HRP20160857T1 - Bispecifična jednolančana protutijela, specifična za visokomolekulske ciljne antigene - Google Patents

Bispecifična jednolančana protutijela, specifična za visokomolekulske ciljne antigene Download PDF

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HRP20160857T1
HRP20160857T1 HRP20160857TT HRP20160857T HRP20160857T1 HR P20160857 T1 HRP20160857 T1 HR P20160857T1 HR P20160857T T HRP20160857T T HR P20160857TT HR P20160857 T HRP20160857 T HR P20160857T HR P20160857 T1 HRP20160857 T1 HR P20160857T1
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Peter Kufer
Claudia BLÜMEL
Roman Kischel
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Amgen Research (Munich) Gmbh
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    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
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    • C07—ORGANIC CHEMISTRY
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    • C07K16/2851—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the lectin superfamily, e.g. CD23, CD72
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    • C07—ORGANIC CHEMISTRY
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    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2863—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against receptors for growth factors, growth regulators
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
    • C07K16/3069—Reproductive system, e.g. ovaria, uterus, testes, prostate
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    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/10—Processes for the isolation, preparation or purification of DNA or RNA
    • C12N15/1034—Isolating an individual clone by screening libraries
    • C12N15/1037—Screening libraries presented on the surface of microorganisms, e.g. phage display, E. coli display
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/39558—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against tumor tissues, cells, antigens
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
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    • C07K2317/00—Immunoglobulins specific features
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    • C07K2317/34—Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues
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    • C07K2317/62—Immunoglobulins specific features characterized by non-natural combinations of immunoglobulin fragments comprising only variable region components
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    • C07K2317/70—Immunoglobulins specific features characterized by effect upon binding to a cell or to an antigen
    • C07K2317/73—Inducing cell death, e.g. apoptosis, necrosis or inhibition of cell proliferation
    • C07K2317/732—Antibody-dependent cellular cytotoxicity [ADCC]

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Claims (13)

