KR20200091682A - Hollow porous silica nanosphere capable of controlling size, number of pores and shell thickness, and manufacturing method thereof - Google Patents

Abstract

The present invention relates to a method for preparing hollow porous silica nanospheres which have voids inside thereof and interstitial holes on the surfaces thereof, and are controllable in size and number of the interstitial holes and in thickness of shells, and hollow porous silica nanospheres which include a drug loaded in the voids thereof, allow the drug to be released slowly, and thus can be used as a drug carrier. The method includes the steps of: preparing a template polymer having a functional group on the surface thereof; depositing a silica precursor to the surface of the template polymer in a basic solution to form porous silica nanospheres; and removing the template polymer through firing to obtain hollow porous silica nanospheres.

Description

Hollow porous silica nanosphere capable of controlling size, number of pores and shell thickness, and manufacturing method thereof

The present invention relates to silica nanospheres, in particular the inside is empty, the surface of the porous silica nanospheres with pores, the size and number of pores in the shell of the porous silica nanospheres and the method of manufacturing the porous silica nanospheres to control the thickness of the shell and The present invention relates to a method for manufacturing hollow porous silica nanospheres that can be utilized as a drug carrier by placing a drug in an empty space inside the porous silica nanospheres and allowing the drug to be slowly released, and the porous silica nanospheres.

When the drug is ingested, biodegradation of the drug occurs in the blood circulation process, and thus an excessive amount of the drug is repeatedly consumed in order to obtain a desired therapeutic effect, thereby causing side effects that affect normal cells.

Therefore, in order to improve this problem, it is important to protect the ingested drug to minimize the amount of the drug that is decomposed and to release the drug steadily to maintain the drug for a long time.

The protection of the drug can be achieved by using hollow porous silica nanospheres, since the hollow nanoparticles can store substances in a central empty space, as a drug delivery system (DDS) and water and air. It can be used to remove pollutants in the air.

Since the first successful synthesis of porous silica nanospheres by the Schacht team in 1996, porous silica nanospheres have been studied extensively because of their unique properties such as large surface area, high thermal mechanical stability, and low density. In particular, these nanospheres are known to have great potential for use as drug carriers and catalyst supports. Porous silica nanospheres are generally made of polymer micelles and polystyrene spheres as molds.

Since silica nanoparticles have recently been approved for biomedical use by the US Food and Drug Administration (FDA), many efforts have been made to develop these nanoparticles into drug carriers.

In using silica nanoparticles as drug carriers, it is most important to control the size and number of crevices in the shell of the porous nanocar nanospheres to minimize the accumulation of unwanted substances into the nanospheres. . Therefore, in order to use the porous silica nanospheres as a drug carrier, it is important to adjust the size of the niche at the nanoscale, but little is known about how to control the size of the niche.

Peptide hormones are important drugs involved in various processes in vivo. One of these peptide-based drugs, Glp-1 (glucagon like peptide-1), is a hormone secreted by the nucleus of L cells in the intestine and isolation of the brain, while stimulating the secretion of insulin from β cells in the pancreas while suppressing the secretion of glucagon. Therefore, it can be used to treat diabetes. In addition, Glp-1 is known to inhibit apoptosis of β cells in the pancreas and stimulate proliferation and division of β cells. In addition, Glp-1 acts on the central nervous system to reduce appetite, so it can be used as a treatment for obesity.

On the other hand, Glp-1 has a half-life of less than 2 minutes in vivo because it is degraded by dipeptidyl peptidase-4 (DPP-4) in the blood circulation process. Therefore, as described above, since the peptide-based drugs that play various roles are easily degraded from proteolytic enzymes in the blood circulation process, it is necessary to develop a drug carrier capable of completely protecting and releasing the peptide-based drugs.

In particular, in order to minimize unwanted substances such as proteolytic enzymes from entering the porous silica nanosphere, it is absolutely necessary to control the size and number of crevices in the shell of the porous nanosphere. In addition, in order to maintain the function of the porous silica nanospheres without disintegration in vivo for a long time, it is essential to maintain a thickness of a certain level or higher.

