HRP20171080T1 - Protutijela s modificiranim affinitetom za fcrn koja potiču uklanjanje antigena - Google Patents
Protutijela s modificiranim affinitetom za fcrn koja potiču uklanjanje antigena Download PDFInfo
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- HRP20171080T1 HRP20171080T1 HRP20171080TT HRP20171080T HRP20171080T1 HR P20171080 T1 HRP20171080 T1 HR P20171080T1 HR P20171080T T HRP20171080T T HR P20171080TT HR P20171080 T HRP20171080 T HR P20171080T HR P20171080 T1 HRP20171080 T1 HR P20171080T1
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- amino acid
- ala
- instead
- acid substitutions
- antibody
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- 239000000427 antigen Substances 0.000 title claims 16
- 108091007433 antigens Proteins 0.000 title claims 16
- 102000036639 antigens Human genes 0.000 title claims 16
- 101150050927 Fcgrt gene Proteins 0.000 title 1
- 125000003275 alpha amino acid group Chemical group 0.000 claims 92
- 238000006467 substitution reaction Methods 0.000 claims 47
- 230000027455 binding Effects 0.000 claims 33
- 108010068617 neonatal Fc receptor Proteins 0.000 claims 19
- 150000001413 amino acids Chemical class 0.000 claims 11
- 108090000623 proteins and genes Proteins 0.000 claims 4
- 238000000034 method Methods 0.000 claims 3
- HNDVDQJCIGZPNO-UHFFFAOYSA-N histidine Natural products OC(=O)C(N)CC1=CN=CN1 HNDVDQJCIGZPNO-UHFFFAOYSA-N 0.000 claims 2
- 238000004519 manufacturing process Methods 0.000 claims 2
- 101710181478 Envelope glycoprotein GP350 Proteins 0.000 claims 1
- 230000003042 antagnostic effect Effects 0.000 claims 1
- 230000035772 mutation Effects 0.000 claims 1
- 239000000825 pharmaceutical preparation Substances 0.000 claims 1
- 238000012216 screening Methods 0.000 claims 1
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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
- C07K16/2803—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 immunoglobulin superfamily
- C07K16/283—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 immunoglobulin superfamily against Fc-receptors, e.g. CD16, CD32, CD64
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Claims (14)
1. Protutijelo, naznačeno time što sadrži domenu za vezanje na antigen i domenu za vezanje na ljudski FcRn, koja ima aktivnost vezanja na ljudski FcRn kod pH 5,5 i kod pH 7,0 i manju aktivnost vezanja na antigen kod pH 5,5 nego kod pH 7,0, gdje je aktivnost vezanja na ljudski FcRn kod pH 7,0 i na 25 °C veća nego KD 3,2 mM, gdje je domena za vezanje na ljudski FcRn Fc domena dobivena zamjenom različite aminokiselina umjesto najmanje jedne aminokiseline u Fc domeni ljudskog IgG1, IgG2, IgG3 ili IgG4.
2. Protutijelo u skladu s patentnim zahtjevom 1, naznačeno time što je omjer između aktivnosti vezanja na antigen kod pH 5,5 i kod pH 7,0 najmanje 2 u vrijednosti KD (kod pH 5,5)/KD (kod pH 7,0).
3. Protutijelo u skladu s patentnim zahtjevom 1 ili 2, naznačeno time što sadrži aminokiselinsku mutaciju u domeni za vezanje na antigen, koja se sastoji u zamjeni histidinom najmanje jedne aminokiseline u domeni za vezanje na antigen ili ubacivanju najmanje jednog histidina.
4. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 3, naznačeno time što se domena za vezanje na antigen dobiva iz biblioteke domena za vezanje na antigen.
5. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 4, naznačeno time što je domena za vezanje na ljudski FcRn domena za vezanje na ljudski FcRn koja sadrži aminokiselinski slijed sa zamjenom različitom aminokiselinom umjesto najmanje jedne aminokiseline koja se bira između onih na položajima 237, 238, 239, 248, 250, 252, 254, 255, 256, 257, 258, 265, 270, 286, 289, 297, 298, 303, 305, 307, 308, 309, 311, 312, 314, 315, 317, 325, 332, 334, 360, 376, 380, 382, 384, 385, 386, 387, 389, 424, 428, 433, 434, te 436 (EU numeriranje) u Fc domeni ljudskog IgG1, IgG2, IgG3 ili IgG4.
6. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 5, naznačeno time što sadrži domenu za vezanje na ljudski FcRn koja sadrži aminokiselinsku zamjenu u Fc domeni ljudskog IgG1, IgG2, IgG3 ili IgG4, koja sadrži najmanje jednu aminokiselinsku zamjenu koju se bira između:
aminokiselinske zamjene Met umjesto Gly na položaju 237;
aminokiselinske zamjene Ala umjesto Pro na položaju 238;
aminokiselinske zamjene Lys umjesto Ser na položaju 239;
aminokiselinske zamjene Ile umjesto Lys na položaju 248;
aminokiselinske zamjene Ala, Phe, Ile, Met, Gln, Ser, Val, Trp ili Tyr umjesto Thr na položaju 250;
aminokiselinske zamjene Phe, Trp ili Tyr umjesto Met na položaju 252;
aminokiselinske zamjene Thr umjesto Ser na položaju 254;
aminokiselinske zamjene Glu umjesto Arg na položaju 255;
aminokiselinske zamjene Asp, Glu ili Gln umjesto Thr na položaju 256;
aminokiselinske zamjene Ala, Gly, He, Leu, Met, Asn, Ser, Thr ili Val umjesto Pro na položaju 257;
aminokiselinske zamjene His umjesto Glu na položaju 258;
aminokiselinske zamjene Ala umjesto Asp na položaju 265;
aminokiselinske zamjene Phe umjesto Asp na položaju 270;
aminokiselinske zamjene Ala, ili Glu umjesto Asn na položaju 286;
aminokiselinske zamjene His umjesto Thr na položaju 289;
aminokiselinske zamjene Ala umjesto Asn na položaju 297;
aminokiselinske zamjene Gly umjesto Ser na položaju 298;
aminokiselinske zamjene Ala umjesto Val na položaju 303;
aminokiselinske zamjene Ala umjesto Val na položaju 305;
aminokiselinske zamjene Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp ili Tyr umjesto Thr na položaju 307;
aminokiselinske zamjene Ala, Phe, Ile, Leu, Met, Pro, Gln ili Thr umjesto Val na položaju 308;
aminokiselinske zamjene Ala, Asp, Glu, Pro ili Arg umjesto Leu ili Val na položaju 309;
aminokiselinske zamjene Ala, His ili Ile umjesto Gln na položaju 311;
aminokiselinske zamjene Ala ili His umjesto Asp na položaju 312;
aminokiselinske zamjene Lys ili Arg umjesto Leu na položaju 314;
aminokiselinske zamjene Ala ili His umjesto Asn na položaju 315;
aminokiselinske zamjene Ala umjesto Lys na položaju 317;
aminokiselinske zamjene Gly umjesto Asn na položaju 325;
aminokiselinske zamjene Val umjesto Ile na položaju 332;
aminokiselinske zamjene Leu umjesto Lys na položaju 334;
aminokiselinske zamjene His umjesto Lys na položaju 360;
aminokiselinske zamjene Ala umjesto Asp na položaju 376;
aminokiselinske zamjene Ala umjesto Glu na položaju 380;
aminokiselinske zamjene Ala umjesto Glu na položaju 382;
aminokiselinske zamjene Ala umjesto Asn ili Ser na položaju 384;
aminokiselinske zamjene Asp ili His umjesto Gly na položaju 385;
aminokiselinske zamjene Pro umjesto Gln na položaju 386;
aminokiselinske zamjene Glu umjesto Pro na položaju 387;
aminokiselinske zamjene Ala ili Ser umjesto Asn na položaju 389;
aminokiselinske zamjene Ala umjesto Ser na položaju 424;
aminokiselinske zamjene Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Ser, Thr, Val, Trp ili Tyr umjesto Met na položaju 428;
aminokiselinske zamjene Lys umjesto His na položaju 433;
aminokiselinske zamjene Ala, Phe, His, Ser, Trp ili Tyr umjesto Asn na položaju 434;
i aminokiselinske zamjene His ili Phe umjesto Tyr na položaju 436 prema EU numeriranju.
7. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 5, naznačeno time što mu domena za vezanje na ljudski FcRn sadrži najmanje jednu aminokiselinu koja se bira između:
Met na aminokiselinskom položaju 237;
Ala na aminokiselinskom položaju 238;
Lys na aminokiselinskom položaju 239;
Ile na aminokiselinskom položaju 248;
Ala, Phe, Ile, Met, Gln, Ser, Val, Trp ili Tyr na aminokiselinskom položaju250;
Phe, Trp ili Tyr na aminokiselinskom položaju 252;
Thr na aminokiselinskom položaju 254;
Glu na aminokiselinskom položaju 255;
Asp, Glu ili Gln na aminokiselinskom položaju 256;
Ala, Gly, Ile, Leu, Met, Asn, Ser, Thr ili Val na aminokiselinskom položaju 257;
His na aminokiselinskom položaju 258;
Ala na aminokiselinskom položaju 265;
Phe na aminokiselinskom položaju 270;
Ala ili Glu na aminokiselinskom položaju 286;
His na aminokiselinskom položaju 289;
Ala na aminokiselinskom položaju 297;
Gly na aminokiselinskom položaju 298;
Ala na aminokiselinskom položaju 303;
Ala na aminokiselinskom položaju 305;
Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Met, Asn, Pro, Gln, Arg, Ser, Val, Trp ili Tyr na aminokiselinskom položaju 307;
Ala, Phe, Ile, Leu, Met, Pro, Gln ili Thr na aminokiselinskom položaju 308;
Ala, Asp, Glu, Pro ili Arg na aminokiselinskom položaju 309;
Ala, His ili Ile na aminokiselinskom položaju 311;
Ala ili His na aminokiselinskom položaju 312;
Lys ili Arg na aminokiselinskom položaju 314;
Ala ili His na aminokiselinskom položaju 315;
Ala na aminokiselinskom položaju 317;
Gly na aminokiselinskom položaju 325;
Val na aminokiselinskom položaju 332;
Leu na aminokiselinskom položaju 334;
His na aminokiselinskom položaju 360;
Ala na aminokiselinskom položaju 376;
Ala na aminokiselinskom položaju 380;
Ala na aminokiselinskom položaju 382;
Ala na aminokiselinskom položaju 384;
Asp ili His na aminokiselinskom položaju 385;
Pro na aminokiselinskom položaju 386;
Glu na aminokiselinskom položaju 387;
Ala ili Ser na aminokiselinskom položaju 389;
Ala na aminokiselinskom položaju 424;
Ala, Asp, Phe, Gly, His, Ile, Lys, Leu, Asn, Pro, Gln, Ser, Thr, Val, Trp ili Tyr na aminokiselinskom položaju 428;
Lys na aminokiselinskom položaju 433;
Ala, Phe, His, Ser, Trp ili Tyr na aminokiselinskom položaju 434; i
His ili Phe kao aminokiselina na položaju 436 (EU numeriranje) u Fc domeni ljudskog IgG1, IgG2, IgG3 ili IgG4.
8. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 7, naznačeno time što ima antagonističku aktivnost.
9. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 8, naznačeno time što se veže na membranski antigen ili topljivi antigen.
10. Protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 9, naznačeno time što se protutijelo bira između kimernog protutijela, humaniziranog protutijela ili ljudskog protutijela.
11. Farmaceutski pripravak, naznačen time što sadrži protutijelo u skladu s bilo kojim od patentnih zahtjeva 1 do 10.
12. Postupak proizvodnje protutijela, naznačen time što obuhvaća korake:
(a) odabir protutijela s većom aktivnošću vezanja na ljudski FcRn kod pH 7,0 nego prije izmjene najmanje jedne aminokiseline u domeni za vezanje na ljudski FcRn protutijela s aktivnošću vezanja na ljudski FcRn kod pH 5,5;
(b) izmjena najmanje jedne aminokiseline u domeni za vezanje na antigen protutijela i odabira protutijela s većom aktivnošću vezanja na antigen kod pH 7,0 nego kod pH 5,5;
(c) dobivanje gena koji kodira protutijelo u kojem su domena za vezanje na ljudski FcRn i domena za vezanje na antigen dobivene u (a) i (b) međusobno povezane; i
(d) proizvodnja protutijela uz upotrebu gena dobivenog u (c).
13. Postupak proizvodnje protutijela, naznačen time što obuhvaća korake:
(a) odabir protutijela s većom aktivnošću vezanja na ljudski FcRn kod pH 7,0 nego prije izmjene najmanje jedne aminokiseline u domeni za vezanje na ljudski FcRn protutijela s aktivnošću vezanja na ljudski FcRn kod pH 5,5;
(b) odabir protutijela s većom aktivnošću vezanja na antigen kod pH 7,0 nego kod pH 5,5;
(c) dobivanje gena koji kodira protutijelo u kojem su domena za vezanje na ljudski FcRn i domena za vezanje na antigen dobivene u (a) i (b) međusobno povezane; i
(d) proizvodnja protutijela uz upotrebu gena dobivenog u (c).
14. Postupak probiranja protutijela, naznačen time što obuhvaća korake:
(a) odabir protutijela s većom aktivnošću vezanja na ljudski FcRn kod pH 7,0 nego prije izmjene najmanje jedne aminokiseline u domeni za vezanje na ljudski FcRn protutijela s aktivnošću vezanja na ljudski FcRn kod pH 5,5; i
(b) izmjena najmanje jedne aminokiseline u domeni za vezanje na antigen protutijela i odabira protutijela s većom aktivnošću vezanja na antigen kod pH 7,0 nego kod pH 5,5.
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