KR20170097840A - Method for manufacturing Fluorochrom combined with Bile acid-Chitosan complex nanoparticle and Composition including this fluorochrom for diagnosis of disease - Google Patents
Method for manufacturing Fluorochrom combined with Bile acid-Chitosan complex nanoparticle and Composition including this fluorochrom for diagnosis of disease Download PDFInfo
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- 210000000941 bile Anatomy 0.000 title claims abstract description 10
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- HSINOMROUCMIEA-FGVHQWLLSA-N (2s,4r)-4-[(3r,5s,6r,7r,8s,9s,10s,13r,14s,17r)-6-ethyl-3,7-dihydroxy-10,13-dimethyl-2,3,4,5,6,7,8,9,11,12,14,15,16,17-tetradecahydro-1h-cyclopenta[a]phenanthren-17-yl]-2-methylpentanoic acid Chemical compound C([C@@]12C)C[C@@H](O)C[C@H]1[C@@H](CC)[C@@H](O)[C@@H]1[C@@H]2CC[C@]2(C)[C@@H]([C@H](C)C[C@H](C)C(O)=O)CC[C@H]21 HSINOMROUCMIEA-FGVHQWLLSA-N 0.000 claims description 8
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- NHBKXEKEPDILRR-UHFFFAOYSA-N 2,3-bis(butanoylsulfanyl)propyl butanoate Chemical compound CCCC(=O)OCC(SC(=O)CCC)CSC(=O)CCC NHBKXEKEPDILRR-UHFFFAOYSA-N 0.000 description 1
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K49/00—Preparations for testing in vivo
- A61K49/04—X-ray contrast preparations
- A61K49/0409—Physical forms of mixtures of two different X-ray contrast-enhancing agents, containing at least one X-ray contrast-enhancing agent which is not a halogenated organic compound
- A61K49/0414—Particles, beads, capsules or spheres
- A61K49/0423—Nanoparticles, nanobeads, nanospheres, nanocapsules, i.e. having a size or diameter smaller than 1 micrometer
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K49/0013—Luminescence
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- C07—ORGANIC CHEMISTRY
- C07J—STEROIDS
- C07J9/00—Normal steroids containing carbon, hydrogen, halogen or oxygen substituted in position 17 beta by a chain of more than two carbon atoms, e.g. cholane, cholestane, coprostane
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- C08—ORGANIC MACROMOLECULAR COMPOUNDS; THEIR PREPARATION OR CHEMICAL WORKING-UP; COMPOSITIONS BASED THEREON
- C08B—POLYSACCHARIDES; DERIVATIVES THEREOF
- C08B37/00—Preparation of polysaccharides not provided for in groups C08B1/00 - C08B35/00; Derivatives thereof
- C08B37/0006—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid
- C08B37/0024—Homoglycans, i.e. polysaccharides having a main chain consisting of one single sugar, e.g. colominic acid beta-D-Glucans; (beta-1,3)-D-Glucans, e.g. paramylon, coriolan, sclerotan, pachyman, callose, scleroglucan, schizophyllan, laminaran, lentinan or curdlan; (beta-1,6)-D-Glucans, e.g. pustulan; (beta-1,4)-D-Glucans; (beta-1,3)(beta-1,4)-D-Glucans, e.g. lichenan; Derivatives thereof
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Abstract
Description
More particularly, the present invention relates to a method for producing a fluorescent substance to which a cholanic acid-chitosan complex nanoparticle is bound, and more particularly, to a method for producing a fluorescent substance comprising a cholanic acid-chitosan (HGC) To a method for producing fluorescent nanoparticles.
Recently, the importance of optical molecular imaging technology has emerged in the early diagnosis of diseases. In particular, the importance of optical molecular imaging technology in the field of early diagnosis and treatment of cancer has been growing, and compared with conventional PET / CT or PET / SPECT diagnostic techniques, cancer tissue imaging by near- , High-resolution images, and low cost.
However, the most effective method for early diagnosis of cancer using near-infrared-visible cancer tissue imaging is to have a maximum absorption and fluorescence within the spectral range of near-infrared wavelength (650 nm ~ 900 nm), high solubility in water, In addition to the development of highly compatible near infrared ray phosphors, such near infrared ray phosphors are selectively transferred to and accumulated in cancer tissues to develop contrast agents exhibiting high fluorescence intensities in cancer diseases.
