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Medicine composition containing interleukin-38 recombinant protein and application of medicine composition

A technology of interleukin and recombinant protein, applied in the field of pharmaceutical compositions containing interleukin-38 recombinant protein, can solve the problem of no treatment means

Inactive Publication Date: 2019-10-11
REYOUNG SUZHOU BIOLOGY SCI & TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, there is still no satisfactory treatment that can specifically target metabolic syndrome-related diseases

Method used

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  • Medicine composition containing interleukin-38 recombinant protein and application of medicine composition
  • Medicine composition containing interleukin-38 recombinant protein and application of medicine composition
  • Medicine composition containing interleukin-38 recombinant protein and application of medicine composition

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1: Expression of IL-38 recombinant protein

[0057] According to Sequence 1 and Sequence 2 in the sequence table, the IL-38 recombinant protein gene fragment was synthesized and connected into the pET-24 expression vector (Suzhou Synbio Biotechnology Co., Ltd.), and the recombinant plasmid was transformed into BL-21 competent strain (Bio -Rad). Wherein, Sequence 2 is the DNA sequence corresponding to the amino acid sequence shown in Sequence 1.

[0058] Inoculate the transformed BL-21 positive clone into 50mL of LB medium, cultivate at 37°C and 220rpm until the OD600 reaches about 0.6, then inoculate 2 bottles of 500ml LB medium with 1% inoculum, and continue to cultivate until the OD600 reaches 0.6, then add IPTG (final concentration 0.5mM) at 30°C, 220rpm to induce expression overnight.

[0059] The total volume of 1 L of BL21 bacterial solution induced to express overnight was centrifuged (8000×g, 15 min, 4° C.), and the bacterial cells were collected....

Embodiment 2

[0076] Embodiment 2: Enzyme-linked immunosorbent assay (ELISA) detects IL-38 protein and IL-36 protein and IL-36R-Fc affinity

[0077] Experimental steps:

[0078] (1) Coat the ELISA plate with IL-38 protein and IL-36 protein at a concentration of 2 μg / mL in a volume of 100 μL, and overnight at 4°C;

[0079] (2) Wash the plate 5 times with 0.1% PBST;

[0080] (3) Block the ELISA plate with 5% BSA solution;

[0081] (4) Wash the plate 5 times with 0.1% PBST;

[0082] (5) Dilute the IL-36R-Fc solution with an initial concentration of 50 μg / mL to 16.7 μg / mL, 5.56 μg / mL, 1.85 μg / mL, 0.62 μg / mL, 0.21 μg / mL according to a 1:3 concentration gradient , 0.07μg / mL, each gradient concentration of IL-36R-Fc solution was added to the ELISA plate, the final volume was 50μL, and incubated at 37°C for 1.5 hours;

[0083](6) Wash the plate 5 times with 0.1% PBST;

[0084] (7) Dilute the HRP-labeled anti-human Fc polyclonal antibody at 1:2000, add 100 μL per well to the ELISA plate, an...

Embodiment 3

[0088] Example 3: ELISA method to detect the competitive binding of IL-38 protein and IL-36 protein to IL-36R-Fc

[0089] Experimental steps:

[0090] (1) Coat the ELISA plate with IL-36 at a concentration of 2 μg / mL in a volume of 100 μL, overnight at 4°C;

[0091] (2) Wash the plate 5 times with 0.1% PBST;

[0092] (3) Block the ELISA plate with 5% BSA solution;

[0093] (4) Wash the plate 5 times with 0.1% PBST;

[0094] (5) Dilute the IL-38 protein solution with an initial concentration of 100 μg / mL to 10 μg / mL, 1 μg / mL, and 0.1 μg / mL in a concentration gradient of 1:10, and add IL-38 to each concentration gradient of IL-38 solution. The final concentrations of 36R-Fc and IL-36R-Fc were both 10 μg / mL, added to the ELISA plate with a final volume of 50 μl, and incubated at 37°C for 1.5 hours;

[0095] (6) Wash the plate 5 times with 0.1% PBST;

[0096] (7) Dilute the HRP-labeled anti-human Fc polyclonal antibody at 1:2000, add 100 μL per well to the ELISA plate, and ...

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Abstract

The invention discloses a medicine composition containing IL-38 recombinant protein and application of the medicine composition. The IL-38 recombinant protein comprises an amino acid sequence shown ina first sequence in a sequence table or a mutant of the amino acid sequence. The IL-38 recombinant protein can be taken as a receptor antagonist of an IL-1 or IL-36 cell factor for inhibiting immuno-inflammatory responses mediated by IL-1 or IL-36 and treating or delaying some metabolic syndrome-related diseases such as diabetes, obesity, the fatty liver disease and hypertension, and some immune-inflammation diseases such as the organ specificity autoimmune disease and the systemic autoimmune disease, wherein the immune-inflammation diseases comprise other tissue or organ damage diseases, such as asthma, caused by inflammatory responses.

Description

technical field [0001] The invention belongs to the field of biomedicine, and in particular relates to a pharmaceutical composition containing interleukin-38 (IL-38) recombinant protein and application thereof. Background technique [0002] Metabolic syndrome is a series of metabolic abnormalities caused by endocrine and cardiovascular diseases such as diabetes, obesity, dyslipidemia, and hypertension. Symptoms of this syndrome include impaired glucose tolerance, diabetes, central obesity, dyslipidemia, hypertension, etc. Metabolic syndrome-related diseases such as fatty liver, cirrhosis, etc., as follows: [0003] The normal liver contains no more than 5% fat. If there is excessive fat deposition in the liver, it is called fatty liver. It is mainly due to excessive fat and fatty acids input into the liver, and lipoprotein synthesis disorders in the liver, or the effects of certain drugs and chemical poisons. , affecting fat metabolism in the liver and exporting to the out...

Claims

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Application Information

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IPC IPC(8): A61K38/20A61P37/02A61P37/08A61P3/00A61P3/04A61P3/06A61P3/10A61P5/50A61P1/04A61P21/04A61P1/16A61P19/02A61P19/08A61P29/00A61P17/00A61P11/06C07K14/54C12N15/70C12N1/21C12R1/19
CPCA61K38/20C07K14/54A61P1/04A61P1/16A61P3/00A61P3/04A61P3/06A61P3/10A61P5/50A61P11/06A61P17/00A61P19/02A61P19/08A61P21/04A61P29/00A61P37/02A61P37/08C12N15/70C12N1/205C12R2001/19
Inventor 陈小波崔文俊姜国栋张佳春郭树华
Owner REYOUNG SUZHOU BIOLOGY SCI & TECH CO LTD
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