L-fucose-modified in-situ lichen hard acid cationic nanoliposome as well as preparation method and application thereof

Abstract

The application provides an L-fucose modified original lichens hard acid cation nanoliposome, adopts an L-fucose / oleylamine coupling compound to modify the surface of the original lichens hard acid cation nanoliposome, and obtains the L-fucose modified original lichens hard acid cation nanoliposome with a particle size in the range of 135nm-165nm after treatment. The nanoliposome has a large specific surface area, a small particle size, and a positive charge on the surface, and the surface of the original lichens hard acid cation nanoliposome is modified by L-fucose to provide more binding sites on the surface, improve the permeability of the nanoliposome to Candida albicans, and help the original lichens hard acid to play a role in inhibiting the Candida albicans biofilm.

Description

Technical Field

[0001] This invention relates to the field of antibacterial drug technology, and in particular to an L-fucose-modified protolichen stearic acid cationic nanoliposome, its preparation method, and its application. Background Technology

[0002] Candida albicans (C. albicans) is an opportunistic pathogen that causes tens of millions of infectious diseases annually in China, making it the fourth most common pathogen in hospitals. The robust biofilm formed by C. albicans and its increasing drug resistance make the treatment of C. albicans infections increasingly difficult. Currently, medication is the main treatment for C. albicans infections; however, traditional antifungal drugs are limited in variety and have drawbacks such as strong toxicity and high cost.

[0003] With the widespread use of antifungal drugs, multidrug resistance in fungi is becoming increasingly serious. Studies have shown that the formation of a Candida albicans biofilm is one of the reasons for its resistance to multiple antifungal drugs. The fungal community binds to receptors on the surface of epithelial cells and secretes extracellular polysaccharides or proteins, forming a membrane-like structure, namely the Candida albicans biofilm. This biofilm encapsulates itself within a spontaneously adhering layer of polysaccharides and proteins, thereby resisting the bactericidal effects of drugs. This is the mechanism by which Candida albicans develops resistance to commonly used therapeutic drugs. Therefore, inhibiting the formation of the Candida albicans biofilm is of great significance for suppressing Candida albicans activity and reversing the development of multidrug resistance.

[0004] Lichenic acid is an organic acid isolated from lichens. Studies have shown that lichenic acid has a good inhibitory effect on Candida albicans and can reverse the development of drug resistance in Candida albicans. However, lichenic acid has poor affinity for Candida albicans and cannot penetrate the Candida albicans capsule to inhibit Candida albicans.

[0005] Therefore, there is an urgent need to modify licheniformic acid to improve its penetration into the Candida albicans membrane, thereby enhancing its inhibitory effect on Candida albicans. Summary of the Invention

[0006] To address the technical challenges in the prior art, this patent provides a cationic nanoliposome modified with L-fucose, its preparation method, and its application. This nanoliposome exhibits high affinity for Candida albicans and demonstrates a good antibacterial effect.

[0007] The technical solution of this invention is as follows:

[0008] This invention provides L-fucose-modified protolichen stearate cationic nanoliposomes, characterized in that the surface of the protolichen stearate cationic nanoliposomes is modified with an L-fucose / oleic acid amine coupler, and after treatment, L-fucose-modified protolichen stearate cationic nanoliposomes with a particle size in the range of 135nm to 165nm are obtained.

[0009] This invention also provides a method for preparing L-fucose-modified lichen-derived acidic cationic nanoliposomes, specifically comprising the following steps:

[0010] S1 Preparation of L-fucose / oleic acid amine coupler:

[0011] Weigh out a certain ratio of L-fucose and oleic acid amine, dissolve them in anhydrous ethanol to obtain a mixture, place the mixture in a round-bottom flask and heat to react, remove the anhydrous ethanol by rotary evaporation to obtain the L-fucose / oleic acid amine couple.

