Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...
Patent
1997-04-15
1999-07-13
Draper, Garnette D.
Chemistry: molecular biology and microbiology
Micro-organism, tissue cell culture or enzyme using process...
Recombinant dna technique included in method of making a...
435 695, 4352523, 4353201, 435 71, 435 72, 536 235, 530351, 424 851, 424 852, 514 2, 930141, C12N 1519, C07K 1454
Patent
active
059225734
DESCRIPTION:
BRIEF SUMMARY
The present invention relates to mutants of the IL-1 receptor antagonist (IL-1ra ), with an improved inhibitory activity compared to the wild type antagonist, means and methods for their preparation, and their use in the therapeutic sector in all pathologies in which IL-1 is thought to be involved.
BACKGROUND OF THE INVENTION
IL-1 (interleukin-1) is the cytokine that the body produces, in response to infections, various kinds of attack or antigenic stimulation, to initiate a defence reaction of the inflammatory or immune type. IL-1 is a polypeptide of approx. 17.5 kDa in its mature form, produced mainly by the macrophages but also by epidermal, lymphoid, vascular and epithelial cells. IL-1 is one of the principal stimulating factors of both the inflammatory and immune responses and, in its circulating form, it is capable of acting as a hormone, inducing a broad spectrum of systemic changes at metabolic, neurological, haematological and endocrinological level. Thus, IL-1 exerts an influence on mesenchymal tissue remodelling, contributing both to destructive processes and to repair processes. Furthermore, IL-1 is an activator of lymphocytes and plays a fundamental role in the initiation and amplification of the immune response. IL-1 also possesses strong activity of the inflammatory type, for example stimulation of the production of prostanoids and of proteases in various cells, including chondrocytes, fibroblasts, synovial cells, and brain cells. Thus, IL-1 is involved in many components of the acute-phase response and is the endogenous mediator of fever (endogenous pyrogen). IL-1 can act in synergy with other cytokines, especially TNF.alpha., significantly amplifying its inflammatory activity.
Cloning of IL-1 has led to the identification of two active forms. The predominant form is IL-1.beta., synthesized as an inactive precursor of 269 amino acids (31 kDa), which is then cut by a protease to give rise to the active mature form (corresponding to amino acids 117-269 of the precursor). A form that occurs about a hundred times less frequently, and is generally associated with the cells, is IL1.alpha., which has about 26% homology with IL1.beta., and which is also synthesized as a precursor of 271 amino acids (which, however, possesses biological activity), which then gives rise to the mature form after proteolysis of the precursor. IL-1 represents a special case among the cytokines (together with fibroblast growth factor=FGF) in that it lacks a signal peptide and so is not secreted via the normal routes. The most abundant extracellular form therefore consists of the mature form of IL1.beta., which is thus responsible for the majority of the biological activities of IL-1, both immunostimulant and inflammatory.
In view of such activities, it has been hypothesized that IL-1 might have a role in the pathogenesis of inflammatory and autoimmune diseases. Thus, in the vast majority of pathologies of acute and chronic inflammation and in many autoimmune pathologies, increased production of IL-1.beta. has been identified as one of the main factors responsible for the pathology (Dinarello C.A. Blood 77: 1, 1991).
The biological activities of IL-1 are inhibited in the presence of specific inhibitors. In view of IL-1's fundamental role in the pathogenesis of many autoimmune diseases and of chronic inflammatory diseases with tissue destruction, it is suggested that inhibition of IL-1 could be useful in the treatment of these pathologies. IL-1ra (IL-1 receptor antagonist) is a cytokine that is structurally very similar to IL-1, but is synthesized with a signal peptide and secreted as mature glycosylated protein. A non-glycosylated intracellular form of IL-1ra , with seven extra amino acids and without a signal peptide, with activity comparable to that of secreted IL-1ra , has also been described. IL-1ra is capable of binding effectively to IL-1R.sub.I and much less well to IL-1R.sub.II. IL-1R.sub.I, the type I IL-1 receptor, is a receptor that belongs to the immunoglobulin superfamily, composed of an extracellular doma
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Boraschi Diana
Bossu Paola
Frascotti Gianni
Frigerio Francesco
Grandi Guido
Dompe S.p.A.
Draper Garnette D.
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