Liposomes co-loading icariin and salvianolic acid B and a preparation method thereof

Abstract

This invention relates to the field of liposome preparation technology, specifically disclosing a liposome co-loaded with icariin and tanshinone B and its preparation method. The liposomes use soybean lecithin and cholesterol as membrane materials, simultaneously encapsulating icariin and tanshinone B, and are prepared using a thin-film hydration-ultrasound method. The liposomes prepared by this invention have an average particle size of 110.49 nm, and the icariin encapsulation efficiency can reach 85.469%. They exhibit relatively uniform particle size distribution, good system stability, and strong controllability of the preparation process. This method can improve the dispersibility and stability of icariin and can serve as a nano-formulation platform for the co-delivery of icariin and tanshinone B.

Description

Technical Field

[0001] This invention relates to the field of liposome preparation technology, specifically to a liposome co-loaded with icariin and tanshinone B and its preparation method. Background Technology

[0002] Flavonoids and their derivatives are the main active components of Epimedium plants, among which icariin is the most representative flavonoid and a key indicator component. Icariin has bidirectional regulatory effects on bone metabolism, including enhancing human immunity, anti-tumor activity, antioxidant activity, promoting osteoblast growth, and inhibiting osteoclast activity. However, as a flavonol glycoside, icariin generally suffers from poor water solubility and insufficient stability, thus limiting its further development and application. Summary of the Invention

[0003] This invention provides liposomes co-loaded with icariin and tanshinone B, and a method for their preparation. The liposomes prepared by this invention have an average particle size of 110.49 nm and an icariin encapsulation efficiency of up to 85.469%. They exhibit characteristics such as relatively uniform particle size distribution, good system stability, and strong controllability of the preparation process. This method can improve the dispersibility and stability of icariin and can serve as a nano-formulation platform for the co-delivery of icariin and tanshinone B.

[0004] This invention provides a liposome co-loaded with icariin and tanshinone B, wherein the liposome uses soybean lecithin and cholesterol as membrane materials and simultaneously encapsulates icariin and tanshinone B.

[0005] This invention utilizes the synergistic potential of icariin and tanshinone B in their pharmacological effects, such as antioxidant and cardiovascular protection, and introduces a co-loaded liposome delivery system, which helps to achieve synergistic pharmacodynamics and pharmacokinetic consistency. The phospholipid bilayer structure of the liposomes can simultaneously encapsulate the hydrophobic icariin and the hydrophilic tanshinone B, forming a stable co-delivery system, which helps to improve the dispersibility and stability of icariin and enhance its application potential.

[0006] Furthermore, the mass ratio of icariin to tanshinone B in the liposomes is 1~4:1~2.

[0007] Furthermore, the mass ratio of cholesterol to soybean lecithin in the liposomes is 1:1~5, and the mass ratio of the total mass of icariin and tanshinone B to soybean lecithin is 1~3:20.

[0008] Furthermore, the liposomes have a particle size of 100 nm to 120 nm, a polydispersity index of 0.2 to 0.3, and a zeta potential of -27.01 mV to -30.77 mV.

[0009] The present invention also provides a method for preparing liposomes co-loaded with icariin and tanshinone B, comprising the following steps: Take soybean lecithin, cholesterol, icariin and tanshinone B according to the mass ratio, add organic solvent and mix well; Organic solvents are removed by rotary evaporation under reduced pressure to form a lipid film; A hydration medium is added, and the lipid film is hydrated by rotary heating. The hydrated system was subjected to ultrasonic treatment to obtain a liposome suspension co-loaded with icariin and tanshinone B.

[0010] Furthermore, the organic solvent is methanol.

[0011] Furthermore, the ultrasonic processing power is 90 W to 110 W, and the time is 2 min to 10 min.

[0012] Furthermore, the hydration temperature is 35℃~45℃, and the hydration time is 4 min~6 min.

