CN108379227B

CN108379227B - Rutin-entrapped polymer micelle and preparation method thereof

Info

Publication number: CN108379227B
Application number: CN201810525567.9A
Authority: CN
Inventors: 许海棠; 周菊英; 赵彦芝; 雷福厚; 卢建芳
Original assignee: Guangxi University for Nationalities
Current assignee: Guangxi University for Nationalities
Priority date: 2018-05-28
Filing date: 2018-05-28
Publication date: 2021-01-12
Anticipated expiration: 2038-05-28
Also published as: CN108379227A

Abstract

A rutin-loaded polymer micelle is prepared from rutin and a rutin-loaded carrier according to the weight ratio of 1: 3-100 mass ratio; a method for preparing a rutin-entrapped polymer micelle solution comprises (1) dissolving rutin and a carrier in an organic solvent, and then dropwise adding the organic solvent in which the carrier and the rutin are dissolved into distilled water under high-speed stirring; (2) volatilizing the organic solvent in the product in the step (1), filtering by a filter membrane, and collecting filtrate to obtain the product. The invention can improve the solubility of rutin which is an insoluble active ingredient of the traditional Chinese medicine and improve the bioavailability.

Description

Rutin-entrapped polymer micelle and preparation method thereof

Technical Field

The invention relates to a rutin-entrapped polymer micelle and a preparation method thereof, belonging to the field of medicines.

Background

Rutin (rutin), also known as rutin and quercitrin, is a flavonoid compound with wide sources, and many plants in the nature such as sophora flower bud, tartary buckwheat, dandelion and the like are rich in rutin, and has various biological activities of oxidation resistance, antibiosis, anti-inflammation, anticonvulsant, neuroprotection, cardiovascular and cerebrovascular protection and the like. Due to its wide biological activity, high safety and abundant sources, rutin is often used in medical care. At present, the domestic rutin is only available in the market of tablets, and because the rutin has poor water solubility, low gastrointestinal dissolution rate and extremely low oral bioavailability, the exertion of the efficacy of the rutin and the clinical application are limited. In order to improve the solubility of rutin in water and improve the bioavailability, the development of a novel rutin preparation is an effective way for improving the treatment effect of rutin, and has important clinical significance.

In order to expand the clinical application of rutin and improve the drug effect of rutin, pharmaceutical workers try various methods to solve the problem of difficult solubility of rutin, such as preparing solid dispersion, preparing liposome, preparing a self-microemulsion drug delivery system and the like, and the problem of difficult solubility of rutin is solved to a certain extent. However, these methods have problems, respectively, such as relatively few latent solvents or co-solvents available, toxic side effects (such as solubilizers and clathrates), and aging phenomena (such as solid dispersion); the surfactant applied in a large amount in the formula of the microemulsion has potential toxicity, and although the liposome increases the drug absorption, the cost is too high, so that the application prospect is short. Therefore, other solubilization methods are needed to be found, wherein the polymeric micelles are more and more concerned due to the advantages of stable properties, good biocompatibility, strong solubilization capacity and the like of the polymeric micelles, the carrier materials capable of forming the polymeric micelles are usually amphiphilic copolymers, the lengths of hydrophilic chains and hydrophobic chains of the polymers are proper, the polymers are self-assembled in water, the hydrophobic sections form hydrophobic cores of the micelles, and the hydrophilic sections form hydrophilic fence shells outside the micelles, so that the micelles have spherical core-shell structures. The research finds that no related report that the solubility of rutin is increased by using an amphiphilic copolymer as a carrier exists at present.

Disclosure of Invention

The invention aims to provide a rutin-entrapped polymer micelle and a preparation method thereof, which can improve the solubility of rutin which is an indissolvable active ingredient of a traditional Chinese medicine and improve the bioavailability.

In order to achieve the purpose of the invention, the invention adopts the technical scheme that:

a rutin-loaded polymer micelle is prepared from rutin and a rutin-loaded carrier according to the weight ratio of 1: (3-100) in mass ratio.