Accordingly, the present inventors have completed the present invention by developing a method for producing a novel polymer nanoparticle-type compound in which a near infrared ray fluorescent substance capable of near-infrared light penetration is chemically bonded to a polymer selectively accumulating in a cancer tissue by a high transmittance of a neovascularization of cancer tissue Respectively.
It is an object of the present invention to provide a method for preparing near infrared fluorescent nanoparticles conjugated with cholanic acid-chitosan complex to improve the low specificity and low fluorescence intensity of cancer tissue which is a disadvantage of conventional fluorescence contrast agents.
In order to solve the above problems, the present invention has developed a method for producing phosphor nanoparticles conjugated with a cholanic acid-chitosan complex, which comprises the steps of: a) purifying chitosan, b) synthesizing cholanic acid-chitosan nanoparticles, c ) Binding the cholanic acid-chitosan nanoparticles to the fluorescent substance.
More particularly, the present invention relates to a method for preparing a cholanic acid-chitosan (HGC) complex by adsorbing bile acid to chitosan, which is a polymer, and binding the complex to a near infrared ray fluorescent substance to obtain fluorescent nanoparticles having a polymer derivative bound thereto , The fluorescent substance of the present invention prepared by the above-described method can be used as a contrast agent composition for early diagnosis of diseases such as cancer.
Structure of Fluorescent Bonded Cholanic Acid-Chitosan (HGC) Nanoparticles
The structure of the cholanic acid-chitosan (HGC) complex is a part excluding the D in the formula (1), and D represents a phosphor bound to the cholanic acid-chitosan (HGC) complex, and the phosphor (D) The phosphor to be displayed may be used.
In
The cyanine-based compound represented by Formula 2 is a cyanine-based compound or a salt thereof.
In
In particular, in the method for producing fluorescent nanoparticles of the present invention, reaction conditions such as temperature, concentration, and time optimized for binding a polymer to a chitosan derivative are provided to the fluorescent substance, and thus, It is possible to synthesize a fluorescent nanoparticle composition having improved fluorescence intensity and long-term storage ratio.
Specific examples of the compound of Formula 3 include the following benzimidazole compounds or salts thereof.
The present invention relates to a method for producing a fluorescent nanoparticle complex in which cholanic acid-chitosan polymer nanoparticles are bonded to a near infrared ray fluorescent substance. The complex is excellent in biocompatibility and biodegradability and can be used for labeling biomolecules. Has a long accumulation rate, and has a high fluorescence intensity.
1 is a graph showing the particle sizes of the respective compounds synthesized in Examples 1, 2, 3 and 4.
Fig. 2 is a graph comparing absorbance and fluorescence intensity of each compound synthesized in Examples 1, 2, 3 and 4. Fig.
FIG. 3 is an image and a graph showing in vivo behavior of each compound synthesized in Examples 1, 2, 3 and 4. FIG.
FIG. 4 is an image obtained by intravenous administration of each compound synthesized in Examples 2 and 3 and a control drug. FIG.
FIG. 5 is an image of a long-term tissue extracted after injection of each compound synthesized in Examples 2 and 3 and a control drug, respectively.
The present invention relates to a method for preparing fluorescent nanoparticles conjugated with a cholanic acid-chitosan complex, which comprises the steps of: a) purifying chitosan, b) synthesizing cholanic acid-chitosan nanoparticles, c) - < / RTI > chitosan nanoparticles.
Particularly, in the method for producing fluorescent nanoparticles of the present invention, the reaction conditions such as temperature, concentration and time optimized for binding the chitosan derivative polymer to the near infrared ray fluorescent substance are provided, And high specificity for cancer sites.
More specifically, the structure of the cholanic acid-chitosan (HGC) complex is as shown in the following formula (4), and the process of synthesizing the cholanic acid-chitosan (HGC) complex includes a step of purifying the chitosan, a step of binding bile acid to the purified chitosan polymer Step, and can be described in more detail as follows.
Structure of bile acid-chitosan (HGC) nanoparticles
First, the process of purifying chitosan to be used in the present invention will be described.