[0013] Compared with the prior art, the beneficial effects of the present invention are as follows: This invention addresses the common technical challenges of poor water solubility and insufficient stability inherent in icariin, a flavonol glycoside. Leveraging the synergistic potential of icariin and tanshinone B in their pharmacological effects, such as antioxidant and cardiovascular protection, a co-loaded liposome delivery system is introduced. By co-loading icariin and tanshinone B into liposomes, a nano-co-delivery system is constructed, which helps achieve synergistic pharmacodynamics and pharmacokinetic consistency. The phospholipid bilayer structure of liposomes can simultaneously encapsulate the hydrophobic icariin and the hydrophilic tanshinone B, forming a stable co-delivery system that helps improve the dispersibility and stability of icariin. Simultaneously, the vesicle structure of liposomes forms a physical barrier for both components, effectively preventing direct exposure of the drug to the external environment, making it a suitable liposome formulation platform for the co-delivery of icariin and tanshinone B. This synergistic co-loading strategy fully utilizes the structural advantages of liposomes in co-delivery, laying a formulation foundation for achieving synergistic effects between the two.

[0014] This invention systematically investigated the effects of key process parameters such as alcohol-liposome ratio, drug-liposome ratio, drug-to-mass ratio, and sonication time on encapsulation efficiency through single-factor experiments combined with response surface methodology (Box-Behnken design). A high-fit nonlinear regression equation was established. After optimization, the optimal preparation conditions were determined (alcohol-liposome ratio 1:3.28, drug-liposome ratio 1.98:20, drug-to-mass ratio 1.27:1), resulting in a measured encapsulation efficiency of 85.469% and a drug loading of 2.59%. This demonstrates that the process established in this invention has the advantages of high encapsulation efficiency, process stability, and strong controllability, enabling efficient encapsulation of two drugs in liposomes. Attached Figure Description

[0015] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0016] Figure 1 The effect of alcohol-to-ester ratio on the encapsulation efficiency of ICA-SaIB@Lip.

[0017] Figure 2 The effect of drug-to-lipid ratio on the encapsulation efficiency of ICA-SaIB@Lip.

[0018] Figure 3 The effect of drug mass ratio on the encapsulation efficiency of ICA-SaIB@Lip.

[0019] Figure 4 The effect of ultrasound time on the encapsulation efficiency of ICA-SaIB@Lip.

[0020] Figure 5 3D plots and contour plots were generated to optimize the preparation process of ICA-SaIB@Lip using response surface methodology. In the figures, A is a 3D plot showing the effect of drug-lipid ratio and alcohol-lipid ratio on encapsulation efficiency; B is a contour plot showing the effect of drug-lipid ratio and alcohol-lipid ratio on encapsulation efficiency; C is a 3D plot showing the effect of drug-lipid ratio and alcohol-lipid ratio on encapsulation efficiency; D is a contour plot showing the effect of drug-lipid ratio and alcohol-lipid ratio on encapsulation efficiency; E is a 3D plot showing the effect of drug-lipid ratio on encapsulation efficiency; and F is a contour plot showing the effect of drug-lipid ratio on encapsulation efficiency.

[0021] Figure 6 The figure shows the particle size distribution, PDI, and potential of Lip and ICA-SaIB@Lip. In the figure, A is the particle size distribution of Lip and ICA-SaIB@Lip; B is the PDI and potential of Lip and ICA-SaIB@Lip.

[0022] Figure 7 This is a transmission electron microscope image of ICA-SaIB@Lip.

[0023] Figure 8 UV absorption spectra of ICA and ICA-SaIB@Lip.

[0024] Figure 9 Here are the XRD patterns for each sample.

[0025] Figure 10The images show the appearance of each sample; in the images, A is ICA-SaIB@Lip, B is lyophilized and reconstituted ICA-SaIB@Lip, and C is the lyophilized product of ICA-SaIB@Lip.

[0026] Figure 11 The figures show the particle size distribution, PDI, and potential of ICA-SaIB@Lip before and after freeze-drying; in the figures, A represents the particle size distribution of ICA-SaIB@Lip before and after freeze-drying; and B represents the PDI and potential of ICA-SaIB@Lip before and after freeze-drying. Detailed Implementation

[0027] The specific embodiments of the present invention are described in detail below, but it should be understood that the scope of protection of the present invention is not limited to the specific embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention. Unless otherwise specified, the experimental methods described in the embodiments of the present invention are conventional methods, and the materials and reagents used in the following embodiments are commercially available unless otherwise specified.

[0028] Example 1: A liposome co-loaded with icariin and tanshinone B and its preparation method.