Further, the carrier is an amphiphilic polymer maleated rosin-polyethylene glycol ester, the hydrophobic group of the carrier is a maleated rosin group, and the hydrophilic group is polyethylene glycol with the molecular weight of 200-6000.

A preparation method of a rutin-entrapped polymer micelle solution comprises the following steps:

dissolving the carrier material and rutin in an organic solvent, and then dropwise adding the organic solvent in which the carrier material and rutin are dissolved into distilled water under high-speed stirring;

and step two, volatilizing the organic solvent in the product obtained in the step one, filtering the product through a filter membrane, and collecting filtrate, namely the rutin-entrapped polymer micelle solution.

Further, the organic solvent in the step one is one or a mixture of methanol, ethanol, acetone, ethyl acetate and dimethyl sulfoxide.

Furthermore, the volume-to-mass ratio of the organic solvent to the polymeric micelle material is (2-20 mL):100 mg.

Further, the volume ratio of the organic solvent to the distilled water is 1: (2-10).

Further, the distilled water in the first step can be replaced by physiological saline.

Further, the stirring in the step one is magnetic stirring in a water bath, and the temperature of the water bath is 25-50 ℃.

Further, the operation of volatilizing the organic solvent in the second step is carried out in a water bath kettle at 50 ℃; filtration was performed using a 0.22 μm microfiltration membrane.

The invention has the beneficial effects that:

according to the invention, a solvent volatilization method is adopted, maleated rosin-polyethylene glycol is taken as an amphiphilic polymer drug-carrying material carrier, and rutin is entrapped in a hydrophobic core of the maleated rosin-polyethylene glycol to prepare a polymer micelle entrapping rutin; the hydrophobic group is a maleated rosin group and is used as a core, the hydrophilic group is polyethylene glycol and contains a plurality of hydroxyl groups, when the concentration is higher than the critical micelle concentration, a hydrophilic shell can be formed to contain more drug molecules and entrap more rutin, the solubility of the rutin in water is improved by about 10 times of that of the bulk drug, the defect that the rutin is difficult to dissolve in water is overcome, and the release degree is improved.

By the form of the polymer micelle, the sustained-release effect is obvious, the dissolving rate of rutin is improved, and the oral bioavailability is improved; meanwhile, the used amphiphilic polymer drug-loaded material also has better biodegradability.

Drawings

Fig. 1 is a schematic diagram of in vitro release of rutin and rutin polymer micelles.

Figure 2 is a graph of rutin and rutin polymer micelles orally administered in rat blood.

Detailed Description

The invention is further illustrated by the following examples.

Example 1

100mg of rutin and 500mg of maleated rosin polyethylene glycol (2000) ester are weighed, dissolved in 10mL of hot absolute ethanol, and added dropwise into 30mL of distilled water which is stirred rapidly. Stirring in 50 deg.C water bath, volatilizing ethanol, filtering the obtained solution with 0.22 μm filter membrane, and collecting filtrate to obtain polymer micelle solution containing rutin.

Example 2

90mg of rutin and 510mg of maleated rosin polyethylene glycol (1000) ester are weighed, dissolved in 10mL of hot absolute ethanol, and added dropwise to 30mL of distilled water with rapid stirring. Stirring in 50 deg.C water bath, volatilizing ethanol, filtering the obtained solution with 0.22 μm filter membrane, and collecting filtrate to obtain polymer micelle solution containing rutin.

Example 3

80mg of rutin and 520mg of maleated rosin polyethylene glycol (200) ester are weighed, dissolved in 10mL of hot absolute ethanol, and added dropwise into 30mL of distilled water which is stirred rapidly. Stirring in 50 deg.C water bath, volatilizing ethanol, filtering the obtained solution with 0.22 μm filter membrane, and collecting filtrate to obtain polymer micelle solution containing rutin.

The High Performance Liquid Chromatography (HPLC) method is adopted to measure the drug encapsulation efficiency and the drug loading capacity of the rutin-encapsulated polymer micelle solution prepared according to the different prescriptions, and the particle size is measured by a laser particle size analyzer, and the results are shown in Table 1. The stability of the solutions was investigated, and the three solutions all had a stabilization time of greater than 24h at room temperature.

