CN106943351A - A kind of method that hypergravity technology prepares nano liposomes - Google Patents

Abstract

The invention discloses a method for preparing nano liposomes by applying the supergravity technology. The method comprises the steps of mixing an organic solution containing a lipid material and an aqueous solution containing a carrier by using a high-gravity rotating bed, and then performing freeze-drying treatment to obtain nano liposomes. The invention utilizes the excellent mass transfer capacity of the high-gravity rotating packed bed, so that the preparation process of the nano-liposome is carried out under highly microscopic and uniform conditions, and the nano-liposome particle with controllable particle size, narrow distribution and high stability is obtained; At the same time, by adding a freeze-drying protective agent, the nano-liposomes are prevented from breaking and aggregating during the freeze-drying process, and the process of hydrating the freeze-dried product to form nano-liposomes is accelerated; the freeze-drying treatment method is used to process organic solvents, and the operation is simple and the organic solvent The removal effect is good; the process is simple, the energy consumption is low, the efficiency is high, the cost is low, and it is easy to enlarge to achieve the invention goal of industrial production. The average particle diameter of the obtained nanoliposome is 20-200nm, the PDI value is 0.1-0.3, and is suitable for large-scale production.

Description

A kind of method that applies supergravity technology to prepare nano liposome

technical field

The invention relates to the technical fields of biomedicine and cosmetics. More specifically, it relates to a method for large-scale application of high-gravity technology to prepare nano-liposomes.

Background technique

Liposomes are closed vesicle-like structures formed by phospholipid bilayers. According to their structure, they can be divided into unilamellar vesicles (SUV), multilamellar vesicles (LUV) and multivesicular vesicles (MIV). Liposomes were first discovered by the British Alec D.Bangham in 1965. Since then, it has been found that liposomes have great application value as material carriers, especially drug carriers, so liposomes have been systematically and extensively studied. .

After more than 20 years of exploration, researchers have proposed many valuable liposome preparation methods. At present, liposomes are mainly prepared by dispersion technique, which can be divided into three categories: 1) Based on mechanical dispersion technique. Such as the film dispersion method (film dispersion), patent CN 103110931A discloses a method for preparing romidolipid liposomes, using cholesterol as a stabilizer, sucrose as a freeze-drying protective agent, and vitamin E as an antioxidant. The romiglutide liposome was prepared by the film dispersion method, and the particle size of the liposome was smaller than 120nm by the high-pressure homogenization method. Although this method is the most classical and widely used method, there are some disadvantages: the organic solvent used is very toxic; industrialized production cannot be realized; The drug distribution between layers is not uniform, and it must be processed by repeated freezing and thawing. 2) Based on surfactant dispersion technology. In this method, lipids and surfactants are stirred together in an aqueous solution to obtain micelles, and then the surfactants are removed from the micelles by dialysis. The advantage is that the prepared liposome is relatively uniform, and because the phospholipid is operated at a temperature lower than its phase transition temperature, the treatment method is mild. However, the concentration of nanoliposomes prepared by this method is low, and there will be a small amount of surfactant residue at the same time, and it takes a long time to balance the micelles in the aqueous phase, which is time-consuming and difficult to produce on a large scale. 3) Based on solvent or anti-solvent dispersion technology. For example, the reverse phase evaporation method (REV), this preparation method has a relatively high encapsulation efficiency and drug loading capacity for water-soluble drugs; in the patent CN 101912388A, a preparation of a medium-chain fatty acid-vitamin C compound liposome is disclosed METHODS: Medium-chain fatty acid-vitamin C compound liposomes were prepared by reverse evaporation-high-pressure microfluidic method through dissolving, mixing, vacuum evaporation and high-pressure microfluidic treatment. Though the average particle diameter of this compound liposome can reach 90nm-200nm, after this method makes liposome, still need to reduce particle diameter through high-pressure micro-jet treatment, the process is loaded down with trivial details. In addition, the emulsion method can also be used to prepare multivesicular liposomes. At present, large-scale production can be realized, but it is limited to the preparation of micron-sized multivesicular liposomes with sustained release function.

Therefore, in view of the above problems, it is necessary to provide a simple and easy method for preparing nano-sized liposomes on a large scale.

Contents of the invention

An object of the present invention is to provide a method for preparing nano-liposomes using the hypergravity technique. The method utilizes the high-gravity rotating bed technology, which can greatly enhance mass transfer and micro-mixing, facilitate amplification, short production time, and is easy to produce on a large scale, and the prepared nano-liposomes have a narrow particle size distribution, which is a good solution to the problem of nano-liposomes. The problem that the body is difficult to industrialize production.

