KR20130104534A - Preparation method for gliadin nanoparticles by electrospray - Google Patents

Abstract

PURPOSE: A gliadin nanoparticle prepared by electrospray is provided to have an average particle size which is smaller than the particle size of gliadin particles prepared by desolvation and to increase zeta potential, thereby reducing aggregation of the nanoparticles. CONSTITUTION: A method for preparing gliadin nanoparticles comprises the steps of: dissolving gliadin in 65-80 (v/v)% ethanol solution to prepare a gliadin solution; and performing electrospray of the gliadin solution. The concentration of gliadin in the gliadin solution is 5-10 wt%. The flux and voltage difference of the gliadin solution during electrospray is 0.1-1 ml/hr and 12-18 kV, respectively. The diameter of a nozzle is 0.1-0.5 mm.

Description

Preparation method for gliadin nanoparticles by electrospray using electrospray drying [

The present invention relates to a method for preparing gliadin nanoparticles or gliadin and gelatin composite nanoparticles usable as drug delivery materials, nanoparticles prepared therefrom, and drug delivery vehicles containing the same.

Glyadine is a vegetable natural protein isolated from wheat gluten, known as a protein rich in neutral and fat soluble amino acids. Since the neutral amino acid of gliadin promotes hydrogen bonding in vivo mucosal tissues and the lipid soluble moiety promotes the interaction in living tissues, the gliadin nanoparticle has a unique hydrophobicity and limited solubility, and bioadhesion ) Capability, particularly as a polymer that can be attached to the mucos layer, has attracted attention as a drug delivery vehicle having biodegradability and biocompatibility.

Gliadin nanoparticles having an average particle size of greater than 500 nm have been prepared for regulated release of retinoic acid and gliadin particles containing scarcelycillin exhibiting anti-Helicobacter pylori activity have been produced, All using a desolvation method.

This desolvation method has a problem in that it is difficult to separate nanoparticles produced in a solution state and has a problem that a separate surfactant is required in the manufacturing process, and that the produced nanoparticles exhibit neutrality, There was a limit to occur.

On the other hand, electrospray is a method of producing fine particles by injecting a polymer solution having a certain degree of electric conductivity and viscosity into a capillary and then applying an electrostatic force, so that the shape and structure of the device are simple and easy to manufacture. The generated particles are not only capable of having a monodisperse distribution but also are useful for producing particles of various sizes, and studies are currently under way to apply them to drug-containing particle production in the field of drug delivery (Xie et al, Biomaterials, 27: 3321 (2006)).

Korean Patent Laid-Open No. 10-2010-0134368 discloses a method for producing composite particles of a cationic polymer and an anionic polymer by using electrospray drying. However, a method of preparing composite particles of a cationic polymer and an anionic polymer by using electroless spray drying, And there is a limit to the technology for providing particles having a size such that the size of the produced particles can not be ingested intracellularly at a micro level.

The present invention relates to a method for preparing gliadin nanoparticles or gliadin and gelatin composite nanoparticles which are useful as drug delivery materials because of their small and uniform nanoparticle size and high zeta potential, Method.

The present invention relates to a process for preparing a gliadin solution by dissolving gliadin in an aqueous solution of 65 to 80 (v / v)% ethanol; And a step of electrospraying the gliadin solution.

In the method for producing the gliadin nanoparticles of the present invention, the gliadin concentration of the gliadin solution is preferably 5 to 10 wt%. When the concentration of the gliadin solution is less than the lower limit, the uniformity of the size of the nanoparticles is lowered. If the concentration exceeds the upper limit, the nanoparticles do not form spherical particles and begin to form a fibrous network.

In the method for producing the gliadin nanoparticles of the present invention, the flow rate of the solution in the electrospray may be 0.1 to 1 ml / hr and the voltage difference may be 12 to 18 kV. This is because the flow rate is very slow compared with the conventional electric spraying and a high voltage is applied. When the flow rate is out of the above range, the nanoparticles of the desired size can not be formed in the present invention.

The method for producing the gliadin nanoparticles of the present invention is characterized in that the diameter of the nozzle in the electrospray is 0.1 to 0.5 mm.

The present invention provides a gliadin nanoparticle which is produced by an electrospray method and has a zeta potential of 15 to 30 mV and an average particle size of 200 to 350 nm. The gliadin nanoparticles of the present invention are significantly smaller in size than the gliadin particles prepared by the conventional desolvation method and have a high zeta potential so that the prepared nanoparticles can be dispersed well without aggregation.

