CN114404368B

CN114404368B - Polymethoxyflavone liposome and preparation method thereof

Info

Publication number: CN114404368B
Application number: CN202210111968.6A
Authority: CN
Inventors: 郑国栋; 韦敏燕; 蔡轶; 王康慧; 宁金容; 杨婉玲; 刘梦诗
Original assignee: Guangzhou Medical University
Current assignee: Guangzhou Medical University
Priority date: 2022-01-29
Filing date: 2022-01-29
Publication date: 2023-02-03
Anticipated expiration: 2042-01-29
Also published as: CN114404368A

Abstract

The invention discloses a polymethoxyflavone liposome and a preparation method thereof, wherein the liposome consists of soybean lecithin, cholesterol and polymethoxyflavone. The encapsulation rate of the dried orange peel polymethoxylated flavone liposome of some examples of the invention reaches more than 80%, the solubility, the bioavailability and the anti-lipase activity of the dried orange peel polymethoxylated flavone are effectively improved, the dried orange peel polymethoxylated flavone liposome has the advantages of accurate content, small particle size, stable property of functional substances, difficult oxidation, high bioavailability, good slow release and the like, and the defects of poor water solubility, low bioavailability and the like of the polymethoxylated flavone are effectively overcome.

Description

Polymethoxyflavone liposome and preparation method thereof

Technical Field

The invention belongs to the field of natural products, and particularly relates to a polymethoxylated flavone liposome and a preparation method thereof.

Background

Polymethoxyflavonoids (PMFs) are a class of flavonoids with strong biological activity containing multiple methoxy groups, low polarity and planar structure, and have 4 or more than 4 methoxy groups at the positions of 3, 4, 5, 6, 7, 8, 2', 3', 4', 5', 6' and the like on phenylchromone structure, mainly come from Citrus of Rutaceae, and are abundant in pericarpium Citri Reticulatae.

PMFs are considered as potential lipase inhibitors in functional foods and have been widely used to reduce the risk of obesity in dietary treatments. Recent studies have found that dried orange peel PMFs show profound effects in preventing hyperlipidemia, obesity and type II diabetes. Pericarpium citri reticulatae PMFs can effectively combat the obesity problem by reducing the absorption of dietary fat through their good anti-lipase ability. The use of PMFs for obesity management may contribute well to the valence value and good sustainability of dried orange peel. PMFs, however, are poorly water soluble, difficult to absorb, and have low bioavailability, making it difficult to exert their efficacy effectively. In order to better perform the function of PMFs, it is necessary to effectively improve the solubility thereof.

Liposomes, which are spherical lipid vesicles surrounded by a lipid bilayer structure, are regarded as safe carriers and have been widely used in the fields of food and medicine. Due to biocompatibility and the lipid bilayer structure, liposomes provide a good system for delivery of hydrophobic compounds. Hydrophobic bioactive compounds may be encapsulated in the lipid bilayer of liposomes to improve their solubility, stability, bioavailability, and pharmacological activity. In addition, liposomes have been used in many medical and functional food fields (e.g., the use of paclitaxel liposomes has been approved by the FDA) due to their superior biocompatibility, high encapsulation efficiency and good stability. The liposome can be a good carrier for delivering the dried orange peel PMFs extract, and is expected to overcome the defects of poor water solubility, low bioavailability and the like of the PMFs.

Research shows that the high encapsulation efficiency of the liposome may cause the tendency of drug leakage in subsequent storage to reduce the encapsulation efficiency of the drug, thereby affecting the stability and subsequent utilization of the liposome. In addition, studies have found that liposomes having a particle size of less than 200 nm may promote oral absorption of bioactive compounds, and excessively high particle sizes may affect the absorption efficiency of liposomes and thus the bioavailability and pharmacological activity of encapsulated drugs, and thus optimal formulations need to be studied in order to solve the above disadvantages.

Disclosure of Invention

The invention aims to overcome at least one defect of the prior art and provides a polymethoxylated flavone liposome and a preparation method thereof.

The technical scheme adopted by the invention is as follows:

in a first aspect of the present invention, there is provided:

a polymethoxyflavone liposome comprises the following components by mass: 70-85% of soybean lecithin, 14-25% of cholesterol and 1-5% of polymethoxylated flavone.

In some examples of the liposome of polymethoxylated flavones, the polymethoxylated flavones are pericarpium citri reticulatae-derived polymethoxylated flavones.

In some examples of the liposomes of polymethoxylated flavones, the liposomes have a particle size of no greater than 100nm.