KR 10-2018-0117920 A (2018.10.30) KR 10-1068474 B1 (2011.09.22) KR 10-1719988 B1 (2017.03.21) KR 10-1791656 B1 (2017.10.24)

Jong-Tak Lee, et al. Preparation of hollow silica nanoparticles with different sizes. Applied Chemistry, 2010, 14.2: 9-12. SCHACHT, S., et al. Oil-water interface templating of mesoporous macroscale structures. Science, 1996, 273.5276: 768-771. POPAT, Amirali, et al. Mesoporous silica nanoparticles for bioadsorption, enzyme immobilisation, and delivery carriers. Nanoscale, 2011, 3.7: 2801-2818. MAHONY, D., et al. In vivo delivery of bovine viral diahorrea virus, E2 protein using hollow mesoporous silica nanoparticles. Nanoscale, 2014, 6.12: 6617-6626. WANG, Peng, et al. Bifunctionalized Hollow Nanospheres for the One-Pot Synthesis of Methyl Isobutyl Ketone from Acetone. ChemSus Chem, 2012, 5.12: 2390-2396. FANG, Xiaoliang, et al. Self-templating synthesis of hollow mesoporous silica and their applications in catalysis and drug delivery. Nanoscale, 2013, 5.6: 2205-2218. GAO, Jinsuo, et al. Azide-functionalized hollow silica nanospheres for removal of antibiotics. Journal of colloid and interface science, 2015, 444: 38-41. LIU, Junjie, et al. Hollow mesoporous silica nanoparticles facilitated drug delivery via cascade pH stimuli in tumor microenvironment for tumor therapy. Biomaterials, 2016, 83: 51-65. WANG, Jiasheng, et al. Silica-based nanocomposites via reverse microemulsions: classifications, preparations, and applications. Nanoscale, 2014, 6.9: 4418-4437. WU, Si-Han; MOU, Chung-Yuan; LIN, Hong-Ping. Synthesis of mesoporous silica nanoparticles. Chemical Society Reviews, 2013, 42.9: 3862-3875. LI, Xiaobo; YANG, Yan; YANG, Qihua. Organo-functionalized silica hollow nanospheres: synthesis and catalytic application. Journal of Materials Chemistry A, 2013, 1.5: 1525-1535. FANG, Xiaoliang, et al. A cationic surfactant assisted selective etching strategy to hollow mesoporous silica spheres. Nanoscale, 2011, 3.4: 1632-1639. SKIBICKA, Karolina Patrycja. The central GLP-1: implications for food and drug reward. Frontiers in neuroscience, 2013, 7: 181. PRESSWALA, Lubaina; SHUBROOK, Jay H. What to do after basal insulin: 3 Tx strategies for type 2 diabetes. Journal of family practice, 2015, 64: 214-220. MABILLEAU, Guillaume, et al. Optimal bone mechanical and material properties require a functional GLP-1 receptor. Journal of Endocrinology, 2013, JOE-13-0146. TOFT-NIELSEN, M.-B.; MADSBAD, Sten; HOLST, J. J. Determinants of the effectiveness of glucagon-like peptide-1 in type 2 diabetes. The Journal of Clinical Endocrinology & Metabolism, 2001, 86.8: 3853-3860. MEIER, Juris J., et al. Intravenous glucagon-like peptide 1 normalizes blood glucose after major surgery in patients with type 2 diabetes. Critical care medicine, 2004, 32.3: 848-851. KIM, Hong Rok, et al. Preparation and photocatalytic properties of Cr/Ti hollow spheres. Materials Chemistry and Physics, 2008, 108.1: 154-159.

Accordingly, the present inventor puts emphasis on solving the limitations and essential requirements of the existing porous silica nanospheres by comprehensively considering the above-mentioned matters, so that the peptide-based drugs having various roles hydrolyze proteins in the blood circulation process. In order to develop a new porous silica nanosphere that can be used as a drug carrier capable of completely protecting and releasing peptidic drugs so as not to be easily degraded from enzymes, the present invention was invented as a result of continuous research. Was done.

Therefore, the technical problem and object to be solved by the present invention is to protect the drug to minimize the amount of the drug to be decomposed, and to release the drug steadily, so that the effect can be maintained for a long time, the gap in the shell of the porous silica nanospheres It is to provide a porous silica nanosphere and a method for manufacturing the porous silica nanosphere that can control the size and number of crevice pores and the thickness of the shell, by adjusting the size of the pores at the nano scale and using the silica nanosphere particles as a drug carrier.

Here, the technical problems and objects to be solved by the present invention are not limited to the technical problems and objectives mentioned above, and other technical problems and objectives not mentioned will be clearly understood by those skilled in the art from the following description.

Method for producing a hollow porous silica nanosphere according to the present invention for achieving the above object, the step of synthesizing a template polymer having a functional group on the surface; Synthesizing porous silica nanospheres by depositing a silica precursor in a basic solution on the surface of the template polymer; And synthesizing the hollow porous silica nanospheres by firing and removing the template polymer.

At this time, in the method for producing hollow porous silica nanospheres according to the present invention, the step of synthesizing the template polymer has a size of 50-600 nm using styrene monomer and methacrylic acid. It is characterized by synthesizing a polystyrene template polymer.