The present invention relates to a process for preparing a gliadin solution by dissolving gliadin in an aqueous solution of 65 to 80 (v / v)% ethanol; Dissolving gelatin in an aqueous solution of 80 to 99 (v / v)% acetic acid to prepare a gelatin solution; Mixing the gliadin solution and the gelatin solution to prepare a mixed solution; And electrospraying the mixed solution of the gliadin and the gelatin. The present invention also provides a method for producing the gliadin and gelatin composite nanoparticles.

The method for producing the gliadin and gelatin composite nanoparticles of the present invention is characterized in that 0.5 to 5 parts by weight of a crosslinking agent is further added to 100 parts by weight of the solid content of the mixed solution of gliadin and gelatin.

In the method for producing the gliadin and gelatin composite nanoparticles of the present invention, the gliadin concentration of the gliadin solution is 5 to 10 wt%, and the gelatin concentration of the gelatin solution is 0.1 to 6 wt% .

The present invention provides a gliadin and gelatin composite nanoparticle which is produced by an electrospray method and has a zeta potential of 15 to 30 mV and an average particle size of 300 to 500 nm.

The present invention relates to the aforementioned gliadin nanoparticles or said gliadin and gelatin composite nanoparticles; And a pharmaceutically active ingredient.

In the present invention, 'pharmaceutically active ingredient' includes all biological and chemical substances used for prevention, treatment, alleviation or alleviation of disease, and a substance which assists the pharmacological effect of other drugs, Active ingredient ". For example, anticancer agents, anti-inflammatory agents, antimicrobial agents, antiviral agents, hormones, and antioxidants may be used as the 'pharmaceutically active ingredients' of the present invention.

In addition to the above components, conventional additives known in the art can be included, such as excipients, stabilizers, pH adjusters, antioxidants, preservatives, binders or disintegrants. At this time, the carrier may further include other additives, solvents, and the like known in the art.

In addition, the drug delivery vehicle may be formulated in the form of oral or parenteral preparations, and may be manufactured into intravenous, muscular, or subcutaneous injections.

In the sustained release drug delivery system of the present invention, the pharmacologically active ingredient is cyclophosphamide, and the sustained drug delivery system can be used for the treatment of breast cancer by inducing natural history of breast cancer cells.

The gliadin nanoparticles or the gliadin and gelatin composite nanoparticles produced by the electrospray of the present invention have an average particle size smaller than that of the gliadin particles produced by the desolvation method and the zeta potential The nanoparticles of the present invention can be used as a drug delivery system. When the nanoparticles of the present invention are used as a drug delivery system, nonspecific toxicity including low solubility, instability, and selectivity of existing drugs can be alleviated .

