WO2010111807A1 - A polysaccharide liposome, the preparation method and use of it - Google Patents

Abstract

A polysaccharide liposome, in particular a lentinan liposome, a method for its preparation, a pharmaceutical composition comprising the liposome, and the use of the liposome for manufacturing antineoplastic or immunoregulatory medicaments are disclosed. Polysaccharide, phospholipid, cholesterol, surfactants and polyethylene glycol are used as starting materials for producing the liposome.

Description

 Polysaccharide liposome, preparation method and use thereof

 The invention relates to a polysaccharide liposome, in particular to a lentinan liposome; the invention also relates to a preparation method and use of a polysaccharide liposome. Background technique

 Polysaccharides are widely found in nature and are an important part of living organisms and are associated with the many functions required to sustain life. Polysaccharides have unique effects in anti-tumor, anti-infection, cardiovascular disease, glucose metabolism, anti-aging, etc., and can improve the body's immune function.

 However, in actual clinical applications, in many cases, polysaccharide drugs cannot achieve the desired ideal pharmacological activity in vivo. This is mainly due to the fact that polysaccharide drugs are mostly metabolized before they reach the lesion site in the body, and the drugs do not have targeting for the lesion site. Therefore, how to make the polysaccharide drug be as little as possible before reaching the lesion, and "targeting" to reach the lesion site to obtain the ideal clinical use effect, has become a universal problem that urgently needs to be solved in the technical field.

 The liposome delivery system has received wide attention due to its good clinical application. However, its preparation, especially the preparation of polysaccharide liposomes with high encapsulation efficiency, is still a recognized problem. It is more difficult to obtain a polysaccharide liposome having a higher drug encapsulation efficiency and a smaller particle diameter. Summary of the invention

 The inventors have obtained a lipid body encapsulated with a polysaccharide drug and a preparation method thereof by repeated research. Meanwhile, the inventors have unexpectedly found that the polysaccharide liposome of the present invention has a high encapsulation ratio and a small particle diameter.

 To this end, the present invention provides a liposome comprising a polysaccharide, a phospholipid, cholesterol, a surfactant, and polyethylene glycol.

 The invention also provides a method of preparing a liposome of the invention, the method comprising the steps of:

 a. Dissolving the raw materials including the above phospholipids, cholesterol and polyethylene glycol in an organic solvent

1 Confirmation In the middle, the oil phase mixture;

 b. dissolving the raw material comprising the above polysaccharide and surfactant in water or an aqueous solution to obtain an aqueous phase mixture;

 C. injecting the aqueous phase mixture into the oil phase mixture for emulsification;

 d. The solvent is removed by evaporation under constant pressure under reduced pressure in a protective gas;

 e. Add water or aqueous solution and incubate in a water bath.

 The present invention also provides a composition comprising the above liposome.

 The present invention also provides the use of the above liposome or a composition comprising the above liposome for the preparation of a medicament for antitumor or for regulating the immune function of a human or animal.

 The polysaccharide liposome of the present invention can have a particle size below Ι μ ηη and a uniform distribution. The encapsulation efficiency of the liposome of the present invention is 65% or more. The administration of the polysaccharide liposome of the present invention in vivo is markedly improved as compared with the administration of a polysaccharide which is not encapsulated into a liposome. DRAWINGS

 Fig. 1 is a view showing the particle size distribution of the liposome of Example 1 of the present invention.

 Fig. 2 is a transmission electron micrograph of the liposome of Example 1 of the present invention. detailed description

 The compounds, compositions, methods, and reagents disclosed herein are not limited to those specifically listed herein, and they may be variously modified. The description of the specific embodiments, particularly the embodiments of the present application, are intended to be illustrative of the invention, and not to limit the scope of the invention.

 The invention will now be described in detail by way of example with reference to specific embodiments. As used herein, "liposome" refers to a closed vesicle composed of a lipidoid bilayer containing a pharmaceutically active ingredient. The liposome may be a single-chamber liposome or a multi-chamber liposome. The liposomes described in this application also include liposomes that have been surface modified.

The polysaccharide described in the present application may be selected from the group consisting of lentinan, astragalus polysaccharide, tremella polysaccharide, glucose, ginseng polysaccharide, ganoderma lucidum polysaccharide, porcini polysaccharide, etc.; preferably, the polysaccharide is fragrant Mushroom polysaccharide or Astragalus polysaccharide; most preferably, the polysaccharide is lentinan.

 The "phospholipid" described herein may be selected from the group consisting of soybean phospholipids, lecithin, hydrogenated soybean phospholipids, hydrogenated lecithin, phosphatidylethanolamine, distearoylphosphatidylcholine, polyethylene glycol derivative phospholipids, mixtures thereof and the like.

 The polyethylene glycol described in the present application may be selected from the group consisting of polyethylene glycol 800-5000, phospholipidized polyethylene glycol derivatives, mixtures thereof and the like; preferably, the polyethylene glycol is polyethylene glycol 1000- More preferably, the polyethylene glycol is polyethylene glycol 2000.

