CN108272818B

CN108272818B - Anti-tumor inorganic selenium-containing nano-particles and preparation method and application thereof

Info

Publication number: CN108272818B
Application number: CN201810393436.XA
Authority: CN
Inventors: 冯刚; 肖东琴; 周玉川; 邓丽; 向小聪; 刘康; 陈竹
Original assignee: Nanchong Central Hospital
Current assignee: Nanchong Central Hospital
Priority date: 2018-04-27
Filing date: 2018-04-27
Publication date: 2020-08-07
Anticipated expiration: 2038-04-27
Also published as: CN108272818A

Abstract

The invention provides a preparation method of an anti-tumor inorganic selenium-containing nano particle, which comprises the following steps: (1) adding Na to the mixed aqueous solution of calcium chloride and phytic acid₂SeO₃·5H₂Adjusting the pH value to 2.3, clarifying the solution, and placing the clarified solution in a closed container with a pore at the top; (2) putting the strong ammonia water into another closed container with a pore on the top; (3) and (3) placing the substances obtained in the step (1) and the step (2) in the same sealed dryer, reacting at room temperature until a precipitate is generated at the bottom of the sealed container with the top provided with the fine hole in the step (1), and then taking the obtained precipitate, washing, centrifuging and drying to obtain the anti-tumor inorganic selenium-containing nano-particles. The inorganic selenium-containing nano-particles obtained by the invention can effectively control the release of selenium and can obviously inhibit the proliferation of tumor cells; the preparation process is simple, and the raw materials are easy to obtain and safe.

Description

Anti-tumor inorganic selenium-containing nano-particles and preparation method and application thereof

Technical Field

The invention belongs to the technical field of nanotechnology and antitumor drugs, and particularly relates to an antitumor inorganic selenium-containing nanoparticle and a preparation method and application thereof.

Background

Selenium, one of the essential trace elements for human and animal life, is widely distributed in the liver and kidney of human, and secondly in muscle, bone and blood, with minimal content in adipose tissue. In organisms, selenium is the only trace element regulated by genes, and selenium and compounds thereof play an important role in improving immunity, resisting oxidation, resisting tumors and the like. A large number of research data and experimental studies have shown that the in vivo selenium content is significantly negatively correlated with the incidence of cancer (Kim I W, Bae S M, Kim YW, et al, serum selenium levels in Korean hepatoma properties [ J].Biological traceelement research,2012,148(1):25-31.

-Bergman L,Torndal U B,Eken S,etal.Selenium prevents tumor development in a rat model for chemicalcarcinogenesis[J].Carcinogenesis,2005,26(1):125-131.)。

In view of the positive effect of selenium in the anti-tumor process, selenium-containing nano materials become a hotspot for research in the field of anti-tumor nano materials. The biological activity of selenium is related to the chemical form and absorbed dose of selenium. Sodium selenite, as a common selenium supplement preparation suggested by WHO, has a killing effect on a plurality of tumor cells, but the dosage of the sodium selenite is difficult to control, and the sodium selenite is easy to cause toxic and side effects on organisms. In order to reduce the toxic effect, a large amount of organic selenium with anticancer activity is synthesized, such as selenoether, selenocyanide, selenium heterocyclic compound, and the like. But all have the problems of difficult accurate control of dosage, easy generation of toxic and side effects and the like. Therefore, the research on the novel selenium form, the antitumor activity and the action mechanism becomes a research hotspot in the multidisciplinary field.

In recent years, with the development of nanotechnology and the discovery of biological activity of novel nano-selenium preparations, research and development of nano-selenium materials have attracted attention of researchers. In the research aspect of inorganic nano selenium, the research of nano simple substance selenium has great advantages in the aspects of tumor prevention and treatment and the like. However, the conventional concept that the zero-valent selenium is difficult to absorb and utilize, and the nano elemental selenium is easy to aggregate and precipitate, so that the antitumor activity of the nano elemental selenium is greatly reduced due to the change of the nano scale, and the nano elemental selenium is limited to be developed into a novel antitumor drug.

Therefore, the development of selenium-containing substances with novel forms, in particular the development of selenium-containing inorganic nano microspheres between the chemical forms of sodium selenite and nano elemental selenium, has better prospects and also faces higher challenges.