Another object of the present invention is to provide a nano-liposome prepared by adopting a high-gravity technique. The nano liposome has small particle size, narrow distribution, good dispersibility and wide application range.

In order to achieve the above-mentioned first object, the present invention adopts the following technical solutions:

A method for preparing nano-liposomes using high-gravity technology, the method comprising mixing an organic solution containing a lipid material and an aqueous solution containing a carrier using a high-gravity rotating bed, followed by freeze-drying to obtain a nano-liposome .

Preferably, the method includes the following specific steps:

1) dissolving lipids and membrane softeners forming nanoliposomes in an organic solvent to obtain an organic solution, and dissolving the water-soluble carrier in pure water to obtain an aqueous solution;

2) At 20-70°C, inject the organic solution and the aqueous solution obtained in step 1) into a high-gravity rotating bed for thorough mixing to obtain a mixed solution;

3) Adding a lyoprotectant to the mixed solution prepared in step 2), followed by lyophilization to obtain lyophilized nanoliposomes.

Preferably, in step 1), the lipids forming nanoliposomes are selected from the group consisting of phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, nerve sheath One or more of phospholipids and hydrogenated lecithin; the membrane softening agent is cholesterol; the organic solvent is selected from ethanol, propanol, isopropanol, glycerol, tert-butanol, acetone, N, N- One or more of dimethylacetamide and dimethyl sulfoxide. The purpose of doing this optimization is to screen out green and safe solvents and reduce solvent toxicity. Moreover, volatile solvents are screened out, which is easy to freeze-dry.

Preferably, in step 1), the water-soluble carrier is selected from sodium monohydrogen phosphate, potassium monohydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate, phosphoric acid, sodium hydroxide, potassium hydroxide, sodium chloride and chlorine One or more of potassium chloride; the water-soluble carrier is dissolved in pure water to adjust the pH value of the aqueous solution to 3-8. The preferred purpose of doing this is to configure the buffer solution. The phospholipids in the liposome are easily hydrolyzed, which will reduce the pH value of the solution and change the particle size and shape of the liposome. The buffer solution will provide a more stable environment, and the pH of the solution will be reduced. The value does not change significantly.

Preferably, in step 1), the weight ratio of the lipid to the membrane softener is 3-10:1; the concentration of the lipid in the organic solution is 5-80 mg/ml. Phospholipids and cholesterol form liposomes, and the purpose of adding cholesterol is to regulate the fluidity of phospholipids. Adding according to the ratio range defined in the present invention can make the formed liposome structure more stable.

Preferably, in step 2), the organic solution and the aqueous solution are respectively injected into the high-gravity rotating bed through a peristaltic pump.

Preferably, in step 2), the feed volume ratio of the organic solution to the aqueous solution is 1:5-30; the injection speed of the organic solution to the high-gravity rotating bed is 1ml/min-8ml/min; The injection speed of the aqueous solution to the high-gravity rotating bed is 5ml/min-240ml/min. And, by changing the feed ratio of the organic phase and the water phase, the particle size of the finally formed liposomes can be changed.

Preferably, in step 2), the rotating speed of the high-gravity rotating bed is 500rmp-2800rmp.

Preferably, in step 3), the lyoprotectant is selected from one or more of lactose, mannitol, xylitol, sucrose, trehalose, dextran and polyvinylpyrrolidone; The weight ratio of the amount of lyoprotectant added to the lipids forming nanoliposomes is 2.5-15:1; the specific operation of the lyophilization process is to freeze the mixed solution in liquid nitrogen or a cryogenic device and then freeze freeze-dry in a dryer, or directly freeze-dry the mixed solution in a freeze dryer.

The purpose of adding a lyoprotectant is to improve the state of liposomes after lyophilization. Freeze-drying is one of the effective methods to improve the long-term stability of liposome preparations, but the processes of pre-freezing and drying involved in lyophilization are not conducive to liposomes. In order to stabilize the plastid structure and function, using the type of lyoprotectant defined in the present invention and adding the lyoprotectant in the amount defined in the present invention can effectively reduce the adverse effects of the lyophilization process on liposomes.