1 is a schematic diagram of an electrospray device consisting of an injection pump, a spray nozzle, a high voltage supplier and a collector.
2 is an electron micrograph of the gliadin nanoparticles prepared using the 7 wt% gliadin solution of Preparation Example 2, Fig. 2B is a 7 wt% gliadin solution of Preparation Example 3, and Fig. 4% by weight gelatin solution. The micrograph of the gliadin and gelatin composite nanoparticles is shown in Fig.
3 is a transmission electron micrograph of the gliadin nanoparticles prepared using the 7 wt% gliadin solution of Preparation Example 2, Fig. 3B is a 7 wt% gliadin solution of Preparation Example 3, and Fig. 4% by weight gelatin solution. The results are shown in Table 1. < tb >< TABLE > Columns = 2 < tb >
FIG. 4A is an electron micrograph of the gliadin nanoparticles prepared by using the 14% by weight gliadin solution of Preparation Example 2 , FIG. 4B is the 7% by weight glyandin solution of Preparation Example 3 and FIG. FIG. 4C is an electron micrograph of the gliadin and gelatin composite nanoparticles prepared using the mixed solution of 8 wt% gelatin solution , and FIG. 4C is a photograph of 14 wt% glyadin solution and 8 wt% gelatin solution of Preparation Example 3 Of a mixed solution of gliadin and gelatin nanoparticles .
FIG. 5A is a graph showing the results of measurement of the activity of the glialazine nanoparticles prepared using the 7% by weight gliadin solution of Preparation Example 2, B, the mixed solution of 7% by weight of the glyandin solution of Preparation Example 3 and 4% by weight of the gelatin solution , And C was a mixture of gliadin and gelatin composite nanoparticles prepared by using a mixed solution of 7 wt% of glyandin solution and 8 wt% of gelatin solution of Preparation Example 3 Particle size distribution.
FIG. 6A is a graph showing the results of measurement of the activity of the gliadin nanoparticles prepared by using the 14% by weight gliadin solution of Preparation Example 2 , and B being the mixed solution of 14% by weight of the glyandin solution of Preparation Example 3 and 4% by weight of the gelatin solution a gliadin and gelatin composite nanoparticles, and C is a particle of a glycidyl Adin and gelatin composite nanoparticles prepared using a mixture solution of 14% by weight of gliadin solution and 8% by weight of the preparation 3 gelatin solution prepared using FIG.
7 is a graph showing the cross-linking polymerization between gliadin and gelatin in the gliadin and gelatin composite nanoparticles prepared from the mixed solution of the 7 wt% gliadin solution and the 4 wt% gelatin solution of Production Example 3 X-ray diffraction analysis graph for confirmation.
FIG. 8 is a graph showing the results of measurement of the activity of the gliadin nanoparticles prepared by using the 7% by weight gliadin solution of Preparation Example 2, a mixed solution of 7% by weight of the glyandin solution of Preparation Example 3 and 4% by weight or 8% FIG. 2 is a graph showing drug release patterns of gliadin and gelatin composite nanoparticles prepared using the method of the present invention.
9 is a graph showing the results of a comparison between the control group 1 (Fig. 9A) in which MCF7 breast cancer cells were cultured for 24 hours, the control group 2 (Fig. 9A) in which only gliadin nanoparticles not containing cyclophosphamide B), a control group 3 (FIG. 9C) in which only cyclophosphamide was added to the medium, and an example (FIG. 9D) to which cyclophosphamide-impregnated gliadin nanoparticles were added (FIG. 9D) , 24 hours, and 48 hours, respectively, and stained with Annexin V and propidium iodine, respectively, to observe cellular apoptosis and cell necrosis.
10 is a Western blot photograph showing the expression levels of Bcl-2 protein in Control Groups 1 to 3 and Examples.

Hereinafter, the present invention will be described in detail by way of examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Preparation Example 1 : Gliadin  Produce

Lyophilized and ground gluten extracted from wheat flour was used. The gluten and dichloromethane were extracted twice at 20 DEG C for 2 hours at a weight ratio of 1:10, followed by degreasing. The extract was filtered and dried under reduced pressure at 20 ° C. The dried gluten powder (100 g) was mixed with a 70 (v / v)% ethanol aqueous solution at a weight ratio of 1:10, gently stirred at 20 ° C for 4 hours, and the suspension was centrifuged at 4600 rpm. Gliadin was present in the supernatant, and the supernatant was again lyophilized. However, since the lyophilized product of the supernatant still contains some glutenin protein, it was washed several times with distilled water to remove glutenin protein, filtered, and the filtered solid was treated with 0.05 M acetic acid, Dean powder was prepared.

Production Example 2 : Gliadin  Preparation of Nanoparticles

The gliadin powder of Production Example 1 was insoluble in water in the aqueous alcohol solution of less than 65 (v / v)% or 80 (v / v)% in the aqueous ethanol solution.

To prepare the gliadin nanoparticles from the gliadin powder, the gliadin powder was mixed in an aqueous 70% (v / v) ethanol solution to a concentration of 14% by weight, stirred at room temperature for 2 hours, Lt; / RTI > to prepare a gliadin solution.

The 14 wt% gliadin solution was diluted to a gliadin concentration of 2, 7, and 14 wt%, respectively, and electrosprayed with each gliadin solution using the electrospray shown in the schematic diagram of Fig.

In an electrospray device composed of a gliadin injection pump (Nano NC, Korea), a spray nozzle, a high voltage supplier and a grounded gliadin particle collector, each gliadin solution was filled into a 10 ml syringe, The solution was fed into a stainless steel nozzle having a diameter of 32 GA (about 0.0097 inch) connected to a high voltage supply at a feed rate of 0.5 ml / hr. The voltage was applied to the high voltage supply so that the potential difference between the nozzle and the grounded stainless steel plate was 12 to 14 kV.

Production Example 3 : Gliadin  And Gelatin Composite Nanoparticles

A gliadin solution was prepared in the same manner as in Production Example 2, and an electric atomizing apparatus with the same apparatus and conditions was used.

