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Yeast cell wall nanoparticle and its preparation method and application

A yeast cell wall and nanoparticle technology, applied in the field of yeast cell wall nanoparticles and its preparation, can solve the problems of systemic toxicity, systemic infection in patients, low tumor regression rate, etc., and achieve good safety, low cost, and good repeatability Effect

Active Publication Date: 2022-04-05
喜丽生物科技(苏州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Although a large number of studies have shown that microbial-based therapeutic strategies provide new directions for cancer immunotherapy, there are still limitations
First, live microbiota may cause systemic infection and severe systemic toxicity in patients, causing the immune system to attack healthy cells
Second, the tumor regression rate caused by bacterial therapy is low, and the efficiency of anti-tumor immunity still needs to be improved; third, it is difficult to guarantee the large-scale production, quality control, and stability of the above-mentioned live bacteria and viruses; Receptivity is also an issue worth considering

Method used

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  • Yeast cell wall nanoparticle and its preparation method and application
  • Yeast cell wall nanoparticle and its preparation method and application
  • Yeast cell wall nanoparticle and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0058] Example 1: Preparation of Yeast-Derived Nanoparticle System

[0059] Weigh 100g of Saccharomyces cerevisiae powder and suspend in 1M NaOH solution, and heat at 80°C for 1 hour. After cooling, centrifuge at 2000 g for 10 minutes to collect insoluble matter containing yeast cell walls. The insoluble matter was suspended in 1 L of ultrapure water adjusted to pH 4-5 with HCl, and incubated at 55°C for 1 hour. After cooling to room temperature, centrifuge at 2000 g for 10 minutes to collect the insoluble matter, and wash once with 1 L of ultrapure water, four times with 200 mL of isopropanol, and twice with 200 mL of acetone. The obtained insoluble matter is placed in a container, and dried at room temperature to obtain a fine white powder, which is the micron-scale yeast cell wall. Subsequently, yeast cell walls of different nanometer sizes were prepared by gradient centrifugation. Specifically, weigh 25 mg of the white powder and dissolve it in 8 mL of PBS (pH=7.4), use...

Embodiment 2

[0060] Example 2: Characterization of yeast-derived nanoparticle systems

[0061] Transmission electron microscopy (TEM) characterized the internal structure of yeast cell walls at the micron level and three nanometer-sized yeast cell walls, and the results are as follows figure 1 As shown, the particle diameters are about 3-4 μm, 500nm, 200nm, and 50nm, respectively, and they all present a spherical shape and are evenly distributed. Dynamic light scattering (DLS) analysis of the particle size and the particle size change of three nanometer-sized yeast cell walls over time under different storage conditions, the results are as follows figure 2 , image 3 As shown, it is stable for at least 2 weeks at both room temperature and 4°C storage conditions. Using BCA protein quantification kit and SDS-PAGE gel electrophoresis to analyze its surface protein, the results are as follows Figure 4 , Figure 5 As shown, the three nanometer-sized yeast cell walls contain the same prote...

Embodiment 3

[0062] Example 3: Immunological effects of yeast-derived nanoparticle systems on dendritic cells

[0063] (1) In vitro cell uptake analysis

[0064] First, the three nanometer-sized yeast cell walls were stained with Cy5.5, and after co-incubating with DC2.4 for 24 hours, the cells were collected by centrifugation at 300g for 3 minutes, and the effect of dendritic cells phagocytosis of the three nanoparticles was analyzed by flow cytometry. At the same time, three nanometer-sized yeast cell walls stained with Cy5.5 were co-incubated with dendritic cells for 24 hours, fixed with 4% paraformaldehyde, stained with DAPI to locate the cell nucleus, and analyzed by confocal microscopy . The result is as Figure 6 , Figure 7 As shown, the yeast cell walls of the three nanometer sizes can be phagocytosed effectively by dendritic cells, and the phagocytosis effect is better as the particle size decreases.

[0065] (2) Analysis of the immunological effects of bone marrow-derived de...

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Abstract

The present invention relates to a yeast cell wall nanoparticle and its preparation method and application. The yeast cell wall nanoparticle of the present invention is obtained by removing the content of yeast and adopting differential centrifugation to obtain a particle diameter of 10 ~1000nm nanoparticles, the potential of yeast cell wall nanoparticles is -1mV ~ -50mV. The nanometer particle system of the present invention is prepared by crushing and differential centrifugation of Saccharomyces cerevisiae. As a probiotic, Saccharomyces cerevisiae is beneficial to the body and therefore has good safety. The nanometer-sized yeast-derived particle system of the present invention has a strong ability to deliver to tumor-draining lymph nodes, can effectively regulate the microenvironment of tumor-draining lymph nodes, and induce immune responses.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a yeast cell wall nanoparticle and a preparation method and application thereof. Background technique [0002] Cancer is a serious threat to human health. According to the latest statistics from the National Cancer Center, the death caused by malignant tumors accounts for 23.91% of all deaths. At present, the incidence and mortality of malignant tumors are on the rise. It is urgent to take measures to prevent Control the severe development trend. In recent years, tumor immunotherapy, a treatment method that utilizes the host immune system to achieve anti-tumor purposes, has received extensive attention and achieved certain results. Even so, the clinical response rate of tumor immunotherapy to solid tumors is low. More and more studies have shown that the inflammatory tumor microenvironment will make tumors sensitive to immune checkpoint inhibitors, and non-inflam...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/06A61K36/064A61K9/14A61P35/00B82Y5/00B82Y40/00C12N13/00
CPCA61K36/064A61K9/14A61P35/00B82Y5/00B82Y40/00C12N1/063C12N1/066C12N13/00A61K2236/15A61K2236/13A61K2236/53
Inventor 汪超徐嘉潞
Owner 喜丽生物科技(苏州)有限公司
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