[0029] I. Experimental Methods 1. Preparation of co-loaded liposomes Icariin and tanshinone B co-loaded liposomes were prepared using a thin-film hydration-ultrasound method. 20 mg of soybean lecithin, 10 mg of cholesterol (soybean lecithin and cholesterol as membrane materials), 1 mg of icariin (ICA), and 1 mg of tanshinone B (SaIB) were used as initial components and placed in a flask. 10 mL of methanol was added, and the mixture was ultrasonicated at 400 W for 5 min to ensure complete mixing. The organic solvent was then removed by rotary evaporation at 40 °C for 15 min to form a lipid film. 5 mL of pure water was added, and the mixture was rotary heated at 40 °C for 5 min to fully hydrate the lipid film. Finally, the mixture was ultrasonicated at 100 W in an ice-water bath for 6 min in a cell disruptor to obtain a suspension of icariin and tanshinone B co-loaded liposomes (ICA-SaIB@Lip).

[0030] 2. Determination of encapsulation efficiency and drug loading Encapsulation efficiency reflects the ability of liposomes to encapsulate drugs. Both encapsulation efficiency and drug loading were determined and calculated using a methodology established by HPLC. Free drug and drug-loaded liposomes in the ICA-SaIB@Lip suspension were separated by centrifugation. 2 mL of ICA-SaIB@Lip suspension was centrifuged at 12000 rpm for 30 min at 4℃, and the free drug in the supernatant was collected. The supernatant was then demulsified with 8 mL of methanol, and the free ICA content was calculated under the following chromatographic conditions: InertSustain C18 column (4.6×250, 5 μm), column temperature 30℃, gradient elution settings as shown in Table 1, flow rate 1 mL / min, injection volume 20 μL, and detection wavelength 270 nm. Encapsulation efficiency and drug loading were calculated using the following formulas:

[0031] ; ; Among them W 总 For ICA dosage, W 游 Free ICA content; W 药 For W 总 -W 游 W 膜 The quality of soybean lecithin and cholesterol.

[0032] Table 1 Gradient elution settings 3. Single-factor experiment Single-factor experiments were used to optimize the preparation process. With other conditions fixed, the encapsulation efficiency of icariin was the main evaluation index. The effects of alcohol-lipid ratio (cholesterol:soy lecithin, w:w), drug-lipid ratio (ICA+SaIB:soy lecithin, w:w), drug-to-mass ratio (ICA:SaIB), and ultrasonic time on the encapsulation efficiency of ICA-SaIB@Lip were investigated (Table 2). The three factors with the greatest impact on the encapsulation efficiency were selected for subsequent response surface methodology optimization of the preparation process.

[0033] Table 2 Single-factor experiment table 4. Response surface optimization Based on the Box-Behnken experimental design principle, and building upon single-factor experiments, three factors that significantly affect encapsulation efficiency were selected: alcohol-to-lipid ratio (A), drug-to-lipid ratio (B), and drug-to-mass ratio (C). A total of 17 experimental groups were designed, and the factor levels are shown in Table 3.

[0034] Table 3. Design of Factors in the Preparation Process of ICA-SaIB@Lip 5. Morphological study of ICA-SaIB@Lip 5.1 ICA-SaIB@Lip particle size, Zeta potential, and dispersion index (PDI) The ICA-SaIB@Lip suspension was analyzed using a Brookhaven light scattering particle size analyzer to obtain the average particle size and PDI; another portion of the ICA-SaIB@Lip suspension was placed into a sample cell for measuring zeta potential, and the zeta potential was measured after equilibration.

[0035] The smaller the PDI value, the more uniform the particle size distribution; the larger the absolute value of the Zeta potential, the better the electrostatic stability of the system.

[0036] 5.2 Transmission Electron Microscopy (TEM) Observation TEM was used to observe the microstructure and dispersion of ICA-SaIB@Lip. 5 mL of ICA-SaIB@Lip suspension was placed in a petri dish (9 cm in diameter, 2 cm in height) and freeze-dried in a freeze dryer (freeze-drying gradient program shown in Table 4) to obtain ICA-SaIB@Lip lyophilized powder. 2 mg of ICA-SaIB@Lip lyophilized powder was dissolved in deionized water to a final volume of 5 mL, then added dropwise onto a hydrophilically treated copper grid. After drying for 2 min, the solution on the copper grid surface was absorbed with filter paper, and the copper grid was then placed in a transmission electron microscope to observe the morphology of ICA-SaIB@Lip.

