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IL-4R antagonist and microorganism for generating the IL-4R antagonist

A technology of antagonists and microorganisms, which is applied in the preparation of the antagonists of the novel interleukin-4 receptor, the preparation of medicines for the treatment of asthma or tumors, and the fields of pharmaceutical compositions containing the antagonists, which can solve problems such as application limitations and the like

Inactive Publication Date: 2008-10-15
MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] At present, several methods have been tried to treat asthma, including the use of sIL-4R, mutant IL-4 (which can specifically bind to IL-4R but also does not induce biological activity), blocking IL-4 with IL-4 antibody Activity, etc., but the above-mentioned substances are all proteins, and the application is subject to certain restrictions

Method used

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  • IL-4R antagonist and microorganism for generating the IL-4R antagonist
  • IL-4R antagonist and microorganism for generating the IL-4R antagonist
  • IL-4R antagonist and microorganism for generating the IL-4R antagonist

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] Embodiment 1, IL-4R antagonist screening model and method

[0067] IL-4R antagonist assay activity model

[0068] The establishment of IL-4R antagonist screening model is based on the principle of competitive binding between antagonists and receptor ligands. Its principle model is as figure 1 shown.

[0069] If the sample to be tested contains an IL-4R antagonist, the antagonist can inhibit the combination of IL-4 and IL-4R, and reduce the binding of IL-4 and sIL-4R coated on the microtiter plate. In response, the light absorption value of the sample at 450nm decreased compared with the control, and according to the change of the light absorption value, it can be preliminarily determined whether the sample to be screened contains an IL-4R antagonist.

[0070] IL-4R antagonist screening method

[0071] IL-4 was dissolved in PBS solution, added to a 96-well plate and placed at 4°C for 8-12 hours, the coating solution was sucked off, 3% PBST (PBS with 0.1% Tween20) was...

Embodiment 2

[0072] Embodiment 2, the cultivation of IL-4R antagonist producing microorganisms

[0073] Streptomyces 4849 cultured on slant medium at 28°C for 7 days was used as the strain, inoculated in seed medium (100mL / 500mL), cultured with shaking at 28°C for 48h, then transferred to fermentation medium, and cultured with shaking at 28°C for 96h , using the screening method described in Example 1 to measure the activity of IL-4R antagonists in the fermentation broth samples obtained at each culture time.

Embodiment 3

[0074] Example 3, Isolation and identification of novel IL-4R antagonists derived from microorganisms

[0075] According to the method described in Example 2, 5 L of the fermented liquid (pH=8.5) was obtained and filtered, and the filtrate was adjusted to pH 7.0 and then passed through an X5 type macroporous adsorption resin (45 mm × 350 mm), discarding the effluent, and washing with 1000 ml of 50% acetone The eluate was collected and the acetone was evaporated, extracted with ethyl acetate and concentrated to obtain a crude product. Dissolve the crude product with a small amount of eluent, namely chloroform-ethyl acetate (5:1), load the sample on a silica gel H column (25mm×300mm), and elute with 500ml of eluent to collect the yellow band I. The yellow band I is concentrated and loaded on a silica gel H column (20mm×200mm), eluted with 100ml of cyclohexane-ethyl acetate (2:1) as the eluent, and the yellow part is collected in separate tubes, and yellow crystals can be obtaine...

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Abstract

The invention relates to a new interleukin-4 receptor (IL-4R) antagonist (shown in formula I) and a pharmaceutical composition containing the antagonist. The invention also relates to an application of the IL-4R antagonist in preparing medicines for treating asthma or tumor. The invention further relates to microorganism for generating the new IL-4R antagonist (shown in formula I) and a method for preparing the new IL-4R antagonist by using the microorganism.

Description

technical field [0001] The present invention relates to a novel interleukin 4 receptor (IL-4R) antagonist and a pharmaceutical composition containing the antagonist. The present invention also relates to the use of the antagonist of interleukin 4 receptor (IL-4R) in the preparation of medicines for treating asthma or tumors. The present invention also relates to a microorganism producing the novel interleukin 4 receptor (IL-4R) antagonist and a method for preparing the novel interleukin 4 receptor (IL-4R) antagonist using the microorganism. Background of the invention [0002] As a multifunctional cytokine, interleukin-4 (IL-4) plays a key role in allergic inflammatory reactions such as asthma. IL-4 exerts its biological effects by binding to the interleukin-4 receptor (IL-4R) on the cell surface. Soluble interleukin 4 receptor (sIL-4R) lacks transmembrane and intracellular domains, and cannot produce signal transduction to mediate the biological activity of IL-4 after bin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/498A61P35/00A61P11/06C12N1/20C12P17/12C12R1/465
Inventor 李元吴剑波王柯郭连宏张洋
Owner MEDICINE & BIOENG INST OF CHINESE ACAD OF MEDICAL SCI
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