0.5 to 1.5 g of Glycol Chitosan (GC) is dissolved in 50 to 120 mL of ultrapure water and then dialyzed. After completion of dialysis, the mixture is heated at 60 to 120 ° C for 10 to 18 hours, preferably 10 To 15 hours, followed by lyophilization. The yield of purified glycol chitosan (GC) was over 45%.
Next, the process of binding bile acid to the purified chitosan (GC) polymer will be described.
300 to 700 mg of purified glycol chitosan (GC) is added to 30 to 100 mL of ultrapure water, and then 30 to 100 mL of methanol is added and stirred. Bile acid in an amount of 80 to 250 mg, preferably 100 to 180 mg, is added to 80 to 150 mL of methanol to be used. 100 to 200 mg of N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (EDC) are added to 800 to 1500 mL of methanol and stirred, followed by stirring in a glycol chitosan (GC) solution. 0.5 to 2 mL of methanol is added to 50 to 100 mg of N-hydroxysuccinimide (NHS), and the mixture is added to a bile acid solution. The mixture is stirred into a glycol chitosan (GC) mixed solution, The temperature is maintained at 20 to 30 DEG C and the reaction time is set to 20 to 30 hours. The reaction solution is filtered, dialyzed for 4 days, dialyzed, dispersed at 20 to 30 ° C for 20 to 60 seconds, and freeze-dried. The yield of the prepared chitosan-chitosan (HGC) nanoparticles was 65% or more.
Finally, a method of binding the cholanic acid-chitosan complex prepared above to the phosphor will be described.
In the present invention, the phosphor represented by Formula 1 or Formula 2 may be used as a fluorescent material to be a target.
Hereinafter, a method of binding cholanic acid-chitosan (HGC) nanoparticles to the phosphor will be described.
20 to 60 mg of the cholanic acid-chitosan (HGC) nanoparticles prepared as described above are dissolved in 10 to 20 mL of a carbonate buffer solution, and the phosphor represented by the general formula (2) or (3) is dissolved in DMSO at a concentration of 8 to 12 mg / mL Then, the solution is added to the cholanic acid-chitosan (HGC) solution and reacted. The temperature is maintained at 20 to 30 ° C., the reaction time is set to 20 to 30 hours, and the dialysis is continued for 5 days after the reaction . The dialyzed compound is lyophilized to obtain a fluorescent nanoparticle in which a cholanic acid-chitosan (HGC) complex is bound to the phosphor. The yield of the fluorescent nanoparticles was 85% or more.
Preferred embodiments are provided 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 examples.
Synthesis of cholanic acid-chitosan (HGC) nanoparticles
1) Purification of chitosan
1 g of Glycol Chitosan (GC) (100 kDa) was dissolved in 80 mL of ultra-pure water in a 250 mL Erlenmeyer flask and dialyzed several times using a dialysis membrane (
2) Synthesis of cholanic acid-chitosan (HGC) nanoparticles
500 mg of purified glycol chitosan (GC) is dissolved in 60 mL of ultrapure water, and 60 mL of methanol is further added thereto. 150 mg of 5? -Cholanic acid was added to 120 mL of methanol and the mixture was thoroughly stirred. 1000 mL of methanol was added to 150 mg of N- (3-dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride (N- (3- (Dimethylaminopropyl) -N'-ethylcarbodiimide hydrochloride) And the mixture was stirred. 72 mg of N-Hydroxysuccinimide (NHS) was added to 1 mL of methanol, stirred, and then added to the bile acid solution and stirred. The bile acid mixture solution was added to the GC mixture solution and stirred at room temperature overnight. The reaction solution was filtered using a syringe filter (pore size: 80 탆), and then dialyzed for 4 days in a dialysis membrane (MWCO 12-14 kDa). After completion of the dialysis, the dispersion was dispersed for 30 seconds on a waterbath sonicator, followed by lyophilization to obtain HGC nanoparticles. (0.492g, 75.7%) The dialysis solution for 1-2 days is a 3: 1 v / v solution of methanol and deionized water. The dialysis solution for 3 days is a 1: 1 v / v solution of methanol and deionized water , And the fourth dialysis solution is ultrapure water.