In addition, in the method for manufacturing hollow porous silica nanospheres according to the present invention, the step of depositing the silica precursor on the surface of the template polymer is hydrolyzed the silica precursor on the surface of the template polymer in a basic solution to form a uniform silica layer. It is characterized by forming.

In addition, in the method for producing hollow porous silica nanospheres according to the present invention, the basic solution is characterized in that the pH is 9-12.

In addition, in the method for producing hollow porous silica nanospheres according to the present invention, the basic solution is preferably an ammonia solution.

In addition, in the method for producing hollow porous silica nanospheres according to the present invention, the silica precursor is a compound containing silicon, alkoxysilane, tetramethyl silicate, tetraethyl silicate and tetraethyl silicate Characterized in that the combination of any one or more of propyl silicate (tetrapropyl silicate).

In addition, in the method for manufacturing hollow porous silica nanospheres according to the present invention, the step of synthesizing the hollow porous silica nanospheres is performed by heating the porous silica nanospheres on which the silica layer is formed in an electric furnace at 300-700° C. for 6 hours. It is characterized by removing the template polymer by firing in air.

In addition, in the method for producing hollow porous silica nanospheres according to the present invention, it is preferable to remove the template polymer while slowly progressing the temperature change from 300°C to 2 hours, 450°C to 2 hours and 600°C to 4 hours. .

In addition, in the method for manufacturing hollow porous silica nanospheres according to the present invention, it is characterized in that the amount of the pores formed on the surface of the nanospheres is controlled by changing the addition amount of the silica precursor to control the thickness of the silica layer. .

In addition, in the method for manufacturing hollow porous silica nanospheres according to the present invention, when the addition amount of the silica precursor is increased, the size of the pores formed on the surface of the nanospheres increases, the number decreases, and the thickness of the silica layer increases. It is characterized by losing.

In addition, in the method for preparing hollow porous silica nanospheres according to the present invention, the amount, number, and size of the pores formed on the surface of the nanospheres by changing the pH of the basic solution from 10.15 to 11.01 by varying the addition amount of the basic solution Method for manufacturing a hollow porous silica nanosphere, characterized in that to control the thickness of the silica layer.

In addition, in the method for producing hollow porous silica nanospheres according to the present invention, when the amount of the basic solution is increased, the size of the pores formed on the surface of the nanospheres increases, the number decreases, and the thickness of the silica layer increases. It is characterized by losing.

In addition, the hollow porous silica nanospheres according to the present invention for achieving the above object is characterized by being produced by the above-described method for producing hollow porous silica nanospheres.

In particular, the hollow porous silica nanosphere according to the present invention is characterized in that it is used as a drug carrier by loading the drug in the hollow inside.

In addition, the hollow porous silica nanospheres according to the present invention is characterized in that the drug is mounted on the hollow is made by adding the hollow porous silica nanospheres to the solution in which the drug is dissolved, and then irradiating with ultrasound.

In addition, the hollow porous silica nanospheres according to the present invention is characterized in that the drug mounted on the hollow is slowly released (slow release) over time.

As described above, the present invention can control the size and number of crevices in the shell and the thickness of the shell in the process of manufacturing porous silica nanospheres with hollow insides and pores in the surface shell, and accordingly, the present invention provides porous silica. There is an effect that can be usefully used as a drug carrier capable of effectively carrying a drug including a peptide-based drug by continuously and slowly releasing the peptide-based drug mounted inside the nanosphere.

In addition, the present invention can prevent side effects caused by normal cells by not repeatedly ingesting an excessive amount of drugs in order to produce a desired therapeutic effect, and minimizes the amount of drugs ingested and releases the drug steadily, thereby improving drug efficacy. It has a long lasting effect.

In addition, the present invention provides a porous nanosphere made of silica, which is a natural mineral component and non-toxic to the human body, so that the nanospheres are self-destructed and discharged outside the body over time, and thus can be utilized as the best drug carrier. .

1 is a schematic view showing a method of manufacturing a hollow porous silica nanosphere according to the present invention,
Figure 2 is a FESEM photo of a template polymer prepared using polystyrene-methacrylic acid,
3A to 3E are FESEM photographs of hollow porous silica nanospheres synthesized according to the change in the amount of tetraethyl silicate added.
4A to 4D are FESEM photographs of hollow porous silica nanospheres synthesized according to the change in the amount of ammonia solution added;
Figure 5 is an SDS-PAGE gel photograph showing a state in which Glp-1 mounted in the interior space of the hollow porous silica nanospheres according to the present invention is slowly released over time.

