KR20140135571A - Preparing method of microcapsule of oral delivery of active substance using natural polymer - Google Patents

Abstract

The present invention relates to a microcapsule for oral delivery of active ingredients using natural polymers comprising the steps of: (a) preparing an electrospraying solution by dissolving the natural polymers; (b) adding active ingredients to the electrospraying solution; (c) forming microcapsules by electrospraying when the electrospraying solution with the active ingredients receives voltages; and (d) coating the surface of the microcapsules with a coating solution in which polymethyl methacrylate is dissolved in a solvent.

Description

TECHNICAL FIELD [0001] The present invention relates to a microcapsule for oral delivery of an effective substance using a natural polymer,

The present invention relates to a method for producing oral microcapsules, and more particularly, to a method for producing microcapsules for oral delivery of an effective substance such as a functional protein or an anti-cancer substance using a natural polymer.

The success of chemotherapy depends on the effective killing of cancer tissues by chemotherapy, because the chemotherapy and radiation therapy that are administered when the cancer develops in the body has serious side effects that are highly toxic to normal tissues, not cancer tissues. In particular, cancer in the gastrointestinal system is treated by radiotherapy after the incision of the cancer tissue through the surgical procedure, but the digestive system tissues are easily damaged by radiation compared with other tissues or organs, There is a serious side effect accompanied by the damage to the shape and function of the tissue.

In order to solve the problems of such radiation therapy, research on a method of delivering an effective substance such as a functional protein capable of inducing tissue regeneration of injured digestive system into the body has become a subject of interest, (Korean Patent Publication No. 10-2011-0095590). Such a capsule-shaped carrier may have advantages such as protection of volatile liquids and active substances, odor, tactile shielding, and timing and speed of release of the encapsulated substance to the outside.

However, the conventional capsule-shaped carrier uses a synthetic polymer, which results in a problem of poor long-tackiness and biocompatibility, and it is difficult and complicated to manufacture the capsule-shaped carrier using the synthetic polymer, .

Disclosure of Invention Technical Problem [8] Accordingly, the present invention has been made keeping in mind the above problems occurring in the prior art, and an object of the present invention is to provide a method for preparing an electrospray solution comprising: (a) dissolving a natural polymer in a solvent to prepare an electrospray solution; (b) adding an effective substance to the electrospray solution; (c) forming a microcapsule by electrospraying the electrospray solution to which the effective material is added in a state of applying a voltage; And (d) coating the surface of the microcapsule with a coating solution in which polymethylmethacrylate is dissolved in a solvent. The present invention also provides a method for producing microcapsules for oral delivery of an effective substance.

It is still another object of the present invention to provide a microcapsule for oral delivery of an effective substance prepared by the above method.

However, the technical problem to be solved by the present invention is not limited to the above-mentioned problems, and other matters not mentioned can be clearly understood by those skilled in the art from the following description.

According to the present invention,

(a) preparing an electrospray solution by dissolving a natural polymer in a solvent;

(b) adding an effective substance to the electrospray solution;

(c) forming a microcapsule by electrospraying the electrospray solution to which the effective material is added in a state of applying a voltage; And

(d) coating the surface of the microcapsule with a coating solution in which polymethylmethacrylate is dissolved in a solvent, to prepare a microcapsule for oral delivery of an effective substance.

In one embodiment of the present invention, in the step (a), the natural polymer is selected from the group consisting of alginic acid, carboxymethylcellulose, carboxymethylchitin, carboxymethylchitosan, glycochitanium, gelatin, fibrin and mixtures thereof And at least one natural polymer.

In another embodiment of the present invention, in the step (b), the effective substance is selected from the group consisting of contrast agents, anticancer agents, cells or bacteria, and siRNA.

In another embodiment of the present invention, in the step (b), the active substance is a cytolysin or a peptide consisting of the amino acid sequence of SEQ ID NO: 1.