In some examples of the polymethoxylated flavone liposomes, the D50 of the liposomes is 70 to 88nm.

In some examples of the polymethoxyflavone liposomes, the D90 of the liposomes is from 40 to 90 nm.

In a second aspect of the present invention, there is provided:

a method for preparing a polymethoxylated flavone liposome, wherein the mass composition of the polymethoxylated flavone liposome is as described in the first aspect of the invention, comprises the following steps:

s1) dissolving a liposome raw material in a solvent, and removing the solvent to form a film;

s2) hydrating the film to obtain a hydration liquid;

s3) homogenizing the hydration liquid to obtain the polymethoxylated flavone liposome.

In some examples of the preparation method, the solvent is a mixture of ethanol and dichloromethane.

In some examples of the preparation method, the volume of ethanol and dichloromethane in the solvent is (2 to 3): (1-2).

In some examples of the preparation process, the homogenization pressure is not lower than 800 bar.

In some preparation examples, the water solution has a soybean phospholipid mass concentration of 24-48 mg/mL.

In some examples of the preparation method, the ratio of polymethoxylated flavone in the aqueous solution: the mass ratio of the soybean phospholipid and the cholesterol is 1 (28-128).

In some preparation examples, the hydration solution has a mass concentration of soybean phospholipid of 24-48 mg/mL, and the mass concentration of the polymethoxylated flavone: the mass ratio of the soybean phospholipid and the cholesterol is 1 (28-128).

In some examples of the preparation method, the hydration solution has a soybean phospholipid mass concentration of 24 mg/mL, and the ratio of polymethoxylated flavone: the mass ratio of the soybean phospholipids is 1.

In some examples of the preparation method, the hydration solution has a soybean phospholipid mass concentration of 24 mg/mL, and the ratio of polymethoxylated flavone: the mass ratio of soybean phospholipids is 1: the mass ratio of (soybean phospholipid + cholesterol) is 1.

In some examples of the preparation method, the hydration solution has a mass concentration of soybean phospholipids of 48mg/mL, and the ratio of polymethoxyflavone: the mass ratio of the soybean phospholipids is 1.

In some examples of the preparation method, the hydration solution has a mass concentration of 48mg/mL of soybean phospholipids, and the ratio of polymethoxylated flavones: the mass ratio of soybean phospholipids is 1: the mass ratio of (soybean phospholipid + cholesterol) is 1.

The invention has the beneficial effects that:

the encapsulation efficiency of the dried orange peel polymethoxylated flavone liposome of some embodiments of the invention reaches more than 80 percent, and the solubility, the bioavailability and the anti-lipase activity of the dried orange peel polymethoxylated flavone are effectively improved.

The dried orange peel polymethoxylated flavone liposome provided by the invention has the advantages of accurate content, small particle size, stable property of functional substances, difficult oxidation, high bioavailability, good slow release and the like, and effectively overcomes the defects of poor water solubility, low bioavailability and the like of polymethoxylated flavone.

Drawings

FIG. 1 is a graph showing the particle size comparison of dried orange peel polymethoxylated flavone liposome composition before and after high pressure homogenization

FIG. 2 is a diagram showing the physicochemical properties of liposome composition of pericarpium Citri Tangerinae-polymethoxylated flavone;

FIG. 3 is a graph showing the change in stability of a liposome composition of citrus peel polymethoxylated flavones stored at 4 and 25 deg.C for 15 days;

FIG. 4 is a graph of in vitro simulated release of liposomal composition of citrus peel polymethoxylated flavones and free polymethoxylated flavones from gastrointestinal tract;

FIG. 5 is a graph of in vitro inhibition of lipase activity by the dried orange peel polymethoxylated flavone liposome composition and free polymethoxylated flavone suspension;

FIG. 6 is a plot of mean plasma concentration versus time for the liposomal composition of citrus peel polymethoxylated flavones and for the suspension of free polymethoxylated flavones following a single gavage of 50mg/kg in rats.

Detailed Description

Experimental data of the invention show that the ratio is favorable for obtaining the liposome with high entrapment rate, high drug-loading capacity and high stability.

In some examples of the polymethoxylated flavone liposomes, the D90 of the liposomes is between 40 and 90 nm.

In a second aspect of the present invention, there is provided:

a method for preparing a liposome of polymethoxyflavone, wherein the liposome of polymethoxyflavone has a composition according to the first aspect of the present invention, comprises the following steps:

s2) hydrating the film to obtain a hydration liquid;

When the liposome is hydrated, a buffer solution close to neutrality is used for hydration, and the particle size, the entrapment rate and the like of the liposome are not obviously influenced.