[0012] S2 Preparation of L-fucose-modified lichen stearic acid cationic liposome suspension:

[0013] Weigh out a certain ratio of soybean lecithin, cationic lipids, sodium cholesterol sulfate, L-fucose / oleic acid amine coupling compound and lichenic acid, dissolve them in anhydrous ethanol to obtain a mixture, place the mixture in a round bottom flask for rotary evaporation to form a film, add buffer to wash the film to obtain an L-fucose modified lichenic acid cationic liposome suspension.

[0014] S3 preparation of L-fucose-modified lichen stearate cationic nanoliposomes:

[0015] The L-fucose-modified protolichen stearate cationic liposome suspension was treated with an ultrasonic cell disruptor, filtered through a microporous membrane, and freeze-dried for preservation to obtain L-fucose-modified protolichen stearate cationic nanoliposomes.

[0016] Specifically, in step S1, L-fucose and oleic acid amine are mixed in a mass ratio of 1:0.5 to 1:2.

[0017] Specifically, the heating reaction described in step S1 has a reaction time of 4 to 6 hours and a heating temperature of 80°C to 90°C.

[0018] Specifically, in step S2, soybean lecithin, cationic liposomes, sodium cholesterol sulfate, L-fucose-oleic acid amine coupler and lichenic acid are mixed in a mass ratio of 1:0.01:0.05:0.05:0.01 to 1:0.01:0.05:0.05:0.1.

[0019] Specifically, the cationic liposomes in step S2 are either DOTMA (1,2-bisoctadecenoxy-3-methylammonium propane) or DOTAP ((1,2-dioleoxypropyl)trimethylammonium chloride).

[0020] Specifically, the buffer solution mentioned in step S2 is either PBS buffer or Tris-HCl buffer, and the pH value of the buffer solution is 6.0-8.0.

[0021] Specifically, the ultrasonic cell disruptor treatment time in step S3 is 1-5 minutes.

[0022] The present invention also provides the application of L-fucose-modified lichen stearate cationic nanoliposomes in a skin spray drug for inhibiting Candida albicans.

[0023] The present invention also provides the application of L-fucose-modified protolichen stearate cationic nanoliposomes in cavity-mixed gels or ointments for inhibiting Candida albicans.

[0024] This invention employs an L-fucose-oleic acid amine coupling compound, loaded onto the surface of cationic liposomes containing licheniformic acid. L-fucose, a polysaccharide, can bind to the squalane lipid layer on the surface of Candida albicans to form a unique complex. This complex provides more binding sites for both compounds, significantly improving their affinity and permeability to Candida albicans. Furthermore, the formation of this complex leads to denaturation of the lipid layer on the surface of Candida albicans, thereby increasing its permeability to the bacteria. Oleic acid amine, on the other hand, exhibits good permeability to the Candida albicans membrane. By using the L-fucose-oleic acid amine coupling compound as an adhesion receptor for Candida albicans, its modification on the surface of licheniformic acid cationic nanoliposomes enhances the permeability of the nanoliposomes to Candida albicans, thus helping licheniformic acid exert its inhibitory effect on the Candida albicans membrane and strengthening its antibacterial efficacy.

[0025] The beneficial effects of this invention are:

[0026] (1) The L-fucose-modified protolichen stearic acid cation nanoliposomes provided by the present invention have a particle size of about 150 nm, a large specific surface area, small particle size, and positive surface charge, which can attract each other with the negative charge on the surface of Candida albicans, making it easier to penetrate into the cell to exert the effect of inhibiting Candida albicans.