In another embodiment of the present invention, the solvent in step (a) is selected from the group consisting of dimethyl sulfoxide, phosphate buffered saline, Hanks buffered saline (HBSS), normal cellulase (0.9 wt. % Sodium chloride aqueous solution) and a mixed solvent thereof.

In another embodiment of the present invention, the applied voltage in step (c) is 5 to 20 kV, and the rate of pushing the electrospray solution is 0.1 to 5 ml / hr.

In another embodiment of the present invention, in the step (d), the polymethylmethacrylate is Eudragit.

In yet another embodiment of the present invention, the solvent in step (d) is selected from the group consisting of water, methanol, ethanol, chloroform, dichloromethane, .

The present invention provides microcapsules for oral delivery of an effective substance prepared by the above method.

According to the method for preparing microcapsules for oral delivery of an effective substance using a natural polymer according to the present invention, it is possible to provide a microcapsule for oral delivery, which comprises alginic acid, carboxymethylcellulose, carboxymethylchitin, carboxymethylchitosan, glycholchitosan, gelatin, fibrin, (Or an anti-cancer substance) capable of inducing the regeneration of the damaged tissue by radiation, and then preparing EUCD-coated microcapsules by electrospray method to prepare reinforced microcapsules It is possible to produce microcapsules having various diameters in accordance with the production conditions, but it is also a process of short-time processing, and thus it is very advantageous to maintain the activity of active substances such as functional proteins and / or anti-cancer substances contained therein .

In addition, the prepared natural polymer microcapsules can maintain the activity of the internal functional protein (or anti-cancer substance) in the harsh environment of the stomach / intestine when orally administered and can induce healing of the injured intestinal tissue, It is possible to deliver cells, drugs, anticancer agents, contrast agents and the like in the intestines in addition to functional proteins of various compositions according to diagnosis and treatment purpose.

BRIEF DESCRIPTION OF DRAWINGS FIG. 1 is a schematic view of an electrospray apparatus for producing alginic acid microcapsules in Example 1-1. FIG.
Fig. 2 is a view showing an image taken by an optical microscope of alginic acid microcapsules prepared by using the electrospray method in Example 1-1. Fig.
Fig. 3 is a graph showing the average diameter of alginic acid microcapsules prepared by the electrospray method in Example 1-1. Fig.
FIG. 4 is a graph showing the results of analysis of changes in average diameter before and after enhancement of physical properties of capsules by Eudragit in Example 1-2. FIG.
Fig. 5 is a view showing an image obtained by observing the surface morphology of alginic acid microcapsules in Examples 1-3 through an electron microscope (SEM). Fig.
6 is a diagram showing the results of analysis using an infrared spectroscope (FTIR) to analyze chemical changes of alginic acid microcapsules in Examples 1-3.
7 is a graph showing the results of analysis of the behavior of the substance P emitted from alginic acid microcapsules in Example 2-2.
FIG. 8 is an optical image of bone marrow-derived mesenchymal stem cells taken in order to examine the cell effect of substance P released from alginate microcapsules in Example 2-3. FIG.
FIG. 9 is a graph showing the results of CCK-8 assay of bone marrow-derived mesenchymal stem cells cultured to examine the cell activity change by substance P released from alginate microcapsules in Example 2-3. FIG.
10 is a photograph showing a live and dead assay staining of E. coli in alginic acid microcapsules in Example 3-1.
11 is a view showing a result of analysis of the survival rate of E. coli in alginate microcapsules in Example 3-1 through a live and dead assay.
12 is a graph showing the results of MTT assay in which the cytolysin secreted by the genetically engineered E. coli in Example 3-2 was analyzed for the influence of tumor and normal cells.

The inventors of the present invention have studied microcapsules for oral delivery of an effective substance using a natural polymer and found that they are composed of alginic acid, carboxymethylcellulose, carboxymethylchitin, carboxymethylchitosan, glycochitanium, gelatin, fibrin, The present inventors have found that when a microcapsule made of at least one natural polymer selected from the group consisting of a functional protein or a bacterial cell contains a functional protein released in the form of a sustained release to the outside, The present invention has been completed.