 The surfactant of the present invention may be selected from the group consisting of deoxycholate, Poloxamer, Tween 80, mixtures thereof and the like; preferably, the surfactant is deoxycholate, such as deoxycholic acid. Sodium or potassium deoxycholate.

 The starting material for preparing the liposome of the present invention may further comprise other substances which can be used for the preparation of liposomes. For example, in one embodiment of the liposome of the present invention, the starting material may further comprise an antioxidant to enhance storage of the liposome. stability. In still another embodiment of the liposome of the present invention, the starting material may further comprise a freeze-protecting agent to facilitate the formation of a lipid solution into the frozen powder.

 The antioxidant of the present invention may be any pharmaceutically acceptable antioxidant such as vitamin E, vitamin C, cysteine, methionine, sodium hydrogen sulfite or a mixture thereof.

 The lyoprotectant of the present invention may be any pharmaceutically acceptable cryoprotectant such as mannitol, trehalose, sucrose, glycine or a mixture thereof and the like. '

 In one embodiment of the liposome of the present invention, the raw material comprises the following components: 1 part of lentinan, 30-100 parts of phospholipid, 2-20 parts of cholesterol; preferably, 1 part of lentinan, phospholipid 40-90 parts, cholesterol 5-15 parts; more preferably, 1 part of lentinan, 50-70 parts of phospholipids, 5-15 parts of cholesterol.

 Unless otherwise stated, the "parts" used in this application are all "parts by weight".

 In one embodiment of the lipid of the present invention, the surfactant is from 3 to 12 parts. In another embodiment of the liposomes of the present invention, the polyethylene glycol in the starting material is from 1 to 8 parts.

 In still another embodiment of the liposome of the present invention, the starting material contains a suitable amount of an antioxidant, which can be readily determined by those skilled in the art based on conventional knowledge, for example, 1-8 parts of antioxidant can be used. .

The liposomes of the invention may be presented in any dosage form suitable for administration to a subject, for example The liposome of the present invention can be prepared as an injection (for example, an injection or a powder injection), an oral preparation (e.g., a tablet, a capsule, or a granule). Preferably, the liposome of the invention is an injection or a powder injection. The liposome of the present invention can be obtained by the above-described production method including the aforementioned ae step.

 Wherein, the liposome of the present invention is required to be formulated into a powder injection, and the preparation method of the liposome of the present invention may further comprise the step of lyophilizing the solution obtained in step e after filtration. The solvent used in the step a may be Any organic solvent suitable for dissolving the starting materials, such as chloroform or chloroform-nonanol or diethyl ether. The organic solvent is used in an amount which enables the raw material to be dissolved, and the amount varies depending on the raw material, and can be easily determined by those skilled in the art based on conventional knowledge.

 The aqueous solution in the step b may be a buffer solution such as a phosphate buffer solution, a citrate buffer solution, an acetate buffer solution, a sodium hydrogencarbonate buffer solution, a tartaric acid buffer solution, or the like; or a chlorination solution; An aqueous sodium solution, such as physiological saline. The water or water solution used in the step b is used in an amount which enables the raw material to be dissolved, and the amount varies depending on the raw material, and can be easily determined by those skilled in the art based on conventional knowledge.

 The emulsification treatment of the step c in the preparation method of the present invention may be a sonication treatment or a high pressure emulsification treatment, which is usually carried out under low temperature conditions, for example, at a temperature below ιοχ, for a time of emulsification to form a mixture into suitable granules. The diameter is a degree, and a person skilled in the art can determine a suitable emulsification time for different particle diameters by any known method, and the emulsification time can be, for example, 3 to 10 min. The ultrasonic treatment of the present invention is preferably intermittent ultrasonic treatment; the high pressure emulsification treatment is preferably a cyclic high pressure emulsification treatment.

 The protective gas of the step d in the production method of the present invention may be an inert gas such as nitrogen.

 The step d in the preparation method of the present invention can be carried out by rotary evaporation at a suitable temperature, preferably at a constant temperature in the temperature range of 35-45 TC.

The step e in the preparation method of the present invention is preferably carried out in a water bath incubation at a temperature ranging from 35 to 50 C, more preferably at 40 in a water bath. The incubation time of the step e of the preparation method of the present invention is preferably 1-2 hours.

 The aqueous solution of the step e of the preparation method of the present invention may be an aqueous solution of sodium chloride, a buffer solution such as a phosphate buffer solution, a citrate buffer solution, an acetate buffer solution, a sodium hydrogencarbonate buffer solution, or tartaric acid. Buffer, etc.