[0027] (2) In this invention, L-fucose and oleic acid amine coupling compound is used as the adhesion receptor of Candida albicans. It is modified on the surface of the proto-lichen stearic acid cationic nanoliposomes, providing more binding sites on the surface, improving the permeability of nanoliposomes to Candida albicans, and helping proto-lichen stearic acid to play an inhibitory role in the Candida albicans membrane. Attached Figure Description

[0028] To more clearly illustrate the technical solutions of the embodiments of this application, the accompanying drawings used in the embodiments of this application will be briefly introduced below. It should be understood that the following drawings only show some embodiments of this application and should not be regarded as a limitation of the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0029] Figure 1 A schematic diagram of the structure of L-fucose-modified protolichen stearic acid cationic nanoliposomes;

[0030] Figure 2 The dynamic growth curves of Candida albicans containing different nanoliposomes prepared in Example 1, Comparative Example 1 and Comparative Example 2 are shown.

[0031] Figure 3 The graph shows the inhibitory effect of different nanoliposomes prepared in Example 1 and Comparative Examples 1 and 2 on the Candida albicans biofilm. Detailed Implementation

[0032] To make the objectives, technical solutions, and advantages of the embodiments of this application clearer, the technical solutions in the embodiments of this application will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0033] Please see Figure 1 , Figure 1 This is a schematic diagram of the structure of L-fucose-modified protolichen stearic acid cationic nanoliposomes provided by the present invention. The nanoliposomes are obtained by modifying the surface of protolichen stearic acid cationic nanoliposomes with an L-fucose / oleic acid amine coupling agent to obtain L-fucose-modified protolichen stearic acid cationic nanoliposomes.

[0034] Example 1

[0035] This embodiment provides an L-fucose-modified protolichen stearate cationic nanoliposome, the preparation method of which includes the following steps:

[0036] (1) Preparation of L-fucose / oleic acid amine coupler:

[0037] Weigh 2.0 g of L-fucose and 3.26 g of oleic acid amine and dissolve them in 300 ml of anhydrous ethanol. Place the mixture in a round-bottom flask and heat it at 80 °C for 6 h. After removing the anhydrous ethanol by rotary evaporation, the L-fucose / oleic acid amine coupler is obtained.

[0038] (2) Preparation of L-fucose-modified lichen stearic acid cationic liposome suspension:

[0039] Weigh out 1.0g of soybean lecithin, 0.01g of DOTMA, 0.05g of sodium cholesterol sulfate, 0.05g of L-fucose / oleic acid amine coupling compound, and 0.1g of licheniformic acid. Mix them and dissolve them in 10ml of anhydrous ethanol. Place the mixture in a round-bottom flask and evaporate by rotary evaporation to form a thin film. Add 20ml of Tris-HCl buffer with a pH of 6.8 to wash the film to obtain an L-fucose-modified licheniformic acid cationic liposome suspension.

[0040] (3) Preparation of L-fucose-modified lichen stearate cationic nanoliposomes:

[0041] The above suspension was treated with an ultrasonic cell disruptor for 1 min, filtered through a 0.22 μm microporous membrane, and freeze-dried for preservation to obtain L-fucose-modified proto-lichen stearic acid cationic nanoliposomes.

[0042] Determination of particle size and distribution, and zeta potential

[0043] The nanoliposomes obtained in Example 1 were diluted 100 times with water, and the particle size distribution of the nanoliposomes in Example 1 was detected by laser dynamic light scattering method (Malvern laser particle size analyzer Mastersizer 3000). The results showed that the particle size of the nanoliposomes was in the range of 150±12nm. The nanoliposomes obtained in Example 1 were diluted 100 times with water, and their Zeta potential was detected by Zeta potential analyzer. The results showed that the Zeta potential of the nanoliposomes was in the range of 32.5±5.2mV.

[0044] Example 2

[0045] This embodiment provides an L-fucose-modified protolichen stearate cationic nanoliposome, the preparation method of which includes the following steps:

[0046] (1) Preparation of L-fucose / oleic acid amine coupler:

[0047] Weigh 1.0 g of L-fucose and 0.5 g of oleic acid amine and dissolve them in 150 ml of anhydrous ethanol. Place the mixture in a round-bottom flask and heat it at 90 °C for 4 h. After removing the anhydrous ethanol by rotary evaporation, the L-fucose / oleic acid amine coupler is obtained.

