KR101823737B1 - Composite of hyaluronic acid-catechol for drug carrier and method for preparing the same - Google Patents
Composite of hyaluronic acid-catechol for drug carrier and method for preparing the same Download PDFInfo
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Abstract
The present invention relates to a drug delivery complex for delivering a drug such as an anti-cancer drug and a method for preparing the drug delivery complex. More specifically, the present invention relates to a drug delivery complex comprising a shell made of hyaluronic acid substituted with catechol; And an inner hollow surrounded by the outer shell, and a method for manufacturing the same.
According to the present invention, there is provided a hyaluronic acid-catechol drug delivery complex and a method for preparing the same, wherein the drug-releasing complex can be further selectively released from the weak acid environment, which is a cancer cell environment, by controlling the oxidation- .
Description
The present invention relates to a drug delivery complex for delivering a drug such as an anti-cancer drug and a method for producing the drug delivery complex.
The byssal thread is a substance produced in the body of fish and shellfish, which makes it possible to attach fish and shellfish to the surface of rocks and the like. The catechol residues present in the pine needles have adhesiveness to various substances, Thereby imparting cohesiveness. The mechanism by which the catechol moiety imparts adhesion is because the catechol moiety binds to a variety of organic and inorganic surfaces via metal-catechol coordination bonds, hydrogen bonds and covalent bonds. Due to this property, It is used for the preparation of antifouling, which is resistant to cell attachment, as immobilization of particles and for biomedical uses. Additionally, catechol moieties play an important role in exhibiting coherence. For example, cohesive biomaterials such as hydrogels and peptide films are produced through intermolecular bonding by catechol coupling.
The foregoing properties represent a pH-dependent phenomenon involving the oxidation-reduction transition of catechol, and the reduced catechol stored in acidic pH exhibits a stronger adhesion capacity when compared to the oxidized quinone structure due to the increase in pH. Cohesion is also achieved by coupling between reduced catechol and quinone type catechol at basic pH. Therefore, attempts have been made to biologically apply catechol having a bifunctional property according to pH, because the catechol substance can attach to the target surface and improve the structural strength by agglomeration.
On the other hand, hyaluronic acid is a type of polysaccharide composed of N-acetylglucosamine and glucuronic acid. It is widely distributed in all tissues of an animal, especially in hepatic insect tissues, and is found in vitreous, amniotic fluid, umbilical cord, joint fluid, pleural fluid, skin, There are many. Physically, it exhibits hygroscopicity as an amorphous solid and easily forms a gel in the presence of metal ions or the like, and chemically forms glucuronic acid, glucosamine and acetic acid by hydrolysis. Hyaluronic acid has inherent targeting properties for biocompatibility, biodegradability and cancer cells, and has recently received considerable attention as a drug delivery candidate. Particularly, specific receptors such as CD44 and RHAMM, which are overexpressed in cancer cells, selectively react with hyaluronic acid, and therefore, the availability of hyaluronic acid has been receiving a great deal of attention in selectively delivering chemotherapeutic drugs to cancer cells. At this time, it is considered that strong hydrophilicity and intramolecular hydrogen bonding problems considered to be obstacles to the production of hyaluronic acid particles can be solved to some extent by functionalizing them using catechol or the like.
In connection with this, a number of techniques for producing a complex having a predetermined biological use using hyaluronic acid substituted with a catechol group have been reported. For example, a mixture of hyaluronic acid and dopamine with EDC (1-ethyl-3- [ (3-dimethylaminopropyl) carbodiimide hydrochloride) and NHS (N-hydroxysuccinimide) to prepare a hyaluronic acid-catechol conjugate, which is then subjected to surface modification of the medical implant or immobilization of a bioactive substance Have been reported (Patent Document 1).
There is also a document that reports that hyaluronic acid-catechol complexes differ in adsorption and flocculation characteristics according to the external environment (Non-Patent Document 1), and in particular, hyaluronic acid-catechol conjugate is adhered There is also a paper describing a process that shows sex and coherence (Non-Patent Document 2).