Hereinafter, the present invention will be described in detail.

According to the present invention,

(a) preparing an electrospray solution by dissolving a natural polymer in a solvent;

(b) adding an effective substance to the electrospray solution;

(c) forming a microcapsule by electrospraying the electrospray solution to which the effective material is added in a state of applying a voltage; And

(d) coating the surface of the microcapsule with a coating solution in which polymethylmethacrylate is dissolved in a solvent, to prepare a microcapsule for oral delivery of an effective substance.

In the step (a), the natural polymer is dissolved in a solvent to prepare an electrospray solution. It is preferable to use at least one natural polymer selected from the group consisting of alginic acid, carboxymethylcellulose, carboxymethylchitin, carboxymethylchitosan, glycholchitosan, gelatin, fibrin, and mixtures thereof, Alginic acid can be used. Alginic acid in the form of an aqueous solution can be easily prepared into ion-crosslinked capsules using divalent cations such as Ca ^{2 +} , Ba ^{2 +} and Sr ^{2 +} . By using a natural polymer having excellent adhesiveness and biocompatibility for producing microcapsules, it is possible to local release and transfer of an effective substance (functional protein) only in a damaged tissue, thereby maximizing the tissue regeneration effect.

Examples of the solvent used in step (a) include dimethyl sulfoxide, phosphate buffered saline, Hanks buffered saline (HBSS), nasal selenium (0.9 wt% aqueous sodium chloride solution) A mixed solvent, and the like, but is not limited thereto.

The active substance of step (b) may be a functional protein and / or an anti-cancer agent, and is preferably selected from the group consisting of a contrast agent, an anticancer agent, a cell or a bacterium, and siRNA, (Substance P, Calbiochem, USA), which is a peptide consisting of the amino acid sequence of SEQ ID NO: 1, but is not limited thereto.

In the step (c), the electrospray solution prepared by the step (a) is injected with a voltage to form microcapsules. At this time, the applied voltage is 5 to 20 kV, the rate of pushing the electrospray solution Is preferably 0.1 to 5 ml / hr, but is not limited thereto. Microcapsules can be formed by the electrospray method. According to the electrospray method, it is possible to produce microcapsules having various diameters according to the production conditions. Since the process is a short-time process, the functional protein and / But also has a great advantage in maintaining the activity of the active substance.

In the step (d), the surface of the microcapsule formed by step (c) is coated with a coating solution in which polymethylmethacrylate is dissolved in a solvent. That is, the microcapsule formed by the step (c) has a weak physical property and is easily decomposed by the pH change of the stomach / intestinal environment and the protection of the effective substance contained therein is weak due to intestinal motility. To solve this problem, the surface of the microcapsule is coated with polymethylmethacrylate to enhance the physical properties. In this case, the polymethylmethacrylate is preferably EUDRAGIT, but the present invention is not limited thereto. The solvent used in step (d) may be a solvent selected from the group consisting of water, methanol, ethanol, chloroform, dichloromethane, and mixed solvents thereof. This is not restrictive. By coating the surface of the microcapsule with the step (d), the physical properties are enhanced, and the effective substance contained in the capsule can be perfectly protected from the pH change or the movement of the intestines.

In one embodiment of the present invention, a microcapsule is formed by a spraying method using a solution of a substance P, which is a functional protein, and alginic acid, and alginic acid microcapsules having enhanced physical properties are prepared through Eudragit coating (Example 2- 1). As a result of analyzing the release behavior of the functional protein, it was confirmed that the sustained-release behavior of the substance P existing in the living body under bio-similar conditions can be controlled (see Example 2-2). In another embodiment of the present invention, a microcapsule is prepared by a spraying method of a solution of a genetically engineered bacterium, E. coli, which secretes an anti-cancer substance, cytolysin, and alginic acid. Then, microcapsules are prepared by coating with Eudragit, Microcapsules were prepared, and the survival rate of the genetically engineered bacteria present in the capsule was evaluated. As a result, it was confirmed that the survival rate of genetically engineered bacteria was maintained (see Example 3-1).

