ME02505B - Heterodimerni vezujući proteini i njihove upotrebe - Google Patents

Heterodimerni vezujući proteini i njihove upotrebe

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Publication number
ME02505B
ME02505B MEP-2016-186A MEP18616A ME02505B ME 02505 B ME02505 B ME 02505B ME P18616 A MEP18616 A ME P18616A ME 02505 B ME02505 B ME 02505B
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Montenegro
Prior art keywords
polypeptide
region
immunoglobulin
heterodimer
chain polypeptide
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MEP-2016-186A
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English (en)
French (fr)
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John W Blankenship
Philip Tan
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Aptevo Res & Development Llc
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Publication of ME02505B publication Critical patent/ME02505B/me

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    • C07K16/18—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2818—Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against CD28 or CD152
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Claims (21)

1. Polipeptidni heterodimer, naznačen time, što sadrži: (a) prvi jednolančani polipetid (SCP-I) koji sadrži jedan do četiri vezujuća domena koja se specifično vezuju za jedan do četiri cilja, zglob (H-I), imunoglobulinski heterodimerizacioni domen (HD-I) i imunoglobulinski CH2 i CH3 domen IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD ili bilo koju njihovu kombinaciju (CH2CH3-I); i (b) drugi jednolančani polipetid (SCP-II) koji sadrži nula do četiri vezujuća domena koja se specifično vezuju za nula do četiri cilja, zglob (hinge, H-II), imunoglobulinski heterodimerizacioni domen (HD-II) i imunoglobulinski CH2 i CH3 domen IgG1, IgG2, IgG3, IgG4, IgA1, IgA2, IgD ili bilo koju njihovu kombinaciju (CH2CH3-II); pri čemu (i) imunoglobulinski heterodimerizacioni domen prvog jednolančanog polipeptida (HD-I) i imunoglobulinski heterodimerizacioni domen drugog jednolančanog polipeptida (HD-II) preferencijalno se udružuju jedan sa drugim da formiraju polipeptidni heterodimer sačinjen od prvog jednolančanog polipeptida (SCP-I) i drugog jednolančanog polipetida (SCP-II), i (1) imunoglobulinski heterodimerizacioni domen prvog jednolančanog polipeptida (HD-I) sadrži prvi imunoglobulinski CH-1 region, i imunoglobulinski heterodimerizacioni domen drugog jednolančanog polipeptida (HD-II) sadrži prvi imunoglobulinski CL region, ili (2) imunoglobulinski heterodimerizacioni domen prvog jednolančanog polipeptida (HD-I) sadrži prvi imunoglobulinski CL region, i imunoglobulinski heterodimerizacioni domen drugog jednolančanog polipeptida (HD-II) sadrži prvi imunoglobulinski CH-1 region; (ii) pri čemu je prvi imunoglobulinski CL region izmenjeni humani imunoglobulinski Cκ region sa jednom ili više amino-kiselina humanog Cκ regiona divljeg tipa, supstituisanih na N29, N30, Q52, V55, T56, S68 ili T70, uz uslov da polipeptidni heterodimer sadrži najmanje dva vezujuća domena koja se specifično vezuju za najmanje dva različita cilja.
2. Polipeptidni heterodimer iz zahteva 1, naznačen time, što su vezujući domeni jednolančani Fv (scFv) polipeptidi.
3. Polipeptidni heterodimer iz zahteva 1, naznačen time, što polipeptidni heterodimer sadrži dva vezujuća domena (BD1 i BD2), pri čemu je poželjno prvi vezujući domen (BD1) na prvom jednolančanom polipeptidu (SCP-I), a drugi vezujući domen (BD2) je na drugom jednolančanom polipeptidu (SCP-II).
4. Polipeptidni heterodimer iz zahteva 3, naznačen time, što se prvi vezujući domen (BD1) nalazi aminoterminalno u odnosu na CH2CH3-I prvog jednolančanog polipeptida, a drugi vezujući domen (BD2) se nalazi aminoterminalno u odnosu na CH2CH3-II drugog jednolančanog polipeptida.
5. Polipeptidni heterodimer iz zahteva 1 ili 2, naznačen time, što polipeptidni heterodimer sadrži tri vezujuća domena (BD1, BD2 i BD3).
6. Polipeptidni heterodimer iz zahteva 1, naznačen time, što polipeptidni heterodimer sadrži četiri vezujuća domena (BD1, BD2, BD3 i BD4), pri čemu su poželjno HD-I i CH2CH3-I smešteni između BD1 i BD2 i HD-II i CH2CH3-II su smešteni između BD3 i BD4.
7. Polipeptidni heterodimer iz zahteva 1 ili 2, naznačen time, što polipeptidni heterodimer sadrži pet do osam vezujućih domena.