Disclosure of Invention

In view of the disadvantages of the prior art, an object of the present invention is to provide a method for preparing anti-tumor inorganic selenium-containing nanoparticles, the method comprising the steps of:

(1) mixing with calcium chloride and phytic acidAdding Na into the water solution₂SeO₃·5H₂Adjusting the pH value to 2.3, clarifying the solution, and placing the clarified solution in a closed container with a fine hole at the top;

(2) putting the strong ammonia water into another closed container with a pore on the top;

(3) and (3) placing the substances obtained in the step (1) and the step (2) in the same sealed dryer, reacting at room temperature until precipitates are generated at the bottom of the sealed container with the top provided with the fine hole in the step (1), and then taking the obtained precipitates, washing, centrifuging and drying to obtain the anti-tumor inorganic selenium-containing nano particles.

According to the invention, after experimental investigation, the organic micromolecule inositol hexaphosphate is finally adopted as a template agent, the organic micromolecule inositol hexaphosphate is adopted as a regulating agent, the selenium-containing inorganic nano-particles are prepared, the release of selenium can be effectively controlled, the inhibition effect on tumor cells is realized, and the toxic and side effects of the selenium-containing particles on normal cells are reduced.

Phytate (i.e., phytic acid), which is found in natural whole grain foods such as rice, oats, corn, wheat, and green beans. Phytic acid widely exists in plant seeds in the form of salt, is a nutrient beneficial to human bodies, can be hydrolyzed into inositol and phospholipid in human bodies, the inositol has the function of resisting aging, and the phospholipid is an important component of human cells. The invention adopts the template agent, so that the raw materials of the invention have the advantages of easy obtainment and safety.

In a preferred embodiment of the present invention, in the step (1), the concentration of calcium chloride is 0.01 mol/L-0.1 mol/L.

As a further preferred embodiment of the present invention, in the step (1), the calcium chloride is mixed with Na₂SeO₃·5H₂The mass ratio of O is 1.

In a preferred embodiment of the present invention, in step (1), the volume fraction of phytic acid in the mixed aqueous solution is 0.03 to 0.3%.

In the course of the research of the present invention, the inventors found that when calcium ions and selenite ions are present in the aqueous solution and no phytic acid is present, the obtained calcium selenite is in the form of lamellar nanoparticles; when phytic acid is added, the product morphology changes. With the increase of the addition of the phytic acid, the shape of the inorganic selenium-containing nano-particles is transited from a flake shape to a sphere shape. When the volume fraction of phytic acid added is less than 0.17%, the product is co-present as lamellar and spherical nanoparticles. When the volume fraction is 0.17% or more, the product is spherical particles.

In actual practice, to facilitate the reaction, in step (1), the solution was clarified and allowed to stand overnight to remove air bubbles from the solution.

And (3) reacting for 2-48 hours at room temperature.

In a preferred embodiment of the present invention, in the step (3), the drying is carried out at a drying temperature of 80 ℃.

Another object of the present invention is to provide the antitumor inorganic selenium-containing nanoparticles prepared by the above method, which are lamellar nanoparticles, a mixture of lamellar and spherical nanoparticles, or spherical nanoparticles.

The invention also aims to provide application of the anti-tumor inorganic selenium-containing nano-particles in preparing anti-tumor drugs, wherein tumors comprise mastadenoma, hepatoma and esophageal tumor.

The invention has the beneficial effects that:

1. the form of the inorganic selenium-containing nano particles obtained by the invention is between the chemical forms of sodium selenite and nano elemental selenium, and the effective control release of selenium can be realized;

2. the inorganic selenium-containing nano-particles obtained by the invention can obviously inhibit the proliferation of tumor cells;

3. the inorganic selenium-containing nano-particles have the advantages of simple preparation process, easily obtained and safe raw materials and good practical application prospect.

Drawings

FIG. 1 is a scanning electron microscope image of inorganic selenium-containing microspheres obtained in example 3 according to the present invention; wherein phytic acid was not added in (a), phytic acid in (b) was added in an amount of 0.03% (v/v), and phytic acid in (c) was added in an amount of 0.17% (v/v);

FIG. 2 shows the release profiles of selenium ions in phosphate buffered saline for different samples; wherein A: buffer pH 7.4, B: the pH of the buffer solution was 5.8; p0 is a product group without phytic acid, P1 is a product group with the phytic acid addition amount of 0.17% (v/v);

FIG. 3 is a graph showing the effect of CCK-8 method for detecting the inhibition of proliferation of normal cells (Het-1A, human esophageal epithelial cells) and tumor cells (Eca-109, human esophageal cancer cells); wherein, control is a blank control group without material addition, no IP6 is a product group without phytic acid addition, IP 6-0.03% is a product group with phytic acid addition of 0.03% (v/v), IP 6-0.17% is a product group with phytic acid addition of 0.17% (v/v), and IP 6-0.3% is a product group with phytic acid addition of 0.3% (v/v);

FIG. 4 is a graph showing the staining results of live and dead cells in co-culture with tumor cells in different samples; wherein, (a) is a blank control group without material addition, (b) is a group without phytic acid addition, and (c) the phytic acid addition amount is 0.17% (v/v);

Detailed Description

The present invention is described in detail below by way of examples, and it should be noted that the following examples are only for illustrating the present invention and should not be construed as limiting the scope of the present invention.