Hereinafter, embodiments of the present invention will be described in more detail with reference to the accompanying drawings.

Prior to this, terms to be described later are defined in consideration of the functions in the present invention, which clearly indicates that they should be interpreted in terms of concepts consistent with the technical spirit of the present invention and commonly recognized or commonly recognized in the art.

In addition, if it is determined that a detailed description of known functions or configurations related to the present invention may obscure the subject matter of the present invention, the detailed description will be omitted.

Here, the accompanying drawings are exaggerated or simplified for a convenience and clarity of description and understanding of the structure and operation of the technology, and it is revealed that each component does not exactly match the actual size and shape.

In addition, when a part includes a certain component, this means that other components may be further included instead of excluding other components unless otherwise specified.

The present invention provides a method for controlling the size and number of crevices in the shell and the thickness of the shell in the process of manufacturing porous silica nanospheres with hollow inside and pores in the surface shell. In addition, the present invention shows that the porous silica nanospheres are loaded with Glp-1, one of the peptide drugs, into the inside to release the drug slowly and steadily. Therefore, the hollow porous silica nanospheres according to the present invention can be usefully used as a drug carrier including a peptide drug.

In addition, since the hollow porous silica nanospheres according to the present invention are natural mineral components and nanoparticles using silica that are not toxic to the human body, they are destroyed by time and discharged outside the body, so it can be utilized as the best drug carrier.

In the present invention, the method for preparing the hollow porous silica nanospheres is composed of the following three steps. The first step is to synthesize a template polymer having a functional group on the surface for the synthesis of the porous silica nanospheres, and the second step is basic. The step of synthesizing the silica layer by depositing the silica precursor in the solution on the surface of the template polymer and the third step consist of the step of synthesizing the porous silica nanospheres after emptying by removing the template polymer.

In the first step of the method for producing hollow porous silica nanospheres according to the present invention, the template polymer having a functional group on the surface is a polystyrene template polymer synthesized using styrene monomer and methacrylic acid. The size of the template polymer may be 50-600 nm, and in one embodiment according to the present invention, a size of 300 nm was used.

In the second step of the method for producing hollow porous silica nanospheres according to the present invention, a uniform silica layer is formed by depositing a silica precursor on the surface of the template polymer synthesized in the first step. The process of depositing the silica precursor on the mold surface can be carried out in a solution having a pH of 9-12, and in one embodiment according to the present invention was performed in a solution having a pH of 10-11. During the synthesis process, the pH of the solution is maintained using an ammonia solution, which is a basic solution, in which silica precursor is hydrolyzed and condensation polymerization occurs. Silica precursors may include silicon-containing compounds, alkoxysilanes, for example, tetramethyl silicate, tetraethyl silicate and tetrapropyl silicate, or these. It can be a combination of.

In the third step of the method for manufacturing hollow porous silica nanospheres according to the present invention, the template polymer is removed by firing in air at 300-700° C. for 6 hours in an electric furnace. In the present invention, in order to prevent the destruction of the porous silica nanospheres in the firing process, the temperature was changed slowly at 300°C for 2 hours, at 450°C for 2 hours, and at 600°C for 4 hours.

The present invention also provides information on the size and number of pores formed on the surface of the nanosphere and the thickness of the shell while changing the addition amount of the tetraethyl silicate silica precursor in the process of synthesizing the porous silica nanosphere from 1 ml to 3 ml. .

The present invention also changes the addition amount of the ammonia solution, which is a basic solution in the process of synthesizing porous silica nanospheres, from 0.5 ml to 2 ml, that is, the size and number of crevice pores formed on the surface of the nanosphere while changing the pH from 10.15 to 11.01 It provides information on the change in thickness.

The present invention also provides information on how steadily and slowly the drug is released from the porous nanospheres over time by mounting ultrasonic waves to mount Glp-1, one of the peptide drugs, into the porous silica nanospheres over time. That is, it also provides the possibility that the synthesized porous silica nanospheres can be used as a drug carrier.

Hereinafter, a method of manufacturing porous silica nanospheres according to an embodiment of the present invention will be described in detail with reference to FIGS. 1 to 5.

First, Figure 1 is a schematic view showing a method for manufacturing hollow porous silica nanospheres according to the present invention, the present invention is a polystyrene as a template (1, hereinafter referred to as a template polymer) to synthesize the hollow porous silica nanospheres (3) It provides a method of controlling the size and number of holes in the gap formed in the shell 2 of the nanosphere and also controlling the thickness of the shell 2. And by loading the peptide drug inside the nanosphere 4, confirming that the drug is slowly released, so that the hollow porous silica nanosphere 3 according to the present invention can be utilized as a drug delivery carrier.