[0037] Table 4 Freeze-drying gradient program 5.3 UV Analysis Take 2 mg of ICA and 2 mg of ICA-SaIB@Lip lyophilized powder, dissolve them in 5 mL of methanol respectively, and scan them in the range of 190 nm to 450 nm using a UV-Vis spectrophotometer.

[0038] 5.4 X-ray diffraction (XRD) analysis XRD analysis can be used to study the microstructure of crystalline and amorphous substances and to determine the encapsulation of ICA. X-ray diffraction was used to characterize ICA, SaIB, a physical mixture (2 mg each of ICA and SaIB), blank liposomes (Lip, prepared in the same manner as ICA-SaIB@Lip, but without ICA and SaIB), and ICA-SaIB@Lip. The experimental conditions were: voltage 45 kV, current 40 mA, scan range 5° to 80°, and scan rate 10° / min.

[0039] 6. Stability assessment of ICA-SaIB@Lip 6.1 Stability Study of ICA-SaIB@Lip Suspension The prepared ICA-SaIB@Lip was stored in a sealed, light-protected container at 4°C. Observe for any precipitation. Samples were taken on days 0, 1, 2, 3, 4, 5, 6, and 7. The particle size distribution, PDI, and Zeta potential were measured using a Brookhaven light scattering particle size analyzer to evaluate its short-term storage stability.

[0040] 6.2. Stability Study of ICA-SaIB@Lip Freeze-Drying Weigh 2 mg of ICA-SaIB@Lip lyophilized powder, add purified water to reconstitute to 5 mL, and detect changes in particle size, PDI, and potential.

[0041] II. Test Results 1. Determination of encapsulation efficiency and drug loading Encapsulation efficiency and drug loading were determined using centrifugation. Under the initial formulation conditions, the encapsulation efficiency of ICA reached 68.27%, and the drug loading was 2.59%.

[0042] 2. Results of single-factor experiments 2.1 Investigation of alcohol-to-ester ratio Depend on Figure 1 It can be seen that the encapsulation efficiency first increases and then decreases with changes in the ratio. The highest encapsulation efficiency, 71.71%, is achieved when the alcohol-liposome ratio is 1:3. This may be because when the membrane material composition is suitable, the liposome membrane structure is more stable and has a more suitable drug loading space; when the membrane material ratio is unsuitable, the loading capacity may decrease.

[0043] 2.2 Investigation of drug-lipid ratio Depend on Figure 2 It can be seen that as the ratio of ICA+SaIB to soybean lecithin increases, the encapsulation efficiency first increases and then decreases, indicating that there is a certain limit to the drug loading capacity of liposomes. When the amount of drug added exceeds a certain range, drug leakage may occur due to steric hindrance or changes in membrane structure stability, which in turn leads to a decrease in the encapsulation efficiency.

[0044] 2.3 Drug quality ratio assessment Depend on Figure 3 It can be seen that as the proportion of ICA increases, the encapsulation efficiency first increases and then decreases. The encapsulation efficiency is highest at 83.45% when the mass ratio of ICA to SaIB is 2:1.

[0045] 2.4 Ultrasonic Time Assessment Depend on Figure 4It can be seen that the encapsulation efficiency gradually increases with the extension of ultrasound time; when the ultrasound time is 8 minutes, the encapsulation efficiency can reach 70.33%. When the ultrasound time is further extended to 10 minutes, the encapsulation efficiency decreases, possibly because prolonged ultrasound disrupts the interaction between ICA and cholesterol and phospholipids, causing particle disintegration and reduced dispersibility, thus affecting the encapsulation efficiency. Therefore, an ultrasound time of 8 minutes is more suitable.

[0046] 3. Optimization results of response surface methodology Based on the single-factor experiments, the Box-Behnken design was used in conjunction with Design-Expert software for data analysis. The effects of alcohol-lipid ratio (A), drug-lipid ratio (B), and drug-mass ratio (C) on the encapsulation efficiency were studied with encapsulation efficiency (Y) as the response value. The results are shown in Table 5.

[0047] Table 5 Experimental design and results of response surface methodology for ICA-SaIB@Lip preparation Using Design-Expert software, regression analysis was performed on the data in Table 3 to obtain the nonlinear regression equations between the alcohol-lipid ratio (A), drug-lipid ratio (B), drug-to-mass ratio (C), and encapsulation efficiency (Y): Y=85.0815+0.291113×A-0.019525×B-1.42664×C-0.878325×AB-0.38375×AC-0.088825×BC-0.780375×A 2 -0.54185×B 2 -1.09652×C 2 ; The significance, variance, and regression equation of the model are analyzed based on the quadratic polynomial regression equation.

