HRP20191337T1 - Proizvodnja heterodimernih proteina - Google Patents

Proizvodnja heterodimernih proteina Download PDF

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Publication number
HRP20191337T1
HRP20191337T1 HRP20191337TT HRP20191337T HRP20191337T1 HR P20191337 T1 HRP20191337 T1 HR P20191337T1 HR P20191337T T HRP20191337T T HR P20191337TT HR P20191337 T HRP20191337 T HR P20191337T HR P20191337 T1 HRP20191337 T1 HR P20191337T1
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full
vitro method
length
range
antibody
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HRP20191337TT
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Michael Gramer
Amitava Kundu
Ewald T.J. Van Den Bremer
Muriel VAN KAMPEN
Patrick PRIEM
Aran Frank Labrijn
Joyce I. MEESTERS
Joost J. Neijssen
Janine Schuurman
Paul Parren
Patrick Van Berkel
Werner L. VOS
Amout Gerritsen
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Genmab A/S
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    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
    • C07K1/107—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides
    • C07K1/113—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure
    • C07K1/1133—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length by chemical modification of precursor peptides without change of the primary structure by redox-reactions involving cystein/cystin side chains
    • 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
    • C07K16/2863—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
    • C07K16/2887—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against CD20
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/46—Hybrid immunoglobulins
    • C07K16/468—Immunoglobulins having two or more different antigen binding sites, e.g. multifunctional antibodies
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/20—Immunoglobulins specific features characterized by taxonomic origin
    • C07K2317/24—Immunoglobulins specific features characterized by taxonomic origin containing regions, domains or residues from different species, e.g. chimeric, humanized or veneered
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/30—Immunoglobulins specific features characterized by aspects of specificity or valency
    • C07K2317/31—Immunoglobulins specific features characterized by aspects of specificity or valency multispecific
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/40—Immunoglobulins specific features characterized by post-translational modification
    • C07K2317/41—Glycosylation, sialylation, or fucosylation
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/524—CH2 domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/526—CH3 domain
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/52—Constant or Fc region; Isotype
    • C07K2317/53—Hinge
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/50—Immunoglobulins specific features characterized by immunoglobulin fragments
    • C07K2317/55—Fab or Fab'
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • 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]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • 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
    • C07K2317/734—Complement-dependent cytotoxicity [CDC]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2317/00—Immunoglobulins specific features
    • C07K2317/90—Immunoglobulins specific features characterized by (pharmaco)kinetic aspects or by stability of the immunoglobulin

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • General Health & Medical Sciences (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Biochemistry (AREA)
  • Biophysics (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Peptides Or Proteins (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)

Claims (21)