When the precipitation of the silica precursor on the surface of the template polymer (1) rapidly occurs while minimizing the bond between the template polymer (1) and the silica precursor under basic conditions, a gap hole is formed in the shell (2) made of a silica layer.

In the present invention, the prepared polystyrene-methylacrylic acid particles are used as a template to synthesize porous silica nanospheres, and different amounts of tetraethyl silicate or ammonia solution are added as silica precursors in the synthesis process (different pH). ), the size and number of crevices formed in the shell of the nanosphere and the change in the thickness of the shell are controlled.

The template polymer (1) having a functional group was prepared using styrene monomer and methacrylic acid, and its form was observed using FESEM (Field Emission Scanning Electron Microscope, Scanning Electron Microscope, hereinafter referred to as ``SEM''), and this is shown in FIG. 2. Did. 2 is a FESEM photograph of a polymer sphere prepared using polystyrene-methacrylic acid, and the polymer sphere of FIG. 2 is used as a template for the synthesis of hollow porous silica nanospheres according to the present invention. The prepared mold polymer (1) has a ball-like structure and may have a diameter of 60 nm, 100 nm, 200 nm, 300 nm, 500 nm, 600 nm, etc., and the diameter of the mold polymer used in the present invention is in the range of 322-334 nm.

3A to 3E are FESEM photographs of hollow porous silica nanospheres synthesized according to the change in the addition amount of tetraethyl silicate, and show hollow porous silica nanospheres formed by varying the addition amount of tetraethyl silicate as a silica precursor. The diameter of the synthesized silica nanospheres is in the range of 360-547 nm, which is larger than the diameter of the polystyrene template polymer. These results indicate that the porous silica nanospheres successfully coated the polystyrene polymer particles as a template polymer.

In the experimental example to be described below, when the amount of tetraethyl silicate increased from 1 ml to 3 ml, the size of the pores formed on the surface of the porous silica nanospheres increased, but the number decreased. And the thickness of the shell shows a tendency to thicken. Exceptionally, when 3 ml of tetraethyl silicate is added, the crevice pores appear microporous in the finest form.

Specifically, when 1 ml of tetraethyl silicate is added as shown in FIG. 3A, the size of the pores on the surface of the porous silica nanospheres is a mesoporous gap between 2 nm and 50 nm and a macroporous gap of 50 nm or more. A large number of holes are formed, and the thickness of the shell is thin.

When 1.5 ml of tetraethyl silicate is added, the size of the crevice hole is mesoporous and macroporous, as shown in FIG. 3B, the number of pores is large, and the thickness of the shell is thin. When 2 ml of tetraethyl silicate is added, the size of the crevice hole is macroporous, the number of holes is small, and the thickness of the shell is thick, as shown in FIG. 3C. When 2.5 ml of tetraethyl silicate is added, the size of the crevice hole is macroporous, the number of holes is small, and the thickness of the shell is thick, as shown in FIG. 3D. When 3 ml of tetraethyl silicate is added, as shown in Fig. 3e, the size of the pores is microporous with a size of 2 nm or less, the number of pores is few, and the thickness of the shell is thick.

As shown in FIGS. 3A to 3E, Table 1 below summarizes the size and number of pores formed on the surface of the porous silica nanospheres tested while varying the amount of tetraethyl silicate, and the tendency to change the thickness of the shell.

* If the number of gaps is 10 or more, it is large, and if it is less than 10, it is small.

** If the thickness of the shell is more than 20nm, it is thick and if it is less than 20nm, it is thin.

On the other hand, in order to prevent destruction of the formed porous silica nanospheres by sudden temperature change, the temperature change during firing for the removal of the template polymer 1 was changed from 300°C to 2 hours, from 450°C to 2 hours, and from 600°C to 4 hours. Progress slowly. At this time, the porous silica nanospheres having a large number of pores in the shell were shrunk to about 5% after being calcined and reduced in size.

Porous silica nanospheres synthesized using 1 ml of tetraethyl silicate have a gap size and number of a suitable size for the shell, so different amounts of ammonia solution are used to vary the size and number of gap holes formed in the shell and the thickness of the shell. The thickness was investigated.

4A to 4D are FESEM photographs of porous silica nanospheres synthesized according to changes in the amount of ammonia solution added, and show SEM images of hollow porous silica nanospheres formed by varying the amount of ammonia solution from 1 ml to 2 ml.

When the amount of the ammonia solution is increased, that is, when the pH is increased from 10.55 to 11.01, the size of the pores formed in the shell increases, and the number tends to decrease, while the thickness of the shell increases.