However, gelatin was mixed with an aqueous solution of 90 (v / v) acetic acid so as to be 4% by weight and 8% by weight, stirred at room temperature for 2 hours and aged for 2 days to prepare a gelatin solution.

7% by weight of gliadin solution or 14% by weight of gliadin solution and 4% by weight of gelatin solution or 8% by weight of gelatin solution were mixed at a weight ratio of 1: 1 to prepare a mixed solution, Electrospraying was carried out under the same conditions using the electrospraying apparatus of Production Example 2. 2 parts by weight of glutaraldehyde was added to 100 parts by weight of the sum of the glycidol and the gelatin solids, and polymerization was carried out at 25 DEG C for 60 minutes.

Experimental Example 1 : Identification of morphological characteristics

The shapes of the gliadin nanoparticles prepared in Preparation Example 2 and the gliadin and gelatin composite nanoparticles of Preparation Example 3 were confirmed by an electron microscope (SEM, TESCAN Model: VEGA / SBH Motorize). The nanoparticles were platinum coated using a turbo-sputter coater and SEM images were obtained at 20 kV.

FIG. 2 (A) shows the results of the gliadin nanoparticles prepared using the 7% by weight gliadin solution of Preparation Example 2, FIG. 2 (B) shows the 7% by weight gliadin solution of Preparation Example 3 and 4% Spherical nanoparticles were formed by scanning electron microscopy (SEM) images of gliadin and gelatin composite nanoparticles prepared using a solution mixture.

TEM images were also obtained using a transmission electron microscope (TEM, JEM-2100F) at a voltage of 200 kV, a negative current of 95, and an emission current of 126 ㎂.

Fig. 3 (A) shows the results of the gliadin nanoparticles prepared using the 7% by weight gliadin solution of Preparation Example 2, Fig. 3 (B) shows the 7% by weight gliadin solution of Preparation Example 3 and 4% The TEM images of the gliadin and gelatin composite nanoparticles prepared using the mixed solution showed that the spherical nanoparticles were formed more clearly than the SEM photographs and the average particle size was smaller in the gliadin nanoparticles can confirm.

Also, in the case of the gliadin nanoparticles prepared using the 2 wt% gliadin solution of Preparation Example 2, the particle size was uneven and it was difficult to obtain monodispersed spherical nanoparticles when the concentration of the gliadin solution was low In the case of the gliadin nanoparticles prepared by using the 14% by weight gliadin solution of Production Example 2 , the particles did not aggregate to form nano-sized particles, and micro-sized particles and nano-particles were mixed together, (Fig. 4A).

Also Manufacturing example  3, 7 wt% Gliadin  Solution and a 8 wt% gelatin solution Gliadin  And gelatin composite nanoparticles aggregated with each other to form non-uniformly shaped particles (Fig. 4B).

Further, 14 wt% Gliadin  Solution and a 8 wt% gelatin solution Gliadin  And in the case of gelatin composite nanoparticles, the sizes of the formed nanoparticles are not uniform Fibrous  Thereby forming a network (FIG. 4C).

Experimental Example 2 : Particle size distribution and Zeta potential  Confirm

The gliadin nanoparticles prepared using the 7% by weight gliadin solution of Preparation Example 2, the 7% by weight gliadin solution of Preparation Example 3 and the 4% by weight gelatin solution, And the gelatin composite nano-particles, and the mixed solution of the 7 wt% glyadin solution and the 8 wt% gelatin solution of Preparation Example 3, the particle size distribution and zeta potential of the gliadin and gelatin composite nanoparticles were confirmed .

The nanoparticles were first dispersed in deionized water, sonicated for 3 minutes at 20% amplitude using a tip sonicator (Sonics, Vibrcell), and analyzed by dynamic light scattering Zetasizer 4 (Malvern Instrument, France) The particle size distribution and the zeta potential were measured, and the results are shown in Table 1 and FIG.

division Average particle size Average zeta potential 7% by weight gliadin 218.66 ± 5.15 18.46 ± 8.29 7% by weight gliadin + 4% by weight gelatin 398.56 + - 4.20 19.00 ± 3.62 7% by weight gliadin + 8% by weight gelatin 450.10 + - 9.70 14.20 ± 3.72

The gliadin nanoparticles prepared using the 7 wt% gliadin solution of Preparation Example 2 had a positive charge with a uniform size (218.66 nm) at a zeta potential of 18.46 mV and a positive charge of 7 wt% % Gelatin mixed solution, the average particle size increased to about 2 times and the zeta potential remained similar. However, in the mixed solution in which the concentration of gelatin was increased to 7 wt% of gliadin by 8 wt%, the average particle size was increased, but the zeta potential was rather decreased.

