CN113797177B - Quercetin oral sustained-release preparation modified by ionic emulsifier chitosan nanoparticles and preparation method thereof - Google Patents

Abstract

The invention relates to an oral sustained-release preparation of quercetin modified by ionic emulsifier chitosan nanoparticles and a preparation method thereof. Specifically, quercetin is used as a packaged drug, and an anionic surfactant with an oral penetration-promoting effect is prepared. Sodium lauryl sulfate and positively charged chitosan are prepared by emulsification ion cross-linking method, the preparation process is simple, low energy consumption, easy to operate, and quercetin with a size of less than 200 nanometers and high encapsulation efficiency is obtained Oral sustained-release preparations to improve the water solubility of quercetin, longer sustained-release effect and promote the penetration of its preparations to enhance quercetin absorption and enhance oral bioavailability, and the present invention uses additional dextran-70 and chitosan nanoparticles to The stable reaction is carried out through the hydrogen bond interaction, the stability of the quercetin oral sustained-release preparation is improved, and it is suitable for the clinical promotion of the natural product quercetin in the field of medicine, and has great application value.

Description

A kind of quercetin oral sustained-release preparation modified by ionic emulsifier chitosan nanoparticles and preparation method thereof

technical field

The invention relates to an oral sustained-release preparation of quercetin modified with chitosan nanoparticles by an ionic emulsifier and a preparation method thereof, belonging to the technical field of oral quercetin sustained-release preparations.

Background technique

Quercetin is a natural polyhydroxy flavonoid compound, which is widely found in fruits, vegetables and various Chinese herbal medicines. It has pharmacological effects, and has the advantages of wide source, low price, no side effects, high biocompatibility, etc., and has great development value in clinical research and application. However, due to its low solubility, low intrinsic activity, poor gastrointestinal absorption, and low bioavailability, its clinical application effect is not ideal, which seriously limits its application in the field of medicine. Therefore, a strategy to increase the solubility and oral bioavailability of quercetin is urgently needed to improve the effective development and medical application value of quercetin resources.

Nanotechnology is a very effective method to enhance drug solubility and improve efficacy. Compared with traditional drugs, nano preparations can change the pharmacokinetics and tissue distribution of drugs by reducing particle size, increasing specific surface area, and improving solubility. And intrinsic pharmacological activity, in order to achieve the effect of reducing adverse reactions, improving drug efficacy and bioavailability. At present, researchers have conducted extensive research on quercetin nano-preparations, and tried to prepare a variety of nano-preparations, such as quercetin nanocrystals, quercetin nanoliposomes, quercetin nanoemulsions, etc. However, how to ensure the stability of quercetin nanocrystals in the preparation process is still a difficult problem faced by researchers. At the same time, quercetin nanoliposomes are prone to deterioration, poor stability, and unsatisfactory controlled and sustained release effects. Limit its application in clinical practice. In addition, quercetin nanoemulsion technology also has disadvantages such as high pharmaceutical cost, short shelf life and impact on standard testing.

The polymer nanoparticle drug-loading system is to dissolve or disperse drugs into nanoparticles. It has been proven that the system as a drug carrier can improve drug solubility and biological activity, and has the advantages of stable drug-loading system and high drug encapsulation efficiency. Among many drug carriers, polybutylcyanoacrylate and chitosan have attracted much attention because of their good biocompatibility and biodegradability, but polybutylcyanoacrylate has the disadvantage of being relatively expensive. Chitosan, as a biocompatible, positively charged natural polymer in solution, has good penetration enhancement drug absorption properties, and has been widely used to form nanocomposite carrier structures, which can significantly improve drug absorption. bioavailability, but chitosan is insoluble in water, and it is difficult to form a stable nano-preparation when used alone. It has been reported that sodium tripolyphosphate is used as a crosslinking agent to prepare stable chitosan nanoparticles, but the use of sodium tripolyphosphate increases the preparation cost, which is not conducive to industrialization, and the prepared chitosan nanoparticles are poor in stability. , large particle size, early drug release due to particle degradation and poor intestinal absorption limit their application.

Contents of the invention

The present invention aims at the low stability of the existing chitosan nanotechnology, the shortcoming of sudden release and low gastrointestinal absorption rate, and provides a combination of chitosan and sodium lauryl sulfate prepared with a small particle size, dispersed It is a quercetin nano oral sustained-release preparation with strong properties, good stability, slow drug release, and effective promotion of drug cell penetration and intestinal absorption.