The solvent need only be effective to dissolve the lipids and PMFs. In some examples of the preparation method, the solvent is a mixture of ethanol and dichloromethane. The mixed solvent can meet the requirements and has better safety.

In some examples of the preparation process, the homogenization pressure is not lower than 800 bar. The relatively higher homogenization pressure is beneficial to obtaining liposome with smaller particle size. In a specific example, the homogenization pressure is 1000 bar. The pressure for homogenization can be adjusted as desired.

In some examples of the method of preparation, the ratio of polymethoxylated flavone: the mass ratio of the soybean phospholipid and the cholesterol is 1 (28-128).

In some examples of the preparation method, the hydration solution has a soybean phospholipid mass concentration of 24 mg/mL, and the ratio of polymethoxylated flavone: the mass ratio of the soybean phospholipids is 1. Experimental data show that under such conditions, encapsulation efficiencies as high as 89.9% can be unexpectedly obtained, while good stability of the liposomes is obtained.

In some examples of the preparation method, the hydration solution has a soybean phospholipid mass concentration of 24 mg/mL, and the ratio of the polymethoxyflavone: the soybean lecithin has the mass ratio of 1: the mass ratio of (soybean phospholipid + cholesterol) is 1.

In some examples of the preparation method, the hydration solution has a mass concentration of 48mg/mL of soybean phospholipids, and the ratio of polymethoxylated flavones: the mass ratio of the soybean phospholipids is 1. Experimental data show that under the condition, the entrapment rate of about 88.0% can be unexpectedly obtained, the drug loading rate is 3.0%, and the obtained liposome has good stability.

The technical scheme of the invention is further explained by combining the examples.

The dried orange peel polymethoxylated flavones (PMFs) used in the following examples may be prepared according to conventional methods or purchased from commercial products, or prepared as follows:

1. petroleum ether reflux extraction and enrichment of dried orange peel polymethoxylated flavones (PMFs)

Dried orange peel (500 g) was pulverized and passed through a 20 mesh screen, and 2L petroleum ether was added to conduct extraction at 50 ℃ under reflux for 1 hour for 5 times until the color of the extraction solvent gradually became colorless. The filtered extract was concentrated on a rotary evaporator until a solid precipitated.

The optimization research is carried out on the optimal condition for enriching the PMFs, the PMFs of the dried orange peels are enriched through different extraction solvents, and then the extraction result is analyzed through HPLC. The extraction rates of different solvents are shown in table 1, and it can be seen from table 1 that nobiletin, which is a characteristic component of pericarpium citri reticulatae PMFs, can be enriched by different extraction solvents, but is interfered by hesperidin. When the orange peel PMFs are extracted by ethyl acetate or petroleum ether, the interference of hesperidin is minimum. However, the pigment components in the dried orange peel can be extracted together during the extraction of the ethyl acetate, and although the extraction rate of the PMFs by the petroleum ether is not the highest, in order to obtain the high-purity dried orange peel PMFs, the petroleum ether is selected as the extraction solvent for enriching the PMFs. Therefore, the petroleum ether enriched PMFs can effectively overcome the interference of other flavonoids (hesperidin and other polar flavonoids) and pigments, and the high-purity pericarpium citri reticulatae PMFs can be obtained.

TABLE 1 different extractionsExtraction rate of hesperidin and nobiletin by solventn=3）

2. Preparation of dried orange peel polymethoxylated flavone freeze-dried powder

After obtaining PMFs by petroleum ether extraction, petroleum ether can be removed by rotary evaporation to obtain PMFs yellow viscous solid. At this point the PMFs obtained by rotary evaporation may adhere to the bottom of the round bottom flask, which may not facilitate the subsequent preparation of liposomes. The extract is lyophilized to obtain dry yellow powder, which is convenient for weighing and improves the convenience of liposome preparation. Meanwhile, the PMFs extract is frozen and dried to form PMFs freeze-dried powder which is not easy to oxidize, so that the stability of the PMFs extract is ensured. Therefore, the PMFs viscous solid obtained by rotary evaporation is pre-frozen at ultralow temperature of-70 to-80 ℃ for 8 to 12 hours, and then the PMFs viscous solid is taken out and dried in a freeze dryer at the temperature of-20 to-50 ℃ for 24 to 72 hours to obtain the dried orange peel polymethoxylated flavone freeze-dried powder.