Accordingly, the present invention provides a microcapsule for oral delivery of an effective substance prepared by the above-described method. The microcapsule may have one or more uses selected from the group consisting of a contrast agent carrier, an anticancer agent carrier, a cell or a bacterial carrier, and an siRNA carrier.

Hereinafter, preferred embodiments of the present invention will be described in order to facilitate understanding of the present invention. However, the following examples are provided only for the purpose of easier understanding of the present invention, and the present invention is not limited by the following examples.

[Example]

Example 1. Preparation and physical and chemical analysis of alginic acid microcapsules

1-1. Preparation of alginic acid microcapsules by electrospray method

In the present embodiment, an electrospray method, which is a system for producing capsules of micro size by applying an electric field to an alginic acid aqueous solution falling freely to make alginic acid microcapsules, was applied. The alginic acid microcapsules prepared by the electrospray method can easily control the size and size distribution of the capsules according to the applied voltage and the speed of pushing the solution.

More specifically, alginic acid (Sigma Aldrich, USA) was dissolved in a 0.9% sodium chloride solution (NaCl, Sigma Aldrich, USA) at a concentration of 2 wt% for about 6 hours to prepare alginic acid microcapsules, , And a schematic schematic view of the electrospray device is shown in FIG. Thereafter, a solution of 50 mM calcium chloride (CaCl ₂ , Sigma aldrich, USA) was prepared as a solution for crosslinking alginic acid. The distance between the syringe needle and the alginic acid cross-linking solution was about 10 cm. A copper wire having a diameter of 10 cm The concentration of alginic acid solution was adjusted to 0.1 ~ 1ml / hr by using a syringe pump (KD science, USA), and the applied voltage was measured using a high voltage power generator (Wookyung Tech, Korea ) To prepare alginic acid microcapsules having various diameters by adjusting to 8 to 20 kV. The alginic acid microcapsules thus prepared were washed five times with 0.9% sodium chloride solution for 30 minutes in order to remove calcium chloride as a residual ion crosslinking agent.

The alginic acid microcapsules prepared above were analyzed using an optical microscope for the diameter, and the results are shown in FIG. 2 and FIG.

As shown in FIG. 2 and FIG. 3, it was confirmed that the alginic acid microcapsules obtained by converting various conditions of the electrospray method were formed in a uniform size, and it was confirmed that the microcapsules And the average straightness of.

1-2. Alginic acid microcapsule reinforced coating

The capsule surface was coated with EUDRAGIT (Evonik Rhm GmbH, Pharma Polymers, Germany) to enhance the physical properties of alginic acid microcapsules prepared by the electrospray method of Example 1-1. More specifically, the EUDRAGIT used was dissolved in an aqueous solution of ethanol (ethanol: water ratio = 9: 1) at a concentration of 10 wt% at L100-55, L100 and S100 for about 1 hour at room temperature. Alginic acid microcapsules with surface water removed were placed on the paper filter, and negative pressure was applied from the bottom of the filter. Each EUDRAGIT solution was poured into capsules. The microcapsules were coated with EUDRAGIT solution and coated with EUDRAGIT. The microcapsules were coated with the EUDRAGIT coating solution before and after coating, The results are shown in Fig.

As shown in FIG. 4, after the EUDRAGIT coating, dehydration of alginic acid microcapsules was induced in the solvent of EUDRAGIT by alcohol, and the average diameter of the microcapsules was reduced by about 40%.

