Cell apoptosis inducer, drug-loaded vesicle and application thereof

A technique of apoptosis induction and cell vesicles, which is applied in the direction of cell culture active agents, animal cells, tumor/cancer cells, etc., can solve the problems of low yield and poor uniformity of cell vesicles, and achieve enhanced innate immunity and uniform The effect of good sex and enhanced therapeutic effect

Active Publication Date: 2021-03-26
HUBEI SOUNDNY BIOLOGICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Aiming at the above problems of low yield and poor uniformity of cell vesicles in the prior art, the present invention provides a cell apoptosis inducer, and provides the cell apoptosis inducer to induce cell apoptosis to prepare drug-loaded vesicles The method, and the drug-loaded vesicle prepared therefrom and the pharmaceutical composition containing the drug-loaded vesicle

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  • Cell apoptosis inducer, drug-loaded vesicle and application thereof
  • Cell apoptosis inducer, drug-loaded vesicle and application thereof
  • Cell apoptosis inducer, drug-loaded vesicle and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0065] Example 1 Brady's yeast mannan and calcium ions synergistically induce apoptosis

[0066] Tumor cell culture: the purified tumor cells were inoculated in RPMI 1640 medium containing 10% fetal bovine serum (v / v), 100 U / mL penicillin, 100 mg / mL streptomycin, at a temperature of 37 ° C, gas The atmosphere is 5% CO 2Cultured under certain conditions, regularly observed and subcultured, digested when the tumor cells grew to the logarithmic phase, and then seeded the dispersed tumor cells in a 24-well plate and incubated overnight for later use.

[0067] Tumor cell apoptosis: Divide the tumor cells in the 24-well plate into 2 groups—experimental group and control group. Add 0, 50, 100, 200, 400, 800 μmol / L CaCl to each group 2 . The control group was also divided into 6 groups, and 0, 50, 100, 200, 400, and 800 μmol / L of CaCl were added to each group, respectively. 2 , without adding Brady's yeast mannan. The tumor cells in the experimental group and the control group we...

Embodiment 2

[0072] The optimization of embodiment 2 brady's yeast mannan concentration

[0073] The method of Example 1 was used for tumor cell culture, and then the tumor cells in the 24-well plate were divided into different groups, and 100 μmol / L and 400 μmol / L of CaCl were added respectively 2 And after gradient concentrations of Brady's yeast mannan (0, 5, 10, 15, 20 mg / L), the tumor cells were placed in a temperature of 37 ° C and a gas atmosphere of 5% CO 2 After culturing for 1 hour, replace with fresh RPMI 1640 medium, and perform apoptosis detection and calculate the apoptosis rate after culturing overnight. For the test results, see image 3 .

[0074] Depend on image 3 It can be seen that the efficiency of mannan of Saccharomyces bradyi and calcium ions entering tumor cells depends on the concentration of mannan of Saccharomyces bradyi within a certain range. This is because the increase of calcium ionophores accelerated the accumulation of intracellular free calcium ion c...

Embodiment 3

[0075] The optimization of embodiment 3 calcium ion concentration

[0076] The method of Example 1 was used for tumor cell culture, and then the tumor cells in the 24-well plate were divided into different groups, and the following treatments were carried out respectively:

[0077] Experimental group: Add 10 mg / L Brady's yeast mannan and gradient concentrations of CaCl to tumor cells 2 After (0, 25, 50, 100, 150, 200 μmol / L), the tumor cells were placed in a temperature of 37 ° C and a gas atmosphere of 5% CO 2 After culturing for 1 hour, replace with fresh RPMI 1640 medium, culture overnight, and undergo density gradient centrifugation to obtain drug-free cell vesicles. The number and particle size of cell vesicles were characterized by Malvern NS300 particle tracking analyzer, and the morphology of cell vesicles was characterized by Hitachi HT7800 120kV transmission electron microscope. The characterization results are shown in Figure 4-6 .

[0078] Control group 1: the ...

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Abstract

The invention provides a cell apoptosis inducer, a drug-loaded vesicle and application thereof, and belongs to the technical field of biology. The apoptosis inducer comprises an effective amount of yeast mannan and an effective amount of calcium ions. The yeast mannan is used as a calcium ion carrier, the permeability of cell membranes to the calcium ions can be improved, high-concentration free calcium ions can be gathered in cells within a short time, then a cell apoptosis procedure is triggered, cell vesicles are generated, and the obtained cell vesicles are high in yield, good in uniformity and small in average cell vesicle diameter. When the cell vesicle is used for wrapping a drug, the cell vesicle can efficiently pass through the barrier of tumor cells and is endocytosed by the tumor cells, so that the drug delivery efficiency is improved.

Description

technical field [0001] The present invention relates to the field of biotechnology, in particular to an apoptosis-inducing agent, and also to a method for preparing drug-loaded vesicles by using the apoptosis-inducing agent to induce apoptosis, and the drug-loaded vesicle prepared thereby Vesicles and pharmaceutical compositions containing the drug-loaded vesicles. Background technique [0002] In recent years, the incidence of tumors has increased year by year. Although scientists have been exploring various ways to overcome tumors, the clinical cure rate of tumors has not yet shown a significant increase. Cancer treatment methods mainly include surgery, radiotherapy and drug therapy. However, these traditional cancer treatment methods are poorly targeted and highly toxic. While killing tumor cells, they will also damage normal cells and cause adverse reactions. has certain limitations. In order to overcome the toxic and side effects of traditional cancer treatment metho...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09A61K9/50A61K47/46A61K45/00A61K31/704A61P35/00
CPCC12N5/0693A61K9/5068A61K45/00A61K31/704A61P35/00C12N2501/90C12N2500/14
Inventor 童雅琪陈彬张一
Owner HUBEI SOUNDNY BIOLOGICAL TECH
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