Binding members against IL-1R1

An IL-6, member technology, applied in the direction of antibodies, anti-inflammatory agents, anti-viral agents, etc., can solve problems such as non-response and no clear denial of IL-1RII

Inactive Publication Date: 2012-01-04
MEDIMMUNE LLC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, at this time it was not definitively negated that IL-1RII is able to recruit another signaling chain, although cells expressing high levels of IL-1RII have been shown to become unresponsive to IL-1β (25)

Method used

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  • Binding members against IL-1R1
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  • Binding members against IL-1R1

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0426] Example 1 Antibody Lead Isolation

[0427] 1.1 Screening

[0428] A large single-chain Fv (scFv) human antibody library based on the filamentous phage M13 cloned into a phage vector was used for screening (106, 107). Anti-IL-1R specific scFv antibodies were isolated from phage display libraries using a series of selection loops on biotinylated recombinant human IL-1R-Fc essentially as described above (108). Briefly, scFv-phage particles were incubated with 100 nM recombinant biotinylated human IL-1R-Fc fusion protein in solution (huIL-1R1-Fc and internal biochemistry as described in Materials and Methods). Following the manufacturer's instructions, antigen-binding ScFv-phages were then captured on streptavidin-coated paramagnetic beads ( M-280). The collected scFv phage particles were then rescued as above (109), and the selection process was repeated for a second round.

[0429] A representative number of individual clones from the second round of screening were g...

Embodiment 2

[0460] Example 2: Antibody Optimization

[0461] 2.1 Compatibility maturation

[0462] Antibody 4 and Antibody 1 were optimized using affinity-based phage and ribosome display screens.

[0463] Large scFv phage libraries derived from lead clones were constructed by mutating the heavy chain (V H ) and light chain (V L ) Complementary defined region 3 (CDR3) oligosaccharide nucleic acid-directed mutagenesis. The library was subjected to affinity-based phage display screening to select variants with high affinity for human IL-1R-Fc. In results, these are expected to show enhanced inhibitory activity of human IL-1β binding IL-1R1. Screening was performed essentially as described previously (108). Briefly, scFv-phage particles were incubated with recombinant biotinylated human IL-1R-Fc in solution. Then, ScFv phage-bound antigens were captured on streptavidin-coated paramagnetic beads following the manufacturer's instructions ( M-280). The selection process was then repe...

Embodiment 4

[0505] Example 4 provides comparable data for Antibody 6 with a different Fc format, Antibody 6 IgG1™ (Triple Mutant 234F, 235E and 331S)

[0506] Analytical Materials and Methods

[0507] Receptor-ligand (IL1-β) binding analyze

[0508] Receptor-ligand (Homogeneous Time-Resolved Fluorescence, Cis-Bio, Bedford, MA, USA) assay format for inhibition of HIS FLAG-tagged human IL-1β (in-house, expressed in E. coli; HIS FLAG IL-1β) was conjugated to the human histidine tag (HIS) IL 1RFc fusion protein (expressed in-house HEK EBNA, as described below). more specifically Analysis can be found in Mathis (1995) Clinical Chemistry 41(9), 1391-1397.

[0509] Results from the lead isolation process were screened without dilution and crude scFv containing periplasmic extracts were prepared in 200 mM HEPES buffer pH 7.4, 0.5 mM EDTA and 0.5 M sucrose. 5 μl of crude scFc samples were added to a 384-well low volume assay plate (Costar 3676). Then 2nM human HISIL 1RFc was added ...

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Abstract

This invention relates to binding members, especially antibody molecules, specific for interleukin 1 receptor 1 (IL-1R1). For example, isolated binding members specific for IL-1R1 which competes with IL-I and IL-1Ra for binding to IL-1R1 and binds I1-1R1 with a KD of 10pM or less when measured by KinexaTM. The binding members are useful for, inter alia, treatment of disorders mediated by IL-1R1 including rheumatoid arthritis, asthma and chronic obstructive pulmonary disease (COPD).

Description

technical field [0001] The present invention relates to binding members, in particular antibody molecules, for interleukin 1 receptor-1 (IL-1R1). The binding members are useful in the treatment of disorders mediated by IL1-R1, including rheumatoid arthritis, asthma, chronic obstructive pulmonary disease (COPD). The invention also relates to methods for the preparation of such binding members, methods of using the binding members of the invention for the treatment of disorders mediated by IL-1R1 and the use of the binding members of the invention for the preparation of medicaments for the treatment of disorders mediated by IL-1R1 . Background technique [0002] Interleukin 1 (IL-1) is a multifunctional cytokine that plays a major role in inflammatory responses during immune-mediated diseases and infections. IL-1 is produced by various cell types upon stimulation by bacterial products, cytokines or immune complexes. IL-1 exhibits autocrine and paracrine activity on various ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/28A61K39/395A61P11/00
CPCC07K16/2866C07K2317/92C07K2317/622C07K2316/96A61P3/10A61P9/10A61P11/00A61P11/06A61P19/02A61P19/10A61P25/28A61P29/00A61P31/18A61P35/00A61P35/02C07K2317/76C07K16/2803C07K16/2809C07K2317/565
Inventor J·I·坎贝尔D·J·科克伦S·C·克鲁威D·K·芬奇M·A·T·格罗夫斯D·C·洛
Owner MEDIMMUNE LLC
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