nukleinske kiseline koja kodira fuzijski protein prema bilo kojem od zahtjeva 1-13.
15. Vektor koji sadrži molekulu nukleinske kiseline prema zahtjevu 14.
16. Vektor prema zahtjevu 15, pri čemu je vektor viralni vektor; izborno retroviralni vektor ili lentiviralni vektor.
17. Stanica domaćin, koja sadrži molekulu nukleinske kiseline koja kodira fuzijski protein prema bilo kojem od zahtjeva 1-13, izborno pri čemu stanica domaćin je T stanica.
18. In vitro postupak aktivacije stanice, koji sadrži dovođenje u kontakt stanice koja sadrži fuzijski protein prema bilo kojem od zahtjeva 1-13 i/ili molekulu nukleinske kiseline prema zahtjevu 14 s veznom domenom specifičnom za jednu ili više od jedne ili više znakovnih kazeta.
19. In vitro postupak za poticanje razmnožavanja stanica, koji sadrži dovođenje u kontakt ne-prirodne stanice koja sadrži fuzijski protein prema bilo kojem od zahtjeva 1-13 i/ili molekulu nukleinske kiseline prema zahtjevu 14 s veznom domenom specifičnom za jednu ili više od jedne ili više znakovnih kazeta i faktorom rasta citokinom na vrijeme dovoljno da se omogući rast stanica.
20. In vitro postupak identifikacije stanice, postupak sadrži: dovođenje u kontakt uzorka koji sadrži stanicu koja sadrži fuzijski protein prema bilo kojem od zahtjeva 1-13 i/ili molekulu nukleinske kiseline prema zahtjevu 14 s veznom domenom specifičnom za jednu ili više od jedne ili više znakovnih kazeta, pri čemu vezna domena specifična za znakovnu kazetu sadrži prepoznatljivi dio, te prepoznavanje prisutnosti stanice u uzorku.
21. In vitro postupak za sortiranje ili odabir stanice ili populacije stanica, postupak sadrži: dovođenje u kontakt uzorka koji sadrži T stanicu koja sadrži fuzijski protein prema bilo kojem od zahtjeva 1-13 i/ili molekulu nukleinske kiseline prema zahtjevu 14, s veznom domenom specifičnom za jednu ili više od jedne ili više znakovnih kazeta, te odabir ili sortiranje od drugih stanica stanicu/e koje se specifično vežu za veznu domenu, čime se postiže odabir ili sortiranje stanice ili populacije stanica od drugih stanica.
22. In vitro postupak za obogaćivanje ili izoliranje stanice ili populacije stanica, postupak sadrži dovođenje u kontakt uzorka koji sadrži stanicu koja sadrži molekulu nukleinske kiseline prema zahtjevu 14 i/ili fuzijski protein prema bilo kojem od zahtjeva 1-13 s veznom domenom specifičnom za jednu ili više od jedne ili više znakovnih kazeta te obogaćivanje ili izoliranje stanice od drugih stanica koje ne ekspresiraju fuzijski protein ili receptor u uzorku.
23. Stanica domaćin prema zahtjevu 17 za uporabu u postupku crpljenja stanica za liječenje raka ili zarazne bolesti, koji sadrži dovođenje u kontakt stanice koja sadrži molekulu nukleinske kiseline prema zahtjevu 14 i/ili fuzijski protein prema bilo kojem od zahtjeva 1-13 s veznom domenom specifičnom za znakovnu kazetu, pri čemu vezanje vezne domene specifične za znakovnu kazetu vodi do smrti stanica domaćina koje ekspresiraju fuzijski protein, izborno pri čemu postupak dalje sadrži praćenje razina citokina u subjektu nakon primjene vezne domene specifične za jednu ili više od jedne ili više znakovnih kazeta.
24. Stanica domaćin prema zahtjevu 17 za uporabu u liječenju bolesti ili stanja.
25. Farmaceutski sastav koji sadrži stanicu domaćin prema zahtjevu 17 i farmaceutski prihvatljivi nosač, razrjeđivač ili ekscipijens.
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