In order to solve the above-mentioned technical problems, the present invention adopts the following technical scheme: a quercetin oral sustained-release preparation modified by an ionic emulsifier chitosan nanoparticle, including the wrapped drug quercetin and the wrapped material chitosan and twelve Sodium Alkyl Sulfate.

Further, the particle diameter of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles is 160-180 nanometers. In implementing the process of the present invention, the inventors found that under this particle diameter, quercetin sustained release Released formulations are more efficiently absorbed orally.

Further, the encapsulation efficiency of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles is 71.97% to 75.83%. Oral bioavailability of sustained-release preparations of corticosteroids is high, in which quercetin is the encapsulated drug, and chitosan and sodium lauryl sulfate are the encapsulating materials.

The present invention provides a preparation method of quercetin oral sustained-release preparation modified by the above-mentioned ionic emulsifier chitosan nanoparticles, comprising the following steps:

(1) Raw material preparation

Prepare cationic cross-linking raw materials: uniformly disperse chitosan in glacial acetic acid aqueous solution;

Preparation of anionic emulsified and crosslinked raw materials: uniformly disperse anionic emulsifier or oral penetration enhancer in ultrapure water;

Prepare stable reaction raw materials: uniformly disperse stabilizer in ultrapure water;

(2) Cross-linking reaction

The cross-linking reaction selects the emulsified ion cross-linking reaction;

The process of the emulsified ion cross-linking reaction is as follows: add quercetin to the cationic cross-linking raw material that has been stirred overnight and filtered through a filter membrane, and after it is fully dissolved by ultrasound, the pH is adjusted to be neutral with sodium hydroxide to obtain a mixture I, and then the anion The cross-linking raw materials were slowly added to the mixture I drop by drop to carry out the emulsified ion cross-linking reaction.

Further, the emulsifier, ionic cross-linking agent and oral penetration enhancer are all sodium lauryl sulfate. In the process of implementing the present invention, the inventors have found that sodium lauryl sulfate can not only be used as anionic surfactant Guarantee the emulsification ion cross-linking reaction with chitosan to form chitosan nanoparticles with small particle size, and because it is an excipient for oral preparations approved by FDA, it can also be an effective penetration enhancer and gastrointestinal Protease inhibitors ensure that the prepared chitosan nanoparticles can effectively penetrate intestinal epithelial cells, promote drug absorption, improve its oral bioavailability, and then improve the curative effect of quercetin.

Further, 0.3-0.6 grams of chitosan is dispersed in every 40 milliliters of 0.8-1.1% glacial acetic acid aqueous solution as the cationic emulsified cross-linking raw material. In the process of implementing the present invention, the inventors have found that the encapsulation efficiency of quercetin is not high for chitosan nanoparticles prepared below or above this range, mainly because chitosan is processed below this range. The amount of emulsified ionic cross-linking reaction is not enough. If it is higher than this range, the cross-linking reaction will be incomplete, which will waste chitosan raw materials, and use glacial acetic acid in this range to subtly dissolve quercetin, which is insoluble in water, to avoid the use of toxic organic reagents to dissolve quercetin.

Further, for the anionic emulsified crosslinking raw material, 0.3-0.6 grams of anionic emulsifiers are dispersed in every 40 milliliters of ultrapure aqueous solution. In the process of implementing the present invention, the inventors found that within this range, the prepared chitosan nanoparticles have smaller diameters, uniform distribution, the highest encapsulation efficiency, and the highest oral bioavailability. Mainly because sodium lauryl sulfate acts as an emulsifier and anionic cross-linking agent in the present invention, but below this range, the particle size is too large and easy to aggregate and very unstable, and the penetration-promoting effect of nanoparticles is poor. Above this range, the electromotive force of the formed nanoparticle is lower than 30 millivolts, which also makes the whole nanosystem very unstable, and the encapsulation efficiency of quercetin is not high.

Further, configure stable reaction raw materials, be dispersed with stabilizer 0.8～1.1 gram in every 20 milliliters of ultrapure aqueous solutions, in implementing the process of the present invention, the contriver finds, within this scope, the chitosan nanoparticle stability that makes Higher, the sustained release effect is slower.

Further, the optimal dosage volume ratio of cationic and anionic emulsifying crosslinking materials in the emulsifying ion crosslinking reaction is 1:1, and the process of the present invention is carried out. The inventors found that using this ratio, the reaction efficiency is higher and the reaction is fully guaranteed. and easy to operate.

