KR20240021590A - Flufenamic Acid-loaded Chitosan Nanoparticle - Google Patents

Abstract

The present invention relates to chitosan nanoparticles loaded with flufenamic acid, and more specifically, by modifying low molecular weight water-soluble chitosan (LMWSC) by combining lauric acid (LA), flufenamic acid. It has the advantage of being able to effectively carry anti-cancer drugs such as and deliver them stably into cells.

Description

Chitosan nanoparticles loaded with flufenamic acid {Flufenamic Acid-loaded Chitosan Nanoparticle}

The present invention relates to chitosan nanoparticles loaded with flufenamic acid, and more specifically, by modifying low molecular weight water-soluble chitosan (LMWSC) by combining lauric acid (LA), flufenamic acid. This relates to drug delivery nanoparticles that can effectively carry anticancer drugs such as and stably deliver them into cells.

Currently, humanity's interest in health and medical care is increasing due to aging, and cancer treatment and research are gaining ground in modern medicine (References 1-3). Representative anticancer drugs for cancer treatment include doxorubicin and paclitaxel, and many cancer treatment studies using them are being conducted (References 4-6).

However, although the anticancer drugs currently in use are effective, they have the disadvantage of being highly toxic and causing numerous side effects, so research is actively underway to solve these problems (References 7 and 8).

Meanwhile, Patent Document 1 below discloses a water-soluble chitosan nanoparticle as a delivery vehicle for an anticancer drug and a method for manufacturing the same, but it had the limitation of not being able to effectively carry an anticancer drug such as flufenamic acid and stably deliver it into cells.

Therefore, there is an urgent need for the development of new modified nanoparticles that can effectively carry anti-cancer drugs such as flufenamic acid and stably deliver them into cells.

Patent Document 1: Republic of Korea Patent No. 10-0578382 (May 11, 2006)

Accordingly, in order to solve the above problem, the present invention

It was discovered that it could be done, and the present invention was completed based on this.

Nanoparticles for drug delivery according to one embodiment of the present invention are characterized in that they are modified by combining lauric acid (LA) with low molecular weight water-soluble chitosan (LMWSC).

The low molecular weight water-soluble chitosan refers to water-soluble chitosan with a molecular weight of 3 kDa to 10 kDa.

Nanoparticles for drug delivery according to one embodiment of the present invention are characterized by having the following formula (1).

[Formula 1]

Nanoparticles for drug delivery according to one embodiment of the present invention use 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) as a crosslinking agent. It is characterized by being manufactured using.

The anti-cancer treatment agent according to one embodiment of the present invention is characterized in that it is manufactured by loading flufenamic acid (FA) on the drug delivery nanoparticles according to the various embodiments above.

The anti-cancer treatment agent according to one embodiment of the present invention is characterized in that it is manufactured by carrying out a process of loading flufenamic acid (FA) on the drug delivery nanoparticles according to the various embodiments above by dialysis.

The anti-cancer treatment agent according to one embodiment of the present invention is characterized as a bladder cancer treatment agent.

The drug delivery nanoparticles according to the present invention are modified by combining lauric acid (LA) with low molecular weight water-soluble chitosan (LMWSC), thereby effectively delivering anticancer drugs such as flufenamic acid. It has the advantage of being able to be carried and stably delivered into cells.

Figure 1 is a diagram showing an overview of synthesizing LMWSC-g-LA (LMLA) using EDC.
Figure 2 is a picture showing the 1H NMR spectrum of LMLA ((a) LMWSC; (b) LA; (c) LMLA 20%; (d) LMLA 30%).
Figure 3 (a) is a diagram showing the pyrene emission spectrum in a 30% aqueous solution of LMLA, and Figure 3 (b) shows the intensity ratio (I1/) of the pyrene emission spectrum as a function of LMLA concentration in distilled water. This is a picture showing I3).
Figure 4 (A) shows the particle size distribution, and Figure 4 (B) shows the morphological images of LMLA and LMLAF ((a) LMLA 20%; (b) LMLA 30%; (c) LMLAF 20%; ( d) LMLAF 30%).
Figure 5 is a figure showing the total cumulative FA release amount of LMLAF according to pH for each time period (0.5-96h).
Figure 6 is a diagram showing the evaluation of cytotoxicity and anticancer activity of LMLA and LMLAF using the MTT assay ((a) HEK293 cell line; (b) T24 cell line).