Preparation of dried orange peel polymethoxylated flavone liposome composition (PLS)

Preparing dried orange peel polymethoxylated flavone liposome composition (PLS) by adopting a film hydration-high pressure homogenization method. Lipids and citrus peel PMFs extracts were dissolved in a solvent and then dried by evaporation on a rotary evaporator to form a film. And then, hydrating the uniform film by using a buffer solution, carrying out homogenization circulation by using a high-pressure homogenizer to obtain dispersed liposome, and carrying out characterization on the physicochemical property of the prepared PLS.

Effect of Liposome composition and the like on the encapsulation efficiency of PMFs

Lipids and citrus peel PMFs extracts were dissolved in ethanol/dichloromethane (2,v/v) and then dried by rotary evaporator at 45 ℃ under vacuum evaporation to form a film. The homogeneous film was then hydrated with HEPES buffer (20 mM HEPES, 144 mM NaCl, pH 7.4) and the PLS suspension was homogenized by a high pressure homogenizer at a high pressure of 1000 bar for 20 cycles. As shown in FIG. 1 (formulation 5), the particle size of PLS was greatly reduced after high pressure homogenization to obtain liposomes having uniform and stable dispersion of particle size (the particle size of polymethoxyflavone liposomes before high pressure homogenization was 1200-1380 nm, and the particle size after high pressure homogenization was 70-88 nm).

In order to obtain the maximum encapsulation efficiency of PMFs in liposomes, the phospholipid concentration and the PMFs extract/lipid ratio (P/L ratio, w/w) in PLS liposome formulations were investigated. Table 2 shows the encapsulation efficiency and drug loading results of different formulations of the dried orange peel polymethoxylated flavone liposome composition, and the encapsulation efficiency of PMFs in PLS is remarkably increased with the decrease of P/L ratio when the phospholipid concentration in the formulation is 24 mg/mL. However, as the P/L ratio decreased, the loading efficiency of PMFs decreased significantly (formulations 1-4). When the P/L ratio is 1. When the P/L ratio was 1. These results also suggest that liposome loading capacity is primarily dependent on phospholipid concentration in the liposome composition.

To increase the loading capacity of PMFs, the phospholipid concentration in PLS was increased. When the prescription concentration of PMFs is 2 mg/mL, the entrapment rate of PMFs is increased from 35.6% to 88.0%, and the phospholipid concentration is increased from 24 mg/mL to 48mg/mL (formula 5). The results demonstrate that after formulation optimization, soybean phospholipids 70-85%, cholesterol 14-25%, polymethoxylated flavones 1-5%, PLS showed high encapsulation efficiency (over 80%) and good drug loading (3.0%) (fig. 2D).

TABLE 2 encapsulation efficiency and drug loading of the tangerine peel polymethoxyflavone liposome composition under different formulations: (n=3)

The physical and chemical property investigation of the prepared dried orange peel polymethoxylated flavone liposome composition (formula 5) shows that PLS has the characteristics of small particle size, negative surface charge, good dispersing ability, high entrapment rate and the like (figures 2A and B), and the physical and chemical property characterization results of different formula polymethoxylated flavone liposome compositions are shown in Table 3. In addition, the solubility of PMFs in PLS and aqueous solutions was also determined, with PLS having a concentration of 1575.60. + -. 8.14. Mu.g/mL, which is 16.70 times that of water (94.30. + -. 2.62. Mu.g/mL), indicating that the solubility and dispersion of PMFs after liposome encapsulation is significantly improved (FIG. 2E). These results also suggest that PLS may be an advantageous vehicle for improving the solubility of PMFs, and may be used for further applications.

There are studies that show that too high an encapsulation efficiency may affect the stability of PLS in case of drug leakage during drug storage. The inventor further tests the stability of the prepared dried orange peel polymethoxylated flavone liposome.

Three batches of PLS were stored at 4 and 25 ℃ for 15 days respectively. The results (table 4, fig. 3) show that during storage, the formulation: 70-85% of soybean phospholipid, 14-25% of cholesterol and 1-5% of polymethoxylated flavone, and the PLS still maintains higher encapsulation efficiency (all reaching more than 80%), thereby effectively overcoming the defects of instability and the like of liposome with overhigh encapsulation efficiency and ensuring the stability and efficacy of the product.