1-3. Physical and chemical analysis of alginate microcapsules

After the EUDRAGIT coating, the capsule surface was analyzed using an electron microscope to analyze the physical shape changes of the alginic acid microcapsules. The alginic acid microcapsules prepared in Examples 1-1 and 1-2 were naturally dried at room temperature for 12 hours, and then analyzed before and after EUDRAGIT coating and after coating, respectively. The results are shown in FIG.

As shown in FIG. 5, there was no change in the diameter of the capsules before and after coating, and it was confirmed that the center portion of the microcapsules was depressed by the EUDRAGIT coating.

The alginate microcapsules were analyzed by Fourier transform infrared spectroscopy (Tensor 27, Germany) for chemical change analysis before and after EUDRAGIT coating. For this purpose, the microcapsules prepared in Examples 1-1 and 1-2 were vacuum-dried for about 12 hours and then mixed with potassium bromide to prepare specimens. The spectroscopic characteristics of the samples in the wavelength range of 4000-500 cm ^-1 were measured And the results are shown in Fig.

As shown in FIG. 6, it was confirmed that characteristic peaks of alginic acid and each EUDRAGIT appeared well before and after coating.

Example 2 Preparation and Performance Evaluation of Alginic Acid Microcapsules Containing Functional Protein

2-1. Preparation of functional protein-containing microcapsules by electrospray method

Substance P is a neuropeptide composed of 11 amino acids as neurotransmitter and neurotransmitter and has the amino acid sequence of SEQ ID NO: 1 (Arg-Pro-Lys-Pro-Gln-Gln-Phe-Phe- -Met). Substance P is known to promote the migration and growth of mesenchymal stem cells to the affected area in the vicinity of injured tissue in the human body and has a regenerating effect on damaged tissues (Reid TW, Murphy CJ, Iwahashi CK, Foster BA, Mannis MJ 1993), "Stimulation of epithelial cell growth by the neuropeptide substance P", Journal of Cellular Biochemistry 52 (4): 476-85.

The alginic acid microcapsules capable of releasing the functional protein were prepared using the electrospray method set forth in Example 1 above. More specifically, 2 wt% of alginic acid solution was immersed in a 0.9 wt% aqueous solution of sodium chloride, and then substance P (Substance P, Calbiochem, USA) was dissolved at a concentration of 100 ng / ml. Alginic acid microcapsules containing the substance P, which is a functional protein, were prepared by using the thus-prepared solution using an electrospray (see Fig. 1). At this time, the applied voltage was 8 to 15 kV, the rate of pushing the alginic acid solution was 0.5 to 1.0 ml / hr, the distance between the syringe needle and the alginic acid cross-linking solution was about 10 cm, and the copper wire was positioned to generate a potential difference therebetween . The alginic acid microcapsules thus prepared were washed with 0.9 wt% sodium chloride solution for 5 minutes for 30 minutes to remove residual crosslinking agent. The physical properties of the alginate microcapsule containing the substance P were enhanced through the EUDRAGIT coating method conducted in Example 1-2.

2-2. Functional protein Release behavior  analysis

Immunoenzymatic immunoassay (R & D system, USA) was performed to analyze the behavior of substance P released from alginate microcapsules. The microcapsules containing the substance P prepared in Example 2-1 were divided into the same number and immersed in 10 ml of PBS, stirred at 100 rpm at a temperature of 37 ° C, and the behavior of the substance P released to the outside of the capsule for 1 week Respectively. In order to analyze the release behavior, the PBS solution was collected at 3, 7, 12, 24, 72, and 168 hours after the start of the experiment to perform immunoenzymatic analysis, and the results are shown in FIG.

As shown in FIG. 7, a relatively large amount of the substance P was initially released from the alginate capsule without the physical property-enhancing coating, but the coated capsule could confirm the sustained release of the substance P.