Further, 0.04-0.10 g of quercetin was added to the cationic cross-linking raw material that was stirred overnight and filtered, and the pH was adjusted to be neutral with sodium hydroxide to obtain mixture I. The process of the present invention was implemented. The inventors found that using this range It can not only ensure that the encapsulation rate of quercetin is 71.97-75.83%, but also ensure its utilization rate. A 0.45 micron filter membrane is used to filter the cationic cross-linked raw material, which can guarantee to obtain a sustained-release preparation with a nanometer of 160-180. The mixture is adjusted to neutral to ensure that the acidity and alkalinity of the prepared quercetin nano-preparation is close to the physiological pH value absorbed by the small intestine.

Further, during the emulsified ion crosslinking reaction, magnetic stirring is performed at a rotational speed of 900-1100 revolutions per minute. In the process of implementing the present invention, the inventors found that, by adopting this stirring rate, the prepared nanoparticles can be prepared with a smaller particle size and a more uniform distribution.

Further, the reaction time of the emulsified ion crosslinking reaction is 1.5-2.5 hours. In the process of implementing the present invention, the inventors found that this range can ensure that the anion and cation cross-linking reactions are more complete, and a more sufficient quercetin-loaded chitosan nanosuspension can be obtained.

Further, the present invention also has the following steps:

(3) Stable reaction

The stabilization reaction process is: slowly add the stabilizer dispersed in ultrapure water dropwise into the solution prepared by the cross-linking reaction for stirring and stabilization reaction, and the polymer-wrapped quercetin nano buffer with high stability can be obtained. The suspension of the oral preparation for release, or the freeze-dried powder of the quercetin nano-sustained-release oral preparation prepared by freeze-drying technology.

Further, the stabilizer is dextran-70, although this raw material is a commonly used nano-preparation stabilizer, but in the process of implementing the present invention, the inventors found that the different order of addition to the nano-preparation improves the stabilizing effect of The difference is very different. When it is added at the end, chitosan nanoparticles have been formed, which can combine with the prepared nano-particles through hydrogen bonds to form a protective shell, making the prepared quercetin nano-preparations more stable. During the delivery process from the stomach to the small intestine, it is ensured that the nanoparticle skeleton does not aggregate or fall apart, until it reaches the destination, it penetrates the cells and slowly releases the drug to exert a therapeutic effect.

Further, in step (3), a magnetic stirrer with a rotating speed of 900-1100 rpm is used for stirring treatment, and the time is 0.5-1 hour. After implementing the process of the present invention, the inventors found that the quercetin nano-preparation obtained in this range has better stability and sustained release effect and oral bioavailability.

The present invention also provides an oral medicine. Including quercetin sustained-release preparations and excipients. The oral medicine can be made into common preparations of oral medicine such as capsules, tablets, pills, powders, granules, syrups, etc., all of which belong to the protection category of the present invention. The specific preparation method can be prepared by referring to the conventional method in the pharmaceutical field, and the auxiliary materials used can be selected according to different dosage forms. In specific implementation, appropriate medical or edible auxiliary materials can be directly added to the suspension and lyophilized powder of the prepared quercetin oral sustained-release preparation, so that any dosage form or food that can be used for medicine or food can be obtained. Material.

The beneficial effects of the present invention are reflected in:

The quercetin oral sustained-release preparation modified by the above-mentioned ionic emulsifier chitosan nanoparticles provided by the invention utilizes the low toxicity of chitosan and sodium lauryl sulfate, the advantages of high biocompatibility and biodegradability, etc. Advantages to prepare non-toxic and harmless, good biocompatibility, biodegradable nanoparticles, and sodium lauryl sulfate is an excipient for oral preparations approved by the FDA, which has the effect of promoting intestinal penetration and absorption, so that the preparation The nano-preparation of quercetin not only enables slow release of quercetin during oral administration, but also enables it to effectively enter intestinal epithelial cells to promote absorption and enhance curative effect.

The quercetin sustained-release preparation provided by the present invention is an oral preparation, which can greatly improve patient compliance compared with injection preparations and suppositories, and is easier to promote patients to take for the treatment of various diseases, thereby obtaining wider biomedical applications .

The quercetin nano-sustained-release preparation provided by the present invention is a kind of nano-preparation, which can increase the contact area of the preparation with intestinal epithelial cells by taking advantage of the small particle size of nano-sized particles, further promote its entry into intestinal epithelial cells, and promote drug Absorption, enhancing the therapeutic effect of quercetin, provides a more convenient prospect for the full development and application of quercetin.

