FC-REGION VARIANTS WITH IMPROVED PROTEIN A-BINDING

• US 20170044246A1
• Filed: 07/14/2016
• Published: 02/16/2017
• Est. Priority Date: 01/15/2014
• Status: Abandoned Application

• Current Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Original Assignee: Hoffmann-La Roche Incorporated (Roche Holding AG)
• Inventors: Schlothauer, Tilman
• Application Number:
  US15/210,464
• Publication Number:
  US 20170044246A1
• Time in Patent Office: Days
• CPC Class Codes: A61K 2039/505 comprising antibodies
• CPC Class Codes: A61K 39/3955 against proteinaceous materials, e.g. enzymes, hormones, lymphokines
• CPC Class Codes: A61K 39/40 bacterial
• CPC Class Codes: A61K 45/06 Mixtures of active ingredients without chemical characterisation, e.g. antiphlogistics and cardiaca
• CPC Class Codes: A61K 9/0019 Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner non-active ingredients are additionally classified in A61K47/00
• CPC Class Codes: A61K 9/0048 Eye, e.g. artificial tears
• CPC Class Codes: A61P 27/02 Ophthalmic agents
• CPC Class Codes: A61P 27/06 Antiglaucoma agents or miotics
• CPC Class Codes: C07K 16/1271 from Micrococcaceae (F), e.g. Staphylococcus
• CPC Class Codes: C07K 16/22 against growth factors ; against growth regulators
• CPC Class Codes: C07K 2317/31 multispecific
• CPC Class Codes: C07K 2317/33 Crossreactivity, e.g. for species or epitope, or lack of said crossreactivity
• CPC Class Codes: C07K 2317/524 CH2 domain
• CPC Class Codes: C07K 2317/526 CH3 domain
• CPC Class Codes: C07K 2317/53 Hinge
• CPC Class Codes: C07K 2317/54 F(ab')2
• CPC Class Codes: C07K 2317/64 comprising a combination of variable region and constant region components
• CPC Class Codes: C07K 2317/71 Decreased effector function due to an Fc-modification
• CPC Class Codes: C07K 2317/92 Affinity (KD), association rate (Ka), dissociation rate (Kd) or EC50 value
• CPC Class Codes: C07K 2317/94 Stability, e.g. half-life, pH, temperature or enzyme-resistance

Abstract

Herein is reported a polypeptide comprising a first polypeptide and a second polypeptide each comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain and an immunoglobulin CH3-domain, wherein the first, the second, or the first and the second polypeptide comprise the mutation Y436A (numbering according to the EU index).

Claims

1. A polypeptide comprising

a first polypeptide and a second polypeptide each comprising in N-terminal to C-terminal direction at least a portion of an immunoglobulin hinge region, which comprises one or more cysteine residues, an immunoglobulin CH2-domain, and an immunoglobulin CH3-domain,

wherein the first, or the second, or the first and the second polypeptide comprise the mutation Y436A (numbering according to the EU index).

2. The polypeptide according to claim 1, characterized in that the first and the second polypeptide comprise the mutation Y436A.

3. The polypeptide according to claim 1, characterized in that the polypeptide does not specifically bind to the human FcRn and does specifically bind to Staphylococcal protein A.

4. The polypeptide according to claim 1, characterized in that

a) the first polypeptide further comprises the mutations Y349C, T366S, L368A and Y407V and the second polypeptide comprises the mutations S354C and T366W, and/or

b) i) the first and the second polypeptide comprise the mutations H310A, H433A and Y436A, or

ii) the first and the second polypeptide comprise the mutations L251D, L314D and L432D, or

iii) the first and the second polypeptide comprise the mutations L251S, L314S and L432S, or

iv) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations H310A, H433A and Y436A, or

v) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251D, L314D and L432D, or

vi) the first polypeptide comprises the mutations I253A, H310A and H435A and the second polypeptide comprises the mutations L251S, L314S and L432S.

5. The polypeptide according to claim 1, characterized in that the immunoglobulin hinge region, the immunoglobulin CH2-domain and the immunoglobulin CH3-domain are of the human IgG1 subclass.

6. The polypeptide according to claim 1, characterized in that the first polypeptide and the second polypeptide further comprise the mutations L234A and L235A.

7. The polypeptide according to claim 1, characterized in that the immunoglobulin hinge region, the immunoglobulin CH2-domain and the immunoglobulin CH3-domain are of the human IgG2 subclass.

8. The polypeptide according to claim 1, characterized in that the immunoglobulin hinge region, the immunoglobulin CH2-domain and the immunoglobulin CH3-domain are of the human IgG4 subclass.

9. The polypeptide according to claim 1, characterized in that the first polypeptide and the second polypeptide further comprise the mutations S228P and L235E.

10. The polypeptide according to claim 1, characterized in that the first polypeptide and the second polypeptide further comprise the mutation P329G.

11. The polypeptide according to claim 1, characterized in that the polypeptide is a bispecific antibody.

14. A pharmaceutical formulation comprising a polypeptide according to claim 1 and a pharmaceutically acceptable carrier.

20. A method of treating an individual having an ocular disease comprising administering to the individual an effective amount of a polypeptide according to claim 1.

23. The method of claim 20, wherein the administering comprises intravitreal injection of the polypeptide to the individual.

24. The method of claim 20, further comprising administering to the individual an additional therapeutic agent selected from the group consisting of Tryptophanyl-tRNA synthetase, anti-VEGF PEGylated aptamer, squalamine, anecortave acetate for depot suspension, Combretastatin A4 Prodrug, pegaptanib, mifepristone-ru486, subtenon triamcinolone acetonide, intravitreal crystalline triamcinolone acetonide, prinomastat, fluocinolone acetonide, a VEGFR inhibitor, a VEGF-Trap, 4-(4-bromo-2-fluoroanilino)-6-methoxy-7-(1-methylpiperidin-4-ylmethoxy)quinazoline, 4-(4-fluoro-2-methylindol-5-yloxy)-6-methoxy-7-(3-pyrrolidin-1-ylpropoxy)quinazoline, vatalanib, sunitinib, linomide, an inhibitor of integrin vβ

3 function, and angiostatin.

25. A method for transporting a soluble receptor ligand from the eye into the blood circulation in an individual comprising administering to the individual an effective amount of a polypeptide according to claim 1 to transport a soluble receptor ligand from the eye over the blood-ocular-barrier into the blood circulation.

26. A method for the removal of one or more soluble receptor ligands from the eye in an individual comprising administering to the individual an effective amount of a polypeptide according to claim 1 to remove one or more soluble receptor ligands from the eye.

12. (canceled)

13. (canceled)

15-19. -19. (canceled)

21. The method of claim 21, wherein the ocular disease is an ocular vascular disease.

22. The method of claim 21, wherein the ocular vascular disease is selected from the group consisting of wet age-related macular degeneration, dry age-related macular degeneration, diabetic macular edema, cystoid macular edema, non-proliferative diabetic retinopathy, proliferative diabetic retinopathy, cystoid macular edema, vasculitis, papilloedema, retinitis, conjunctivitis, uveitis, choroiditis, multifocal choroiditis, ocular histoplasmosis, blepharitis, Sjogren'"'"'s disease, and eye diseases associated with ocular neovascularization, vascular leakage, and/or retinal edema.