1. Postupak odabira bispecifičnih jednolančanih protutijela, koja sadrže prvu vežuću domenu, koja se može vezati na epitop CD3, i drugu vežuću domenu, koja se može vezati na izvanstaničnu domenu membranskog antigena specifičnog za prostatu (PSMA), naznačen time što obuhvaća korake: (a) osiguravanje najmanje tri tipa stanica domaćina koje eksprimiraju (i) ljudsku izvanstaničnu domenu PSMA divljeg tipa sa SEQ ID:447 na površini stanice; (ii) mutirani oblik ljudskog PSMA divljeg tipa na površini stanice, gdje su aminokiselinski ostaci na položajima 140, 169, 191, 308, 334, 339, 344, 624, 626, 716, 717 i 721 mutirani u odgovarajuće aminokiselinske ostatke PSMA glodavca divljeg tipa; i (iii) izvanstaničnu domenu PSMA glodavca divljeg tipa na površini stanice; (b) stavljanje u kontakt svakog od tipova stanica domaćina (i), (ii) i (iii) u koraku (a) s bispecifičnim jednolančanim protutijelima i efektorskim T-stanicama; i (c) identifikacija i izdvajanje bispecifičnih jednolančanih protutijela koja posreduju kod lize stanica domaćina koje eksprimiraju ljudsku izvanstaničnu domenu PSMA divljeg tipa na površini stanice u skladu s (b)(i) i stanica domaćina koje eksprimiraju mutirani oblik ljudskog PSMA divljeg tipa na površini stanice u skladu s (b)(ii), no ne i stanica domaćina koje eksprimiraju t izvanstaničnu domenu PSMA glodavca divljeg tipa na površini stanice u skladu s b(iii).
2. Postupak odabira bispecifičnih jednolančanih protutijela, koja sadrže prvu vežuću domenu, koja se može vezati na epitop CD3, i drugu vežuću domenu, koja se može vezati na izvanstaničnu domenu fibroblast-aktivirajućeg proteina α (FAPα), naznačen time što obuhvaća korake: (a) osiguravanje najmanje tri tipa stanica domaćina koje eksprimiraju (i) ljudsku izvanstaničnu domenu FAPα divljeg tipa sa SEQ ID:448 na površini stanice; (ii) mutirani oblik ljudskog FAPα divljeg tipa na površini stanice, gdje su aminokiselinski ostaci na položajima 144, 185, 186, 229, 267, 273, 274, 278, 284, 301, 328, 329, 331, 335 i 362 mutirani u odgovarajuće aminokiselinske ostatke FAPα glodavca divljeg tipa; i (iii) izvanstaničnu domenu FAPα glodavca divljeg tipa na površini stanice; (b) stavljanje u kontakt svakog tipa stanica domaćina (i), (ii) i (iii) u koraku (a) s bispecifičnim jednolančanim protutijelima i efektorskim T-stanicama; i (c) identifikacija i izdvajanje bispecifičnih jednolančanih protutijela koja posreduju kod lize stanica domaćina koje eksprimiraju ljudsku izvanstaničnu domenu FAPα divljeg tipa na površini stanice u skladu s (b)(i) i stanica domaćina koje eksprimiraju mutirani oblik ljudskog FAPα divljeg tipa na površini stanice u skladu s (b)(ii), no ne i stanica domaćina koje eksprimiraju izvanstaničnu domenu FAPα glodavca divljeg tipa na površini stanice u skladu s b(iii).
3. Postupak odabira bispecifičnih jednolančanih protutijela, koja sadrže prvu vežuću domenu, koja se može vezati na epitop CD3, i drugu vežuću domenu, koja se može vezati na izvanstaničnu domenu receptora čimbenika rasta hepatocita (c-MET), endosijalina (TEM1) i tip 1 receptora inzulinu sličnog čimbenika rasta (IGF-1R), naznačen time što se sastoji u koracima: (a) identificiranja membranskih proksimalnih 640 aminokiselinskih ostataka u ljudskom i glodavačkom homologu izvanstanične domene c-MET, TEM1 ili IGF-1R; (b) osiguravanja stanica domaćina koje eksprimiraju (i) ljudske izvanstanične domene divljeg tipa izvanstanične domene c-MET, TEM1 ili IGF-1R na površini stanice; (ii) fuzijski protein koji sadrži ljudske membranske proksimalne 640 aminokiselinskih ostataka identificiranih u koraku (a) i glodavačke aminokiselinske ostatke >640 u c-MET, TEM1 ili IGF-1R; i (iii) izvanstaničnu domenu c-MET, TEM1 ili IGF-1R divljeg tipa glodavca; (c) stavljanja u kontakt stanica domaćina u skladu s korakom (b) s bispecifičnim jednolančanim protutijelima i efektorskim T-stanicama; i (d) identificiranja i izdvajanja bispecifičnih jednolančanih protutijela koja posreduju kod lize stanica domaćina u skladu s (b)(i) i (b)(ii), no ne i stanica domaćina u skladu s b(iii).
4. Postupak u skladu s bilo kojim od patentnih zahtjeva 1 do 3, naznačen time što prva vežuća domena se veže na CD3 epsilon (CD3ε) kod čovjeka, te Callithrix jacchus, Saguinus oedipus ili Saimiri sciureus.
5. Postupak u skladu s patentnim zahtjevom 4, naznačen time što prva vežuća domena sadrži područje VL koje sadrži CDR-L1, CDR-L2 i CDR-L3, koje se bira između: (a) CDR-L1, kao što je opisano u SEQ ID NO. 27, CDR-L2, kao što je opisano u SEQ ID NO. 28 i CDR-L3, kao što je opisano u SEQ ID NO. 29; (b) CDR-L1, kao što je opisano u SEQ ID NO. 117, CDR-L2, kao što je opisano u SEQ ID NO. 118 i CDR-L3, kao što je opisano u SEQ ID NO. 119; i (c) CDR-L1, kao što je opisano u SEQ ID NO. 153, CDR-L2, kao što je opisano u SEQ ID NO. 154 i CDR-L3, kao što je opisano u SEQ ID NO. 155.