However, conventionally, pH is controlled during the production process of hyaluronic acid-catechol complex by using catechol whose adsorption and flocculation properties vary depending on the pH, and a drug carrier is prepared, and the prepared drug carrier is selectively No technology has been proposed to release the drug.
In the present invention, it is intended to produce a drug delivery vehicle using a hyaluronic acid-catechol complex, and in particular, by controlling the oxidation-reduction competition reaction of catechol through pH control, more selective drug release is possible in a weakly acidic environment Hyaluronic acid-catechol drug delivery complex and a method for producing the same.
In order to solve the above problems,
A shell comprising hyaluronic acid substituted with catechol; And
A hyaluronic acid-catechol drug delivery complex comprising an inner hollow surrounded by the outer shell.
According to one embodiment of the present invention, the hyaluronic acid-catechol drug delivery complex may have a hydration radius of 160 to 190 nm.
According to another embodiment of the present invention, the hyaluronic acid-catechol drug delivery complex may have 38 to 39% of the carboxyl groups in the entire carboxyl groups in the hyaluronic acid are substituted with catechol.
Further, in order to solve the above-mentioned other problems,
a) preparing a mold particle having an OH group on its surface;
b) preparing hyaluronic acid-catechol complex by reacting hyaluronic acid with catechol;
c) coating the hyaluronic acid catechol complex on the surface of the template particles by reacting the template particles with the hyaluronic acid catechol complex; And
and d) removing the template particles to prepare a hollow hyaluronic acid-catechol drug delivery complex.
According to one embodiment of the present invention, the template particles may be at least one particle selected from the group consisting of silica particles, Ag particles, Fe 3 O 4 particles, CdSe-ZnS particles and Au particles.
According to another embodiment of the present invention, the step c) comprises: stirring the mixed solution of the template particles and the hyaluronic acid catechol complex for 1.5 hours to 3 hours under the condition of pH 4.5 to 5.6; And stirring the mixed solution under the conditions of pH 7.2 to pH 7.6 for 20 hours to 30 hours.
According to another embodiment of the present invention, the reaction may be performed for 20 hours to 30 hours under the condition of pH 7.2 to pH 7.6, followed by repeatedly performing the sonication and stirring.
Further, in order to solve the above-described problems,
A shell comprising hyaluronic acid substituted with catechol; And
And a hyaluronic acid-catechol drug complex comprising an inner hollow-supported drug surrounded by the outer shell.
According to one embodiment of the present invention, the drug is at least one selected from the group consisting of doxorubicin, 5-fluorouracil, floxiuridine and mitomycin C It may be a hydrophilic anticancer drug.
According to another embodiment of the present invention, the release characteristics of the drug loaded on the drug complex may vary depending on the surrounding pH.
According to the present invention, there is provided a hyaluronic acid-catechol drug delivery complex and a method for preparing the same, wherein the drug-releasing complex can be further selectively released from the weak acid environment, which is a cancer cell environment, by controlling the oxidation- .
1 is a schematic view illustrating a process for preparing a hyaluronic acid-catechol drug delivery complex according to the present invention.
FIG. 2 is a graph showing the results of 1 H-NMR spectroscopy to examine the degree of substitution of catechol in the hyaluronic acid-catechol complex prepared according to an embodiment of the present invention.
FIG. 3 is a photograph comparing the particle size and morphology of HA-CA sheaths by atomic force microscopy for silica @ HA-CA at pH 5.5, 7.4 and 8.5.
FIG. 4 is a graph showing the results of chemical analysis of FT-IR analysis of silica @ HA-CA at pH 5.5, 7.4 and 8.5. FIG.
FIG. 5 is a graph showing the results of XPS analysis of silica @ HA-CA at pH 5.5, 7.4, and 8.5, where the carbon species are measured. FIG.
FIG. 6 is a diagram schematically showing each form in which mold particles (silica @ HA-CA) in the state of coating a hyaluronic acid catechol complex at pH 5.5, 7.4 and 8.5 are present.
FIG. 7 shows SEM images of silica @ HA-CA at pH 5.5, 7.4 and 8.5, respectively after agitation for 24 hours, after two sonication, and before removal of the finally prepared template silica particles.