[0048] (2) Preparation of L-fucose-modified lichen stearic acid cationic liposome suspension:

[0049] Weigh out 0.5g of soybean lecithin, 0.005g of DOTAP, 0.025g of sodium cholesterol sulfate, 0.025g of L-fucose / oleic acid amine coupling compound, and 0.025g of lichenic acid stearic acid. Dissolve them in 5ml of anhydrous ethanol, place them in a round-bottom flask, and evaporate by rotary evaporation to form a thin film. Add 10ml of Tris-HCl buffer with a pH of 7.3 to wash the film and obtain a suspension.

[0050] (3) Preparation of L-fucose-modified lichen stearate cationic nanoliposomes:

[0051] The above suspension was treated with an ultrasonic cell disruptor for 3 minutes and then filtered through a 0.22 μm microporous membrane to obtain fucose-modified protolichen stearic acid cationic nanoliposomes.

[0052] The particle size of the L-fucose-modified protolichen stearic acid cationic nanoliposomes obtained in Example 2 was determined to be in the range of 148±14 nm and the Zeta potential was in the range of 30.5±6.3 mV, using the same method as in Example 1.

[0053] Example 3

[0054] This embodiment provides an L-fucose-modified protolichen stearate cationic nanoliposome, the preparation method of which includes the following steps:

[0055] (1) Preparation of L-fucose / oleic acid amine coupler:

[0056] Weigh 1.5g of L-fucose and 2.6g of oleic acid amine and dissolve them in 150ml of anhydrous ethanol. Place the mixture in a round-bottom flask and heat it at 85℃ for 5h. After removing the anhydrous ethanol by rotary evaporation, the L-fucose / oleic acid amine coupler is obtained.

[0057] (2) Preparation of L-fucose-modified lichen stearic acid cationic liposome suspension:

[0058] Weigh out 1.0g of soybean lecithin, 0.01g of DOTMA, 0.05g of sodium cholesterol sulfate, 0.05g of L-fucose / oleic acid amine coupling compound, and 0.08g of licheniformic acid. Mix them and dissolve them in 10ml of anhydrous ethanol. Place the mixture in a round-bottom flask and evaporate by rotary evaporation to form a thin film. Add 20ml of Tris-HCl buffer with a pH of 6.8 to wash the film to obtain an L-fucose-modified licheniformic acid cationic liposome suspension.

[0059] (3) Preparation of L-fucose-modified lichen stearate cationic nanoliposomes:

[0060] The above suspension was treated with an ultrasonic cell disruptor for 5 minutes and then filtered through a 0.22 μm microporous membrane to obtain fucose-modified protolichen stearic acid cationic nanoliposomes.

[0061] The particle size of the L-fucose-modified protolichen stearic acid cationic nanoliposomes obtained in Example 2 was determined to be in the range of 148±14 nm and the Zeta potential was in the range of 30.5±6.3 mV, using the same method as in Example 1.

[0062] Comparative Example 1

[0063] This comparative example provides an L-fucose-modified lichen stearic acid nanoliposome, the main difference in its preparation method compared to Example 1 is that cationic lipids were not used.

[0064] (1) Preparation of L-fucose / oleic acid amine coupler:

[0065] Weigh 2.0 g of L-fucose and 3.26 g of oleic acid amine and dissolve them in 300 ml of anhydrous ethanol. Place the mixture in a round-bottom flask and heat it at 80 °C for 6 h. After removing the anhydrous ethanol by rotary evaporation, the L-fucose / oleic acid amine coupler is obtained.