The gliadin and gelatin nanoparticles prepared from the mixed solution of 4% by weight and 8% by weight of gelatin in 14% by weight of gliadin of Production Example 3, respectively, had particle sizes of 471.6 ± 7.8 nm and 516.1 14.8 nm, and the zeta potentials were as high as 19.3 mV and 21.8 mV, respectively (Figs. 6B and 6C) .

From the above results, it can be seen that the average particle size of the nanoparticles increases in proportion to the concentration of the gliadin solution, and that the fibrous structure becomes more than 14% by weight. When the concentration of the gliadin solution is constant, The average particle size increases proportionally, and when the concentration of the gliadin solution is 7 wt%, the fibrous network is formed when the concentration of the gelatin solution is 8 wt% or more.

Experimental Example 3 : Gliadin  And cross-linking of gelatin

Confirmation of cross-linking polymerization between gliadin and gelatin in the gliadin and gelatin composite nanoparticles prepared from the mixed solution of 7% by weight of gliadin solution and 4% by weight of gelatin solution in Production Example 3 X-ray diffraction analysis was performed using an X-ray diffractometer (Rigaku Corporation, Japan), and the results are shown in Fig.

Gelatin had main peaks at 18 degrees, and gliadin particles showed main peaks at 12 and 18 degrees. In the case of gliadin and gelatin composite nanoparticles, a peak at 12 degrees and a main peak at 18 degrees were observed together because gelatin was bound to the surface of gliadin.

Experimental Example 4 : Drug Release Analysis

Cyclophosphamide, an anticancer agent, was used to analyze drug release patterns of the nanoparticles of Production Examples 2 and 3.

The nanoparticles injected with the drug Gliadin  Solution, or Gliadin  And gelatin For mixing In the liquid Cyclophosphamide  One mg Of ml Were mixed and prepared by electrospray.

The cyclophosphamide eluted from the nanoparticles injected with cyclophosphamide was prepared by measuring the absorbance at 325 nm after the production of the nitroso derivative of cyclophosphamide, and the amount of the released cyclophosphamide Is shown in FIG. 8 as a percentage.

Using a 7% by weight gliadin solution of Preparation Example 3 and a 4% by weight or 8% by weight of gelatin solution mixed solution of the gliadin nanoparticles prepared using the 7% by weight gliadin solution of Preparation Example 2 Both of the prepared gliadin and gelatin composite nanoparticles exhibited a two - step emission pattern.

The gliadin and gelatin composite nanoparticles prepared by using the mixed solution of 7 wt% of glyandin solution and 8 wt% of gelatin solution of Preparation Example 3 in Step 1 were eluted with 46% cyclophosphamide within 1 hour . However, the gliadin and gelatin composite nanoparticles prepared by using the mixed solution of the gliadin nanoparticles of Production Example 2, the 7 wt% glyandin solution of Production Example 3 and the 4 wt% gelatin solution, 25% and 30%, respectively. The fast release of the first step was judged to be due to the cyclophosphamide attached to the surface of the nanoparticles. Especially, as the gelatin content was increased, the faster release in the first step was thought to be due to the high solubility of the gelatin.

Cyclophosphamide was gradually released up to 48 hours in the second stage after the first stage of rapid release, and 80% of the gliadin nanoparticles prepared by using the 7% by weight gliadin solution was released at 48 hours. In the case of the gliadin and gelatin composite nanoparticles prepared by using the mixed solution of the 7 wt% gliadin solution and the 4 wt% gelatin solution, a mixture of 90%, 7 wt% gliadin solution and 8 wt% gelatin solution And 95% in the case of the gliadin and gelatin composite nanoparticles prepared using the solution.

From the above results, the drug release pattern is affected by the wettability of the surface of the nanoparticles. In the case of the hydrophilic gliadin nanoparticles, the drug is gradually released. However, as the hydrophilic gelatin is included in the nanoparticles, Thus, it has been found that the release rate of the drug can be controlled by controlling the concentration of the gelatin added to the gliadin nanoparticles. However, in the case of the gliadin and gelatin composite nanoparticles prepared by using the mixed solution of the 7 wt% gliadin solution and the 8 wt% gelatin solution, adverse effects occurred in the in vivo administration, It was judged that it was not appropriate.