FIGS. 8A and 8B show the results of silica @ HA-CA before removal of the template silica particles prepared at pH 7.4 and Bio-TEM of the hollow HA-CA finally prepared, after removing the internal silica particles FIG.
FIG. 9 is a graph showing the results of DLS analysis of the silica @ HA-CA before the removal of the template silica particles prepared under the pH 7.4 condition and the hollow HA-CA finally prepared by removing the internal silica particles.
FIG. 10 is a chart showing the results of TGA analysis of silica @ HA-CA before removal of the template silica particles prepared at pH 7.4 and hollow HA-CA finally prepared by removing the internal silica particles.
11 is a graph showing the loading and loading efficiency of doxorubicin loaded into a hollow drug delivery complex under various inlet weight ratios.
12 is a graph showing the rate at which doxorubicin is released over time at 37 DEG C, pH 5.5 and pH 7.4.
Hereinafter, the present invention will be described in more detail with reference to the drawings and examples.
In the present invention, a drug delivery system was prepared using a complex of bifunctional catechol and hyaluronic acid. In view of the fact that catechol-substituted hyaluronic acid can be self-assembled on the template particles, the surface of the template particles is coated with catechol-substituted hyaluronic acid, and then the inner mold particles are removed to prepare a hollow drug carrier Respectively. The catechol-substituted hyaluronic acid coated on the template particles competitively exhibits adsorption and flocculation characteristics as the ambient pH conditions are changed. As a result, the drug release characteristics are varied depending on various pH conditions, To cancer cells.
Therefore, in the present invention,
A shell comprising hyaluronic acid substituted with catechol; And
A hyaluronic acid-catechol drug delivery complex comprising an inner hollow surrounded by the outer shell.
As described above, the drug delivery complex according to the present invention is different in shape and size depending on pH conditions, and exhibits adhesiveness remarkably under an acidic pH condition (approximately pH 5.5 or less), but under basic pH conditions Approximately pH 8.5 or more), the cohesiveness becomes prominent. In addition, the adsorbability and cohesiveness are appropriately balanced under substantially neutral conditions. Due to these properties, the drug delivery complex according to the present invention can selectively release the drug carried on the inner hollow in a pH-dependent manner.
In the drug delivery system according to the present invention, the hyaluronic acid-catechol complex forms a shell to form a hollow in which the drug is carried, the hydration radius is 160 nm to 190 nm, and the catechol content in the shell is, Can be included at levels such that from 38% to 39% of the total carboxyl groups in the lactic acid are replaced with catechol.
Hereinafter, a method of preparing a drug delivery complex according to the present invention will be described in detail with reference to the drawings. FIG. 1 schematically illustrates a process for preparing a hyaluronic acid-catechol drug delivery complex according to the present invention. Referring to FIG. 1,
a) preparing a mold particle having an OH group on its surface;
b) preparing hyaluronic acid-catechol complex by reacting hyaluronic acid with catechol;
c) coating the hyaluronic acid catechol complex on the surface of the template particles by reacting the template particles with the hyaluronic acid catechol complex; And
d) removing the template particles to prepare a hollow hyaluronic acid-catechol drug delivery complex.
In the present invention, first, a step of producing hyaluronic acid substituted with catechol-substituted template particles and OH groups on the surface is carried out (a) and b).
As the template particles, at least one particle selected from the group consisting of silica particles, Ag particles, Fe 3 O 4 particles, CdSe-ZnS particles and Au particles may be used. Depending on the type of the selected template particles, The step of removing the mold particles in the mold is different.
The next step is to self-assemble the catechol-substituted hyaluronic acid on the surface of the template particles, which is believed to be due to the fact that hyaluronic acid, initially substituted with catechol, , And performing the reaction under neutral conditions such that the adhered hyaluronic acid catechol complex may become competitive in terms of adhesiveness and cohesiveness.
Specifically, the step c) may include stirring the mixed solution of the template particles and the hyaluronic acid catechol complex under a condition of pH 4.5 to pH 5.6 for 1.5 hours to 3 hours; And stirring the mixed solution under the conditions of pH 7.2 to pH 7.6 for 20 hours to 30 hours.