Preferably, it also includes the step of hydrating the lyophilized nanoliposomes prepared in step 3) to obtain a nanoliposome solution; the hydration treatment refers to redissolving the lyophilized nanoliposomes to A hydrated nanoliposome solution is formed, and the solvent used for reconstitution is pure water. Water is the cheapest, environmentally friendly and safe solvent with good biocompatibility, and the prepared liposomes are mainly used in the fields of drug carriers and cosmetics, so the preferred reconstitution solvent is pure water. In addition, it can also be completed at a suitable temperature by using a suitable buffer solution. In order to accelerate the hydration, it can also be completed under the action of mechanical force, such as shaking, stirring or ultrasonication.

Further, the present invention also protects the nanoliposomes prepared by the method for preparing nanoliposomes using the above-mentioned high gravity technology, the average particle diameter of the nanoliposomes is 20-200nm, and the PDI value is 0.1-0.3.

Preferably, the average particle diameter of the nanoliposome is 20-120nm.

In the prior art, the nanoliposomes prepared on the basis of mechanical dispersion technology use organic solvents that are highly toxic and cannot be industrialized, and when hydrated with a drug-containing aqueous solution, the formed multilayer liposome layer and layer Inhomogeneous distribution of drugs among patients must undergo repeated freezing and thawing treatments; nanoliposomes prepared based on surfactant dispersion technology have low concentration and residual surfactants are difficult to completely remove; solvent or anti-solvent dispersion technology is used as the basis The basic preparation of liposomes is cumbersome and limited to the preparation of micron-sized multivesicular liposomes with sustained release function. Aiming at the deficiencies in the prior art, the present invention proposes for the first time to combine the preparation of nanoliposomes with the supergravity technology, by selecting suitable lipids and membrane softeners, and matching the corresponding water-soluble carriers, to adjust the ratio between the raw materials. The proportion relationship, through the appropriate injection speed into the high-gravity rotating bed for mixing, and then freeze-drying with the protection of a suitable freeze-drying protective agent, can effectively remove organic solvents and other residues, and ensure the purity of liposomes. And dry liposomes are good for preservation. Finally, the hydrated nano-liposome product with uniform particle size and good stability is obtained by simple reconstitution and hydration for application. In the present invention, by optimizing all steps and process parameters in the preparation method, they coordinate and cooperate with each other to form the technical solution of the present invention and achieve good technical effects, that is, the present invention utilizes the excellent mass transfer capacity of the high-gravity rotating packed bed , combine it with the preparation of liposomes to strengthen the transfer and mixing of solvent-anti-solvent, so that the preparation process of nano-liposomes can be carried out under highly microscopic and uniform conditions, and the particle size can be controlled, the distribution is narrow, and the stability High nanoliposome particles, and can achieve large-scale production.

The beneficial effects of the present invention are as follows:

1. The present invention utilizes the excellent mass transfer capacity of the high-gravity rotating packed bed to strengthen the transfer and mixing of solvent-anti-solvent, so that the preparation process of nano-liposomes is carried out under highly microscopic and uniform conditions, and the particle size can be controlled and distributed. Narrow, highly stable nanoliposomal particles.

2. The present invention avoids nano-liposome rupture and aggregation during the freeze-drying process by adding a freeze-drying protective agent, and accelerates the process of hydration of freeze-dried products to form nano-liposomes.

3. The present invention adopts the freeze-drying treatment method to process the organic solvent, which is easy to operate and has good removal effect of the organic solvent.

4. The process of the present invention is simple, easy to realize, less energy consumption, high in efficiency, low in cost, and very easy to scale up, so as to achieve the invention goal of industrialized production.

Description of drawings

The specific implementation manners of the present invention will be further described in detail below in conjunction with the accompanying drawings.

Figure 1 shows the scanning electron micrograph of the nanoliposome prepared in Example 1 of the present invention.

FIG. 2 shows the scanning electron micrograph of the nanoliposome prepared in Comparative Example 1 of the present invention.

Fig. 3 shows the scanning electron micrograph of the nanoliposome prepared in Example 5 of the present invention.

Fig. 4 shows a scanning electron micrograph of the nanoliposome prepared in Example 6 of the present invention.

detailed description

In order to illustrate the present invention more clearly, the present invention will be further described below in conjunction with preferred embodiments and accompanying drawings. Similar parts in the figures are denoted by the same reference numerals. Those skilled in the art should understand that the content specifically described below is illustrative rather than restrictive, and should not limit the protection scope of the present invention.