[0066] (2) Preparation of L-fucose-modified lichen stearate liposome suspension:

[0067] Weigh out 1.0 g of soybean lecithin, 0.05 g of sodium cholesterol sulfate, 0.05 g of L-fucose / oleic acid amine coupling compound and 0.1 g of lichen stearic acid, mix them and dissolve them in 10 ml of anhydrous ethanol. Place the mixture in a round-bottom flask and evaporate by rotary evaporation to form a thin film. Add 20 ml of Tris-HCl buffer with a pH of 6.8 to wash the film to obtain an L-fucose-modified lichen stearic acid liposome suspension.

[0068] (3) Preparation of L-fucose-modified lichen stearic acid nanoliposomes:

[0069] The above suspension was treated with an ultrasonic cell disruptor for 1 min, filtered through a 0.22 μm microporous membrane, and freeze-dried for storage to obtain L-fucose-modified lichen stearic acid nanoliposomes.

[0070] The particle size of the L-fucose-modified lichen stearic acid nanoliposomes obtained in Comparative Example 1 was determined to be in the range of 159 ± 10 nm, and the Zeta potential was in the range of 1.5 ± 3.2 mV, using the same method as in Example 1.

[0071] Comparative Example 2

[0072] This comparative example provides a cationic nanoliposome of lichen stearate, the main difference in its preparation method compared to Example 1 is that it does not use L-fucose / oleic acid amine coupling agent to modify it.

[0073] (1) Liposome suspension of lichen stearate:

[0074] Weigh out 1.0g of soybean lecithin, 0.01g of DOTMA, 0.05g of sodium cholesterol sulfate and 0.1g of licheniformic acid, mix them and dissolve them in 10ml of anhydrous ethanol. Place the mixture in a round-bottom flask and evaporate by rotary evaporation to form a thin film. Add 20ml of Tris-HCl buffer with a pH of 6.8 to wash the film to obtain a licheniformic acid cationic liposome suspension.

[0075] (2) Preparation of cationic nanoliposomes of lichen stearic acid:

[0076] The above suspension was treated with an ultrasonic cell disruptor for 1 min, filtered through a 0.22 μm microporous membrane, and freeze-dried for preservation to obtain lichen stearic acid cationic nanoliposomes.

[0077] The cationic nanolipids of lichen stearate obtained in Comparative Example 2 were determined using the same method as in Example 1.

[0078] Application examples

[0079] Weigh 2g of the L-fucose-modified protolichen stearic acid cationic nanoliposomes prepared in Example 1, dissolve them in 100mL of sterile water, shake and mix well to prepare a skin spray drug with a drug loading of 2% to inhibit Candida albicans.

[0080] A water-soluble natural polymer material such as sodium alginate is dissolved in sterile water, and L-fucose-modified proto-lichen stearic acid cationic nanoliposomes prepared in Example 1 are added. The mixture is stirred and mixed evenly at a stirring speed of 4000 rpm to ensure that the liposomes are evenly dispersed in the matrix, thus preparing an ointment for inhibiting Candida albicans in cavities.

[0081] Weigh chitosan, add acetic acid solution, and allow it to swell at room temperature for 12-24 hours to form a homogeneous and viscous solution. Add deionized water to make up the volume to obtain a gel. Mix the L-fucose-modified proto-lichen stearic acid cationic nanoliposomes prepared in Example 1 with the gel. After 2-5 hours, a homogeneous and viscous solution will be formed. Continue stirring and add deionized water to make up the volume to prepare a mixed gel agent for use as a drug to inhibit Candida albicans in cavities.

[0082] Inhibition effect determination

[0083] In this application example, the nanoliposomes obtained in Example 1, Comparative Example 1, and Comparative Example 2 were used to determine their inhibitory effects on Candida albicans and Candida albicans biofilm.

[0084] (1) Determination of inhibitory ability against Candida albicans

[0085] The growth curve method was used to determine the inhibitory effect of the nanoliposomes prepared in Example 1, Comparative Example 1, and Comparative Example 2 on the growth of Candida albicans, with no drugs added as a blank control.