2-3. Cellular effect evaluation of alginic acid microcapsules containing functional protein

A cell culture insert (Corning, USA) was used to analyze the cellular effects of substance P released from alginate microcapsules. Five thousand bone marrow-derived mesenchymal stem cells (passage number: 10, Lonza, USA) were inoculated on the bottom of a tissue culture plate (TCP, Corning, USA), and the microcapsules prepared in Example 2-1 culture inserts were injected onto each TCP and the effect of substance P released from capsules on bone marrow-derived mesenchymal stem cells was assessed by optical microscopy and cell counting kit (CCK-8, Dojindo Molecular Technologies , JAPAN). The cell culture medium used was 10 v / v% Fetal bovine serum (FBS, Welgene, Korea) and 1 v / v% antibiotics (Welgene, Korea) in Dulbecco's Modified Eagle Medium (DMEM, low glucose, Invitrogen, USA) The cells were cultured under the condition of 5% CO 2 at 37 ° C for 1 week, and the cells were observed and analyzed 1, 4, and 7 days after the start of culture. The results are shown in FIG. 8 And Fig. 9.

As shown in FIGS. 8 and 9, in the microcapsules without EUDRAGIT coating, it was confirmed that the substance P was rapidly released earlier than the other groups, thereby promoting the proliferation of bone marrow-derived mesenchymal stem cells. On the other hand, it was confirmed that the substance P released from the alginate capsule coated with the physical property enhances stem cell proliferation continuously for 1 week as it is gradually released over a week.

Example  3. Preparation and performance evaluation of alginate microcapsules containing anticancer substance-secreted genetically engineered bacteria

3-1. Of the genetically engineered bacteria present in the capsule Survival rate  evaluation

In this Example, alginic acid microcapsules containing genetically engineered bacteria were prepared for the purpose of delivering a genetically engineered bacterium ( E. coli ) which secretes an anticancer substance, Cytolysin, to intestinal / ductal tissues. Using Example 1 Genetic manipulation of E. coli 1.0 × 10 ⁵ CFU / ml was dispersed at a concentration of (see Fig. 1) electrospray devices at a alginate solution of 2 wt% using an electrical spraying established for this purpose in E The alginic acid microcapsules containing . coli were prepared. At this time, the applied voltage was 8 to 15 kV, the rate of pushing the alginic acid solution was 0.5 to 1.0 ml / hr, the distance between the syringe needle and the alginic acid crosslinking solution was about 10 cm and the copper wire to generate the potential difference therebetween . The alginic acid microcapsules thus prepared were rapidly washed with sodium chloride solution of 0.9 wt% for 5 minutes for 10 minutes in order to remove residual crosslinking agent. The physical properties of alginate microcapsules containing E. coli were enhanced by EUDRAGIT coating method in Example 1-2.

The survival rate of E. coli in the prepared alginate microcapsules was measured by Live & Dead kit (Invitrogen, USA). Prior to the analysis, the same amount of microcapsules were placed in each tube to remove the E. coli present in the prepared alginate microcapsules. After the ion-crosslinked alginate was pulverized by scissors, alginate was eluted by immersing it in a mixed solution of 55 mM sodium citrate (Sigma Aldrich, USA), 30 mM EDTA (ethylenediaminetetraacetic acid, Sigma aldrich, USA) and 0.15 M NaCl. Then, bacteria were precipitated at 4000 rpm for 5 minutes using a centrifuge and then stained with Live & Dead kit. After staining, the bacterial solution was analyzed using a fluorescence microscope and a miroplate reader (Molecular Devices, USA), and the results are shown in FIGS. 10 and 11.

As shown in FIGS. 10 and 11, the survival rate of the bacteria was found to be about 90% in the alginate microcapsules as compared with the control group, and the survival rate of the bacteria in the capsules Was about 60%.