In the preparation method of the quercetin nano-sustained-release preparation provided by the present invention, both quercetin and chitosan are cleverly dissolved in glacial acetic acid, no organic solvent is used, and the problems of post-treatment and solvent residue after removing the solvent are avoided. The method has the advantages of simple process, low cost, low requirements on reaction conditions, and easy industrialization, which not only brings economic benefits and environmental protection significance, but also has great social benefits and has high application prospects.

The preparation method of the quercetin oral sustained-release preparation modified by chitosan nanoparticles of an ionic emulsifier according to the present invention can be prepared in the form of milky white suspension or white freeze-dried powder, spherical or ellipsoidal in shape, granular The diameter is less than 200 nanometers, the dispersion index of the suspension in ultrapure water is 0.128-0.213, the electromotive force is 30.10-45.01 millivolts, the encapsulation rate is 71.97-75.83%, and the slow drug release can last up to 72 hours. , uniform dispersion without adhesion, good stability, easy penetration and easy absorption, controlled release and other advantages.

The present invention uses chitosan and sodium lauryl sulfate to promote absorption at the same time, and the emulsifying properties of sodium lauryl sulfate increase the solubility of quercetin and its nano-preparation, which inhibits enzyme activity and promotes penetration, Effectively protect the long-term delivery of quercetin from the stomach to the small intestine from degradation, further enhance the penetration of chitosan nanoparticles into intestinal cells, and promote the therapeutic activity of quercetin in cells.

In the preparation method of quercetin oral sustained-release preparation modified by ionic emulsifier chitosan nanoparticles, the stabilizing reaction is carried out at last, and the hydroxyl group of dextran-70 is combined with the hydroxyl group of chitosan nanoparticles by hydrogen bonding , forming a layer of barriers to prevent the degradation of the coated crispy nanoparticles, better prevent the degradation of quercetin nano-preparations in the harsh environment of the gastrointestinal tract, and improve the stability of its oral preparations, thus prolonging the drug effect time of the preparations , the pharmacodynamic effect increases, and the bioavailability increases.

Description of drawings

Fig. 1 is the flow chart of the preparation scheme of quercetin oral sustained-release preparation modified by a kind of ionic emulsifier chitosan nanoparticles prepared in Example 1.

Fig. 2 is the quercetin oral slow-release preparation that a kind of ionic emulsifier chitosan nanoparticle modification that embodiment 1 makes and particle diameter are less than the transmission electron microscope picture of 200 nanometers;

Fig. 3 is the zeta potential figure that the quercetin oral slow-release preparation of a kind of ionic emulsifier chitosan nanoparticle modification that embodiment 1 makes efficiently improves preparation stability;

Fig. 4 is the UV scanning spectrogram that proves that quercetin is effectively encapsulated by the quercetin oral sustained-release preparation modified by a kind of ionic emulsifier chitosan nanoparticles prepared in embodiment 1;

Fig. 5 is that embodiment 6 proves that the oral sustained-release preparation of quercetin modified by ionic emulsifier chitosan nanoparticles significantly improves the sustained-release research figure of quercetin in vitro;

Figure 6 is a diagram illustrating that the oral sustained-release formulation of quercetin modified by ionic emulsifier chitosan nanoparticles in vivo is significantly higher than the oral absorption and effective bioavailability of quercetin alone in its blood concentration-time curve.

Detailed ways

The present invention will be described in detail below in conjunction with examples.

Example 1

As shown in Figure 1, this implementation provides the preparation of quercetin oral sustained-release preparations modified by ionic emulsifier chitosan nanoparticles

The preparation method comprises the following steps:

(1) Raw material preparation

Prepare cationic cross-linking raw materials: uniformly disperse 0.6 g of chitosan in 40 ml of 1.1% glacial acetic acid aqueous solution;

Preparation of anionic emulsified crosslinking raw materials: disperse 0.6 g of sodium lauryl sulfate in 40 ml of ultrapure aqueous solution;

Preparation of stable reaction raw materials: uniformly disperse 1.1 g of dextran-70 in 20 ml of ultrapure aqueous solution.

(2) Emulsion ion cross-linking reaction

Accurately weighed 0.04 g of quercetin and added it to the cationic cross-linking raw material which was stirred overnight at room temperature at a magnetic stirring rate of 1000 rpm and filtered through a 0.45 μm filter membrane, and the pH was adjusted to 6.9 with sodium hydroxide to obtain a mixture I. Then at a magnetic stirring rate of 1100 rpm, the anionic crosslinking raw material was slowly added dropwise to the mixture I to carry out the emulsified ion crosslinking reaction. The total reaction time was 2.5 hours, and quercetin was preliminarily loaded into chitosan. nanosuspensions.