Example 1

I. Adding CaCl₂Dissolving in distilled water, adding a certain amount of phytic acid, and stirring.

II. Adding Na into the step I₂SeO₃.5H₂O, after dissolution, by addition of HNO₃Or NaOH to adjust the pH of the solution to 2.3. Stirring, clarifying, transferring into conical flask, standing overnight to remove bubbles, sealing with aluminum foil, and needling several small holes. On the other hand, the strong ammonia water is taken and placed in a conical flask, and a plurality of small holes are punched after the aluminum foil is sealed. Drying the two conical flasks simultaneouslyThe reaction vessel was sealed and allowed to react at room temperature for 2 days.

III, after the reaction is finished, pouring the supernatant, taking the lower-layer precipitate, washing the precipitate, centrifuging, and drying at 80 ℃ to obtain the inorganic selenium-containing nano particles.

In the above scheme, CaCl₂The concentration is 0.1 mol/L, Na₂SeO₃.5H₂The O concentration was 0.1 mol/L the volume percentage of phytic acid (50% concentration) in the aqueous solution was 0.17%.

Example 2

Except for CaCl₂The concentration is 0.05 mol/L, Na₂SeO₃.5H₂The O concentration was 0.05 mol/L, and the concentration of phytic acid (50%) in the aqueous solution was 0.17% by volume, which was the same as that of example 1.

Example 3

Except for CaCl₂The concentration is 0.01 mol/L, Na₂SeO₃.5H₂The O concentration was 0.01 mol/L, and the concentration of phytic acid (50%) in the aqueous solution was 0.17% by volume, which was the same as that of example 1.

Comparative examples 1 to 3

The same procedures as in examples 1 to 3 were repeated except that phytic acid was not added in step (1).

Experimental example 1

The inorganic selenium-containing nanoparticles obtained in example 3 were subjected to scanning electron microscopy, ion release, cytotoxicity, and tumor cell inhibition test, and the results are shown in fig. 1 to 4.

As shown in fig. 1, when the volume fraction of phytic acid in the aqueous solution is 0.17%, the obtained inorganic selenium-containing nanoparticles have a spherical shape, which may facilitate the preparation of corresponding formulations.

As shown in fig. 2, the selenium-containing nanoparticles are released faster in a slightly acidic environment than in a neutral environment, and the addition of phytic acid is more beneficial to control the release rate of selenium ions.

As can be seen from fig. 3 and 4, the inorganic selenium-containing nanoparticles obtained by the present invention have significant inhibitory effect on tumor cells.

Claims

1. A preparation method of an anti-tumor inorganic selenium-containing nanoparticle is characterized by comprising the following steps:

(1) adding Na to a mixed aqueous solution of calcium chloride and phytic acid₂SeO₃·5H₂Adjusting the pH value to 2.3, clarifying the solution, and placing the clarified solution in a closed container with a pore at the top;

(3) placing the substances obtained in the step (1) and the step (2) in the same sealed dryer, reacting at room temperature until a precipitate is generated at the bottom of the sealed container with the top provided with the fine hole in the step (1), and then taking the obtained precipitate, washing, centrifuging and drying to obtain the anti-tumor inorganic selenium-containing nano-particles;

the volume fraction of the phytic acid in the mixed aqueous solution is 0.17%;

in the step (1), the concentration of the calcium chloride is 0.01 mol/L-0.1 mol/L, and the calcium chloride and Na₂SeO₃·5H₂The mass ratio of O is 1.

2. The method according to claim 1, wherein in the step (1), the solution is left to stand overnight after being clarified to remove air bubbles in the solution.

3. The method according to claim 1, wherein the reaction is carried out at room temperature for 2 to 48 hours in the step (3).

4. The production method according to claim 1, wherein in the step (3), the drying is performed at a drying temperature of 80 ℃.

5. The antitumor inorganic selenium-containing nanoparticles prepared by the method of any one of claims 1 to 4, which are spherical nanoparticles.

6. The use of the anti-tumor inorganic selenium-containing nanoparticles of claim 5 in the preparation of anti-tumor drugs, wherein the tumors comprise breast tumor, liver tumor and esophageal tumor.