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Nanovesicles derived from genus bacillus bacteria and use thereof

A technology of Bacillus and Bacillus subtilis, applied in the direction of medical preparations containing active ingredients, applications, preparations for toiletry, etc., can solve the problem of vesicle-free prevention or treatment of inflammatory diseases

Active Publication Date: 2019-05-21
MD HEALTHCARE INC
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no case where the quantification of genes in vesicles secreted from bacteria of the genus Bacillus or Bacillus genus is used for the diagnosis of cancer, chronic inflammatory diseases, or metabolic diseases, and there are no cases where vesicles are used for the prevention or treatment of inflammatory diseases Case

Method used

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  • Nanovesicles derived from genus bacillus bacteria and use thereof
  • Nanovesicles derived from genus bacillus bacteria and use thereof
  • Nanovesicles derived from genus bacillus bacteria and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0105] Example 1. Analysis of the Body Absorption, Differentiation and Secretion Patterns of Bacteria and Bacteria-Derived Vesicles

[0106] In order to evaluate whether bacteria of the genus Bacillus and vesicles derived from the bacteria are absorbed into the whole body through the gastrointestinal tract, experiments were performed by the following method. Fluorescence-labeled bacteria or bacteria-derived vesicles were administered gastrointestinally into mouse stomachs at a dose of 50 μg, and fluorescence was measured after 0 min, 5 min, 3 h, 6 h or 12 h. By observing whole images of mice, bacteria were not taken up systemically, but vesicles derived from bacteria were taken up systemically at 5 minutes after administration, and strong fluorescence was observed in liver and kidney at 30 minutes after administration, which indicated that Vesicles are secreted into the liver and urinary system. Furthermore, it can be seen that vesicles are present in vivo until 12 hours afte...

Embodiment 2

[0108] Example 2. Metagenomic Analysis of Bacteria-Derived Vesicles in Clinical Samples

[0109] A clinical sample such as blood is first added to a 10 ml tube and centrifuged (3,500×g, 10 minutes, 4° C.) to pellet suspended material, and then only the supernatant is transferred to a new 10 ml tube. After removing bacteria and cell debris using a 0.22-μm filter, the resulting product was transferred to a Centriprep tube (centrifugal filter 50 kD) and centrifuged at 1500 × g and 4 °C for 15 min to discard material smaller than 50 kD. The resulting product was then concentrated to 10 ml. Again, bacteria and cellular debris were removed using a 0.22-μm filter, and vesicles were isolated and dissolved in phosphate-buffered saline (PBS).

[0110] 100 μL of vesicles isolated by the method described above were boiled at 100 °C to release the internal DNA from the lipids, and then cooled on ice for 5 min. In addition, to remove residual suspended matter, the vesicles were centrifuge...

Embodiment 3

[0114] Example 3. Confirmation of reduced Bacillus-derived vesicles by metagenomic analysis of bacteria-derived vesicles in the blood of patients with liver cancer

[0115] According to the method described in Example 2, by extracting genes from vesicles present in the blood, blood samples from 91 patients with liver cancer and 99 normal persons matched with the patient's sex and age as normal controls were subjected to Metagenomic analysis then assessed the distribution of vesicles derived from bacteria of the genus Bacillus. The results of the evaluation confirmed that vesicles derived from bacteria of the genus Bacillus were significantly reduced in the blood of gastric cancer patients compared with the blood of normal people (ratio of normal people to liver cancer patients: 0.37% vs. 0.13%; fold change: 0.37 ; p=0.0002) (see image 3 ).

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Abstract

The present invention relates to vesicles derived from the genus Bacillus bacteria and a use thereof. The present inventors have confirmed that the vesicles are significantly reduced in samples of patients with cancers, such as liver cancer, bladder cancer, breast cancer, and ovarian cancer; inflammatory diseases, such as asthma and atopic dermatitis; or metabolic diseases, such as diabetes and cirrhosis, compared with normal persons, and the vesicles inhibited the secretion of inflammatory mediators by pathogenic vesicles, such as E. coli-derived vesicles, which are causative factors of inflammatory diseases, diabetes, and the like. Therefore, the vesicles derived from the genus Bacillus bacteria according to the present invention can be advantageously used for the purpose of developing adiagnostic method for cancer, inflammatory diseases, and metabolic diseases, and a preventive or therapeutic composition therefor.

Description

technical field [0001] The present invention relates to nanovesicles derived from bacteria of the genus Bacillus and uses thereof, more particularly, to methods for diagnosing cancer, inflammatory diseases, or metabolic diseases using nanovesicles derived from bacteria of the genus Bacillus, and using Compositions comprising nanovesicles for the prevention or treatment of cancer, inflammatory diseases or metabolic diseases. Background technique [0002] Entering the 21st century, while acute infectious diseases, which in the past were considered epidemics, have become less important, disease patterns have changed such that chronic diseases resulting from incompatibility between humans and the microbiome are considered to be Major diseases that determine quality of life and longevity in humans. Chronic diseases are characterized by chronic inflammation accompanied by immune dysfunction, and cancer, chronic inflammatory diseases, metabolic diseases, etc. are becoming serious ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/742A23L33/135A61K8/99A61K9/00A61Q19/00
CPCA61Q19/00A61K8/99A23L33/135C12Q1/6883C12Q1/6886C12Q2600/158A61K9/0073C12Q1/689A61P37/00A61K35/742A61P35/00A61P3/00A61P29/00Y02A50/30A61P1/16A61P17/14A61P17/00A61P3/10A61Q7/00A23V2200/324A23V2200/328A23V2200/318A23V2200/308A23V2002/00C12Q2531/113C12Q2600/112C12Q1/6851A61K9/007A61K2035/115
Inventor 金润根
Owner MD HEALTHCARE INC
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