Polypeptides, antibody variable domains & antagonists

Abstract

The invention relates to anti-TNFR1 polypeptides and antibody single variable domains (dAbs) that are resistant to degradation by a protease, as well as antagonists comprising these. The polypeptides, dAbs and antagonists are useful for as therapeutics and/or prophylactics that are likely to encounter proteases when administered to a patient, for example for pulmonary administration, oral administration, delivery to the lung and delivery to the GI tract of a patient, as well as for treating inflammatory disease, such as arthritis or COPD.

1 . An anti-TNFα receptor immunoglobulin single variable domain comprising an amino acid sequence that is at least 93% identical to the amino acid sequence shown in SEQ ID NO: 224. 
     
     
         2 . The anti-TNFα receptor immunoglobulin single variable domain of  claim 1 , wherein the anti-TNFα receptor immunoglobulin single variable domain is a TNFα receptor type 1 antagonist. 
     
     
         3 . A TNFα receptor type 1 antagonist that competes with an immunoglobulin single variable domain comprising the amino acid sequence shown in SEQ ID NO: 224 for binding to TNFR1. 
     
     
         4 . A protease resistant immunoglobulin single variable domain comprising a TNFα receptor type 1 binding site, wherein the variable domain is resistant to protease when incubated with
 (i) a concentration (c) of at least 10 micrograms/ml protease at 37° C. for time (t) of at least one hour; or 
 (ii) a concentration (c′) of at least 40 micrograms/ml protease at 30° C. for time (t) of at least one hour, 
 wherein the variable domain comprises an amino acid sequence that is at least 90% identical to the amino acid sequence shown in SEQ ID NO:224. 
 
     
     
         5 . A method of oral delivery or delivery of a medicament to the lung or pulmonary tissue of a patient, wherein the method comprises administering to the patient a pharmaceutically effective amount of the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         6 . A method of treating a respiratory disease or an inflammatory condition in a patient in need thereof, the method comprising administering to the patient a therapeutically effective amount of the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         7 . A pulmonary delivery device containing the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         8 . The device of  claim 7 , wherein the device is an inhaler or an intranasal administration device. 
     
     
         9 . An oral formulation comprising the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         10 . The formulation of  claim 9 , wherein the formulation is a tablet, pill, capsule, liquid or syrup. 
     
     
         11 . A dual-specific ligand comprising the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         12 . An isolated or recombinant nucleic acid encoding a polypeptide comprising the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         13 . A vector comprising the nucleic acid of  claim 12 . 
     
     
         14 . A host cell comprising the nucleic acid of  claim 12 . 
     
     
         15 . A method of producing polypeptide comprising an immunoglobulin single variable domain, the method comprising maintaining a host cell of  claim 14  under conditions suitable for expression of the nucleic acid, whereby a polypeptide comprising an immunoglobulin single variable domain is produced. 
     
     
         16 . A pharmaceutical composition comprising the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         17 . A polypeptide comprising the anti-TNFα receptor immunoglobulin single variable domain of  claim 1 . 
     
     
         18 . A fusion protein comprising the polypeptide of  claim 17 . 
     
     
         19 . An isolated or recombinant nucleic acid encoding the fusion protein of  claim 19 .