Single domain antibodies directed against tumour necrosis factor-alpha and uses therefor

Abstract

The present invention relates to polypeptides derived from single domain heavy chain antibodies directed to Tumor Necrosis Factor-alpha. It further relates to single domain antibodies that are Camelidae VHHs. It further relates to methods of administering said polypeptides. It further relates to protocols for screening for agents that modulate the TNF-alpha receptor, and the agents resulting from said screening.

Description

RELATED APPLICATIONS [0001] This application is a continuation of U.S. patent application Ser. No. 10 / 534,348 filed May 9, 2005, currently pending, which is a National Stage of PCT / BE03 / 00192, filed Nov. 7, 2003, which claims priority to PCT / EP03 / 06581, filed Jun. 23, 2003 and PCT / EP03 / 07313, filed Jul. 8, 2003; this application also claims the benefit of U.S. provisional application Ser. No. 60 / 425,073, filed Nov. 8, 2002 and U.S. provisional application Ser. No. 60 / 425,063, filed Nov. 8, 2002.FIELD OF THE INVENTION [0002] The present invention provides polypeptides comprising one or more single domain antibodies directed towards tumor necrosis factor alpha (TNF-alpha). The present invention further relates to their use in diagnosis and therapy. Such antibodies may have a framework sequence with high homology to the human framework sequences. Compositions comprising antibodies to tumor necrosis factor alpha (TNF-alpha) alone or in combination with other drugs are described. BACKGRO...

AI Technical Summary

Benefits of technology

The present invention relates to an anti-TNF-alpha polypeptide that includes at least one single domain antibody. The polypeptide can be used to treat disorders associated with high levels of TNF-alpha, such as rheumatoid arthritis, Crohn's disease, and ulcerative colitis. The polypeptide can also be used in combination with other single domain antibodies, such as anti-IFN-gamma and anti-TNF-receptor, for more effective treatment. The invention also includes a method for identifying agents that can modulate the binding of the anti-TNF-alpha polypeptide to TNF-alpha.

Problems solved by technology

Yet none of the presently available drugs are completely effective for the treatment of autoimmune disease, and most are limited by severe toxicity.

In addition, it is extremely difficult and a lengthy process to develop a new chemical entity (NCE) with sufficient potency and selectivity to such target sequence.

However, conventional antibodies are difficult to raise against multimeric proteins where the receptor-binding domain of the ligand is embedded in a groove, as is the case with TNF-alpha.

The use of antibodies derived from sources such as mouse, sheep, goat, rabbit etc., and humanised derivatives thereof as a treatment for conditions which require a modulation of inflammation is problematic for several reasons.

Traditional antibodies are not stable at room temperature, and have to be refrigerated for preparation and storage, requiring necessary refrigerated laboratory equipment, storage and transport, which contribute towards time and expense.

Furthermore, the manufacture or small-scale production of said antibodies is expensive because the mammalian cellular systems necessary for the expression of intact and active antibodies require high levels of support in terms of time and equipment, and yields are very low.

Furthermore the large size of conventional antibodies, would restrict tissue penetration, for example, at the site of inflamed tissue.

Furthermore, traditional antibodies have a binding activity which depends upon pH, and hence are unsuitable for use in environments outside the usual physiological pH range such as, for example, in treating gastric bleeding, gastric surgery.

Furthermore, traditional antibodies are unstable at low or high pH and hence are not suitable for oral administration.

Furthermore, traditional antibodies have a binding activity, which depends upon temperature, and hence are unsuitable for use in assays or kits performed at temperatures outside biologically active-temperature ranges (e.g. 37±20° C.).

Another important drawback of conventional antibodies is that they are complex, large molecules and therefore relatively unstable, and they are sensitive to breakdown by proteases.

This means that conventional antibody drugs cannot be administered orally, sublingually, topically, nasally, vaginally, rectally or by inhalation because they are not resistant to the low pH at these sites, the action of proteases at these sites and in the blood and / or because of their large size.

Furthermore, subjects commonly experience physical and psychological stress prior to and upon receiving an injection.

Images