6. Postupak u skladu s patentnim zahtjevom 4, naznačen time što prva vežuća domena sadrži područje VH koje sadrži CDR-H 1, CDR-H2 i CDR-H3, koje se bira između: (a) CDR-H1, kao što je opisano u SEQ ID NO. 12, CDR-H2, kao što je opisano u SEQ ID NO. 13 i CDR-H3, kao što je opisano u SEQ ID NO. 14; (b) CDR-H1, kao što je opisano u SEQ ID NO. 30, CDR-H2, kao što je opisano u SEQ ID NO. 31 i CDR-H3, kao što je opisano u SEQ ID NO. 32; (c) CDR-H1, kao što je opisano u SEQ ID NO. 48, CDR-H2, kao što je opisano u SEQ ID NO. 49 i CDR-H3, kao što je opisano u SEQ ID NO. 50; (d) CDR-H1, kao što je opisano u SEQ ID NO. 66, CDR-H2, kao što je opisano u SEQ ID NO. 67 i CDR-H3, kao što je opisano u SEQ ID NO. 68; (e) CDR-H1, kao što je opisano u SEQ ID NO. 84, CDR-H2, kao što je opisano u SEQ ID NO. 85 i CDR-H3, kao što je opisano u SEQ ID NO. 86; (f) CDR-H1, kao što je opisano u SEQ ID NO. 102, CDR-H2, kao što je opisano u SEQ ID NO. 103 i CDR-H3, kao što je opisano u SEQ ID NO. 104; (g) CDR-H1, kao što je opisano u SEQ ID NO. 120, CDR-H2, kao što je opisano u SEQ ID NO. 121 i CDR-H3, kao što je opisano u SEQ ID NO. 122; (h) CDR-H1, kao što je opisano u SEQ ID NO. 138, CDR-H2, kao što je opisano u SEQ ID NO. 139 i CDR-H3, kao što je opisano u SEQ ID NO. 140; (i) CDR-H1, kao što je opisano u SEQ ID NO. 156, CDR-H2, kao što je opisano u SEQ ID NO. 157 i CDR-H3, kao što je opisano u SEQ ID NO. 158; i (j) CDR-H1, kao što je opisano u SEQ ID NO. 174, CDR-H2, kao što je opisano u SEQ ID NO. 175 i CDR-H3, kao što je opisano u SEQ ID NO. 176.
7. Postupak u skladu s bilo kojim od patentnih zahtjeva 3 do 6, naznačen time što se druga vežuća domena veže na epitope/vežuća mjesta koja se bira iz skupine koju čine: epitopi/vežuća mjesta u četiri Ig domene sa SEQ ID:436, cisteinom bogata domena sa SEQ ID:437 ili beta-lanac sema domene sa SEQ ID:438 izvanstanične domene c-MET, epitopi/vežuća mjesta u mucinskoj domeni sa SEQ ID:440, tri EGF-u slične domene sa SEQ ID:441 ili Sushi/SCR/CCP domena sa SEQ ID:442 izvanstanične domene TEM1, i epitopi/vežuća mjesta u tri fibronektin tip III domene sa SEQ ID:444 i L2 domeni sa SEQ ID:445 izvanstanične domene IGF-1R.
8. Bispecifično jednolančano protutijelo, naznačeno time što sadrži prva vežuća domena, koja se može vezati na epitop CD3 epsilon (CD3ε) kod čovjeka i primata koji nije čimpanza, gdje je epitop dio aminokiselinskog slijeda koji pripada skupini koju čine SEQ ID NO. 2, 4, 6 i 8, a druga vežuća domena se bira iz skupine koju čine: (a) vežuća domena, koja se može vezati na izvanstaničnu domenu mutiranog ljudskog PSMA s aminokiselinskim slijedom kao što je opisan u SEQ ID NO: 447, gdje su aminokiselinski ostaci na položajima 140, 169, 191, 308, 334, 339, 344, 624, 626, 716, 717 i 721 mutirani u odgovarajuće aminokiselinske ostatke PSMA divljeg tipa glodavca, no ne i na izvanstaničnu domenu PSMA glodavca; (b) vežuća domena, koja se može vezati na izvanstaničnu domenu kimere mutiranog ljudskog FAPα s aminokiselinskim slijedom kao što je opisan u SEQ ID NO: 448, gdje su aminokiselinski ostaci na položajima 144, 185, 186, 229, 267, 273, 274, 278, 284, 301, 328, 329, 331, 335 i 362 mutirani u odgovarajuće aminokiselinske ostatke FAPα divljeg tipa glodavca, no ne i na izvanstaničnu domenu FAPα divljeg tipa; (c) vežuća domena, koja se može vezati na četiri Ig domene sa SEQ ID:436, cisteinom bogatu domenu sa SEQ ID:437 ili beta-lanac sema domene sa SEQ ID:438 izvanstanične domene c-MET; (d) vežuća domena, koja se može vezati na mucinsku domenu sa SEQ ID:440, tri EGF-u slične domene sa SEQ ID:441 ili Sushi/SCR/CCP domenu sa SEQ ID:442 izvanstanične domene TEM1; i (e) vežuća domena, koja se može vezati na tri fibronektin tip III domene sa SEQ ID:444 i L2 domenu sa SEQ ID:445 izvanstanične domene IGF-1R.
9. Nukleinskokiselinski slijed, naznačen time što kodira molekulu bispecifičnog jednolančanog protutijela u skladu s patentnim zahtjevom 8.
10. Vektor, naznačen time što sadrži nukleinskokiselinski slijed u skladu s patentnim zahtjevom 9.
11. Stanica domaćin, naznačena time što je se transformira ili transfektira vektorom u skladu s patentnim zahtjevom 10.
12. Postupak proizvodnje molekule bispecifičnog jednolančanog protutijela u skladu s patentnim zahtjevom 8, naznačen time što se navedeni postupak sastoji u uzgoju stanice domaćina u skladu s patentnim zahtjevom 11 u uvjetima koji omogućuju eksprimiranje polipeptida u skladu s patentnim zahtjevom 8 i prikupljanje proizvedenog polipeptida iz kulture.
13. Farmaceutski pripravak, naznačen time što sadrži molekulu bispecifičnog jednolančanog protutijela u skladu s patentnim zahtjevom 8, ili proizvedenog postupkom u skladu s patentnim zahtjevom 12.
HRP20160857TT 2008-10-01 2009-10-01 Bispecifična jednolančana protutijela, specifična za visokomolekulske ciljne antigene HRP20160857T4 (hr)

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US10193308P 2008-10-01 2008-10-01
EP09736398.0A EP2352763B2 (en) 2008-10-01 2009-10-01 Bispecific single chain antibodies with specificity for high molecular weight target antigens
PCT/EP2009/062794 WO2010037837A2 (en) 2008-10-01 2009-10-01 Bispecific single chain antibodies with specificity for high molecular weight target antigens

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