Anti-il-1r3 antibodies for use in inflammatory conditions

A technology for inflammatory diseases and inflammatory diseases, applied in the field of anti-IL-1R3 antibodies for inflammatory diseases, can solve problems such as hindering therapeutic use

Pending Publication Date: 2020-02-21
SANOFI BIOTECH SAS
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] Unfortunately, existing IL-1 pathway inhibitors such as Kineret, ilaris or Arcalyst have certain disadvantages that hinder their therapeutic use

Method used

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  • Anti-il-1r3 antibodies for use in inflammatory conditions
  • Anti-il-1r3 antibodies for use in inflammatory conditions
  • Anti-il-1r3 antibodies for use in inflammatory conditions

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0188] Example 1: Human IL-1R3 biochemical ELISA

[0189] Analysis principle:

[0190] The NUNC Maxisorp 384-well microtiter plate was coated with Fc-tagged human IL-1R3. After the blocking process, the specific antibody from the B cell supernatant binds to the antigen and is subsequently detected with POD-labeled antibody. The sample is tested at 1:2 dilution.

[0191] material:

[0192] Plate: 384-well NUNC Maxisorp plate; catalog number 464718

[0193] Protein: Fc-tagged human IL-1R3 (concentration 1.5mg / ml; analysis concentration 0.5μg / mL)

[0194] Standard Ab: P013-02 (concentration 1mg / ml; initial analysis concentration 2μg / mL)

[0195] Detection Ab: anti-rabbit IgG, peroxidase-linked species-specific complete antibody (from donkey) (ECL); GE; catalog number NA9340; analytical dilution: 1:5000

[0196] PBS: buffer in the box, premixed PBS buffer, 10x; Roche Applied Sciences; catalog number 11666789001

[0197] BSA: Bovine Serum Albumin Fraction V from Bovine Serum; Roche Applied Sci...

Embodiment 2

[0216] Example 2: Characterization of h-IL-1R3 specific antibody that inhibits h-IL-1R3-receptor by luciferase reporter experiment

[0217] Analysis principle:

[0218] Inoculate poly-D-lysine-cell culture plates with 293T / 17-FR cells expressing NF-kB-RE firefly luciferase reporter molecule. After IL-1b stimulation, the 293T / 17-FR lysate was tested for activated NF-kB using Steady-Glo Luciferase Assay Kit. The supernatant containing functional antibodies binds to hIL-1R3 and inhibits NF-kB activation, which is shown by a low signal.

[0219] material:

[0220] Plate: Cell plate: 384-well PDL Costar cell culture plate; catalog number 3844

[0221] Analysis plate: 384-well lumitrac white-plate; Corning; catalog number 3572

[0222] Cells: 293T / 17-FR; analysis concentration 250,000 cells / ml

[0223] Protein: IL-1b (concentration 0.03mg / ml; analytical concentration 115pg / ml; working concentration 230pg / ml)

[0224] Standard Ab: P013_06 (concentration 0.2mg / ml; initial working concentration 6...

Embodiment 3

[0238] Example 3: Inhibition of NFκB expression in A549-NFκB-RE-Luc stably transfected cells after IL-1 (α / β) stimulation

[0239] Analysis principle:

[0240] A549-NFκB-RE-Luc stably transfected cells (Signosis) were aspirated into a 384-well plate and incubated overnight. On the second day, the cells stably transfected with the anti-IL1R3 antibody and bound A549-NFkB-RE-Luc were allowed, and then these cells were stimulated by adding IL-1 (α or β). This leads to the transcription of the luciferase gene due to the activation of the NFκB signaling pathway and can be measured by lysing the cells and adding luciferin.

[0241] Test whether the antibody can inhibit the activation of the NFkB pathway and thus reduce the luminescence signal.

[0242] material:

[0243] Plate: Sterile 384-well low-side white flat-bottomed polystyrene TC-treated microplate; Corning; catalog number 3570

[0244] Protein: IL-1α (P026_09); Recombinant human IL-1α / IL-1F1; 10μg / mL; R&D Systems; Catalog number 200-...

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Abstract

The present invention relates to methods for treating medical conditions and / or disorders characterized by uncontrolled or abnormal expression of members of the IL1R3 signaling pathway such as IL-lalpha, IL-Ibeta, IL-33, IL-36, IL1RA and / or IL1R3, as well as variants thereof. More specifically, the present invention relates to anti-IL1R3 antibodies for use in the treatment of an ILlR3-mediated inflammatory condition and / or disorder in a subject. Such conditions and disorders include but are not limited to inflammatory diseases, immune disorders, fibrotic disorders, eosinophilic disorders, infection, pain, a central nervous system disorder, an ophthalmologic disorder, Hereditary Systemic Inflammatory Diseases, and Systemic and Local Inflammatory Diseases and cancer associated chronic inflammation.

Description

Technical field [0001] The present invention relates to a method for treating medical conditions and / or diseases characterized by uncontrolled or abnormal expression of IL1R3 signaling pathway members such as IL-1α, IL-1β, IL-33, IL-36, IL1RA, and / or IL1R3. More specifically, the present invention relates to methods and uses of anti-IL-1R3 antibodies in inflammatory conditions and / or diseases. Background technique [0002] Interleukin 1 receptor accessory protein (IL1RAP), also known as IL1R3, is a co-receptor for type 1 interleukin 1 receptor (IL1R1) and is indispensable for the transmission of IL-1 signaling. Once IL-1 is bound, IL-1R1 associates with IL-1RAcP to form a functional signaling receptor complex, which stimulates NFkB activity. [0003] IL-33, its receptor ST2 and IL-1RAcP also form a complex (IL-33 / ST2 / IL-1RAcP) whose activity is similar to the IL-1β / IL-1R1 / IL-1RAcP complex in terms of NFkB activation. IL-36 (IL-36α (IL-1F6), IL-36β (IL-1F8) and IL-36γ (IL-1F9)), i...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61P29/00C07K16/28
CPCC07K16/2866A61K2039/505C07K2317/71C07K2317/76A61P29/00C07K2317/565
Inventor S·费舍尔K·贝克曼
Owner SANOFI BIOTECH SAS
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