The reaction in the neutral condition may include periodically repeating the sonication and stirring. Such sonication and stirring may be, for example, repeating the process of sonicating for 20 minutes and stirring for 10 minutes four times. This sonication and agitation process enables dispersion of the agglomerated particles.
The final step in the method according to the present invention is a process (d) of preparing a hollow hyaluronic acid-catechol drug delivery complex by removing the template particles present in the core.
This step d) can be carried out differently depending on the type of the template particles used in step a). For example, when silica particles are used as the template particles, they are treated with HF solution. When Ag particles are used, NH 4 OH treatment, treatment with HCl solution when Fe 3 O 4 particles are used, treatment with CdSe-ZnS particles in an acid or ammonium solution for about 5 days or Au particles using NaCN solution treatment It is possible to remove the mold particles.
The present invention also provides a hyaluronic acid-catechol drug complex in which a drug is supported in the hollow of the hyaluronic acid-catechol drug delivery complex.
Although the drug capable of being carried on the drug delivery complex according to the present invention is not limited thereto, considering that the complex according to the present invention is excellent in the release effect of an internal drug carried in an acidic environment such as cancer cells, may be a hydrophilic anticancer agent such as doxorubicin, 5-fluorouracil, floxiuridine and mitomycin C, and the like.
As described above, the drug complex according to the present invention has different releasing properties depending on the peripheral pH. This is because the drug complex according to the present invention exhibits mainly adherence in an acidic environment and cohesiveness in a basic environment And in a neutral environment, a state in which adhesion and cohesiveness compete with each other are expressed.
EXAMPLES Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are intended to assist the understanding of the present invention and should not be construed as limiting the scope of the present invention.
Manufacturing example
material
Hyaluronic acid (HA, 200 kDa) was purchased from Lifecore Biomedical (Chaska, MN, USA). Tetraethylorthosilicate (TEOS. 99.9%) and 1- (3-dimethylaminopropyl) ethylcarbodiimide hydrochloride (EDC) were purchased from Alfa Aesar. Hydroxybenzotriazole hydrate (HOBt, ≥≥97.0%), dopamine hydrochloride, ammonium hydroxide solution (28.0-30.0% in H 2 O) and hydrofluoric acid (HF, 48.0-51.0% in H 2 O) were purchased from Sigma Aldrich. Dulbecco's Phosphate Buffered Saline (DPBS) was purchased from WELGENE (Daegu, Korea). Doxorubicin hydrochloride (DOX) was purchased from TCI (Tokyo, Japan). Anhydrous ethanol (≥ 99.9%) and dimethylsulfoxide (DMSO, 99.8%) were used, and deionized water was used with a resistance value greater than 18.2 MΩ.
Preparation of hyaluronic acid-catechol complex
Hyaluronic acid (HA-CA) substituted with catechol was prepared by an EDC / HOBt coupling reaction, which is achieved by conjugation between the carboxylic acid group of HA and the amine group of dopamine. Specifically, 100 mg of HA were dissolved in 50 mL of DPBS and the solution was purged with nitrogen for 30 minutes. The pH was adjusted to 5.5 using 1 M HCl. In the solution, 142 mg of EDC and 104 mg of HOBt dissolved in 4 mL of H2O: DMSO = 1: 1 were added and 143 mg of dopamine hydrochloride was added. The pH of the solution was maintained at 5.5 using 1 M HCl and 1 M NaOH for 4 hours. The solution was then dialyzed against de-ionized water for 2 days (MWCO: 6000-8000 g / mol SpectraPor), unreacted reagents and salts for 1 day for pH 5.0 HCl solution. The product was lyophilized for 2 days. The molar ratio of HA: EDC: HOBt: dopamine to carboxylic acid group was 1: 3: 3: 3.
Preparation of mold particles
The silica template particles were prepared by the Stober method (W. Stober, A. Fink, E. Bohn, J Colloid Interf Sci 1968 , 26 , 62- "). During the stirring, 3 mL of TEOS was added to 9 mL of ammonium hydroxide and 100 mL of ethanol mixture. The reaction was continued for 24 hours. The resulting silica colloidal dispersion was purified by centrifugation and redispersed in ethanol. The final purification was repeated in water. The purification step was repeated 3 times in each solvent.