Table 1 quality evaluation of rutin polymer micelle solutions of different prescriptions (n ═ 3)

Name of vector	Encapsulation efficiency (%)	Drug loading (%)	Average particle diameter(nm)
				Maleated rosin polyethylene glycol 2000 ester	84.5％	14.1％	143
Maleated rosin polyethylene glycol 1000 ester	82.6％	12.4％	128
				Malachin polyethylene glycol 200 ester	80.3％	10.7％	106

Example 4

And (3) freeze-drying the micelle solution obtained in the embodiment 1-3 to respectively obtain three rutin-entrapped polymer micelle freeze-dried powders, and then measuring the solubility of rutin and the rutin polymer micelle freeze-dried powders in water by adopting HPLC.

Respectively putting excessive rutin and rutin polymer micelle lyophilized powder into test tubes containing 10mL of purified water, performing ultrasonic treatment until the rutin and rutin polymer micelle lyophilized powder are not dissolved, then putting the test tubes into an oscillator (100r/min), oscillating for 24h at 25 ℃, taking out saturated solution after dissolution balance, putting the saturated solution into a centrifuge, and centrifuging for 6min at 10000 r/min. The supernatant was aspirated with precision and diluted with methanol, determined by HPLC, and the peak area was recorded. The solubility of rutin in the rutin and rutin polymer micelle freeze-dried powder can be obtained by a regression equation.

The solubility of rutin and rutin polymer micelles with different prescriptions is examined in the experiment, and the result is shown in table 2. As shown in Table 2, the solubility of rutin in water is 82.7. mu.g/mL, and the solubility of rutin in water of the drug-loaded polymer micelle prepared according to example 1 is 832. mu.g/mL, which is 10 times higher than that of the bulk drug.

Table 2 solubility in water of rutin and its polymeric micelles (n ═ 3)

Sample name	Rutin	Example 1	Example 2	Example 3
					Solubility (. mu.g/mL)	82.7±2.36	832±23.5	815±20.1	803±20.5

Example 5

In vitro release degree of rutin polymer micelle

Taking a proper amount of rutin polymer micelle freeze-dried agent which is about 20mg of rutin, directly putting into a dissolution cup, determining the dissolution rate and the release degree by referring to a paddle method in the four general rules of Chinese pharmacopoeia 2015 edition, taking artificial gastric juice containing 1% of Tween 80 as a dissolution medium, rotating speed of 100r/min, the temperature of the dissolution liquid of 37 +/-0.5 ℃, operating according to the method, sampling 5mL at a preset time point, and supplementing the release medium with the same volume and temperature. The sample was filtered through a 0.22 μm filter and the subsequent filtrate was taken as a test solution. Measuring rutin in the sample by using a high performance liquid chromatograph, and calculating the cumulative dissolution percentage. The cumulative dissolution percentage was plotted as the ordinate and the time as the abscissa, and the dissolution curve was plotted, and the results are shown in FIG. 1.

As can be seen from figure 1, the drug substance is released rapidly, the in vitro cumulative dissolution percentage is 47.1% in 15min, the in vitro cumulative dissolution percentage reaches 78.2% in 3h, then the release tends to slow, and the in vitro cumulative dissolution percentage is 85.2% in 12 h. The rutin polymer micelle prepared according to the embodiment 1 releases slow medicament in the initial stage of medicament release (0-2 h), and the in vitro cumulative dissolution percentage does not reach 20% in 2 h; the medicine release is quick within 2-6 h, and the in vitro cumulative dissolution percentage reaches 86.3% within 6 h; thereafter, the release was smooth, and the percentage of in vitro cumulative dissolution at 12h was 92.3%. Compared with the bulk drugs, the rutin polymer micelle provided by the invention has obvious slow release characteristics.