(3) Stable reaction

The stabilization reaction process is as follows: slowly add dextran-70 uniformly dispersed in ultrapure water to the suspension prepared by the cross-linking reaction drop by drop, and carry out stirring and stabilization reaction at 1100 rpm for 1 hour to obtain relatively stable High chitosan-encapsulated quercetin nano-oral preparations.

The preparation flow chart of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles obtained in this embodiment is shown in Figure 1, the transmission electron microscope of the prepared preparation is shown in Figure 2, the Zeta potential is shown in Figure 3, and the ultraviolet scanning spectrum See Figure 4.

It can be concluded that the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this embodiment has a simple preparation process, is easy to operate, is green, and has low energy consumption. The raw materials used are economical and easy to obtain, making the preparation The process is easy for industrialization, and the prepared preparation is spherical, with small particle size of 188.73±5.26 nanometers, uniform distribution, good dispersion, no particle adhesion, and an encapsulation rate of 75.83%.

In Fig. 3, 0.04 gram of quercetin is fully dissolved in 100 milliliters of glacial acetic acid aqueous solutions and the obtained solution, the blank nanoparticle solution prepared by the same preparation process as in this example without adding quercetin and the prepared nanoparticle solution of this example Three samples of quercetin oral sustained-release preparation solution modified by ionic emulsifier chitosan nanoparticles were diluted ten times, and 1 ml was drawn respectively to measure the samples with a particle size analyzer. From the results, it can be seen that the zeta potential of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this example is about +36 millivolts, and has good stability. The zeta potential of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this example is positive, which makes it easy to combine with negatively charged intestinal cells and increases the affinity between the nanoparticles and intestinal cells , making the formulation easy to penetrate the intestinal epithelial cells and enhance the oral absorption of quercetin.

In Fig. 4, the solution obtained by fully dissolving 0.04 gram of quercetin in 100 milliliters of glacial acetic acid aqueous solution, the blank nanoparticle solution prepared by the same preparation process without adding quercetin in this example, and the solution prepared in this example The spectrogram results of the spectral measurement of three samples of the ionic emulsifier chitosan nanoparticles modified quercetin oral sustained-release preparation solution. It is known that the maximum absorption wavelength of quercetin is 374 nanometers, so according to the ultraviolet scanning spectrum results of three samples, it can be known that the chitosan nano oral sustained-release preparation prepared in this embodiment can effectively wrap quercetin and increase its solubility. .

Example 2

Preparation of oral sustained-release formulation of quercetin modified with ionic emulsifier chitosan nanoparticles

The preparation method comprises the following steps:

(1) Raw material preparation

Preparation of cationic cross-linking raw materials: uniformly disperse 0.3 g of chitosan in 40 ml of 0.8% glacial acetic acid aqueous solution;

Preparation of anionic emulsified crosslinking raw materials: disperse 0.3 g of sodium lauryl sulfate in 40 ml of ultrapure aqueous solution;

Preparation of stable reaction raw materials: uniformly disperse 0.8 g of dextran-70 in 20 ml of ultrapure aqueous solution.

(2) Emulsion ion cross-linking reaction

Accurately weighed 0.10 g of quercetin was added to the cationic cross-linking raw material stirred overnight at room temperature at a magnetic stirring rate of 800 rpm and filtered through a 0.45 μm filter membrane, and the pH was adjusted to 6.6 with sodium hydroxide to obtain a mixture I. Then, at a magnetic stirring rate of 900 rpm, the anionic crosslinking raw material was slowly added dropwise to the mixture I to carry out the emulsified ion crosslinking reaction. The total reaction time was 1.5 hours, and quercetin was preliminarily loaded into chitosan. nanosuspensions.

(3) Stable reaction

The stabilization reaction process is as follows: slowly add dextran-70 uniformly dispersed in ultrapure water to the suspension prepared by the cross-linking reaction drop by drop, and carry out the stabilization reaction at 900 rpm for 0.5 hours to obtain relatively stable High chitosan-encapsulated quercetin nano-oral preparations.

The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained-release preparation obtained in this example is similar in transmission electron microscope, zeta potential, and ultraviolet scanning spectrogram to Example 1. For the sake of concise description, it will not be repeated. .

The particle size of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this example is 162.31±5.48 nanometers, and the encapsulation efficiency is 80.15%.