Before describing the present invention in more detail, the terms and words used in the specification and claims should not be limited to their common or dictionary meanings, and the concepts of the terms should be appropriately used to explain the invention in the best way. It must be interpreted as meaning and concept consistent with the technical idea of the present invention based on the principle that it can be defined clearly. Therefore, the configuration of the embodiment described in this specification is only one preferred example of the present invention, and does not represent the entire technical idea of the present invention, and therefore, various equivalents and modifications that can replace them at the time of filing the present application You must understand that there may be.

A drug delivery method that can compensate for the shortcomings of current anticancer drugs and maximize their efficacy is a method that uses polymeric mediators in the form of prodrugs and polymeric micelles that can solubilize poorly soluble anticancer drugs through chemical bonding to polymeric materials. Among them, the drug carrier in the form of a polymeric micelle is synthesized through chemical modification of a hydrophobic material on a hydrophilic polymer base. When dispersed in water, core-shell shaped nanoparticles are formed, and the drug is poorly soluble by carrying the drug on the hydrophobic core. Since it is soluble in water, it has the advantage of minimizing side effects and maximizing drug effectiveness (Reference 9-13).

In this study, polymeric micelle-type nanoparticles were prepared by chemically modifying the natural polymer chitosan, a hydrophilic material, with a hydrophobic material, and flufenamic acid (FA), a well-known bladder cancer treatment drug, was loaded onto the nanoparticles to act as an anticancer drug. We wanted to prove the possibility of use.

Chitosan is abundant in nature after cellulose, and is a natural polysaccharide manufactured by deacetylating chitin obtained from crustaceans. It is made up of D-glucomine and N-acetyl-D-glucomine linked by a β-(1,4)-glucosidic bond. It is a natural polysaccharide composed of two monomers (References 14-16).

It has various biological activities such as anticancer effect, cholesterol reduction, immune response enhancement, and antibacterial activity, and has the advantage of being biocompatible, biodegradable, and non-toxic, so it is applied in various fields such as cosmetics, environment, and medicine (Reference 17- 19).

Many researchers have used chitosan in their studies with various organic acids introduced to increase the solubility of chitosan in water. However, when chitosan containing organic acids is applied for medical purposes, it has the disadvantage of causing side effects to the living body due to the organic acids.

To solve this problem, the chitosan used in this study was low molecular weight water-soluble chitosan (LMWSC), which has a free amine group and was developed by removing organic acids by the salt-removal method. (Reference 20).

LMWSC has a free amine group, so it is widely used as a drug and gene delivery vehicle. It has high solubility in water and low toxicity, so it has excellent biocompatibility.

The FA used in this study for the purpose of treating bladder cancer is N-(3-Trifluoro-methylphenyl) anthranilic acid or 2-(3-Trifluoromethylanilino)benzoic acid, which is an aromatic amino acid composed of anthranilic acid with N- (References 21 and 22).

In general, it has properties such as antipyretic, anti-inflammatory, and analgesic effects, so it is mainly used as an analgesic, anti-inflammatory, and cold medicine for pain and inflammation in musculoskeletal and joint disorders, and is also widely used as a non-steroidal anti-inflammatory drug (NID) (References 23 and 24) ).

In recent research, it is being studied as a new treatment for prostate cancer, and its effect on anticancer resistance and metastasis inhibition in bladder cancer, a specific cancer cell, has been studied due to FA's property of inhibiting aldoketo reductase 1C1 (AKR1C1) (Reference 25) .

However, the FA drug itself has a problem in that its solubility in water is significantly low, and the therapeutic effect is halved due to immune response and drug resistance (Reference 26).