Preparation of cholanic acid-chitosan (HGC) 648 nanoparticles
HGC-648 nanoparticles
1) Synthesis of HGC-648-1 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL of 648 Vinylsulfone (0.32 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to the bile acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
2) Synthesis of HGC-648-2 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL of 648 Vinylsulfone (1.28 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to the cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
Preparation of cholanic acid-chitosan (HGC) 675 nanoparticles
HGC-675 nanoparticles
1) Synthesis of HGC-675-1 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL of 675 Vinylsulfone (0.32 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
2) Synthesis of HGC-675-2 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mg of 675 Vinylsulfone (1.28 mu mol) in DMSO at a concentration of 10 mg / ml, and added dropwise to the cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
Production Example of Bile Acid-Chitosan (HGC) 749 Nanoparticles
HGC-749 nanoparticles
1) Synthesis of HGC-749-1 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL of 749 Vinylsulfone (0.32 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to the cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
2) Synthesis of HGC-749-2 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL of 749 Vinylsulfone (1.28 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to the cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
Preparation of cholanic acid-chitosan (HGC) 774 nanoparticles
HGC-774 nanoparticles
1) Synthesis of HGC-774-1 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL 774 Vinylsulfone (0.32 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to the cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
2) Synthesis of HGC-774-2 nanoparticles
Chitosan-chitosan (HGC) was dissolved in 12 mL 774 Vinylsulfone (1.28 mu mol) in DMSO at a concentration of 10 mg / mL, and added dropwise to the cholanic acid-chitosan (HGC) solution. The solution was dialyzed for 5 days using a dialysis membrane (MWCO 12-14 kDa). The dialyzed solution was lyophilized to obtain HGC-648-1 nanoparticles (28.5 mg, 95%). The dialysis solution for 1 to 2 days was a solution of 8.4 mg / L of purified water / ultrapure water, The solution used was deionized water.
Particle size measurement of HGC fluorescent nanoparticles
1 mg of each compound synthesized in Examples 1, 2, 3 and 4 was aliquoted and dispersed in 1 ml of ultrapure water to give a concentration of 1 mg / ml. The resulting solution was dispersed in a probe type ultrasonic disperser (Probe type Sonicator, Sonics Vibracell VCX750, Sonics) was dispersed at intervals of 5 seconds for 1 minute. After the dispersed solution was immersed in a cuvette for a particle size analyzer, the particle size was measured using a particle size analyzer (Zetasizer Nano ZS, Malvern). The results are shown in Table 1.
As shown in Table 1 and FIG. 1, it was confirmed that the synthesized compounds each had a particle size of 300 nm or less, and it was confirmed that the size of the HGC fluorescent nanoparticles was maintained regardless of the capacity and type of each fluorescent Dye.
Optical characterization of HGC fluorescent nanoparticles
1 ml of each compound synthesized in Examples 1, 2, 3 and 4 was prepared at a concentration of 62.5 μg / ml using ultrapure water, and then analyzed with a UV spectrometer (SHIMADZU, UV-2600), a fluorescence spectrometer (SCINCO, FluoroMate FS-2).
As shown in FIG. 2, in the case of HGC-648-1 and HGC-648-2, the absorbance and fluorescence intensity of HGC-648-2 using Flamma 648 Vinylsulfone Dye four times higher than that of HGC-648-1 , And similar performance was confirmed in case of HGC-675,749,774.
Analysis of long-term accumulation trend of HGC fluorescent nanoparticles by small animal experiment
The following experiments were carried out in order to confirm the long-term accumulation tendency of each compound synthesized in Examples 1, 2, 3 and 4.
1) SCC7 cells were injected with 2 * 10 6 cells per sperm on the upper right hind leg of Balb / c nude mouse (male, 6weeks old). After 3 days, the tumor size of the mouse is observed daily, and the time point when the diameter of the cancer becomes 0.7 to 1 cm is referred to as the drug injection timing.
2) 0.1 ml of the synthesized HGC fluorescent nanoparticles were injected into the tail vein at a dose of 5 mg / kg per mouse. The animals were photographed at 1, 3, 6, 9, and 24 hours before and after injection using an optical imaging device (Neoscience, FOBI). After 24 hours, the organs were removed and the degree of residual drug by each organ was measured using an optical imaging device.
As shown in FIG. 3, when the in vivo behavior was confirmed up to day 7, it was confirmed that the substance accumulates in the whole body and tumor part at the highest efficiency at 9 hours. After 24 h in vivo, the organ was excised immediately and the wavelength of each substance was set and the most optimized images were obtained. HGC-648-1, HGC-675-1, HGC-749-1, HGC-774-2, and HGC- 1, the accumulation of substances in the tumor was higher than that in the liver.