Specifically, when 1 ml of ammonia solution is added, that is, when the pH is 10.55, the size of the gap hole is mesoporous and macroporous, the number of holes is large, and the thickness of the shell is thin. When 1.5 ml of ammonia solution was added, that is, when the pH was 10.85, the size of the crevice hole was mesoporous and macroporous and the number of mesoporous pores was small, but the number of macroporous pores was large and the thickness of the shell as shown in FIG. 4C. Is thick. When 2 ml of ammonia solution was added, that is, when the pH was 11.01, the size of the pores was macroporous, the number of pores was small, and the thickness of the shell was thick, as shown in FIG. 4D.

Exceptionally, when 0.5 ml of ammonia solution is added, that is, when the pH is 10.15, the size of the pores formed on the surface of the porous silica nanospheres is microporous, the number of pores is small, and the thickness of the shell is thick as shown in FIG. 4A. . Table 2 below summarizes the size and number of pores formed on the surface of the porous silica nanospheres and the tendency to change the thickness of the shell while varying the amount of ammonia solution added.

* If the number of gaps is 10 or more, it is large, and if it is less than 10, it is small.

** If the thickness of the shell is more than 20nm, it is thick and if it is less than 20nm, it is thin.

In order to obtain data on differentiation of the loading and slow release of the peptide drug according to the difference in the size of the pores in the hollow porous silica nanosphere shell according to the present invention, the number of pores formed at pH 10.15 is small and microporous silica An experiment was conducted to find out which of the mixture of nanospheres and a mixture of mesoporous and macroporous silica nanospheres having a large number of pores synthesized at pH 10.55 is good for Glp-1 loading and slow release.

5 is an SDS-PAGE gel photograph showing a state in which Glp-1 mounted in the inner space of the hollow porous silica nanospheres according to the present invention is slowly released over time, a solution in which Glp-1, one of peptide drugs, is dissolved After adding the hollow porous silica nanospheres according to the present invention and irradiating ultrasound to mount the drug into the silica nanospheres, perform SDS-PAGE (protein electrophoresis) and then stain with Coomassie blue This is a picture of the gel after (staining). Here, the arrow in FIG. 5 indicates the position of Glp-1.

It can be seen that Glp-1 enters better into the mixture of the mesoporous and macroporous silica nanospheres in lane 2 than the microporous silica nanospheres in lane 1 (shown as day 0). This result shows that the mesoporous or macroporous nanospheres having a larger gap than the microporous silica nanospheres having a gap hole that does not significantly differ from the size of Glp-1 having a molecular weight of 3356 and an isoelectric point of 4.49 Glp- It is much more advantageous to mount 1 and indicates that the pore size of the porous silica nanospheres must be larger than the size of the drug to be mounted.

On the other hand, since the porous silica nanospheres having hydroxyl groups (OH) on the surface have a large empty space therein, it was found that there is no difficulty in mounting weak acidic drugs such as Glp-1. If the drug is acidic, the surface of the silica can be modified with a basic compound such as amine (NH ₂ ), and if the drug is basic, the surface of the silica can be modified with a compound having a carboxyl group (COOH).

The above results show that the hollow porous silica nanospheres according to the present invention block a attack from external proteolytic enzymes that are approximately 10 times larger than the mass of the drug by putting a peptide drug such as Glp-1 inside. Indicates that it can be a good candidate.

As shown in FIG. 5, it was observed that until 6, 9, and 21 days of measurement, Glp-1 was continuously released from about 20 ug, i.e., about 10%, from the hollow porous silica nanospheres according to the present invention. It was confirmed that Glp-1 was steadily released until the 35th and 41st days.

As described above, the hollow porous silica nanospheres prepared according to the method for manufacturing hollow porous silica nanospheres according to the present invention, when irradiated with ultrasonic waves, when one of the peptide drugs Glp-1 is loaded into the porous silica nanospheres, It indicates that the drug can be protected from attack by foreign proteolytic enzymes. This indicates that the drug can be prevented from biodegrading in the blood circulation process, and thus the drug efficacy can be maintained for a long time.

In particular, the hollow porous silica nanospheres according to the present invention can control the size and number of crevices on the surface of the nanospheres according to the molecular weight of the drug to be put inside, so that the drug can control the concentration released per hour. It can be, accordingly, the hollow porous silica nanospheres according to the present invention can be utilized as a useful drug carrier.

The hollow porous silica nanospheres according to the present invention loaded with a drug may be suspended in an aqueous solution such as physiological saline and administered into the human body as an injection, or freeze-dried and formulated into an oral dosage form such as powder.