Example 3

Preparation of oral sustained-release formulation of quercetin modified with ionic emulsifier chitosan nanoparticles

The preparation method comprises the following steps:

(1) Raw material preparation

Preparation of cationic crosslinking raw materials: uniformly disperse 0.4 g of chitosan in 40 ml of 0.9% glacial acetic acid aqueous solution;

Preparation of anionic emulsified crosslinking raw materials: disperse 0.4 g of sodium lauryl sulfate in 40 ml of ultrapure aqueous solution;

Preparation of stable reaction raw materials: uniformly disperse 0.9 g of dextran-70 in 20 ml of ultrapure aqueous solution.

(2) Emulsion ion cross-linking reaction

Accurately weighed 0.08 g of quercetin was added to the cationic cross-linking raw material stirred overnight at room temperature at a magnetic stirring rate of 900 rpm and filtered through a 0.45 μm filter membrane, and the pH was adjusted to 6.7 with sodium hydroxide to obtain a mixture I. Then, at a magnetic stirring rate of 900 rpm, the anionic crosslinking raw material was slowly added dropwise to the mixture I to carry out the emulsified ion crosslinking reaction. The total reaction time was 1.5 hours, and quercetin was preliminarily loaded into chitosan. nanosuspensions.

(3) Stable reaction

The stabilization reaction process is as follows: slowly add dextran-70 uniformly dispersed in ultrapure water to the suspension prepared by the cross-linking reaction drop by drop, and carry out the stabilization reaction at 1000 rpm for 0.5 hours to obtain relatively stable High chitosan-encapsulated quercetin nano-oral preparations.

The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained-release preparation prepared in this embodiment has transmission electron microscope images, zeta potential images, and ultraviolet scanning spectrograms similar to those in Example 1. For the sake of concise description, it is not repeated. .

The particle size of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this example is 172.56±4.83 nanometers, and the encapsulation efficiency is 73.57%.

Example 4

Preparation of oral sustained-release formulation of quercetin modified with ionic emulsifier chitosan nanoparticles

The preparation method comprises the following steps:

(1) Raw material preparation

Preparation of cationic crosslinking raw materials: uniformly disperse 0.5 g of chitosan in 40 ml of 1.0% glacial acetic acid aqueous solution;

Preparation of anionic emulsified cross-linking raw materials: disperse 0.5 g of sodium lauryl sulfate in 40 ml of ultrapure aqueous solution;

Preparation of stable reaction raw materials: uniformly disperse 1.0 g of dextran-70 in 20 ml of ultrapure aqueous solution.

(2) Emulsion ion cross-linking reaction

Accurately weighed 0.06 g of quercetin was added to the cationic cross-linking raw material stirred overnight at room temperature at a magnetic stirring rate of 900 rpm and filtered through a 0.45 micron filter membrane, and the pH was adjusted to 6.8 with sodium hydroxide to obtain a mixture I. Then, at a magnetic stirring rate of 900 rpm, the anionic crosslinking raw material was slowly added dropwise to the mixture I to carry out the emulsified ion crosslinking reaction. The total reaction time was 2.0 hours, and quercetin was preliminarily loaded into chitosan. nanosuspensions.

(3) Stable reaction

The stabilization reaction process is as follows: slowly add dextran-70 uniformly dispersed in ultrapure water to the suspension prepared by the cross-linking reaction drop by drop, and carry out the stabilization reaction at 1000 rpm for 1.0 hour to obtain relatively stable High chitosan-encapsulated quercetin nano-oral preparations.

The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained-release preparation prepared in this embodiment has transmission electron microscope images, zeta potential images, and ultraviolet scanning spectrograms similar to those in Example 1. For the sake of concise description, it is not repeated. .

The particle size of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this example is 179.62±4.67 nanometers, and the encapsulation efficiency is 74.92%.

Example 5

Preparation of oral sustained-release formulation of quercetin modified with ionic emulsifier chitosan nanoparticles

The preparation method comprises the following steps:

(1) Raw material preparation

Preparation of cationic crosslinking raw materials: uniformly disperse 0.5 g of chitosan in 40 ml of 1.0% glacial acetic acid aqueous solution;

Preparation of anionic emulsified crosslinking raw materials: disperse 0.4 g of sodium lauryl sulfate in 40 ml of ultrapure aqueous solution;

Preparation of stable reaction raw materials: uniformly disperse 0.9 g of dextran-70 in 20 ml of ultrapure aqueous solution.