Coating of hyaluronic acid catechol complex on the surface of the template particles
10 mg of HA-CA was dissolved in 20 mL of DPBS solution adjusted to pH 5.5. 5 mg of silica particles dissolved in 5 mL of DPBS (pH 5.5) were added dropwise to the above HA-CA solution. The mixture was stirred for 2 hours so that the HA-CA could be fully attached to the silica. Thereafter, the resultant solution was divided into three conditions.
The first case was maintained at pH 5.5 under nitrogen purge to prevent catechol oxidation. The pH of the second case was increased to 7.4 using 1 M NaOH. The pH of the third case was further increased to 8.5. The three solutions at each pH were stirred for one day. Each solution was dispersed by repeating 4 times with a process consisting of sonication-agitation (20 min-10 min). The product was then dialyzed against deionized water for 1 day and lyophilized.
Preparation of hollow hyaluronic acid-catechol drug delivery complex by casting particle removal
6.25 mL of HF (48.0-51.0%) was added to 25 mL of the silica @ HA-CA (hyaluronic acid catechol complex coated solution) solution at pH 7.4 in a 10% v / v HF solution . The mixture was stirred slowly. After 24 hours, the resultant was dialyzed against a sufficient amount of deionized water and lyophilized.
Hollow hyaluronic acid-catechol drug delivery complex inside Doxorubicin Bearing
The doxorubicin loading into the hollow HA-CA particles was carried out with various feed weight ratios (fwr (%) = mass of doxorubicin / mass of polymer in initial solution x 100). Doxorubicin was dissolved in 0.3 mL of DMSO. 2 mg of hollow HA-CA particles (10 mM, pH 7.4) dissolved in 2.7 mL of DPBS were added rapidly to DMSO phase. The mixture was stirred for 1 day. Excess drug and DMSO were removed by dialysis against DPBS buffer (2 mM, pH 7.4) for 1 day (2 L, MWCO: 6000-8000 g / mol SpectraPor) After the dialysis, the concentration of HA-CA hollow particles changed from 0.67 mg / mL to 0.57 mg / mL due to the osmotic effect. The loading amount of doxorubicin was measured by UV-Vis spectroscopy. absorbance at λ max = 485 nm is dialyzed bar The hollow HA-CA particles alone did not exhibit absorption at these wavelengths. The quantification of doxorubicin was performed using a calibration curve of doxorubicin in DPBS .
[Formula 1]
Loading content = (doxorubicin content in loaded HA - CA particle) / (HA content - CA particle) x 100
[Formula 2]
Loading efficiency = (doxorubicin content in loaded HA - CA particles) / initial
sign From bitro pH-dependent Doxorubicin Aging characteristics
The release experiments of doxorubicin were carried out under two pH conditions (pH 5.5 and pH 7.4) at the same temperature (37 ± 1 ° C). A dialysis tube containing a solution of 3.5 mL of doxorubicin-loaded hollow HA-CA particles ([HA-CA particles] = 0.57 mg / mL) was introduced into the release medium in vitro, which contained 50 mL of DPBS mM, pH 7.4). To observe the release characteristics at pH 5.5, the pH of the doxorubicin-loaded hollow HA-CA particle solution was adjusted by the addition of 0.1 M HCl immediately after dialysis. 3.5 mL of the solution was then immersed in an in vitro release medium (2 mM, pH 5.5) containing 50 mL of acetate buffer. In both cases, the immersion conditions were maintained by replacing the release medium with fresh media at each sampling time with 2 mL of in vitro. Doxorubicin release was calculated using the following formula:
[Formula 3]
% Doxorubicin release = (1 - absorbance (t) / absorbance (t 0 )) × 100
Evaluation example
Used equipment
Dynamic light scattering (DLS) experiments were performed with an ALV / DLS / SLS 5000 goniometer system using a fiber-optic SO-SIPD / DUAL detection unit. The numerical radius values were measured by a He-Ne laser (wavelength,? = 632.8 nm, 35 mW) at a scattering angle of 90 °. SEM images of silica @ HA-CA were obtained using SNE-4000 M (SEC Co., Korea) driven at 15 kV and samples were prepared on silica wafers and coated with gold deposition. The Fourier transform infrared (FT-IR) spectra to determine the chemical composition were obtained with FTLA 2000 (ABB Ltd.) using KBr pellets mixed with silica @ HA-CA prepared at each pH. FT-IR analysis was performed within the range of 500 - 4.000 cm -1 ,
Degree of catechol substitution of hyaluronic acid catechol complex
1 H-NMR spectroscopy was performed to determine the degree of substitution of catechol in the hyaluronic acid-catechol complex. The results are shown in FIG. 2, when a peak (1.8-2.2 ppm) of a carboxyl group (indicated by a blue circle in the figure) and a carboxyl group (indicated by a red circle in the figure) of dopamine bonded to hyaluronic acid in the hyaluronic acid without dopamine substitution By comparing the peaks (6.8 to 7.2 ppm), it was confirmed that 39% of carboxyl groups in hyaluronic acid were conjugated with dopamine.