Comparison of long-term accumulation trends with small animal experiments with other products
In order to compare the long-term accumulation tendency with other products, a small animal experiment was conducted on two selected compounds, HGC-675-2 and HGC-749-2, through optical analysis of the compound and analysis of long-term accumulation tendency . The comparison products are PerkimElmer's
1) Animal model: BALB / c-nude (male, 5 weeks old, 20 rats)
2) Construction of Xenograft model
a. Subcutaneous injection of SCC7 cell line (1x106 / 0.1ml) to Balb / c-nude mouse.
3) Test group composition and dose
SCC7 xenograft model
Single
4) Drug administration
a) Drug administration when the volume is 60 ~ 80mm2 (takes about 10 ~ 15 days)
b) Concentration of administration: 100 μg / 100 μl of each drug or 2 nol / 100 μl of each drug
c) Method of administration: i.v injection.
As shown in FIG. 4, by intravenous administration of HGC-675-2 and HGC-749-2 in an SCC7 Xenograft model, opial imaging was taken over time and it was confirmed that it was specifically targeted to accumulate in cancer tissues. The control drug AngioSense 680EX and AngioSense 750EX from PerkinElmer showed higher fluorescence intensity in whole body including tumor tissue compared to HGC-675-2 and HGC-749-2, which seems to be due to the difference in drug injection concentration.
As shown in FIG. 5, ex vivo imaging was performed at 24 hours after injection. As a result, compared with HGC-675-2 and HGC-749-2, AngioSense 680EX and AngioSense 750EX , And the rate of cancer accumulation in liver was higher than that of the control group (HGC-675-2 and HGC-749-2). Especially, the accumulation rate of Kidney was less than that of the control group.
Claims (10)
(2)
In Formula 2, R 1 . Each R 1 'is independently hydrogen, a sulfonic acid group or a sulfonic acid group; R 2 , R 2 ', R 3 and R 3 ' are each independently hydrogen or an alkyl group having 1 to 6 carbon atoms; R 4 is hydrogen, an alkyl group having 1 to 6 carbon atoms, or a carboxy group; B is (CH 2) P, para- ( C 6 H 4) or a meta- (C 6 H 4) and; m is an integer from 1 to 5; m 'is an integer from 5 to 10; P and n are each independently an integer of 1 to 5; (Arrows indicate sites where bile acid-chitosan (HGC) complexes bind).
(3)
In Formula 3, the four R 1 s are the same or different from each other and are independently hydrogen, a sulfonic acid group or a sulfonic acid group; The two R 2 are the same as or different from each other and each independently represent a carbon or hydrogen group of 1 to 6; The two R < 3 > s are the same or different from each other and are independently hydrogen or an alkyl group having 1 to 6 carbon atoms; R 4 is hydrogen, an alkyl group having 1 to 6 carbon atoms, a carboxyl group, a sulfonic acid group, or a sulfonic acid group; Wherein B is (CH 2) P, p- ( C 6 H 4) or m- (C 6 H 4), and wherein n and P is an integer from 1 to 5, wherein the two m have the same as or different from each other, and Each independently an integer of 1 to 10. (Arrows indicate sites where bile acid-chitosan (HGC) complexes bind).
[Chemical Formula 5]
(7)
[Chemical Formula 6]
[Chemical Formula 8]
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KR20190103631A (en) * | 2018-02-28 | 2019-09-05 | (주)바이오액츠 | Fluorescence compounds, complex nanoparticles comprising the same, and preparation method thereof |
KR20210072875A (en) * | 2019-12-09 | 2021-06-18 | 전남대학교산학협력단 | Hydrophobically modified glycol chitosan nanoparticle and renal targeted drug delivery system using the same |
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Cited By (2)
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KR20190103631A (en) * | 2018-02-28 | 2019-09-05 | (주)바이오액츠 | Fluorescence compounds, complex nanoparticles comprising the same, and preparation method thereof |
KR20210072875A (en) * | 2019-12-09 | 2021-06-18 | 전남대학교산학협력단 | Hydrophobically modified glycol chitosan nanoparticle and renal targeted drug delivery system using the same |
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