NZ733959B2

NZ733959B2 - Cysteine protease

Info

Publication number: NZ733959B2
Application number: NZ733959A
Authority: NZ
Inventors: Sofia Jarnum; Christian Kjellman; Emma Nordahl
Original assignee: Hansa Biopharma AB
Filing date: 2016-02-12
Publication date: 2024-10-30

Abstract

The present invention relates to a novel polypeptide which displays IgG cysteine protease activity, and in vivo and ex vivo uses thereof. Uses of the polypeptide include methods for the prevention or treatment of diseases and conditions mediated by IgG, and methods for the analysis of IgG.

Claims

1. A polypeptide having IgG cysteine protease activity and comprising a variant of the sequence of SEQ ID NO:2, which variant: (a) is at least 80% identical to SEQ ID NO: 2; (b) has a cysteine (C) at the position in said variant sequence which corresponds to position 94 of SEQ ID NO: 1; and (c) has, at the positions in said variant sequence which correspond to positions 84, 262, 284 and 286 of SEQ ID NO: 1, a lysine (K), a histidine (H), an aspartic acid (D) and an aspartic acid (D), respectively; wherein said polypeptide is at least 1.5 fold more effective at cleaving IgG than IdeS when measured in the same assay and is less immunogenic than IdeS; and wherein said variant of the sequence of SEQ ID NO: 2: (1) has a positively charged amino acid at the position in said variant which corresponds to position 130 of SEQ ID NO: 1; and/or (2) has a positively charged amino acid at the position in said variant which corresponds to position 131 of SEQ ID NO: 1; and/or (3) does not include the contiguous sequence NQTN; and/or (4) has deleted in its entirety the first twenty residues at the N terminus of SEQ ID NO: 2 comprising the contiguous sequence DSFSANQEIR YSEVTPYHVT (SEQ ID NO: 19).

2. A polypeptide according to claim 1, wherein the positively charged amino acid at the position in said variant which corresponds with position 130 of SEQ ID NO: 1 is arginine (R) or lysine (K), and/or wherein the positively charged amino acid at the position in said variant which corresponds with position 131 of SEQ ID NO: 1 is arginine (R) or lysine (K).

3. A polypeptide according to claim 1 or 2, wherein said variant of the sequence of SEQ ID NO: 2 is at least 80%, 90%, 95% or 99% identical to SEQ ID NO: 2.

4. A polypeptide according to any one of the preceding claims, which comprises or consists of the sequence of any one of SEQ ID NOs: 3 to 5 and 9 to 16.

5. A polypeptide according to any one of the preceding claims, wherein said sequence includes an additional methionine at the N terminus and/or a histidine tag at the C terminus.

6. A polypeptide according to any one of the preceding claims, wherein said polypeptide is at least 2.0 fold, 2.5 fold, 3.0 fold, 4.0 fold, 4.5 fold, 5.0 fold, 6.0 fold, 7.0 fold or 7.5 fold more effective than IdeS at cleaving IgG, when measured in the same assay.

7. A polypeptide according to any one of the preceding claims which is less immunogenic than IdeS.

8. A polypeptide according to claim 7, wherein the immunogenicity of said polypeptide is no more than 85% of the immunogenicity of IdeS when measured in the same assay.

9. A polynucleotide or expression vector which comprises a nucleic acid sequence encoding a polypeptide of any one of the preceding claims.

10. A host cell comprising the polynucleotide or expression vector of claim 9, wherein the host cell excludes a host cell in vivo in a human.

11. A host cell according to claim 10 which is a bacterial cell.

12. A host cell according to claim 10 or claim 11 which is a cell of E. coli.

13. A composition comprising a polypeptide according to any one of claims 1 to 8 and at least one pharmaceutically acceptable carrier or diluent.

14. A use of a polypeptide according to any one of claims 1 to 8 in the manufacture of a medicament for (a) treating a disease or condition mediated in whole or in part by pathogenic IgG antibodies or (b) improving the benefit to a subject of a gene therapy or organ transplant.

15. A use according to claim 14, wherein said disease or condition is selected from Cancer, Addison’s disease, Anti-GBM glomerulonephritis, Anti-neutrophil cytoplasmic antibody-associated vasculitides, Anti-NMDAR Encephalitis, Anti-phospholipid antibody syndrome (APS), Catastrophic APS, Autoimmune bullous skin diseases, Pemphigus foliaceus, fogo selvagem, pemphigus vulgaris, Autoimmune hemolytic anemia, Autoimmune hepatitis, Autoimmune neutropenia, Bullous pemphigoid, Celiac disease, Chronic utricaria, Complete congenital heart block, Diabetes type 1A, Epidermolysis bullosa acquisita, Essential mixed cryoglobulinemia, Goodpasture’s syndrome, Goitre, hyperthyroidism, infiltrative exopthalmos and infiltrative dermopathy, Guillain-Barré syndrome, Acute inflammatory demyelinating polyneuropathy, acute motor axonal neuropathy, Hemophilia, Acquired FVIII deficiency, Idiopathic thrombocytopenic purpura, Lambert-Eaton myasthenic syndrome, Mixed Connective Tissue Disease, Multiple Myeloma, Myasthenia gravis, Myasthenic crisis, Myocarditis, dilated cardiomyopathy, Neuromyelitis Optica, Primary biliary cirrhosis, Primary Progressive Multiple Sclerosis, Rheumatic heart disease, Rheumatoid Arthritis, Serum-sickness, immune complex hypersensitivity (type III), Systemic Lupus Erythematosis, Lupus nephritis, Stiff-person syndrome, Transplant rejection and Thrombotic Thrombocytopenic Purpura.

16. An ex vivo method for the cleavage of IgG, the method comprising contacting a sample containing IgG with a polypeptide according to any one of claims 1 to 8 under conditions which permit IgG cysteine protease activity to occur.

17. A method according to claim 16 which is conducted to generate Fc and Fab fragments and/or wherein the sample is a blood sample taken from a subject suffering from a disease or condition as defined in claim 14 or claim 15.