[0048] The significance analysis of the model (Table 6) showed that P < 0.0001, indicating that the model has good statistical significance and can be used to analyze and predict the preparation process of ICA-SaIB@Lip. The order of influence of each factor on the encapsulation efficiency was: drug mass ratio > alcohol-lipid ratio > drug-lipid ratio.

[0049] Table 6. Analysis of Variance of ICA-SaIB@Lip Response Surface Model Note: "-" indicates that this item is not available.

[0050] Optimization using Design-Expert software yielded the following optimal process conditions for the preparation of ICA-SaIB@Lip: a cholesterol to soybean lecithin mass ratio of 1:3.28, a drug-to-lecithin ratio of 1.98:20, and a drug-to-lipid ratio of 1.27:1. Figure 5 (A~F). The test was repeated 4 times under the same process conditions, and the average encapsulation rate was 85.469%, which is close to the model prediction.

[0051] 4. Characterization results of ICA-SaIB@Lip 4.1 ICA-SaIB@Lip particle size, Zeta potential, and PDI Depend on Figure 6 As shown in A and B, the particle size of ICA-SaIB@Lip is 110.49 nm. The particle size of ICA-SaIB@Lip after drug loading is slightly larger than that of Lip, indicating that the drug was successfully introduced into the liposome system. Its PDI is 0.245±0.014, indicating that the particle size distribution is relatively uniform. The Zeta potential is -28.89±1.88 mV, indicating that the surface of ICA-SaIB@Lip carries a negative charge and the system has good dispersion stability.

[0052] 4.2 Morphological Investigation of ICA-SaIB@Lip The morphology of ICA-SaIB@Lip is as follows Figure 7 As shown. In Figure 7 In the study, ICA-SaIB@Lip exhibits a uniform spherical vesicle structure with relatively clear boundaries, good dispersion, and no obvious aggregation or fusion.

[0053] 4.3 UV Analysis Depend on Figure 8 It can be seen that the UV spectrum of ICA-SaIB@Lip retains the relevant characteristic absorption trends, indicating that ICA did not undergo significant chemical structural changes during the encapsulation process, and ICA-SaIB@Lip mainly exhibits physical encapsulation of the drug.

[0054] 4.4 XRD Analysis Depend on Figure 9It was observed that ICA exhibited distinct characteristic diffraction peaks in the X-ray diffraction pattern, with crystalline peaks between 5° and 80° (2θ = 8.44°, 8.95°, 10.36°, 12.47°, 17.49°, 21.10°, and 22.62°). In contrast, SaIB, Lip, and ICA-SaIB@Lip generally exhibited more pronounced amorphous characteristics. Some characteristic diffraction peaks of ICA were still observed in the XRPD pattern of the physical mixture, but their intensity decreased, showing amorphous characteristics at 2θ = 16.09° and 2θ = 23.63°. In the ICA-SaIB@Lip sample, the characteristic diffraction peaks of ICA essentially disappeared, and the overall peak intensity decreased. These results indicate that the drug does not simply exist in its original crystalline form within the ICA-SaIB@Lip system. Combined with other characterization results, this suggests that ICA and SaIB have been successfully encapsulated within ICA-SaIB@Lip.

[0055] 5. Stability test results of ICA-SaIB@Lip 5.1 Stability Study of ICA-SaIB@Lip Suspension The ICA-SaIB@Lip prepared by the optimal process was stored at 4℃ in the dark for 7 days, and its particle size distribution, PDI and Zeta potential were detected periodically. The results are shown in Table 7.

[0056] Table 7. Stability results of ICA-SaIB@Lip suspension As shown in Table 7, during 7 days of storage at 4℃ in the dark, the particle size of ICA-SaIB@Lip increased slightly, but the PDI and Zeta potentials remained relatively stable, indicating good short-term stability of the system within 7 days. The initial particle size of ICA-SaIB@Lip was 112.706±3.580 nm, and the PDI was 0.244±0.030. After 7 days of storage, the particle sizes became 132.530±5.261 nm and 0.246±0.046 nm, respectively, showing minimal change. The Zeta potential remained negative overall, with some fluctuations. The high negative potential on the surface of ICA-SaIB@Lip provided electrostatic repulsion between particles, which was an important reason for the stability of the system. During storage, the ICA-SaIB@Lip suspension remained clear, and no significant precipitation was observed, indicating that ICA-SaIB@Lip has good physical stability under short-term storage conditions.