In this study, to solve the problem of FA itself, LMWSC-g-LA (LMLA) was synthesized by chemically modifying hydrophilic LMWSC with lauric acid (LA), a hydrophobic material, and FA was loaded into LMLA using dialysis. LMLAF nanoparticles were prepared.

The drug delivery nanoparticle according to the present invention is characterized by modifying low molecular weight water-soluble chitosan (LMWSC) by combining lauric acid (LA).

Nanoparticles for drug delivery according to the present invention have the following formula (1).

[Formula 1]

In addition, the drug delivery nanoparticle according to one embodiment of the present invention is 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC). It may be manufactured using a crosslinking agent.

The anti-cancer treatment agent according to one embodiment of the present invention is prepared by loading flufenamic acid (FA) on the drug delivery nanoparticles according to the various embodiments above.

In addition, the anti-cancer treatment agent according to one embodiment of the present invention may be manufactured by carrying out a process of loading flufenamic acid (FA) on the drug delivery nanoparticles according to the various embodiments above by dialysis. .

In particular, the anti-cancer treatment agent according to one embodiment of the present invention is a bladder cancer treatment agent.

Meanwhile, the present inventor identified the physicochemical properties of LMLAF using various analysis equipment and performed MTT assay on normal cells and bladder cancer cells to confirm toxicity and anticancer activity, thereby demonstrating that the LMLAF nanoparticles developed in this study were used to treat bladder cancer. We would like to suggest that it is an anticancer agent that can maximize .

[Experiment]

1. Reagents and Materials

Low molecular water-soluble chitosan (LMWSC, Mw: 10kDa) used in this study was purchased from KITTOLIFE Co., Korea, and was 1-ethyl-3-(3-dimethylaminopropyl) Carbodiimide hydrochloride (EDC), lauric acid (LA), and flufenamic acid (FA) were purchased from Sigma-Aldrich (St, Louis, MO, USA). In this study, the human embryonic kidney cells (HEK293) cells used to confirm cytotoxicity and anticancer activity were from the American type culture collection (ATCC, Korea), and the human bladder tumor cells (T24) cells used were from the Korean Cell Line Bank (KCLB, Seoul, Korea). Korea) and conducted experiments.

Dulbecco's modified eagle's medium (DMEM) was purchased from Lonza (USA) and RPMI-1640 were purchased from Hyclone (USA), and ethylenediamintetetraacetic acid (EDTA) and fetal bovine serum (FBS) were purchased from Gibco (USA) and used for cell culture. did. In addition, dimethylsulfoxide (DMSO) and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) were purchased from Sigma-Aldrich (St, Louis, MO, USA), and other Reagents and solvents were purchased as GR grade and used in the experiment without purification.

2. Synthesis of LMWSC-g-LA (LMLA)

LMLA was synthesized by introducing hydrophobic LA into the amine group of LMWSC using the cross-linker EDC. 100 mg of LMWSC was stirred at room temperature to completely dissolve in 2mL of MES buffer. To activate the carboxyl group of LA, 20% and 30% of LMWSC were dissolved in DMSO, and EDC cross-linker was added at twice the molar ratio of LA and stirred for 30 minutes. The LA solution with activated carboxyl group was slowly added dropwise to the LMWSC solution and stirred at room temperature for 4 hours. After the reaction was completed, the solution was precipitated in acetone, washed three times, and dialyzed against distilled water using a MWCO 3500 cellulose membrane for 24 hours. The final material, LMLA, was obtained as powder after freeze-drying and used in the experiment.

3. Structural analysis of LMLA

The presence or absence of LMLA synthesis was analyzed using nuclear magnetic resonance (1H NMR, 400 MHz, Bruker, Germany). LMWSC and LMLA were dissolved in D2O and LA in DMSO, respectively, and the presence or absence of synthesis was confirmed by analyzing the characteristic peak and chemical shift in the range of 0-9 ppm.