Example 6

Pharmacokinetics research of rutin polymer micelle in rat body

Test drugs: the rutin polymer micelle freeze-dried agent prepared according to the embodiment 1 is dissolved by normal saline before use to prepare a solution containing rutin 2 mg/mL; rutin bulk drugs are prepared into 0.5 percent carboxymethyl cellulose suspension containing 2mg/mL of rutin, and the suspension is used as a reference preparation.

Dosing and bleeding protocol: SD rats 12 with a body weight of 180. + -.20 g were randomly divided into 2 groups. The rutin raw material and the rutin polymer micelle solution (equivalent to 40mg/kg of raw material medicine) are respectively administered after fasting and free drinking water in the day before the experiment. At predetermined time points, rats were bled from their orbits, placed in plastic centrifuge tubes containing 1% heparin sodium, plasma was centrifuged and the plasma drug concentration was determined by HPLC at each time point. The results were plotted on the abscissa as the administration time and on the ordinate as the blood concentration of rutin, as shown in fig. 2.

As a result: as can be seen from fig. 2, after oral administration, the peak reaching time of rutin drug substance in blood is about 2h, the peak blood concentration (Cmax) is 8.92 μ g/mL, the peak reaching time of rutin polymer micelle prepared in example 1 is about 6h, the Cmax is 36.5 μ g/mL, which is about 4.1 times of the drug substance, the elimination in vivo becomes slow, the average retention time of the drug in vivo is prolonged, the area under the curve during drug administration is significantly increased, which indicates that the oral absorption and bioavailability of rutin are improved.

The polymer micelle solution is an intermediate preparation form, can be prepared into freeze-dried powder for storage, can be further prepared into suitable preparation forms such as injections or oral preparations according to requirements, such as oral liquid, tablets, capsules, granules and the like, uses safe auxiliary materials, can fully ensure the safety and the compliance of clinical use, and can be widely used for preventing and treating related diseases. Can also be made into health food for improving and regulating body function.

Claims

1. A preparation method of a rutin-entrapped polymer micelle solution is characterized by comprising the following steps:

step one, mixing a mixture of 1: dissolving 3-100 parts of rutin and a rutin-coated carrier in an organic solvent, and then dropwise adding the organic solvent in which the carrier and the rutin are dissolved into distilled water under high-speed stirring;

volatilizing the organic solvent in the product obtained in the step one, filtering the product through a filter membrane, and collecting filtrate, namely the rutin-entrapped polymer micelle solution;

the carrier is amphiphilic polymer maleated rosin-polyethylene glycol ester, the hydrophobic group of the carrier is maleated rosin base, and the hydrophilic group is polyethylene glycol with the molecular weight of 200-6000.

2. The preparation method of the rutin-encapsulated polymer micelle solution according to claim 1, wherein the prepared rutin-encapsulated polymer micelle is prepared from rutin and a rutin-encapsulated carrier according to the ratio of 1: 3 to 100 by mass.

3. The method for preparing the polymeric micelle solution encapsulating rutin as in claim 1, wherein the organic solvent in the step one is one or a mixture of methanol, ethanol, acetone and ethyl acetate.

4. The method for preparing the polymeric micelle solution encapsulating the rutin according to claim 1, wherein the volume-to-mass ratio of the organic solvent to the polymeric micelle material is 2-20 mL:100 mg.

5. The method for preparing the polymeric micellar solution entrapped with rutin of claim 1, wherein the volume ratio of the organic solvent to the distilled water is 1: 2 to 10.

6. The method for preparing the polymeric micellar solution entrapped rutin of claim 1, wherein the distilled water in step one can be replaced by physiological saline.

7. The method for preparing the polymeric micelle solution encapsulating the rutin according to claim 1, wherein the stirring in the first step is magnetic stirring in a water bath, and the temperature of the water bath is 25-50 ℃.

8. The method for preparing the polymeric micelle solution encapsulating rutin as in claim 1, wherein the operation of volatilizing the organic solvent in the second step is carried out in a water bath kettle at 50 ℃; filtration was performed using a 0.22 μm microfiltration membrane.