3-2. Evaluation of tumor cell killing effect of anticancer substances secreted by genetically engineered bacteria

To investigate the effect of cytolysin secreted by genetically engineered E. coli on tumor cells, the tumor cells were cultured in Lovo (American Type Culture Collection), HCT-8 (ATCC: American Type Culture Collection), WiDr Cell line bank) and NF-46 (Young Science, KOREA), a normal cell, were cultured in culture plates at 2 × 10 ⁴ / ml cells per well at 37 ° C and 5% CO _2. DMEM or RPMI1640 was added with 10% FBS. Cells were stabilized for 24 hours and then 0, 2.5, 5, 15, 50, and 125 ug of cytolysin were added to each cell culture medium. Thereafter, the cells were cultured for 47 hours and evaluated for 1 hour using 3- (4,5-Dimethylthiazol-2-yl) -2,5-diphenyltetrazolium bromide (MTT) assay. The results are shown in FIG.

As shown in FIG. 12, it was confirmed that the degree of sensitivity to ClyA was in the order of HCT-8, WiDr, NF46 and Lovo on the basis of the group treated with 15 μg. More specifically, the WiDr group showed about 40% cell activity for 2.5 ug of ClyA, and the activity decreased as the concentration increased. In the case of Lovo group, the activity was more than 80% at 5 ug. The HCT-8 group showed more than 80% activity up to 5 ug, but the activity of the HCT-8 group fell to 15% or less in the group of 15 ug or more. NF46, the primary cultured cell, decreased in activity in proportion to ClyA concentration.

It will be understood by those skilled in the art that the foregoing description of the present invention is for illustrative purposes only and that those of ordinary skill in the art can readily understand that various changes and modifications may be made without departing from the spirit or essential characteristics of the present invention. will be. It is therefore to be understood that the above-described embodiments are illustrative in all aspects and not restrictive.

<110> Korea Institute of Radiological Medical Sciences <120> PREPARING METHOD OF MICROCAPSULE OF ORAL DELIVERY OF ACTIVE          SUBSTANCE USING NATURAL POLYMER <130> PB13-11396 <160> 1 <170> Kopatentin 2.0 <210> 1 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> amino acid sequence of Neuropeptide substance P <400> 1 Arg Pro Lys Pro Gln Gln Phe Phe Gly Leu Met   1 5 10

Claims (9)

(a) preparing an electrospray solution by dissolving a natural polymer in a solvent;
(b) adding an effective substance to the electrospray solution;
(c) forming a microcapsule by electrospraying the electrospray solution to which the effective material is added in a state of applying a voltage; And
(d) coating the surface of the microcapsule with a coating solution in which polymethylmethacrylate is dissolved in a solvent.
The method of claim 1, wherein the natural polymer is selected from the group consisting of alginic acid, carboxymethylcellulose, carboxymethylchitin, carboxymethylchitosan, glycochitosan, gelatin, fibrin, and mixtures thereof. Wherein the microcapsules are natural macromolecules of at least two kinds.
2. The method of claim 1, wherein the effective material in step (b) is selected from the group consisting of a contrast agent, an anti-cancer agent, a cell or a bacteria, and an siRNA.
The method of claim 1, wherein the effective material in step (b) is a cytolysin or a peptide consisting of the amino acid sequence of SEQ ID NO: 1.
The method of claim 1, wherein the solvent in step (a) is selected from the group consisting of dimethyl sulfoxide, phosphate buffered saline, Hanks buffered saline (HBSS), nasal cell line (0.9 wt% ) And a mixed solvent thereof. &Lt; RTI ID = 0.0 &gt; 11. &lt; / RTI &gt;
The method of claim 1, wherein the applied voltage in step (c) is 5 to 20 kV, and the rate of pushing the electrospray solution is 0.1 to 5 ml / hr.
2. The method of claim 1, wherein in step (d), the polymethylmethacrylate is EUDRAGIT.
The method of claim 1, wherein the solvent in step (d) is selected from the group consisting of water, methanol, ethanol, chloroform, dichloromethane, By weight.
A microcapsule for oral delivery of a pharmaceutically effective substance prepared by the method of any one of claims 1 to 8.

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