4. Manufacturing and confirmation of loading efficiency of FA-loaded LMLA (LMLAF)

FA-loaded LMLA (LMLAF) was manufactured using the evaporation technique, and the experimental method was as follows. After dissolving 30 mg of LMLA in 7 mL of distilled water, 35 mL of tetrahydrofuran (THF) was added to the LMLA aqueous solution and stirred for 10 minutes. FA was dissolved in 1 mL of THF at a weight ratio of 5% relative to the amount of LMLA, then slowly added dropwise to the LMLA solution and stirred for 1 hour. THF was removed from this solution using a rotary evaporator, dialyzed against a MWCO 3500 cellulose membrane for 24 hours, and then freeze-dried to obtain the final material, LMLAF. The loading efficiency of FA from LMLAF was calculated using an ultraviolet-vis spectrometer (UV-1601, Shimadzu, Japan), and the experimental method was as follows.

2 mg of LMLAF was completely dissolved in 200 μL of 0.1% trifluoroacetic acid (TFA), then 1.8 mL of DMSO was added and shaken for 24 hours. An FA solution was obtained using this solution using an ultra filtration membrane, and the absorbance was confirmed at 289 nm, the maximum absorption wavelength, using an ultraviolet spectrometer, and the loading efficiency was calculated using Equations 1 and 2 below.

[Calculation Formula 1]

[Calculation Formula 2]

5. LMLA and LMLAF nanoparticle preparation and characterization

LMLA and LMLAF nanoparticles were manufactured by dialysis using MWCO 3500 cellulose membrane, and the particle size, critical aggregation concentration (CAC), and morphological image characteristics of the prepared LMLA or LMLAF nanoparticles were analyzed as follows. did.

LMLA has hydrophobic properties due to LA and hydrophilic properties due to LMWSC, so it forms core-shell particles through self-aggregation in aqueous solution. Therefore, the minimum concentration CAC at which particle formation is possible was measured using the following method.

Pyrene is dissolved in acetone to 6×10-5 M and then distilled water is added to make the final concentration 1.2×10-6 M. After removing acetone from this solution under reduced pressure at 40°C for 2 hours, the pyrene solution and various concentrations of nanoparticle solutions (3.0×10?5-2 mg/mL) were mixed to obtain a final concentration of 6.0×10-7. It was made to be M. This solution was stirred in the dark at 60°C for 3 hours, and the optical behavior was observed using a fluorescence photometer (RF-5301PC, Shimadzu, Japan) in the wavelength range of 350-460 nm. CAC was determined using the intensity ratio (I1/I3) at specific wavelengths (I1 and I3) according to concentration.

The particle size of LMLA and LMLAF was measured using dynamic light scattering (DLS) equipment, and DLS was performed using an ELS8000 electro phoretic LS spectrophotometer (Otsuka Electronics Co., Japan). The size and distribution of particles were measured using a dynamic light scattering device, and LMLA and LMLAF were each prepared as aqueous solutions at a concentration of 2 mg/mL. In addition, the zeta potential of LMLA and LMLAF was confirmed by preparing an aqueous solution with a concentration of 1.5 mg/mL and measuring it using the same equipment used to measure particle size.

Morphological analysis of LMLA and LMLAF was confirmed using a transmission electron microscope (TEM, Hitachi, H-7500, Japan) as follows. After preparing LMLA and LMLAF at a concentration of 10 mg/mL each, they were placed on a copper carbon grid and completely dried at room temperature. This was stained with 3% uranyl acetate for 15 minutes, dried for 24 hours, and particle images were observed through TEM.

6. Confirmation of FA release behavior from LMLAF

The emission behavior of FA from LMLAF was confirmed using an ultraviolet spectrometer (UV-Vis spectrometer, UV1601, Shimadzu, Japan). 4 mg of LMLAF was dissolved in 2 mL of pH 5.5 and 7.4 PBS buffer, respectively, added to MWCO 1000 cellulose membrane, and stirred at 140 rpm at 37°C. 2 mL of each solution released over time (0.5-96 hours) was taken, the absorbance was measured at 289 nm through an ultraviolet spectrometer, and the cumulative release amount was calculated as a percentage.