Example 1

Dissolve 80g of phosphatidylcholine and 10g of cholesterol in 2L of ethanol, add 5g of potassium dihydrogen phosphate and 5g of dipotassium hydrogen phosphate into 20L of water and adjust the pH to 6.5, turn on the high-gravity rotating bed and adjust the speed to 2000rpm, and use 2ml of ethanol /min, the water phase is fed at 20ml/min, and the temperature of the control system is 30°C. After the ethanol phase is fed, the high-gravity rotary bed is closed, and 300g sucrose is added to the resulting mixed solution, and the organic solvent is removed after freeze-drying. A dry freeze-dried powder is obtained. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. Figure 1 shows the scanning electron micrograph of the nanoliposome prepared in Example 1 of the present invention. It can be seen from the figure that the average particle size of the nanoliposome is 40nm, and its particle size distribution test shows that its PDI value is 0.18, showing good dispersion. Figure 2 shows the particle size distribution diagram of the nanoliposomes prepared in Example 1 of the present invention. It can be seen from the figure that the average particle diameter of the nanoliposome solution is 40nm, which is consistent with the electron microscope result.

Comparative example 1

80mg of phosphatidylcholine and 10mg of cholesterol were dissolved in 2ml of ethanol, and 20ml of dipotassium hydrogen phosphate buffer solution with a pH of 6.5 was prepared in Example 1. Under magnetic stirring conditions, the ethanol solution was added dropwise to the aqueous solution at 30°C. Add 300 mg of sucrose to the obtained mixed solution, remove the organic solvent after freeze-drying, and obtain dry nano-liposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. FIG. 2 shows the scanning electron micrograph of the nanoliposome prepared in Comparative Example 1 of the present invention. The average particle diameter of the nanoliposome is 500nm, and its particle size distribution test shows that its PDI value is 0.35, and its dispersibility is poor. It can be seen that the use of ordinary mechanical mixing is not only impossible for mass production, but also time-consuming, and the obtained products are lacking in particle size and uniformity.

Example 2

Dissolve 70g of phosphatidylcholine and 10g of cholesterol in 2L of isopropanol solution, dissolve 4g of potassium dihydrogen phosphate and 6g of disodium hydrogen phosphate in 20L of water, adjust the pH of the buffer to 7.4, and turn on the high-gravity rotating bed to adjust the speed to 2000rpm. The isopropanol phase is fed at 3ml/min, the water phase at 30ml/min, and the temperature of the control system is 30°C. After the isopropanol phase is fed, the supergravity rotary bed is closed, and 300g of sucrose is added to the resulting mixed solution. After freeze-drying, the solvent is removed to obtain dry nano-liposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle diameter of the nanoliposome is 80nm, and its particle size distribution test shows that its PDI value is 0.16, showing good dispersibility.

Example 3

Dissolve 70g of phosphatidylcholine and 15g of cholesterol in 2L of tert-butanol solution, dissolve 3g of disodium hydrogen phosphate and 2g of phosphoric acid in 10L of water and adjust the pH of the buffer to 5, turn on the supergravity rotating bed and adjust the speed to 1000rpm, t The butanol phase is fed at 3ml/min, the water phase is fed at 30ml/min, and the temperature of the system is controlled at 30°C. After the organic solution is fed, the high-gravity rotating bed is closed, and 300g trehalose is added to the resulting mixed solution. After drying, the solvent is removed to obtain dry nanoliposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle diameter of the nanoliposome is 65nm, and its particle size distribution test shows that its PDI value is 0.2, showing good dispersion stability.

Example 4

Dissolve 70g of phosphatidylcholine and 10g of cholesterol in 3L of N,N dimethylacetamide, dissolve 2g of sodium chloride and 2g of phosphoric acid in 20L of water and adjust the pH of the buffer to 7, turn on the supergravity rotating bed and adjust the speed to 1000rpm , N, N dimethylacetamide phase is fed at 2ml/min, water phase is fed at 15ml/min, the temperature of the system is controlled at 40°C, after the N, N dimethylacetamide phase is fed, the supergravity rotation is turned off bed, add 300g trehalose to the resulting mixed solution, and remove the solvent after freeze-drying to obtain dry nanoliposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle size of the nanoliposome is 75nm, and its particle size distribution test shows that its PDI value is 0.19, showing good dispersion stability.

