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Composition, comprising beta-glucan-peptide complex and antibiotic, for prevention, alleviation, or treatment of inflammatory disease

A technology of inflammatory diseases and compositions, which is applied in the field of prevention or treatment of inflammatory diseases, can solve problems that have not yet been studied, and achieve the effect of increasing individual survival rate, improving or treating inflammatory diseases

Inactive Publication Date: 2021-02-02
KISHIN BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the complex of β-glucan and peptide combined with schizophyllan, etc., its method of combined use with antibiotics, etc. have not been studied

Method used

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  • Composition, comprising beta-glucan-peptide complex and antibiotic, for prevention, alleviation, or treatment of inflammatory disease
  • Composition, comprising beta-glucan-peptide complex and antibiotic, for prevention, alleviation, or treatment of inflammatory disease
  • Composition, comprising beta-glucan-peptide complex and antibiotic, for prevention, alleviation, or treatment of inflammatory disease

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0085] Experiment preparation

[0086] cells and CLP mice

[0087] As a mouse macrophage cell line (macrophage cell line) RAW2647 (American Type Culture Collection Center, ATCCTIB-71; American Type Culture Collection) cells using 10% fetal bovine serum (fetal bovine serum, FBS; Invitrogen), Sodium pyruvate (sodium pyruvate), non-essential amino acids, penicillin G (100 IU / ml), and streptomycin (100 ug / ml) were cultured in DMEM medium (Invitrogen).

[0088] A mouse model of sepsis induced by appendix ligation and puncture (CLP) was prepared using 6-week-old C57BL / 6 female mice (Samtako, Korea). All animal-related procedures were approved by the Institutional Animal Care and Use Committee of Hanyang University, Korea. The administration substances listed in Table 1 below were intraperitoneally administered to CLP mice three times (3 hours, 6 hours and 12 hours). SPG-SC is a peptide complex having the sequence of SPG-SH3 randomly mixed, and the antibiotic (gentamycin) and c...

Embodiment 2

[0097] ROS production inhibitory effect of SPG-SH3 (in vitro)

[0098] The purpose is to confirm whether ROS generation is inhibited when SPG-SH1, SPG-SH2 and SPG-SH3 are respectively treated in RAW2647 cells. In particular, NADPH oxidase (NADPH oxidase; NOX) activity and mitochondrial ROS level were respectively confirmed.

[0099] Specifically, after stimulating RAW2647 cells with LPS, SPG-SH1, SPG-SH2, and SPG-SH3 were treated at concentrations of 1 μg / mL, 5 μg / mL, and 10 μg / mL, respectively. NADPH oxidase activity was verified by measuring superoxide generation in the RAW2647 cells by lucigenin (bis-N-methylacridinium nitrate)-ECL method. Mitochondrial ROS production was measured by FACS analysis after the RAW2647 cells were stained with 5 μM MitoSOX for 30 minutes and the mean fluorescence intensity (MFI) was measured.

[0100] As a result, such as figure 2 As shown, the NADPH oxidase activity was the lowest compared with the case of SPG-SH3 treatment, but it was conf...

Embodiment 3

[0102] ROS production inhibitory effect of SPG-SH3 and antibiotics (in vivo)

[0103] The purpose was to confirm whether or not the production of ROS is inhibited in the case of combined treatment of SPG-SH3 and antibiotics in CLP mice. In particular, NADPH oxidase (NADPH oxidase; NOX) activity, total ROS (Total ROS) and mitochondrial ROS levels were respectively confirmed.

[0104] Specifically, CLP mice (n=5) administered with the same administration substances as in Example 1-1 were prepared. The measurement of NADPH oxidase activity was measured and verified in the same manner as in Example 2 after obtaining and crushing the spleen (spleen) of CLP mice. In terms of total ROS, the spleens of CLP mice were obtained and crushed, and the reagents of the total ROS activity assay kit (Total ROS Activity Assay kit, CA-R900) were treated with 5 μM for 60 minutes, and measured at 490nm / 525nm Ex / Em (excitation wavelength / emission wavelength). As for the production of mitochondria...

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Abstract

The present invention relates to a composition for use in preventing, alleviating, or treating inflammatory disease, or an anti-inflammatory composition, which comprises: beta-1,3-glucan; and the peptide composed of the amino acid sequence of SEQ ID NO: 4. The present invention also relates to an antibacterial composition against bacteria causative of sepsis, which comprises: beta-1,3-glucan; thepeptide composed of the amino acid sequence of SEQ ID NO: 4; and an antibiotic. The present invention also relates to a method for preventing or treating an inflammatory disease, the method comprisinga step of administering beta-1,3-glucan; and the peptide composed of the amino acid sequence of SEQ ID NO: 4 to a patient. Accordingly, when included, beta-1,3-glucan and the peptide composed of theamino acid sequence of SEQ ID NO: 4 according to the present invention can reduce levels of reactive oxygen, sepsis-causing bacteria, and inflammatory cytokines in sepsis mice and significantly increase the survival rate of sepsis mice, thus being effective for a composition for prevention, alleviation, or treatment of an inflammatory disease such as sepsis, or a preventive or therapeutic method.Particularly, the composition reduces the reactive oxygen generated by phagocytes in a specific pattern and, when used in combination with an antibiotic, exhibits an additionally better effect becausethe composition can significantly increase reactive oxygen and expression levels and survival rates of sepsis-causing bacteria and inflammatory cytokines.

Description

technical field [0001] This application claims the right of priority to the Korean patent application submitted to the Korean Patent Office on June 20, 2018, with application number 10-2018-0070846, the entire contents of which are references of this application. [0002] The present invention relates to a composition comprising β-1,3-glucan (β-1,3-glucan) and a peptide composed of the amino acid sequence of SEQ ID NO: 4 for preventing, improving or treating inflammatory diseases or anti-inflammatory with composition. [0003] The present invention also relates to an antibacterial composition against sepsis-causing bacteria comprising β-1,3-glucan (β-1,3-glucan), a peptide consisting of the amino acid sequence of SEQ ID NO: 4, and an antibiotic. [0004] The present invention also relates to a method for preventing or treating inflammatory diseases comprising the step of administering β-1,3-glucan (β-1,3-glucan) and a peptide consisting of the amino acid sequence of SEQ ID NO...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/716A61K38/10A61K31/545A23L33/10A23L33/18A61K45/06A61P29/00A61P31/00
CPCA61P29/00A23L33/18A61K38/10A61P31/00A61K31/716A61K45/06Y02A50/30A23L33/21A23L2/52A23L2/66A61K2300/00A61K31/545A23L33/10A23V2002/00A23V2200/324
Inventor 阳哲洙金泳均朴秉熙李种大金济京张容晚金成洪朴恩智
Owner KISHIN BIOTECHNOLOGY CO LTD
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