7. Confirmation of cytotoxic and anticancer activity of LMLA and LMLAF

The toxicity and anticancer activity effects of the prepared LMLA and LMLAF were confirmed by performing MTT assay on HEK293 normal cells and T24 bladder cancer cells. HEK293 cells were cultured in DMEM medium containing 10% FBS, and T24 cells were cultured in RPMI medium containing 10% FBS in an incubator with a CO2 concentration of 5% and a temperature of 37°C. Cell seeding was done in a 96 well plate at a concentration of 5 LMLAF solutions prepared at concentrations of 0.25, 0.125, 0.063, 0.031, 0.016, and 0.008 mg/mL were added to T24 cells, respectively, and cultured in the dark in an incubator for 48 hours. After 48 hours, MTT was treated and cultured in the dark for 4 hours. Finally, after MTT was removed, 200 μL of DMSO was treated and absorbance was measured at 560 nm and 670 nm. Using the measured absorbance value, cell viability was calculated using equations 3 and 4 below.

[Calculation Formula 3]

[Calculation Equation 4]

[Experiment result]

1. Synthesis and structural characterization of LMLA

Self-use of FA drugs, which are well-known as treatments for bladder cancer, has the problem that the treatment effect is halved due to immune response and drug resistance. In this study, to solve these problems, we attempted to maximize the effectiveness of FA in treating bladder cancer by developing a drug carrier that can form core-shell self-assembled nanoparticles. To manufacture core-shell type nanoparticles, hydrophilic LMWSC was synthesized with LA, a hydrophobic material, through chemical modification using the cross-linker EDC. The carboxyl group (-COOH) of LA was activated using the cross-linking agent EDC, and this was introduced into the amine group (-NH2) of LMWSC to finally synthesize LMWSC-g-LA (LMLA) (Figure 1).

To determine whether LMLA was synthesized, its structural characteristics were analyzed using nuclear magnetic resonance (1H NMR, 400 MHz, Bruker, Germany). In LA, the hydrogen peak of -CH3 at 0.7-0.9 ppm and the hydrogen peak of -CH2 at 1.5-2.3 ppm can be confirmed, and in LMWSC, the hydrogen peak at carbon position 1 is 4.9 ppm and the hydrogen peak at carbon position 2 is 4.9 ppm. The hydrogen peak at carbon positions 3-6 was confirmed at 2.9 ppm, and the characteristic peak due to the acetyl group of chitin in the chitosan structure was confirmed at 2.1 ppm (Figure 2). In LMLA, the final synthesized material, both the characteristic peaks of LMWSC and LA were confirmed, and it was confirmed that the synthesis was successful through chemical shift of the peak.

2. Characterization of LMLA and LMLAF nanoparticles

LMLA and LMLAF nanoparticles were prepared by dialysis, and the characteristics of the nanoparticles were characterized through particle size, zeta potential, CAC, and morphological analysis. LMLA nanoparticles exhibit an amphipathic structure in which hydrophobic LA is bound to hydrophilic LMWSC, which forms core-shell shaped nanoparticles through self-assembly in an aqueous solution. In this study, two types of drug carriers containing 20% and 30% LMLA were synthesized according to the LA binding ratio, and CAC was confirmed using a fluorescence spectrometer. As a result, when the binding ratio of LA to LMWSC was 20%, the CAC value was 0.22 mg/mL, and when it was 30%, the CAC value was 0.13 mg/mL, which means that when the binding ratio of LA was 30%, the concentration was lower. The CAC value could be confirmed (Table a and Figure 3).

[Table a]

In addition, as a result of performing TEM and DLS to confirm the shape and size of LMLA and LMLAF particles during particle formation, spherical nanoparticles (Figure 4(B)) and monodisperse particle distribution were confirmed for both LMLA and LMLAF. It was confirmed that the particle size was smaller when 30% LA was combined, at 320.1 ± 10.8 nm for 20% and 279.6 ± 12.8 nm for 30% LMLA (Table a and Figure 4(A)).