Experimental Example 5 : Identification of Natural History of Breast Cancer Cells

In Experimental Example 4, the gliadin and gelatin composite nanoparticles also exhibited a controlled release effect of about 88 to 90% of the drug released at 24 hours. However, in order to confirm the natural history of breast cancer cells, cyclophosphamide The gliadin nanoparticles prepared using the 7% by weight gliadin solution of the preparation example 2 were used. This is because the gliadin nanoparticles showed a controlled release effect with a drug release of 75% at 24 hours and an average particle size of 218.7 nm was considered to be capable of endocytosis.

In order to confirm the natural history of the gliadin nanoparticles in breast cancer cells, MCF7 breast cancer cells were cultured for 24 hours, and then treated with control 1 (Control in FIG. 9) in which the cells were not added with cyclophosphamide Control group 2 (NP in Fig. 9) in which only gliadin nanoparticles were added, Control group 3 (CPA in Fig. 9) in which only cyclophosphamide was added to the medium, and gliadin nanoparticles injected with cyclophosphamide (NP + CPA in FIG. 9) were cultured for 12 hours, 24 hours, and 48 hours. The cells were stained with Annexin V to identify cells that had died of the disease (green staining) and stained with propidium iodine (Red staining).

In the case of control 1 and 2, it was not possible to identify naturally occurring cells or cell necrosis. However, in the case of control group 3 in which only cyclophosphamide alone was added, it was confirmed that cell necrosis occurred in culture for 24 hours, and in the case of injecting cyclophosphamide into gliadin nanoparticles, the natural history of breast cancer cells was confirmed I could. It can be seen that the killed cells were significantly increased in the Example compared to the Control 3.

Western blot analysis was performed to confirm the natural history of the cyclophosphamide-injected gliadin nanoparticles on the breast cancer cells. The results are shown in FIG. It was confirmed that the production of Bcl-2 protein was significantly reduced in the gliadin nanoparticles injected with the cyclophosphamide of the example and the breast cancer cells cultured for 24 hours, whereas the control group cultured with addition of the gliadin nanoparticles 2, Bcl- 2 protein decreased in breast cancer cells .

Also, when cultured with the gliadin nanoparticles injected with the cyclophosphamide of the example, the cells did not develop apoptosis until 12 hours. In particular, when cultured with the addition of only the cyclophosphamide of the control 3, But not cell death. This was because the gliadin nanoparticles injected with the cyclophosphamide of the Example were able to improve the regulation of drug release through intracellular ingestion.

Claims (11)

Dissolving gliadin in an aqueous 65 to 80 (v / v)% ethanol solution to prepare a gliadin solution; And electrospraying the gliadin solution.
The method of claim 1, wherein the gliadine concentration of the gliadine solution is 5 to 10% by weight.
The method according to claim 1 or 2, wherein the flow rate of the solution in the electrospray is 0.1 to 1 ml / hr, and the voltage difference is 12 to 18 kV.
4. The method of claim 3, wherein the diameter of the nozzle in the electrospray is 0.1 to 0.5 mm.
Gliadine nanoparticles prepared by electrospray method, zeta potential is 15 to 30 mV, average particle size is 200 to 350 nm.
Dissolving gliadin in an aqueous 65 to 80 (v / v)% ethanol solution to prepare a gliadin solution; Dissolving gelatin in an aqueous solution of 80 to 99 (v / v)% acetic acid to prepare a gelatin solution; Mixing the gliadin solution and the gelatin solution to prepare a mixed solution; And electrospraying the mixed solution of the gliadin and the gelatin. The method for producing the gliadin and gelatin composite nanoparticles according to claim 1,
The method according to claim 6, wherein 0.5 to 5 parts by weight of a crosslinking agent is further added to 100 parts by weight of a solid content of the mixed solution of gliadin and gelatin.
[7] The method according to claim 6 or 7, wherein the gliadin concentration of the gliadin solution is 5 to 10 wt% and the gelatin concentration of the gelatin solution is 0.1 to 6 wt%. A method for producing nanoparticles.
Wherein the zeta potential is 15 to 30 mV and the average particle size is 300 to 500 nm, which is produced by an electrospray method.
Claim 5, the gliadin nanoparticles or claim 9 of the gliadin and gelatin composite nanoparticles; And a pharmaceutically active ingredient.
11. A sustained release drug delivery system according to claim 10, wherein said pharmaceutically active ingredient is cyclophosphamide and is for the treatment of breast cancer patients.

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