MODIFIED PROTEINS AND PEPTIDES
The present invention relates to modified proteins and peptides that have reduced ability to bind to pre-existing antibodies. Such modified protein/peptide molecules can comprise C-terminal additions, extensions or tags and/or certain amino acid substitutions. Such modified molecules (e.g. fusions and conjugates) comprise proteins, peptides, antigen binding molecules, antibodies or antibody fragments such as single variable domains e.g. human immunoglobulin (antibody) single variable domains, and also single variable domains derived from non-human sources such as a llama or camel, e.g. a VHH including a nanobody TM (described in e.g. WO 94/04678 and WO 95/04079 inter alia). The invention further relates to uses, formulations, compositions comprising such modified C terminally extended and/or amino acid substituted molecules and also to methods of production and expression of these molecules.
1 . (canceled)
2 . A method of treating or preventing at least one disease, disorder or condition selected from among an inflammatory disease or disorder, a respiratory disease or disorder, or a pulmonary disease or disorder, comprising:
administering to a subject a therapeutically or prophylactically effective amount of a composition comprising a single immunoglobulin variable domain (dAb), wherein said dAb comprises a modification selected from among one or more of (a) an affinity tag at the C-terminus; (b) an amino acid substitution at any one of positions 14, 41, 108, 110 or 112; and (c) a C-terminal extension which comprises an amino acid extension of from one amino acid to 5 amino acids, whereby one or more undesirable side effects of administering the dAb are prevented or reduced.
3 . The method of claim 2 , wherein said tag is a myc-tag, a FLAG tag, a His-tag, a chemical modification, a protein domain or any combination thereof.
4 . The method of claim 3 , wherein said chemical modification is polyethylene glycol (PEG).
5 . The method of claim 3 , wherein said protein domain is a Fc domain.
6 . The method of claim 2 , wherein said amino acid substitution is selected from among P14A, P14K, P14Q, P14T, P41A, L108A, L108 Q, T110A and S112A.
7 . The method of claim 2 , wherein said C-terminal amino acid extension in (c) is selected from among A, AS, AST, ASTK, ASTKG, AAA and T.
8 . The method of claim 2 , wherein said modified dAb comprises a C-terminal amino acid extension of one amino acid.
9 . The method of claim 8 , wherein said one amino acid is alanine.
10 . The method of claim 2 , wherein said at least one disease, disorder or condition is selected from: psoriasis, arthritis, multiple sclerosis, inflammatory bowel disease, Crohn's disease, ulcerative colitis; von Willebrand disease, chronic obstructive pulmonary disease (COPD), chronic bronchitis, chronic obstructive bronchitis, emphysema, lung inflammation, asthma, pneumonia, hypersensitivity pneumonitis, pulmonary infiltrate with eosinophilia, environmental lung disease, bronchiectasis, cystic fibrosis, interstitial lung disease, primary pulmonary hypertension, pulmonary thromboembolism, disorders of the pleura, disorders of the mediastinum, disorders of the diaphragm, hypoventilation, hyperventilation, sleep apnea, acute respiratory distress syndrome, mesothelioma, sarcoma, graft rejection, graft versus host disease, lung cancer, allergic rhinitis, allergy, asbestosis, aspergilloma, aspergillosis, bronchiectasis, chronic bronchitis, emphysema, eosinophilic pneumonia, idiopathic pulmonary fibrosis, invasive pneumococcal disease, influenza, nontuberculous mycobacteria, pleural effusion, pneumoconiosis, pneumocytosis, pulmonary actinomycosis, pulmonary alveolar proteinosis, pulmonary anthrax, pulmonary edema, pulmonary embolus, pulmonary inflammation, pulmonary histiocytosis X, pulmonary hypertension, pulmonary nocardiosis, pulmonary tuberculosis, pulmonary veno-occlusive disease, rheumatoid lung disease, sarcoidosis, Wegener's granulomatosis, Acute lung injury (ALI), Acute Respiratory Distress syndrome (ARDS) and complications thereof.
11 . The method of claim 2 , wherein said modified dAb is administered in a repeat dosing regimen.
12 . The method of claim 2 , wherein said subject is not screened for anti-drug antibody (ADA) titres prior to administration of said modified dAb.
13 . The method of claim 10 , wherein said method is for prevention or treatment of psoriasis.
14 . The method of claim 13 , wherein said modified dAb is administered in a repeat dosing regimen.
15 . The method of claim 13 , wherein said modified dAb comprises the sequence of SEQ ID NO:16, or a sequence having at least 90% sequence identity to SEQ ID NO:16.
16 . The method of claim 2 , wherein said modified dAb is derived from a Camelid heavy chain (VHH).
17 . The method of claim 10 , wherein said method is for prevention or treatment of arthritis.
18 . The method of claim 17 , wherein said modified dAb comprises the sequence of SEQ ID NO:7, a sequence having at least 90% sequence identity to SEQ ID NO: 7, the sequence of SEQ ID NO:8, or a sequence having at least 90% sequence identity to SEQ ID NO:8.
19 . The method of claim 10 , wherein said method is for prevention or treatment of Von Willebrand's disease.
20 . The method of claim 19 , wherein said modified dAb comprises the sequence of SEQ ID NO:9, or a sequence having at least 90% sequence identity to SEQ ID NO:9.
21 . The method of claim 2 , wherein said administering comprises subcutaneous, intravenous or intramuscular injection.
22 . The method of claim 2 , wherein said administering comprises parenteral, oral, rectal, transmucosal, ocular, pulmonary or GI tract delivery.