Comparison of particle size according to pH condition (dynamic Light scattering (dynamic light scattering) measurement)
Particle sizes were compared by dynamic light scattering measurements on template particles alone, silica @ HA-CA (pH 5.5, 7.4 and 8.5) and hollow hyaluronic acid-catechol drug delivery complexes.
As a result, the particle size of each of the template particles alone> hollow hyaluronic acid-catechol drug delivery complex> silica @ HA-CA was the order of magnitude. In case of silica @ HA-CA, The size decreased with increasing pH and the polydisperse hydration radius appeared at pH 8.5. The measurement results are shown in Table 1 below.
(pH 5.5)
(pH 7.4)
(pH 8.5)
150 ± 10
700 ± 17
Referring to Table 1, it can be seen that the polydispersity of silica @ HA-CA at pH 8.5 is observed by the template particles alone, the layered gel and the small particles.
Comparison of particle sizes according to pH conditions (atomic force microscopy measurement)
Particle sizes were compared by atomic force microscopy for silica @ HA-CA at pH 5.5, 7.4 and 8.5. The results are shown in FIG. 3. Referring to FIG. 3, it can be seen that roughly spherical silica @ HA-CA particles are observed at pH 5.5 and less spherical silica @ HA-CA particles at pH 7.4, It can be seen that the specific shape of the particles can not be found and the layer is of the wide layer type.
FT-IR and XPS Confirmation of chemical composition at each pH and confirmation of oxidation-reduction transition of catechol
The chemical composition was confirmed by FT-IR analysis on silica @ HA-CA at pH 5.5, 7.4 and 8.5, and the results are shown in FIG. Referring to FIG. 4, it can be seen that the specific signal of the catecholator appears at a large value at pH 8.5.
In addition, 1s carbon species were measured by XPS, and the results are shown in Fig. 5, when catechol is in a reduced state at pH 5.5, reduced catechol and reduced catechol are coexisted at pH 7.4 and reduced catechol at pH 5.5 at pH 8.5 (two OH, aromatic Structure) existed in a dominant state.
Through the results of FIGS. 2 to 5, modeling of the particle presence form and chemical composition of silica @ HA-CA at each pH state was performed, and the results are schematically shown in FIG.
Particle characterization by reaction step
In order to investigate the characteristics of the particles according to the reaction step in the process of preparing the hyaluronic acid-catechol drug delivery complex according to the present invention, after stirring for 24 hours at pH 5.5, 7.4 and 8.5, And SEM images of silica @ HA-CA before the finally prepared template silica particles were removed. The results are shown in FIG.
Referring to FIG. 7, it can be seen that the dispersion of HA-CA between pH 7.4 is more easy (this is because of the weak cohesiveness due to the formation of oxidative catechol) It was confirmed that the silica @ HA-CA prior to the removal of the template silica particles is better formed. On the other hand, it was confirmed that the adsorbability was strong at pH 5.5, and it was embedded in viscous HA-CA, and at pH 8.5, the silica was embedded in the cohesive HA-CA,
In order to confirm whether the inner mold silica particles were smoothly removed by the HF treatment, the silica @ HA-CA before the removal of the template silica particles and the Bio @ -TEM were performed, and the results are shown in FIGS. 8A and 8B.