Example 5

Dissolve 100g of sphingomyelin and 15g of cholesterol in 2L of ethanol solution, dissolve 2g of potassium dihydrogen phosphate and 2g of sodium chloride in 20L of water and adjust the pH of the buffer to 6, turn on the supergravity rotating bed and adjust the speed to 1500rpm, and the ethanol phase Feed at 3ml/min, water phase at 20ml/min, control the temperature of the system at 50°C, after the ethanol phase is fed, close the high-gravity rotating bed, add 300g trehalose to the resulting mixed solution, freeze-dry and remove vehicle to obtain dry nanoliposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. Fig. 3 shows the scanning electron micrograph of the nanoliposome prepared in Example 5 of the present invention. The average particle diameter of the nanoliposome is 80nm, and its particle size distribution test shows that its PDI value is 0.12, showing good dispersion stability.

Example 6

Dissolve 100g of phosphatidylcholine and 10g of cholesterol in 2L of propanol solution, dissolve 2g of potassium dihydrogen phosphate and 2g of potassium dihydrogen phosphate in 20L of water, adjust the pH of the buffer to 5.5, and turn on the supergravity rotating bed to adjust the speed to 1000rpm , the propanol phase is fed at 3ml/min, the water phase is fed at 250ml/min, and the temperature of the system is controlled at 40°C. After the propanol phase is fed, the supergravity rotating bed is closed, and 300g trehalose is added to the resulting mixed solution. After freeze-drying, the solvent is removed to obtain dry nano-liposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. Fig. 4 shows a scanning electron micrograph of the nanoliposome prepared in Example 6 of the present invention. The average particle size of the nanoliposome is 90nm, and its particle size distribution test shows that its PDI value is 0.22, showing good dispersion stability.

Example 7

Dissolve 100 g of phosphatidylcholine, 20 g of cholesterol and 0.02 μmol/ml α-tocopherol in 2 L of isopropanol/ethanol solution (1:1), dissolve 1 g of sodium chloride and 2 g of sodium dihydrogen phosphate in 20 L of water and adjust The pH of the buffer solution is 5, turn on the supergravity rotating bed and adjust the speed to 2500rpm, feed the organic solution at 3ml/min, and the aqueous solution at 30ml/min, control the temperature of the system at 30°C, and turn off the supergravity rotation after the organic phase is fed bed, add 300g trehalose to the resulting mixed solution, and remove the solvent after freeze-drying to obtain dry nanoliposome freeze-dried powder. The freeze-dried powder can be preserved for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle size of the nanoliposome is 95nm, and its particle size distribution test shows that its PDI value is 0.25, showing good dispersion stability.

Example 8

Dissolve 80g of phosphatidylcholine and 15g of cholesterol in 2L of glycerol solution, dissolve 6g of sodium hydroxide and 2g of phosphoric acid in 20L of water and adjust the pH to 4.5, turn on the high-gravity rotating bed and adjust the speed to 2000rpm, and the glycerol phase Feed at 2ml/min, water phase at 25ml/min, control the temperature of the system at 60°C, after the glycerol phase is fed, close the high-gravity rotating bed, add 300g trehalose to the resulting mixed solution, freeze-dry Finally, the solvent is removed to obtain dry nanoliposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle size of the nanoliposome is 70nm, and its particle size distribution test shows that its PDI value is 0.23, showing good dispersion stability.

Example 9

Dissolve 80g of phosphatidylethanolamine and 10g of cholesterol in 2L of ethanol/tert-butanol (1:1) solution, dissolve 4g of disodium hydrogen phosphate and 3g of sodium dihydrogen phosphate in 15L of water, adjust the pH to 4, and start the supergravity rotation The bed adjustment speed is 1000rpm, the organic solution is fed at 3ml/min, the aqueous solution is fed at 30ml/min, the temperature of the control system is 30°C, after the organic solution is fed, the supergravity rotating bed is closed, and 300g of seaweed is added to the resulting mixed solution Sugar, after being freeze-dried, the solvent is removed to obtain dry nano-liposome freeze-dried powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle diameter of the nanoliposome is 103nm, and its particle size distribution test shows that its PDI value is 0.212, showing good dispersion stability.

Example 10

Dissolve 80g of hydrogenated phospholipids and 10g of cholesterol in 2L of acetone solution, dissolve 2g of dipotassium hydrogen phosphate and 3g of sodium dihydrogen phosphate in 20L of water and adjust the pH to 3.5, turn on the supergravity rotating bed and adjust the speed to 1500rpm, and use 3ml of acetone phase /min, the water phase is fed at 30ml/min, and the temperature of the system is controlled at 70°C. After the acetone phase is fed, the high-gravity rotary bed is closed, and 300g trehalose is added to the resulting mixed solution, and the solvent is removed after freeze-drying. Obtain dry nano liposome lyophilized powder. The freeze-dried powder can be stored for a long time, and the dried freeze-dried powder is reconstituted and hydrated with pure water before use to obtain a nanoliposome solution. The average particle size of the nanoliposome is 68nm, and its particle size distribution test shows that its PDI value is 0.155, showing good dispersion stability.