TABLE 3 physicochemical Properties of different formulations of Polymethoxyflavone Liposomal compositions: (n=3)

Table 4 different formulations have storage stability at 4 and 25 c for 15 days at the same encapsulation ratio respectively (expressed as encapsulation ratio%,n=3)

in vitro simulated gastrointestinal release investigation

To investigate the release kinetics of PLS, an in vitro simulated gastrointestinal release study was performed. In particular, the in vitro kinetics of PMFs released from liposomes was examined by dialysis. PLS and free PMFs suspensions were placed in dialysis bags (molecular weight [ Mw ]]8,000-14,000 Da), simulated gastric fluid (SGF, 0.5% SDS, 1 mol/L HCl, 10 mg/L pepsin, pH 1.37) and simulated intestinal fluid were added（SIF, 0.5% SDS, 0.5 mol/L KH ₂ PO ₄ 0.4% NaOH, 10 mg/L trypsin, pH 6.8) was used for simulated release, and magnetic stirring was carried out at 37 ℃ for 5 days. At different time points, samples were taken from the release solution and replaced with an equal volume of fresh medium (n= 3). PMFs release was measured by UV-visible spectrophotometer at 330 nm and cumulative release data was fitted to the simulation equation by Origin 9.0 software.

The results (FIG. 4) show that the aqueous solutions of free PMFs rapidly diffused within 4h, releasing rates in gastric and intestinal fluids of 38.66. + -. 0.50% and 35.13. + -. 2.40%, respectively. Meanwhile, compared with the aqueous solution of free PMFs, the corresponding liposome composition of dried orange peel polymethoxylated flavone shows similar release behavior in gastric juice (34.18 +/-0.61%) and intestinal juice (37.25 +/-1.06%) within 4 h. 18 After h, the aqueous solution of free PMFs was rapidly and completely released in both gastric and intestinal fluids, while the release of PLS in gastric fluid increased to 59.57. + -. 2.48% within 18 h, and then reached complete release within 98 h. Similarly, the release of PLS in intestinal fluid increased to 76.90. + -. 5.50% over 18 h, then to full release over 48 h. These phenomena indicate that PLS release has a significant sustained release effect compared to aqueous solutions of free PMFs. After a rapid diffusion of PMFs from liposomes for 4h, PMFs may reach high concentrations in vivo and then be subsequently released on a sustained basis. In vitro release studies show that PLS has a significant sustained release effect in gastrointestinal fluids, the high concentration of PMFs in the digestive tract is maintained, and it can be orally absorbed without frequent administration. Physical and chemical characterization and in vitro release studies have shown that PLS has a significant advantage in oral absorption.

In vitro anti-lipase Activity Studies

To investigate the in vitro anti-lipase activity of PLS, an in vitro inhibition study of pancreatic lipase (PPL) activity was performed and 4-methylumbelliferone oleate (4-MUO) was selected as the substrate. Different concentrations of PLS (25. Mu.L) and free PMFs suspensions (25. Mu.L) were placed in 96-well plates, respectively. To the mixture was added 25 μ L of PPL solution (0.5 mg/mL, tris-HCl, pH 8.0) and incubated at 37 ℃ for 10 minutes. 4-MUO (50. Mu.L) was then added and the mixture incubated at 37 ℃ for 60 minutes. The fluorescence value of the product 4-methylumbelliferone (4-MU) was measured at 37 ℃ with a microfluorescent microplate reader (Em/Ex =460/355 nm). Orlistat was used as a positive control and all experiments were repeated 3 times.

In vitro anti-lipase results and statistical analysis are shown in figure 5. Free PMFs inhibit lipase activity, IC, in a concentration-dependent manner ₅₀ The value (concentration inhibition of PPL activity by 50%) was 913.10. + -. 1.98. Mu.g/mL. The results show that free PMFs exhibit good inhibition of lipase. Considering the anti-lipase effect of soybean phosphatidylcholine, liposome composed of lipid bilayer is adopted to deliver dried orange peel PMFs, so as to improve the anti-lipase effect of PMFs. The results in figure 5 also show that PLS has a stronger anti-lipase effect compared to free PMFs and blank liposomes without PMFs. The concentration of PMFs increased from 385 to 1575. Mu.g/mL, and the inhibition of PLS increased from 42.93. + -. 3.12 to 75.40. + -. 2.06%. Meanwhile, the inhibition rate of free PMFs increased from 35.03 + -2.45 to 68.79 + -4.90% at the same PMFs concentration. When the concentration of PMFs reached 1575. Mu.g/mL, the inhibition rate of PLS on pancreatic lipase (PPL) was 75.40 + -2.06%, which is similar to orlistat (80.35 + -0.48%), while the inhibition rate of free PMFs on PPL was 68.79 + -4.90%, and the inhibition rate of PLS on pancreatic lipase (PPL) was about 10% higher than that of free PMFs. In addition, the concentration (IC) that inhibits the activity of PPL by 50% was measured ₅₀ Value), the results show the IC of PLS ₅₀ A value of 458.08. + -. 7.44. Mu.g/mL, which is about 2 times lower than that of free PMFs (913.10. + -. 1.98. Mu.g/mL) ((C))p< 0.05)。