8. Polipeptidni heterodimer iz bilo kojeg od zahteva 1 do 7, naznačen time, što se najmanje jedan od vezujućih domena specifično vezuje, ili je antagonist, TCR kompleksa, TCRα, TCRβ, CD3γ, CD3δ, CD3ε, CD28, CD79b, hiperIL-6, monoIL-10, CD86, CD20, PSMA, CD19, HLA-DR, Ron, c-Met, CEACAM-6, LIGHT, GITRL, CD40, PDL1, PDL2, HVEM, LTBR, EGFR, EGFRυIII, ErbB2, ErbB3, ErbB4, IGF1R, EphA2, PDGFR, VEGFR1-4, angiopoetina 2, CD64, CD32A, CD16, CD71, TNFR1, TNFR2, TWEAKR, TACI, BAFF-R, BCMA, FAS, CD32B, CD21, CD22, CD30, CD33, CD37, CD38, CD70, TNFα, IL-6, hiperIL-6, IL-2, IL-1, IL-7, IL-8, IL-17A/C, IP-10, IFNγ, IFNα, RANKL, FASL, TGFβ, IL10, IL17A/F, CSF2, IGF1, IGF2, BlyS/APRIL, HGF, MSP, EGF (uključujući epiregulin, herregulin, β-regulin, neuregulin), HIF-1α, VEGFA, VEGFB, VEGFC, VEGFD, TNFα, Wnt, sHH, TGFβ, PDGF, TWEAK, EpCAM, CEA, PCTA-1, STEAP-1, PSCA, ALCAM (CD166), EphA2, CD151, CA-125, MUC-1, MAGE-1, TROP2, CCR5, HER-3, HER-4, EGFR, CEA, MUC2, MUC3, MUC4, MUC5AC, MUC5b, MUC7, βhCG, Lewis-Y, gangliozida GD3, 9-O-acetil-GD3, GM2, globo H, fukozil-GM1, poli SA, GD2, karboanhidraze IX (MN/CA IX), CD44υ6, sonic hedgehog (Shh), Wue-1, plazma-ćelijski antigen, (za membranu vezanog) IgE, sa melanomom udruženog hondroitin-sulfatnog proteoglikana (MCSP), CCR8, TNF-alfa prekursora, STEAP, mezotelina, A33 antigena, antigena matičnih ćelija prostate (PSCA), Ly-6, dezmogleina 4, E-kadherin neoepitopa, fetalnog acetilholinskog receptora, CD25, CA19-9 markera, CA-125 markera i receptora tipa II Muellerian-ove inhibitorne supstance (MIS), sTn (sijalizovani Tn antigen; TAG-72), FAP (fibroblastni aktivacioni antigen), endosijalina, EGFRυIII, LG, SAS, CD63, IGF1R, CD151, TGFBR2, GHRHR, GHR, IL-6R, gp130, TNFR2, OSMRβ, Patched-1, Frizzled, Robo1, CD80, CD81, CD86, OX40, CD40, CD137, LIFRβ, TLR7 ili TLR9.
9. Polipeptidni heterodimer iz bilo kojeg od prethodnih zahteva, naznačen time, što se jedna ili više amino-kiselina humanog Cκ regiona divljeg tipa supstituišu jednom ili više aminokiselinskih supstitucija koje se biraju između Ala (A), Arg (R), Trp (W), Tyr (Y), Glu (E), Gln (Q), Lys (K), Asp (D), Met (M) Ser (S) i Phe (F).
10. Polipeptidni heterodimer iz bilo kojeg od prethodnih zahteva, naznačen time, što izmenjeni humani imunoglobulinski Cκ region sadrži sledeće supstitucije: (i) N30Y V55A T70E; ili N30M V55A T70E.
11. Polipeptidni heterodimer iz bilo kojeg od prethodnih zahteva, naznačen time, što je CH1 region izmenjeni humani imunoglobulinski CH1 region koji sadrži aminokiselinsku supstituciju kojom je Val (V) na poziciji 68 supstituisan Lys (K), Arg (R) ili His (H), i gde je Cκ region izmenjeni humani imunoglobulinski Cκ region koji sadrži aminokiselinsku supstituciju kojom je Leu (L) na poziciji 27 supstituisan Asp (D) ili Glu (E) ili gde je CH1 region izmenjeni humani imunoglobulinski CH1 region koji sadrži aminokiselinsku supstituciju kojom je Val (V) na poziciji 68 supstituisan Asp (D) ili Glu (E), i gde je Cκ region izmenjeni humani imunoglobulinski Cκ region koji sadrži aminokiselinsku supstituciju kojom je Leu (L) na poziciji 27 promenjen u Lys (K), Arg (R) ili His (H).