1. Postupak in vitro za proizvodnju bispecifičnog protutijela pune duljine, naznačen time, da obuhvaća sljedeće korake: a) inkubacija prvog protutijela pune duljine s drugim protutijelom pune duljine pod reduciranim uvjetima koji su dovoljni za omogućavanje redukcije međulančanih disulfidnih veza u zglobnom području, i pritom CH3-područje od navedenog prvog protutijela pune duljine je prvo CH3-područje i posjeduje Arg na poziciji 409, a CH3-područje od navedenog drugog protutijela pune duljine je drugo CH3-područje te posjeduje: (i) Asp, Glu, Gly, Asn, Arg, Ser, Thr, Val ili Trp na poziciji 368; (ii) Phe, His, Lys, Arg ili Tyr na poziciji 399; (iii) Gly, Leu, Met ili Trp na poziciji 407 i Lys na poziciji 409, te pritom prvi CH3 ima Tyr na poziciji 407; ili (iv) aminokiselinu različitu od Phe, Arg ili Gly na poziciji 405 i ima Lys na poziciji 409, te pritom prvi CH3 dalje ima Phe na poziciji 405, tako da su sekvence od navedenih prvog i drugog CH3-područja različite i takve su, da je heterodimerna interakcija između navedenog prvog i drugog CH3-područja, jača nego svaka od homodimernih interakcija navedenih prvog i drugog CH3-područja, pri čemu su pozicije aminokiselina numerirane u skladu s EU-indeksom opisanim u Kabat-u, gdje navedeno prvo i drugo protutijelo pune duljine veže različite epitope, dok svako od navedenih prvog i drugog protutijela pune duljine: 1) posjeduje Fc-područje s aminokiselinskom sekvencom od izotipa IgG1; 2) posjeduje Fc-područje s aminokiselinskom sekvencom od izotipa IgG2; 3) posjeduje Fc-područje s aminokiselinskom sekvencom od izotipa IgG3; ili 4) posjeduje Fc-područje s aminokiselinskom sekvencom od izotipa IgG4; i pri čemu su navedeni prvo i drugo protutijelo humanizirana protutijela ili pri čemu svako od navedenih prvog i drugog protutijela pune duljine, obuhvaća teški lanac pune duljine ljudskog protutijela; te nadalje pri čemu korak a) obuhvaća inkubaciju u trajanju od najmanje 90 minuta na temperaturi od najmanje 20°C u prisutnosti 2-merkaptoetilamina s koncentracijom od najmanje 25 mM; a) podvrgavanje najmanje 10 mL od sastava koji je dobiven u koraku a) uvjetima oksidacije koji su dovoljni za omogućavanje oksidacije cisteina u bispecifičnom protutijelu pune duljine za međulančane disulfidne veze; i b) dobivanje bispecifičnog protutijela pune duljine.
2. Postupak in vitro prema patentnom zahtjevu 1, naznačen time, da oba i prvo i drugo protutijelo pune duljine sadrže sekvencu Cys-Pro-Pro-Cys u zglobnom području.
3. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da korak a) obuhvaća dodavanje kelatnog sredstva za metal, kao što je EDTA, EGTA ili limunska kiselina.
4. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da se korak a) provodi pod reduciranim uvjetima s redoks potencijalom između –150 i –600 mV, kao primjerice između –350 i –450 mV.
5. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da se prvo i drugo protutijelo pune duljine nalaze u puferu koji sadrži fosfat u rasponu od 1 – 100 mM, kao primjerice od 1 – 50 mM fosfata ili u rasponu od 1 – 25 mM, ili u rasponu od 5 – 20 mM.
6. Postupak in vitro prema patentnom zahtjevu 5, naznačen time, da pufer ima pH-vrijednost u rasponu od 4,5 – 8,5, kao primjerice u rasponu od 6,5 – 7,5.
7. Postupak in vitro prema patentnom zahtjevu 5, naznačen time, da je pufer odabran iz skupine koja se sastoji od a) 8,1 mM natrijevog fosfata (Na2HPO4–7H2O), 1,5 mM kalijevog fosfata (KH2PO4), 138 mM natrijevog klorida (NaCl), 2,7 mM kalijevog klorida (KCl) s pH 5,0; b) 8,1 mM natrijevog fosfata (Na2HPO4–7H2O), 1,5 mM kalijevog fosfata (KH2PO4), 138 mM natrijevog klorida (NaCl), 2,7 mM kalijevog klorida (KCl) s pH 7,0; 3) 20 mM Tris-HCl s pH 7,8.
8. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da korak b) obuhvaća pH-vrijednost u rasponu od 6 – 8,5.
9. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da korak b) obuhvaća redoks potencijal od najmanje –300 mV.
10. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da oksidacijski uvjeti u koraku b) obuhvaćaju prisutnost od najmanje 0,05 mM kisika.
11. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da oksidacijski uvjeti u koraku b) obuhvaćaju dodavanje kisika, pri čemu se dodavanje kisika provodi putem postupaka koji sadrže: mehaničko, primjerice pomoću tresenja/mućkanja, miješanja, povećanja tlaka ili stvaranja strujanja, ili pomoću škropljenja kisikom ili zrakom.
12. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da oksidacijski uvjeti u koraku b) obuhvaćaju oksidirajuće sredstvo, kao primjerice dehidroaskorbinsku kiselinu (dhAA).
13. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da korak b) obuhvaća podvrgavanje kromatografiji sastava koji je dobiven u koraku a), kao što je primjerice kromatografija na stupcu, ili podvrgavanje filtraciji, poželjno diafiltraciji kao što je tangencijalna protočna filtracija (TFF-eng. tangential flow filtration) ili filtracija normalnog protoka (NFF-eng. normal flow filtration).
14. Postupak in vitro prema zahtjevu 13, naznačen time, da se diafiltracija provodi putem cirkuliranja sastava kroz šuplju vlaknastu filtersku patronu koja obuhvaća minimalnu razlikovnu vrijednost u rasponu od 10 – 50 kDa, gdje je površinsko područje u rasponu od 0,05 – 1 m2, te s patronskim ulaznim tlakom u rasponu od 70 – 280 kPa, sve dok se ne provede od jednoga do sedam volumena puferske izmjene.
15. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da koncentracija bispecifičnog protutijela pune duljine u sastavu koji je dobiven iz koraka a), leži u rasponu od 1 – 100 g/L.
16. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da oksidacijski uvjeti u koraku b) obuhvaćaju dodavanje metalnog iona, poželjno metalnog iona odabranog iz skupine koja se sastoji od sljedećih: bakar, mangan, magnezij, željezo, nikal i kobalt, pri čemu je koncentracija metalnog iona poželjno u rasponu od 0,1 do 100 μM.
17. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da omjer prvog prema drugom protutijelu pune duljine u koraku a) leži u rasponu od 1:1,01 do 1:2.
18. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da ili prvo protutijelo pune duljine ili drugo protutijelo pune duljine nije u mogućnosti vezati se na Protein A i/ili Protein G.
19. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da prvo protutijelo i drugo protutijelo pune duljine sadrže različite lake lance.
20. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da ukupna koncentracija od prvog protutijela pune duljine i drugog protutijela pune duljine u koraku a) iznosi najmanje 0,25 mg/mL.
21. Postupak in vitro prema bilo kojem od prethodnih patentnih zahtjeva, naznačen time, da navedeno prvo protutijelo pune duljine obuhvaća Phe na poziciji 405 i Arg na poziciji 409, a navedeno drugo protutijelo pune duljine obuhvaća Leu na poziciji 405 i Lys na poziciji 409.
HRP20191337TT 2011-10-27 2019-07-23 Proizvodnja heterodimernih proteina HRP20191337T1 (hr)