It is believed that the reason for showing these CAC and particle size results is that more LA is bound in the case of 30% LMLA, and the hydrophobic density increases due to a strong hydrophobic effect inside the core.

In addition, in the case of LMLAF, it can be seen that the particle size is relatively reduced compared to LMLA (Table a and Figure 4(A)), which means that as the FA drug is supported in the core part of LMLA, there is a gap between the inner core of LMLA and the hydrophobic drug FA. It is believed that the hydrophobic interaction increased to form smaller particles.

In addition, the zeta potentials of LMLAF 20% and LMLAF 30% were 32.8 ± 0.5 mV and 27.8 ± 0.7 mV, respectively, showing a strong positive charge (Table a), which indicates a high charge interaction considering that the surface charge of cancer cells is negative. It is believed that the reason why LMLAF 30% shows a lower zeta potential than LMLAF 20% is because more LA is introduced into the amine (-NH2) group of LMWSC, causing amine This is believed to be due to the relative decrease in the group. Through analysis of the various characteristics of these LMLA nanoparticles, it was confirmed that they can not only be used as carriers for FA, but can also be applied as drug carriers in the medical field.

3. Analysis of FA loading efficiency and release behavior from LMLAF

To improve the shortcomings of FA used as a treatment for bladder cancer and maximize its treatment efficiency, FA was loaded inside the LMLA nanoparticle core by dialysis. The FA loading efficiency according to the binding ratio of hydrophobic LA is summarized in Table a. As a result, the DC, which is the FA loading efficiency per 1 mg of LMLAF, was 3.8% (LMLAF 20%) and 4.5% (LMLAF 30%), respectively, and the total FA loading efficiency, EE, was 66.12% (LMLAF 20%) and 80.46. High loading efficiency was confirmed by % (LMLAF 30%). The reason why LMLA 30% has a higher FA loading efficiency than LMLA 20% is that in the case of LMLA 30%, where more hydrophobic LA is bound, a high hydrophobic density is formed due to strong hydrophobic action inside the core, which increases interaction with hydrophobic FA drugs. It is believed that this is because it was done. In addition, as a result of confirming the drug release behavior of FA from LMLAF according to pH changes, in the case of pH 7.4, LMLAF 20% showed a total cumulative FA release of 29.5% and LMLAF 30% showed a release amount of 23.4% over about 96 hours. In the case of pH 5.5, the total cumulative FA release for 20% LMLAF was 39.7%, and 30% LMLAF was 28.0% (Figure 5).

In the case of LMLAF 30%, it can be seen that the release amount is relatively lower than that of LMLAF 20%. This is because more hydrophobic LA is bound to LMLAF 30%, and the FA drug is released more slowly due to the strong hydrophobic density inside the core. It is believed that In addition, it can be seen that the amount of FA released is greater at pH 5.5 than at pH 7.4, which means that in a slightly acidic environment with the pKa of LMWSC being 5.5-6.4, hydrogen is protonated on the chitosan amine (-NH2) group and converted to -NH3+, resulting in charge repulsion. It is believed that this was induced and a greater amount of FA was released. Based on these results of loading efficiency and release behavior, when LMLAF is used as a treatment for bladder cancer, it induces sustained drug release within cancer cells that have characteristics of an acidic environment, thereby solving the problem of drug resistance and maximizing the treatment effect. It is believed that it can be done.

4. Elucidation of toxicity and anticancer effects of LMLA and LMLAF

To investigate the possibility of using LMLAF as a bladder cancer treatment, the in vivo cytotoxicity of LMLA nanoparticles was confirmed in HEK293 normal cells and the anticancer effect of LMLAF in T24 bladder cancer cells using MTT assay. As a result, it was confirmed that LMWSC, LMLA 20%, and LMLA 30% all showed a cell viability of more than 70% at the highest concentration of 1 mg/mL, showing almost no toxicity (Figure 6(a)). In addition, as a result of confirming the anticancer effect in T24 bladder cancer cells, it was confirmed that at a concentration of 0.25 mg/mL, the FA drug itself showed a cell survival rate of 23%, while in the case of LMLAF 30%, a cell survival rate of 38% was observed (Figure 6(b) )).