(2) Emulsion ion cross-linking reaction

Accurately weighed 0.08 g of quercetin was added to the cationic cross-linking material stirred overnight at room temperature at a magnetic stirring rate of 800 rpm and filtered through a 0.45 μm filter membrane, and the pH was adjusted to 6.7 with sodium hydroxide to obtain a mixture I. Then, at a magnetic stirring rate of 900 rpm, the anionic crosslinking raw material was slowly added dropwise to the mixture I to carry out the emulsified ion crosslinking reaction. The total reaction time was 2.0 hours, and quercetin was preliminarily loaded into chitosan. nanosuspensions.

(3) Stable reaction

The stabilization reaction process is as follows: slowly add dextran-70 uniformly dispersed in ultrapure water to the suspension prepared by the cross-linking reaction drop by drop, and carry out the stabilization reaction at 1100 rpm for 0.5 hours to obtain relatively stable High chitosan-encapsulated quercetin nano-oral preparations.

The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained-release preparation prepared in this embodiment has transmission electron microscope images, zeta potential images, and ultraviolet scanning spectrograms similar to those in Example 1. For the sake of concise description, it is not repeated. .

The particle size of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in this example is 171.83±5.82 nanometers, and the encapsulation efficiency is 74.11%.

Observed under a transmission electron microscope, the quercetin oral sustained-release preparations prepared in Examples 1 to 5 have small particle sizes, spherical particles, uniform distribution, good dispersion, and no particle adhesion. Through particle size analyzer analysis, the size of the quercetin oral sustained-release preparation products prepared in Examples 1 to 5 are all less than 200 nanometers, the dispersion index is 0.128～0.213, and the electromotive force is 30.10～45.01 millivolts, and the results similar to those shown in Figure 4 are obtained. Ultraviolet absorption spectrum, the quercetin in the above examples has been fully encapsulated.

Example 6

Release test of quercetin oral sustained-release preparation modified by ionic emulsifier chitosan nanoparticles

With the phosphate buffer saline (PBS) that pH value is 7.4 as release medium, accurately weigh quercetin 4 milligrams and 20 milliliters of release mediums and evenly mix to obtain contrast quercetin release solution, then make the The quercetin nano-preparation suspension was uniformly mixed with the release medium to obtain a quercetin oral sustained-release preparation release test group with a total volume of 20 ml. Then, transfer them to the dialysis bags soaked in distilled water at the same time, tie the two ends of the dialysis bags tightly to the slurry, place them in dissolution Erlenmeyer flasks filled with 250 milliliters of release medium respectively, and shake them at a constant temperature (37°C). Shake at a constant speed in the bed (120 rpm).

Take 2 ml samples at the set time and store them in a -4°C refrigerator, add 2 ml of release medium solution after each sampling, and centrifuge the samples at -4°C for 15 minutes in a high-speed low-temperature centrifuge at 10,000 rpm , take 1 ml of the supernatant and filter it through a 0.45 micron microporous membrane, and take the supernatant.

Use high-performance liquid chromatography to measure the content of quercetin in the supernatant of samples at different time points, inject 20 microliters each time, measure the peak area of quercetin, and calculate the concentration of quercetin according to the standard curve regression equation , to obtain the cumulative release percentage of quercetin, and draw the release curve, the experimental results are shown in Figure 5.

Figure 5 shows that the ionic emulsifier chitosan nanoparticles can slowly control the release of quercetin, and the release time is as long as 3 days, which effectively proves that the quercetin modified by the ionic emulsifier chitosan nanoparticles has a slow control releasing effect.

Example 7

Oral bioavailability test of quercetin oral sustained-release preparation modified by ionic emulsifier chitosan nanoparticles

Prepare 12 adult healthy SD male rats, divide them into two groups on average: group A and group B, fast for 12 hours, take blood from the retro-orbital sinus as a blank control sample, and then give it to each 100 grams of rat body weight 1 milliliter of medicinal solution was administered to 12 rats. Wherein, group A gave 0.04 gram of quercetin fully dissolved in 100 milliliters of glacial acetic acid (0.1%) gained solution in the aqueous solution, group B gave the quercetin nanometer oral sustained-release preparation solution that the embodiment made, and recorded different gavage respectively time.

At 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 14 hours, 16 hours, 18 hours, 20 hours, 22 hours, and 24 hours after two groups of rats were given oral 0.5 ml of blood was collected from the posterior venous sinus and placed in a 5 ml vacuum blood collection tube containing anticoagulant. Next, add 3 milliliters of methanol to precipitate blood protein components, vortex for 10 minutes, and centrifuge at 1000 rpm for 10 minutes. After the supernatant is filtered through a 0.45-micron filter membrane, 20 microliters of the filtrate is precisely sucked and analyzed by high-performance liquid chromatography. The concentration of quercetin in the filtrate was measured by sample injection. And draw the plasma concentration-time curve of quercetin and quercetin nano-preparation as shown in Figure 6.