Apparently, the above-mentioned embodiments of the present invention are only examples for clearly illustrating the present invention, and are not intended to limit the implementation of the present invention. Those of ordinary skill in the art can also make It is impossible to exhaustively list all the implementation modes here, and any obvious changes or changes derived from the technical solutions of the present invention are still within the scope of protection of the present invention.

Claims (10)

1. A method for preparing nano-liposomes using high gravity technology, characterized in that: the method comprises the use of a high gravity rotary bed to mix the organic solution containing the lipid material and the aqueous solution containing the carrier, and then freeze-drying to obtain The steps of nanoliposomes. 2. a kind of method according to claim 1 is prepared nano liposome using high gravity technology, is characterized in that, comprises following concrete steps: 1) dissolving lipids and membrane softeners forming nanoliposomes in an organic solvent to obtain an organic solution, and dissolving the water-soluble carrier in pure water to obtain an aqueous solution; 2) At 20-70°C, inject the organic solution and the aqueous solution obtained in step 1) into a high-gravity rotating bed for thorough mixing to obtain a mixed solution; 3) Adding a lyoprotectant to the mixed solution prepared in step 2), followed by lyophilization to obtain lyophilized nanoliposomes. 3. a kind of method that application hypergravity technology prepares nano liposome according to claim 2, is characterized in that: in step 1), the described lipid material that forms nano liposome is selected from phosphatidylcholine, One or more of phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidylglycerol, diphosphatidylglycerol, sphingomyelin and hydrogenated lecithin; the membrane softening agent is cholesterol; the organic solvent is selected from One or more of ethanol, propanol, isopropanol, glycerol, tert-butanol, acetone, N,N-dimethylacetamide and dimethyl sulfoxide. 4. a kind of method that application supergravity technology prepares nano liposome according to claim 2 is characterized in that: in step 1), described water-soluble carrier is selected from sodium monohydrogen phosphate, potassium monohydrogen phosphate, phosphoric acid One or more of sodium dihydrogen, potassium dihydrogen phosphate, phosphoric acid, sodium hydroxide, potassium hydroxide, sodium chloride and potassium chloride; the pH value of the aqueous solution is adjusted after the water-soluble carrier is dissolved in pure water 3-8. 5. a kind of method that application supergravity technology prepares nano liposome according to claim 2, is characterized in that: in step 1), the weight ratio of described lipid substance and membrane softening agent is 3-10:1 ; The concentration of the lipid substance in the organic solution is 5-80mg/ml. 6. a kind of method according to claim 2 using high gravity technology to prepare nano liposome, is characterized in that: in step 2), the feed volume ratio of described organic solution and aqueous solution is 1:5-30; The injection speed of the organic solution into the high-gravity rotating bed is 1ml/min-8ml/min; the injection speed of the aqueous solution into the high-gravity rotating bed is 5ml/min-240ml/min. 7. A method for preparing nano-liposomes by applying high-gravity technology according to claim 2, characterized in that: in step 2), the rotating speed of the high-gravity rotating bed is 500rmp-2800rmp. 8. a kind of method according to claim 2 using high gravity technology to prepare nano liposome, is characterized in that: in step 3), described lyoprotectant is selected from lactose, mannitol, xylitol, sucrose , trehalose, dextran and polyvinylpyrrolidone; the weight ratio of the added amount of the lyoprotectant to the lipid substance forming nanoliposomes is 2.5-15:1 ; The specific operation of the freeze-drying process is to freeze the mixed solution in liquid nitrogen or a low-temperature device and then freeze-dry it in a freeze-dryer, or directly freeze-dry the mixed solution in a freeze-dryer. 9. a kind of method that application hypergravity technology prepares nanoliposome according to claim 2, is characterized in that: also comprise step 3) the nanoliposome of lyophilized state of preparing is carried out hydration treatment and obtains nanolipid The step of the plastid solution; the hydration treatment refers to redissolving the freeze-dried nano-liposomes to form a hydrated nano-liposome solution, and the solvent used for re-dissolving is pure water. 10. The nanoliposome prepared by the method for preparing nanoliposome using the high gravity technology as claimed in any one of claims 1-9, characterized in that: the average particle diameter of the nanoliposome is 20-200nm , the PDI value is 0.1-0.3.