<Experimental Example>

Introducing a specific experimental example for the production of hollow porous silica nanospheres according to the present invention.

First, in the process of preparing a polystyrene mold having a diameter of 300 nm or less with a carboxyl group on the surface, 0.2 g of potassium peroxodisulfate (potassium peroxodisulfae) is dissolved in 160 ml of distilled water and heated at 80 rpm while mixing at 300 rpm. To this solution, 50 ml of styrene solution was added, and after 20 minutes, 2 ml of methacrylic acid was added and reacted while mixing for 24 hours. After adding the ethanol to the reaction solution, the water is removed by a rotary evaporator and dried in a vacuum dryer at 60°C for 12 hours.

Next, in the process of synthesizing porous silica nanospheres with a polystyrene mold having a diameter of 300 nm, 1 g of polystyrene particles are added to 100 ml of ethanol and then dispersed by irradiating with ultrasound for 30 minutes. After adding 1 ml of tetraethyl silicate, 3 ml of ammonia solution and 5 ml of distilled water to the dispersed solution, mix at 300 rpm for 24 hours at room temperature. The mixed solution was centrifuged at 3000 rpm for 20 minutes, and then the supernatant was discarded. Then, the precipitate is harvested and washed with distilled water and ethanol (1:1 v/v) solution. After centrifugation, the solids are collected and dried at 60°C for 12 hours.

Subsequently, in the process of synthesizing the porous silica nanospheres while changing the addition amount of tetraethyl silicate, the amount of tetraethyl silicate added in the synthesis process of the porous silica nanospheres was changed to 1 ml, 1.5 ml, 2 ml, 2.5 ml, and 3 ml. Repeat the process.

Next, the process of synthesizing the porous silica nanospheres while changing the addition amount of the ammonia solution, while changing the amount of the ammonia solution added during the synthesis of the porous silica nanospheres to 0.5ml, 1ml, 1.5ml and 2ml, Repeat.

Next, in the process of removing the polystyrene mold, after placing the dry solid in an electric furnace, calcining while slowly raising the temperature from 300°C to 2 hours, 450°C to 2 hours, and 600°C to 4 hours, cooled to room temperature, and then turned white. A powder of porous silica nanospheres is obtained.

Subsequently, in the process of loading Glp-1 into the porous silica nanospheres, 200 µg of Glp-1 was dissolved in 100 ul of a buffer solution (50 mM Tris-HCl, pH 8.0, 10 mM CaCl ₂ ), followed by the present invention. 1 mg of porous silica nanospheres are added, and the mixed solution is irradiated with ultrasonic waves for 30 minutes to allow Glp-1 to enter the silica spheres. After centrifuging them, 10 ul of supernatant was taken and 2.5 ul of 4×SDS-PAGE sample loading solution (20 mM Tris-HCl, pH 6.8, 12 mM β-mercaptoethanol, 2% SDS, 24% glycerol, 0.1% bromophenol blue) was added and stored in a cryogenic freezer at -70°C (labeled 0 days in FIG. 5).

Next, in the process of slowly releasing Glp-1 from the porous silica nanospheres according to the present invention, the mixture solution was left at room temperature for 6 days and centrifuged to take 10ul of the supernatant and 2.5ul of 4 After loading the ×SDS-PAGE sample loading solution, it was stored in a cryogenic freezer at -70°C (indicated by 6 days in FIG. 5 ). After leaving for 9 and 21 days, the same experiment was repeated (indicated by 9 and 21 in FIG. 5). 10 µl of a buffer solution was newly added to the precipitate, and after standing for 14 days, centrifuged to obtain the same amount of supernatant, 2.5 µl of 4×SDS-PAGE sample loading solution was added thereto, and then stored in a cryogenic freezer (35 in FIGS. 5 to 35). In days). The same process was carried out for 6 more days (indicated as 41 in FIG. 5).

Accordingly, in the process of confirming Glp-1 released from the porous silica nanospheres according to the present invention, samples stored in a cryogenic freezer were electrophoresed for 1 hour at 100 volts in SDS-polyacrylamide gel. To separate. The electrophoretic gel was left to stand for 15 minutes in a Coomassie blue staining solution (1.25 g Coomassie Blue R-250, 225 ml methanol, 225 ml distilled water, 50 ml glacial acetic acid) and then stained, followed by a staining solution (300 ml methanol, 100 ml acetic acid, After standing for 20-30 minutes in 600 ml distilled water), check the strip of the separated Glp-1.