[0057] Figure 10 Figures A through C show the appearance of the ICA-SaIB@Lip suspension, the lyophilized and reconstituted sample, and the lyophilized product. As can be seen from the figures, the ICA-SaIB@Lip prepared by the thin-film hydration-ultrasonic method is relatively uniformly dispersed, with no obvious flocculation or stratification observed.

[0058] Depend on Figure 11 As shown in A and B, the lyophilized and reconstituted ICA-SaIB@Lip still has a relatively uniform particle size distribution, with a particle size of approximately 115.0 nm. The PDI and potential changes are both within a reasonable range, indicating that ICA-SaIB@Lip still has good reconstitution performance after lyophilization.

[0059] This invention focuses on icariin and tanshinone B, and uses a thin-film hydration-ultrasound method to prepare ICA-SaIB@Lip. The preparation process was optimized through single-factor experiments and response surface methodology. Using encapsulation efficiency as the evaluation index, the optimal preparation conditions were obtained: an alcohol-to-lipid ratio of 1:3.28, a drug-to-lipid ratio of 1.98:20, and a drug-to-mass ratio of 1.27:1.

[0060] The ICA-SaIB@Lip prepared under the above conditions had an average particle size of 110.49 nm, a PDI of 0.245±0.014, and a Zeta potential of -28.89±1.88 mV, indicating that ICA-SaIB@Lip has a relatively uniform particle size distribution and good dispersion stability. TEM results showed that its morphology was mainly spherical and well dispersed. UV and XRD results showed that the drug did not undergo significant chemical structural changes during the encapsulation process and was successfully encapsulated in the liposome system. The lyophilized and reconstituted samples had a uniform appearance and small changes in particle size, PDI, and potential. The ICA-SaIB@Lip suspension could be stably stored at 4℃ for 1 week without significant precipitation, showing good short-term stability.

[0061] Although preferred embodiments of the invention have been described, those skilled in the art, once they have learned the basic inventive concept, can make other changes and modifications to these embodiments.

[0062] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. Therefore, if these modifications and variations fall within the scope of the claims of this invention and their equivalents, this invention also intends to include these modifications and variations.

Claims

1. A liposome co-loaded with icariin and tanshinone B, characterized in that, The liposomes use soybean lecithin and cholesterol as membrane materials, and simultaneously encapsulate icariin and tanshinone B.

2. The liposomes co-loaded with icariin and tanshinone B according to claim 1, characterized in that, The mass ratio of icariin to tanshinone B in the liposomes is 1-4:1-2.

3. The liposomes co-loaded with icariin and tanshinone B according to claim 1, characterized in that, The mass ratio of cholesterol to soybean lecithin in the liposomes is 1:1~5, and the mass ratio of the total mass of icariin and tanshinone B to soybean lecithin is 1~3:

20.

4. The liposomes co-loaded with icariin and tanshinone B according to claim 1, characterized in that, The liposomes have a particle size of 100 nm to 120 nm, a polydispersity index of 0.2 to 0.3, and a zeta potential of -27.01 mV to -30.77 mV.

5. A method for preparing liposomes co-loaded with icariin and tanshinone B according to any one of claims 1 to 4, characterized in that, Includes the following steps: Take soybean lecithin, cholesterol, icariin and tanshinone B according to the mass ratio, add organic solvent and mix well; Organic solvents are removed by rotary evaporation under reduced pressure to form a lipid film; A hydration medium is added, and the lipid film is hydrated by rotary heating. The hydrated system was subjected to ultrasonic treatment to obtain a liposome suspension co-loaded with icariin and tanshinone B.

6. The preparation method according to claim 5, characterized in that, The organic solvent is methanol.

7. The preparation method according to claim 5, characterized in that, The ultrasonic treatment power is 90 W to 110 W, and the time is 2 min to 10 min.

8. The preparation method according to claim 5, characterized in that, The hydration temperature is 35℃~45℃, and the hydration time is 4min~6min.

Images