[0086] Viable Candida albicans were inoculated into four groups of LB culture media containing 2% NaCl, and all were cultured in a 37°C incubator for 24 hours. The bacterial suspension concentration was then adjusted to 1×10⁻⁶. 6 CFU / mL. The nanoliposomes prepared in Example 1, Comparative Example 1 and Comparative Example 2 were diluted with LB medium to obtain three solutions with a mass concentration of 20 μg / mL, which were prepared and used immediately.

[0087] The above four groups of 100 μL solutions with a concentration of 1×10⁻⁶ were added to the nanoliposomes prepared in Example 1, Comparative Example 1, Comparative Example 2, and the blank control, respectively. 6 Four treatment solutions were obtained by shaking a bacterial suspension of CFU / mL at 150 rpm for 5 min in a 37℃ bacterial incubator. 0.3 mL of each treatment solution was mixed with 0.1 mL of yeast extract peptone glucose agar liquid medium and placed in four 96-well plates. The absorbance at 600 nm was measured every 4 hours using an automated growth curve analyzer to plot the 48-hour dynamic growth curve of the bacteria, i.e., the inhibitory effect of different nanoliposomes on the bacteria within 48 hours.

[0088] Please refer to the results of the assay for inhibition of bacterial cells. Figure 2 ,Depend on Figure 2 As can be seen, under the given dose, the bacteria in the blank control grew rapidly within 0–48 h, and the growth curve showed a rapid upward trend, indicating that it had no significant effect on bacterial growth. The inhibitory effects of Example 1, Comparative Example 1, and Comparative Example 2 on bacterial cells were all higher than those of the blank control. However, Example 1 showed the most significant inhibitory effect on Candida albicans, with an inhibitory effect far exceeding that of Comparative Example 1 and Comparative Example 2. This indicates that L-fucose modification and cationic lipids increased the affinity and permeability of liposomes to Candida albicans, resulting in the best antibacterial ability.

[0089] (2) Determination of the ability to inhibit Candida albicans biofilm formation

[0090] Candida albicans biofilms are aggregated membrane-like structures formed by Candida albicans adhering to the contact surface and secreting polysaccharide matrix, fibrin, lipid proteins, etc., to encapsulate itself. Biofilm formation is an important cause of drug resistance and refractory infections. Therefore, this invention investigated the inhibitory effect of L-fucose-modified proto-lichen stearic acid cationic nanoliposomes on Candida albicans biofilm formation.

[0091] The inhibitory effects of the nanoliposomes prepared in Example 1, Comparative Example 1, and Comparative Example 2 on the biofilm of Candida albicans were determined, with no drugs added as a blank control.

[0092] Viable Candida albicans were inoculated into LB medium containing 2% NaCl and cultured at 37°C for 24 hours. The bacterial suspension concentration was then adjusted to 1×10⁻⁶. 6 The cell suspension was added to four groups of 96-well plates at CFU / mL and cultured at 37°C for about 2 hours to form four groups of membranes.

[0093] The nanoliposomes prepared in Example 1, Comparative Example 1, and Comparative Example 2 were diluted with LB medium to obtain three groups of solutions with a mass concentration of 20 μg / mL, which were prepared and used immediately.

[0094] Example 1, Comparative Example 1, Comparative Example 2 and blank control were added to four groups of membranes and cultured at the same temperature. The absorbance values ​​were measured at 570 nm using an ELISA reader at 2.0, 12.0, 24.0 and 48.0 h, respectively.

[0095] Please refer to the results of the inhibition test on the affected area. Figure 3 ,Depend on Figure 3 As can be seen, the amount of Candida albicans biofilm at different time points after treatment with Examples 1, Comparative Examples 1, and Comparative Examples 2 was lower than that in the blank control. Even after Candida albicans had formed a mature biofilm after 2 hours of culture, Examples 1, Comparative Examples 1, and Comparative Examples 2 all showed a high inhibitory effect on Candida albicans biofilm formation. Example 1 showed the most significant inhibitory effect on Candida albicans, with the strongest inhibitory ability. This indicates that L-fucose modification and cationic lipids increased the affinity and permeability of liposomes for Candida albicans biofilm formation, resulting in a better ability to inhibit bacterial biofilm formation.