In vivo pharmacokinetic Studies

To investigate the bioavailability of PLS, in vivo SD rat pharmacokinetic studies were performed. The specific experimental method is as follows:

male Sprague-Dawley (SD) rats (SPF, 200-220 g) were provided by Beijing Huafukang Biotech, inc. (Beijing, china). Rats were fed and maintained under constant conditions of room temperature, 50 ± 5% humidity and 12h light/dark cycle. All rats were fasted for 12h during the experiment and had free access to water. Rats were randomly divided into two groups (each group)n= 5). Group 1 was intragastrically administered (ig) with 50mg/kg of an aqueous solution of PMFs, while group 2 was treated with 50mg/kg of PLS in the same administration, the aqueous solution andthe concentration of PMFs in the PLS suspension was 1575. Mu.g/mL. Approximately 200 μ L of blood samples were collected from the orbital vein of each rat at 0, 0.25, 0.5, 0.75, 1, 2, 3, 4, 5, 7, 10, and 12h and placed in heparinized tubes. And then immediately centrifuging the sample at the rotating speed of 6000 rpm for 10 minutes, taking 100 mu L of plasma, placing the plasma in a 1.5mL sterile centrifuge tube, adding 10 mu L of daidzein internal standard working solution (20 mu g/mL), uniformly mixing by vortex for 10 s, then adding 250 mu L of acetonitrile, precipitating protein, carrying out vortex oscillation for 30 s, centrifuging at 13000rpm at the temperature of 4 ℃ for 10 min, and taking the supernatant, introducing a sample, detecting and analyzing.

After a series of treatments of the plasma samples, the mean plasma concentration-time curves of PMFs in aqueous solution and PMFs in PLS after a single 50mg/kg dose was plotted in rats are shown in FIG. 6. The nobiletin and the hesperetin in the liposome can be maintained in blood plasma for 12 hours, the nobiletin in the aqueous solution is lower than the limit of quantification after 10 hours of detection, and the hesperetin cannot reach the limit of quantification after 5 hours. Furthermore, at the same time point, the plasma concentrations of PLS were significantly higher than free PMFs, indicating that PLS had higher bioavailability with a 1.52 fold increase in bioavailability (PLS) ((r))p<0.05). The high oral bioavailability of PLS compared to free PMFs is probably due to the small particle size of the drug, the significantly better solubility and the easy absorption of the drug. Secondly, the liposome double-layer structure can be fused with cell membranes, and is more favorable for the absorption of medicines.

The foregoing is a more detailed description of the invention and is not to be taken in a limiting sense. It will be apparent to those skilled in the art that various modifications, additions and substitutions can be made without departing from the spirit and scope of the invention.

Claims

1. A preparation method of a polymethoxyflavone liposome comprises the following steps:

s1) dissolving a liposome raw material in a solvent, removing the solvent to form a film, wherein the solvent is a mixture of ethanol and dichloromethane, and the volume of the ethanol and the dichloromethane is (2-3): (1-2), the liposome raw material comprises the following components in parts by mass: 960mg, cholesterol: 160mg, polymethoxylated flavone: 40mg, wherein the polymethoxylated flavone is pericarpium citri reticulatae polymethoxylated flavone extracted and enriched by petroleum ether reflux;

s2) hydrating the film with 20 mM HEPES, 144 mM NaCl and HEPES buffer solution with pH of 7.4 to obtain a hydration solution, wherein the mass concentration of soybean phospholipid in the hydration solution is 48 mg/mL;

s3) homogenizing the hydration liquid to obtain the polymethoxylated flavone liposome, wherein the homogenizing pressure is 1000 bar.

2. The method of claim 1, wherein: the particle size of the liposome is not more than 100nm.

3. The method of claim 2, wherein: the D50 of the liposome is 70-88 nm.

4. The method of claim 2, wherein: the D90 of the liposome is 40-90 nm.

5. The production method according to claim 3, characterized in that: the D90 of the liposome is 40-90 nm.