Referring to FIG. 8A, it can be seen that a spherical structure is formed by surrounding the silica core (the darker portion of the center) by the HA-CA sheath (the bright portion of the enclosure), and also with reference to FIG. 8B, The hollow structure is formed by etching removal.
On the other hand, the size of the hollow particles was reduced by the removal of the core, and DLS analysis was performed to further confirm this. The results are shown in FIG. Referring to FIG. 9, it can be seen that at pH 7.4, the R h values of the silica @ HA-CA particles decrease and then change to a wider size distribution after the HF treatment. This difference is caused by the space occupied by the cores where the HA-CA envelope is scaled down and removed.
On the other hand, TGA analysis was performed to confirm whether or not the silica core was completely removed, and the results are shown in FIG. Referring to FIG. 10, it was found that at a temperature of 600 ° C, the silica @ HA-CA particles showed a weight loss of 28.9%, whereas the hollow hyaluronic acid-catechol drug delivery complex showed almost 100% . This difference corresponds to the silica core portion of the silica @ HA-CA particles and in the case of the hollow hyaluronic acid-catechol drug delivery complex, the silica core is absent.
Drug loading and release of hyaluronic acid-catechol drug complex
To confirm whether or not the hollow hyaluronic acid-catechol drug delivery complex according to the present invention can be used as a carrier for delivering an anticancer drug, doxorubicin, which is one of widely known anticancer drugs, is loaded on the drug delivery complex and its release characteristics Respectively.
Figure 11 is a graph showing the loading and loading efficiency of doxorubicin loading into a hollow drug delivery complex under various feed weight ratios (fwr (%) = mass of doxorubicin / mass of polymer in initial solution x 100).
Referring to FIG. 11, the linear increase in the loading content with increasing doxorubicin fwr indicates that the number of captured doxorubicin molecules increases. Also, an increase in loading efficiency above 40% fwr is a common phenomenon in polymer carriers that carry doxorubicin.
FIG. 12 is a graph showing the rate at which doxorubicin is released over time at 37 DEG C, pH 5.5 and pH 7.4, with doxorubicin release at pH 5.5 nearly complete before the initial 8 hour period, and 67% of the loaded doxorubicin, Lt; / RTI > However, at pH 7.4, the loaded doxorubicin was slowly released, and after 60 hours, only 25% of doxorubicin was released. As described above, it can be judged that the phenomenon that different drug release profiles appear depending on pH is mainly caused by two factors. One is that the solubility of doxorubicin is increased due to completely hydrogenated amine groups, Is due to a decrease in the surface negative charge of HA-CA, which reduces the electrostatic attraction between doxorubicin and the hollow drug delivery complex HA-CA. As a result, these results show that the hollow hyaluronic acid-catechol drug delivery complex according to the present invention can exhibit the release characteristic triggered by the pH change in cancer cells because of the weak acidity .
Claims (10)
b) preparing hyaluronic acid-catechol complex by reacting hyaluronic acid with catechol;
c) coating the hyaluronic acid catechol complex on the surface of the template particles by reacting the template particles with the hyaluronic acid catechol complex; And
d) removing the template particles to produce a hollow hyaluronic acid-catechol drug delivery complex,
Wherein the step c) comprises stirring the mixed solution of the template particles and the hyaluronic acid catechol complex under a condition of pH 4.5 to pH 5.6 for 1.5 hours to 3 hours; And a step of reacting the stirred mixed solution under the condition of pH 7.2 to pH 7.6 for 20 hours to 30 hours. The method for producing a hyaluronic acid-catechol drug delivery complex according to claim 1,
Wherein the template particles are at least one particle selected from the group consisting of silica particles, Ag particles, Fe 3 O 4 particles, CdSe-ZnS particles, and Au particles.
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Non-Patent Citations (3)
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| Advanced Fuctional Materials. 2013. Vol.23, pp.1774-1780.* |
| Advanced Materials. 2008. Vol.20, pp.4154-4157.* |
| Chemistry of Materials. 2011. Vol.23, pp.3141-3143.* |
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