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Application Number Priority Date Filing Date Title
US201161552272P 2011-10-27 2011-10-27
DKPA201100826 2011-10-27
EP12783931.4A EP2771364B1 (en) 2011-10-27 2012-10-26 Production of heterodimeric proteins
PCT/EP2012/071294 WO2013060867A2 (en) 2011-10-27 2012-10-26 Production of heterodimeric proteins

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US (3) US10344050B2 (hr)
EP (2) EP3674320A3 (hr)
JP (2) JP6475017B2 (hr)
KR (1) KR102049803B1 (hr)
CN (3) CN110964115B (hr)
AU (3) AU2012328322A1 (hr)
BR (1) BR112014010023B1 (hr)
DK (1) DK2771364T3 (hr)
ES (1) ES2739808T3 (hr)
HK (1) HK1201540A1 (hr)
HR (1) HRP20191337T1 (hr)
HU (1) HUE044633T2 (hr)
IL (2) IL232022B (hr)
IN (1) IN2014CN03887A (hr)
LT (1) LT2771364T (hr)
PL (1) PL2771364T3 (hr)
PT (1) PT2771364T (hr)
RS (1) RS59166B1 (hr)
SG (2) SG11201401422VA (hr)
SI (1) SI2771364T1 (hr)
WO (1) WO2013060867A2 (hr)

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