As described above, the porous silica nanospheres according to the present invention and a method for manufacturing the porous silica nanospheres having a hollow inside and having pores in the surface shell control the size and number of gaps in the shell and the thickness of the shell. It can be, can be effectively used as a drug carrier because it can effectively transport the drug containing the peptide-based drug by releasing the peptide-based drug mounted on the porous silica nanospheres steadily and slowly.

In addition, the porous silica nanospheres according to the present invention and a method for manufacturing the same can prevent side effects caused by normal cells by not repeatedly taking an excessive amount of drugs in order to produce a desired therapeutic effect, and minimize the amount of drugs consumed. And the drug can be released steadily to keep the drug for a long time.

In addition, the porous silica nanospheres according to the present invention and its manufacturing method are natural mineral components, and by providing porous nanospheres made of silica that is not toxic to the human body, the nanospheres are destroyed by time and discharged outside the body. It can be used as a drug carrier.

On the other hand, in the detailed description of the present invention has been specifically described with reference to preferred embodiments referenced by the accompanying drawings, various modifications are possible within the limits without departing from the scope of the present invention. Therefore, the scope of the present invention should not be limited to the described embodiments, but should be determined not only by the scope of the claims described below, but also by the scope and equivalents of the claims.

1: Polystyrene template polymer 2: Silica shell
3: hollow porous silica nanospheres 4: hollow

Claims (16)

Synthesizing a template polymer having a functional group on the surface;
Synthesizing porous silica nanospheres by depositing a silica precursor in a basic solution on the surface of the template polymer; And
A method for manufacturing hollow porous silica nanospheres, characterized by comprising; the step of synthesizing the hollow porous silica nanospheres by firing and removing the template polymer.
The method of claim 1, wherein the step of synthesizing the template polymer,
A method for producing hollow porous silica nanospheres, characterized by synthesizing a polystyrene template polymer having a size of 50-600 nm using styrene monomer and methacrylic acid.
The method of claim 1, wherein the step of depositing the silica precursor on the surface of the template polymer,
A method for preparing hollow porous silica nanospheres, wherein a silica layer is hydrolyzed on the surface of the template polymer in a basic solution to form a uniform silica layer.
The method of claim 3, wherein the basic solution,
Method for producing hollow porous silica nanospheres, characterized in that the pH is 9-12.
The method of claim 4, wherein the basic solution,
Method for producing hollow porous silica nanospheres, characterized in that the ammonia solution.
The method of claim 3, wherein the silica precursor,
Hollow characterized by combining any one or two or more of alkoxysilane, alkoxysilane, teramethyl silicate, tetraethyl silicate, and tetrapropyl silicate, which are silicon-containing compounds. Method for preparing porous silica nanospheres.
According to claim 3, The step of synthesizing the hollow porous silica nanospheres,
A method for producing hollow porous silica nanospheres, characterized in that the porous silica nanospheres on which the silica layer is formed are fired in an electric furnace at 300-700° C. for 6 hours to remove the template polymer.
The method of claim 7,
A method for producing hollow porous silica nanospheres, characterized by removing the template polymer while slowly progressing the temperature change from 300°C to 2 hours, from 450°C to 2 hours, and from 600°C to 4 hours.
The method of claim 6,
A method for manufacturing hollow porous silica nanospheres, wherein the amount of the pores formed on the surface of the nanospheres is controlled by changing the amount of the silica precursor added, and the thickness of the silica layer is controlled.
The method of claim 9,
When the amount of the silica precursor is increased, the size of the pores formed on the surface of the nanospheres increases, the number decreases, and the thickness of the silica layer becomes thicker.
The method of claim 4,
Hollow porous silica nano, characterized in that by adjusting the addition amount of the basic solution, the pH of the basic solution is changed from 10.15 to 11.01 to control the size, number of pores formed in the nanosphere surface, and the thickness of the silica layer. Old manufacturing method.
The method of claim 11,
When the amount of the basic solution is increased, the size of the pores formed on the surface of the nanospheres increases, the number decreases, and the thickness of the silica layer becomes thicker.
A hollow porous silica nanosphere prepared by the method of any one of claims 1 to 12.
The method of claim 13,
A hollow porous silica nanosphere characterized in that it is used as a drug carrier by loading a drug inside the hollow porous silica nanosphere.
The method of claim 14,
Loading the drug inside the hollow porous silica nanospheres, the porous porous silica nanospheres, characterized in that made by adding the hollow porous silica nanospheres to the solution in which the drug is dissolved, and then irradiating with ultrasound.
The method of claim 15,
The hollow porous silica nanospheres, characterized in that the drug mounted on the inside of the hollow porous silica nanospheres is slowly released over time (slow release).

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