When calculating the content of FA drug loaded at a concentration of 0.25 mg/mL with LMLAF 30%, it was confirmed to be about 0.0113 mg/mL, which is about 1/22 times less than the amount of FA of the drug itself. It was found that even with a small amount of drug, the anticancer effect can be maximized when loaded inside the manufactured LMLA drug carrier. Through these results, it is believed that by identifying the cytotoxic and anticancer effects, the manufactured LMLA and LMLAF nanoparticles can be used as anticancer agents that are safe in the body and can be expected to have high treatment efficiency for bladder cancer.

[conclusion]

In this study, to improve the problems of FA drugs used as bladder cancer treatments, a core-shell type LMLA drug carrier was synthesized using the cross-linker EDC, and FA was supported inside the hydrophobic core to maximize the therapeutic effect. The presence or absence of LMLA synthesis was confirmed through 1H NMR. In addition, we demonstrated that LMLA can be used as a carrier for FA by analyzing the physical and chemical properties of LMLA and LMLAF nanoparticles through particle size, zeta potential, CAC, and morphological images, as well as the loading efficiency and release behavior of FA from LMLAF. Furthermore, the cytotoxic and anticancer effects of LMLA and LMLAF were confirmed through MTT assay, and the results showed that there was no cytotoxicity at all and at the same time, high anticancer effects were observed even with a small amount of drug. These results suggest that LMLAF developed in this study not only has high safety in the human body but can also be used as an anticancer agent that can maximize cancer treatment efficiency.

[Comparative experiment]

<Example 1>

Nanoparticles for drug delivery (LMLAF) modified by combining lauric acid (LA) with low molecular weight water-soluble chitosan (LMWSC) according to the present invention prepared by the above-described method.

<Comparative Example 1>

Low molecular weight water-soluble chitosan (LMWSC) not modified with lauric acid (LA)

<Comparative Example 2>

Water-soluble chitosan nanoparticles disclosed in Example 1 of Patent Document 1 (10-0578382)

Using the nanoparticles prepared through Example 1 and Comparative Examples 1 and 2, flufenamic acid (FA) was loaded and introduced into T24 bladder cancer cells under the same experimental conditions, and bladder cancer was treated using MTT assay. An experiment was conducted to test the effect (other experimental conditions were set identically), and the results are shown in Table 1 below (each experiment was conducted at a concentration of 0.25 mg/mL).

division cell viability Example 1 38% Comparative Example 1 25% Comparative Example 2 27%

Looking at the results in Table 1 above, in the case of Example 1 according to the present invention, the nanoparticles of Comparative Example 1 (low molecular weight water-soluble chitosan not modified with lauric acid) and Comparative Example 2 (Example 1 of Patent Document 1) It can be seen that the loading efficiency of flufenamic acid is much better than when used.

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Claims (6)

Nanoparticles for drug delivery modified by combining lauric acid (LA) with low molecular weight water-soluble chitosan (LMWSC). In claim 1,
Nanoparticles for drug delivery, characterized by having the following formula (1).
[Formula 1]
In claim 1,
Nanoparticles for drug delivery, characterized in that they are manufactured using 1-ethyl-3-(3-dimethyl aminopropyl) carbodiimide (EDC) as a cross-linking agent. An anti-cancer treatment agent prepared by loading flufenamic acid (FA) on the drug delivery nanoparticle according to any one of claims 1 to 3. In claim 4,
An anti-cancer treatment agent, characterized in that the process of loading flufenamic acid (FA) on the drug delivery nanoparticles is performed by dialysis. In claim 4,
The anti-cancer treatment agent is an anti-cancer treatment agent, characterized in that the anti-cancer treatment agent is a bladder cancer treatment agent.