The results of analysis in Figure 6 show that the plasma peak concentration of quercetin oral sustained-release preparation solution modified by ionic emulsifier chitosan nanoparticles is higher than that of quercetin solution and the time to reach the peak is prolonged, while the ion emulsifier chitosan The area under the curve of the plasma concentration-time curve of the quercetin oral sustained-release preparation modified by nanoparticles was significantly larger than that of quercetin. The results prove that the quercetin oral sustained-release preparation modified by ionic emulsifier chitosan nanoparticles can significantly improve the bioavailability of quercetin and can achieve the purpose of slow and continuous oral drug absorption.

The quercetin in vitro release test and the oral bioavailability (blood drug concentration-time curve) test result of the quercetin oral sustained-release preparation modified by the ionic emulsifier chitosan nanoparticles prepared in embodiment 1 to embodiment 5 are compared with The quercetin sustained-release oral preparation prepared in Example 1 is similar, and for the sake of concise description, it is not repeated here.

The above content is a detailed description of the present invention in conjunction with specific embodiments, and it cannot be assumed that the specific implementation of the present invention is limited to these descriptions. Those skilled in the art can make various modifications or changes based on it, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principles of the present invention shall be included within the protection scope of the present invention.

Claims (5)

1. An ion emulsifier chitosan nanoparticle modified quercetin oral sustained-release preparation is characterized in that: comprises wrapped medicine quercetin, wrapping materials chitosan and lauryl sodium sulfate, the particle diameter of the quercetin nano sustained-release preparation is 160-180 nm, the encapsulation rate is 71.97-75.83%, and the preparation method comprises the following steps:

(1) Preparation of raw materials

Preparing a cation crosslinking raw material: evenly dispersing chitosan in glacial acetic acid aqueous solution, wherein 0.3-0.6 g of chitosan is dispersed in 0.8-1.1% glacial acetic acid aqueous solution per 40 ml of cation crosslinking raw material;

preparing an anionic emulsifying crosslinking raw material: dispersing anionic emulsifier or oral permeation enhancer in ultrapure water, wherein 0.3-0.6 g of anionic emulsifier is dispersed in each 40 ml of ultrapure water solution in the anionic crosslinking raw material;

preparing stable reaction raw materials: dispersing stabilizer in ultrapure water uniformly, and dispersing 0.8-1.1 g of stabilizer in each 20 ml of ultrapure water solution in the stable reaction raw material;

(2) Crosslinking reaction

The crosslinking reaction adopts an emulsifying ion crosslinking reaction;

the emulsifying ionic crosslinking reaction process comprises the following steps: adding quercetin into cation crosslinking raw material which is stirred overnight and filtered by a filter membrane, carrying out ultrasonic treatment to fully dissolve the quercetin, adjusting pH to be neutral by using sodium hydroxide to obtain a mixture I, and then dropwise and slowly adding anion crosslinking raw material into the mixture I to carry out emulsification ion crosslinking reaction.

(3) Stable reaction

The stable reaction process is as follows: the stabilizing agent dispersed in the ultrapure water is slowly and dropwise added into the solution prepared by the cross-linking reaction to be stirred for stabilizing the reaction, and the polymer-coated quercetin nanometer preparation with higher stability can be obtained.

2. The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained release formulation as claimed in claim 1, characterized in that: the emulsifier, the anionic cross-linking agent and the oral penetration enhancer are all sodium dodecyl sulfate, and the stabilizer is dextran-70.

3. The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained-release formulation as set forth in claim 1 or 2, wherein: in the process of the emulsification ion crosslinking reaction, the mass concentration of the quercetin in the mixture I is 1.0-2.5 g/L, and the pH value of the solution is 6.6-6.9 after the pH value of the solution is adjusted to be slightly neutral by sodium hydroxide.

4. The ionic emulsifier chitosan nanoparticle modified quercetin oral sustained-release formulation as set forth in claim 1 or 2, wherein: the emulsion ion crosslinking reaction time is 1.5-2.5 hours, and the stable reaction time is 0.5-1.0 hour.

5. An oral medicament, comprising: an oral sustained-release formulation of quercetin comprising the ionic emulsifier chitosan nanoparticle modified according to any one of claims 1 to 4 and an adjuvant.

Images