12. Polipeptidni heterodimer iz bilo kojeg od zahteva 1 do 11, naznačen time, što je prvi CH1 region izmenjeni humani imunoglobulinski CH1 region, sa cisteinom humanog imunoglobulinskog CH1 regiona divljeg tipa uključenim u formiranje disulfidne veze sa humanim imunoglobulinskim CL regionom divljeg tipa, uklonjenim ili supstituisanim.
13. Polipeptidni heterodimer iz bilo kojeg od prethodnih zahteva, naznačen time, što je Cκ region izmenjeni humani imunoglobulinski Cκ region, sa cisteinskom reziduom humanog Cκ regiona divljeg tipa uključenom u formiranje disulfidne veze sa humanim imunoglobulinskim CH1 regionom divljeg tipa, uklonjenom ili supstituisanom.
14. Polipeptidni heterodimer iz bilo kojeg od zahteva 1 do 13, naznačen time, što je zglob i prvog i drugog jednolančanog polipeptida imunoglobulinski zglobni region.
15. Polipeptidni heterodimer iz bilo kojeg od zahteva 1 do 14, naznačen time, što je zglobni region (a) aminoterminalno u odnosu na CH2CH3-I ili CH2CH3-II, (b) smešten između vezujućeg domena i imunoglobulinskog heterodimerizacionog domena prvog jednolančanog polipeptida (HD-I) i/ili imunoglobulinskog heterodimerizacionog domena drugog jednolančanog polipeptida (HD-II), (c) smešten između imunoglobulinskog heterodimerizacionog domena i CH2CH3-I ili CH2CH3-II, (d) na amino-terminusu prvog ili drugog jednolančanog polipeptida, (e) zglobni region lektina tipa C, (f) NKg2A ili NKg2D peptid ili njihov derivat, (g) smešten između CH2CH3-I i vezujućeg domena, ili (h) na karboksi-terminusu prvog ili drugog jednolančanog polipeptida.
16. Heterodimer iz bilo kojeg od prethodnih zahteva, naznačen time, što prvi jednolančani polipeptid sadrži vezujući domen koji se specifično vezuje za TCR kompleks ili njegovu komponentu, i drugi jednolančani polipeptid sadrži vezujući domen koji se specifično vezuje za tumor-specifični antigen.
17. Smeša, naznačena time, što sadrži polipeptidni heterodimer iz bilo kojeg od zahteva 1 do 16 i farmaceutski prihvatljivi ekscipijent.
18. Polipeptidni heterodimer prema bilo kojem od zahteva 1 do 17, naznačen time, što se upotrebljava u: b)metodu usmeravanja aktivacije T-ćelija, pri čemu polipeptidni heterodimer sadrži vezujući domen koji se specifično vezuje za TCRα, TCRβ, CD3γ, CD3δ, CD3ε ili njihovu kombinaciju, i drugi vezujući domen koji se specifično vezuje za različiti cilj; ili (b) inhibiranju rasta, metastaza ili metastatskog rasta maligniteta, pri čemu polipeptidni heterodimer sadrži vezujući domen koji se specifično vezuje za TCRα, TCRβ, CD3γ, CD3δ, CD3ε, c-Met ili Ron. (c) lečenju autoimunskog ili zapaljenskog stanja, pri čemu polipeptidni heterodimer sadrži vezujući domen koji se specifično vezuje za TCRα, TCRβ, CD3γ, CD3δ, CD3ε ili CD28. (d) lečenju poremećaja ili bolesti vezanih za B-ćelije, pri čemu polipeptidni heterodimer sadrži vezujući domen koji se specifično vezuje za TCRα, TCRβ, CD3γ, CD3δ, CD3ε i drugi vezujući domen koji se specifično vezuje za CD19, CD20, CD79b ili HLA-DR.
19. Ekspresioni vektor, naznačen time, što je sposoban za ekspresiju polipeptidnog heterodimera iz bilo kojeg od zahteva 1 do 16, koji sadrži prvi polinukleotid koji kodira prvi jednolančani polipeptid i drugi polinukleotid koji kodira drugi jednolančani polipeptid.
20. Ćelija-domaćin, naznačena time, što sadrži (i) ekspresioni vektor iz zahteva 19 ili (ii) prvi i drugi ekspresioni vektor sposoban za ekspresiju prvog odnosno drugog jednolančanog polipeptida polipeptidnog heterodimera iz bilo kojeg od zahteva 1 do 16.
21. Metod pravljenja polipeptidnog hterodimera, naznačen time, što uključuje (a) kultivisanje ćelije-domaćina iz zahteva 20 pod uslovima pogodnim za ekspresiju prvog i drugog jednolančanog polipeptida, i (b) opciono izolovanje ili prečišćavanje iz kulture, heterodimera formiranih od prvog i drugog jednolančanog polipeptida.
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