[0096] Based on the above test results, the L-fucose-modified protostolic acid nanoliposomes prepared using the above embodiments of the present invention have a large specific surface area and a positively charged surface, which can attract the negatively charged surface of Candida albicans, making it easier to penetrate into cells and exert an inhibitory effect on Candida albicans. In addition, using the L-fucose-oleic acid amine coupling compound as the adhesion receptor for Candida albicans, which is modified on the surface of the protostolic acid nanoliposomes, provides more binding sites on the surface, improves the permeability of the nanoliposomes to Candida albicans, and enables protostolic acid to better exert its inhibitory effect on the Candida albicans membrane.

[0097] The above are merely embodiments of this application and are not intended to limit the scope of protection of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of protection of this application.

Claims

1. A method for preparing L-fucose-modified protolichen stearic acid cationic nanoliposomes, characterized in that, Includes the following steps: S1 Preparation of L-fucose / oleic acid amine coupler: Weigh out a certain ratio of L-fucose and oleic acid amine, dissolve them in anhydrous ethanol to obtain a mixture, place the mixture in a round-bottom flask and heat to react, remove the anhydrous ethanol by rotary evaporation to obtain the L-fucose / oleic acid amine couple. S2 Preparation of L-fucose-modified lichen stearic acid cationic liposome suspension: Weigh out a certain ratio of soybean lecithin, cationic lipids, sodium cholesterol sulfate, L-fucose / oleic acid amine coupling compound and lichenic acid, dissolve them in anhydrous ethanol to obtain a mixture, place the mixture in a round bottom flask for rotary evaporation to form a film, add buffer to wash the film to obtain an L-fucose modified lichenic acid cationic liposome suspension. S3 Preparation of L-fucose-modified lichen stearate cationic nanoliposomes: The suspension of L-fucose-modified protolichen stearate cationic liposomes was treated with an ultrasonic cell disruptor, filtered through a microporous membrane, and freeze-dried for preservation to obtain L-fucose-modified protolichen stearate cationic nanoliposomes. The surface of protolichen stearic acid cationic liposomes was modified with L-fucose / oleic acid amine coupling agent, and L-fucose modified protolichen stearic acid cationic nanoliposomes with a particle size in the range of 135 nm to 165 nm were obtained after treatment.

2. The method for preparing L-fucose-modified protolichen stearic acid cationic nanoliposomes according to claim 1, characterized in that, In step S1, L-fucose and oleic acid amine are mixed in a mass ratio of 1:0.5 to 1:

2.

3. The method for preparing L-fucose-modified protolichen stearic acid cationic nanoliposomes according to claim 1, characterized in that, The heating reaction described in step S1 takes 4 to 6 hours and is carried out at a temperature of 80°C to 90°C.

4. The method for preparing L-fucose-modified protolichen stearic acid cationic nanoliposomes according to claim 1, characterized in that, In step S2, soybean phospholipids, cationic lipids, sodium cholesterol sulfate, L-fucose-oleic acid amine coupler and lichenic acid are mixed in a mass ratio of 1:0.01:0.05:0.05:0.01 to 1:0.01:0.05:0.05:0.

1.

5. The method for preparing L-fucose-modified protolichen stearic acid cationic nanoliposomes according to claim 1, characterized in that, The cationic liposomes mentioned in step S2 are either DOTMA or DOTAP; the buffer solution is either PBS buffer or Tris-HCl buffer, and the pH value of the buffer solution is 6.0-8.

0.

6. The method for preparing L-fucose-modified protolichen stearic acid cationic nanoliposomes according to claim 1, characterized in that, In step S3, the ultrasonic cell disruptor treatment time is 1-5 minutes.

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