 Preferably, the pH of the reaction step of the present invention is inconsistent with the pH of the reaction environment of the step e. More preferably, the b step of the present invention is opposite to the acidity and alkalinity of the reaction environment of the e step, and the acid alkaline condition is opposite when the reaction environment of one of the steps b and e is acidic. The reaction environment in the other step is alkaline. Particularly preferably, the reaction environment of the step b is basic, and the reaction environment of the step e is acidic.

 In one embodiment of the preparation process of the invention, the feedstock in step a further comprises an antioxidant.

 In still another embodiment of the preparation method of the present invention, the aqueous solution in the step e contains a lyoprotectant. Suitable amounts of the lyoprotectant can be readily determined by one of ordinary skill in the art from routine prior knowledge.

 The polysaccharide liposome of the present invention has significant targeting properties for specific tissues or cells such as liver, bone marrow, tumor tissue, inflammatory tissue, and the like, and has long-circulating properties.

 The polysaccharide liposome of the present invention can be used for treating or assisting in the treatment of tumor diseases such as liver, ovary, intestine, lung, breast, prostate, pancreas, kidney, stomach, endometrium, esophagus, head or neck tumor, and cancerous chest. Tumors such as ascites tumors. The polysaccharide liposome of the present invention also has an immunomodulatory effect, for example, it can be used for improving the immune function of hepatitis, rheumatoid arthritis or AIDS patients. The administration of the polysaccharide after being encapsulated into a liposome is significantly improved as compared with the administration of a simple polysaccharide.

 The polysaccharide liposome of the present invention has a high encapsulation efficiency. Among them, the encapsulation efficiency of the polysaccharide liposome of the present invention is 65% or more; preferably, the encapsulation ratio is 70% or more; more preferably, the encapsulation ratio is 80% or more. The liposome of the invention has a uniform smaller particle size while having a higher encapsulation ratio, and generally the particle size of the liposome is below Ι μηη; a preferred particle size is 100 to 500 nm; more preferred particles The diameter is 140~300 nm.

The encapsulation efficiency of the liposome referred to in the present invention means the weight percentage of the polysaccharide encapsulated in the liposome and the polysaccharide in the raw material for preparing the liposome _β The liposome of the present invention may be formulated into a corresponding pharmaceutical composition with one or more other pharmaceutically active ingredients selected from the group consisting of:

 Antineoplastic agents, such as cytotoxic agents such as Bleomycin, Doxorubicin, Mitomycin, Carmustine, Carboplatin, Cisplatin , Iproplatin, Cyclophosphamide, Gemcitabine, Docetaxel, Etoposide, Fluorouracil

 (Fluorouracil), Melphalan, Methotrexate, Vinorelbine, Vincristine, Paclitaxel, etc., hormones or anti-hormones such as aminoglutethimide ( Aminoglutethimide), Anastrozole, Bicalutamide, Irinotecan, Rituximab, Tamoxifen, Trastuzumab Wait;

 Anti-infective disease drugs, for example, antiviral drugs such as Ganciclovir, Ribavirin, Vidarabine, Famciclovir

 (Famciclovir), Lamivudine, Abacavir, Nevirapine, Indinavir, etc., anti-inflammatory drugs such as Aspirin, Ibuprofen, Indometacin, Piroxicam, Nimesulide, Glucosamine, etc.;

 Immunomodulators such as interferons or interleukin active drugs.

 The liposomes of the present invention or the above compositions may also be used in combination with one or more of the following pharmaceutically acceptable carriers, excipients or adjuvants, such as fillers, absorbents, wetting agents, binders, lubricants, colorants, Flavoring agents, stabilizers, preservatives, buffers, disintegrating agents, coating materials, dispersing agents and suspending agents, and the like. The polysaccharide liposome of the present invention or a composition thereof can be delivered to a subject's site of interest in a variety of ways. For example, the therapeutic site can be targeted to the treatment site by oral administration, injection administration (intravenous injection, intramuscular injection, intraperitoneal injection, subcutaneous injection) in a suitable form to obtain enhanced immunomodulatory effects or antibiotics. Tumor effect, etc.

The liposomes of the invention may also be combined with other active agents such as those described above. In vivo co-administration for the above therapeutic effects. Embodiments of the invention are further illustrated by the following examples. The compounds or reagents used in the following examples are commercially available or can be prepared by conventional methods known to those skilled in the art; the experimental apparatus used is commercially available. Example 1 Prescription of the raw material of liposome 1 :

 Preparation of liposome 1 :

 Weigh 8.0 g of soybean phospholipid, 1.0 g of cholesterol, 0.2 g of polyethylene glycol 2000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0. lg injection grade lentinan and 0.9 g sodium deoxycholate, add 300 ml of water for injection, and add 0.1 mol/L sodium hydroxide solution to dissolve the lentinan, and adjust the pH value of the solution with 0.1 mol/L hydrochloric acid. From 7.0 to 8.0, an aqueous mixture is obtained. After cooling in an ice water bath, the aqueous phase mixture was slowly added to the oil phase mixture, and then intermittently emulsified for about 5 minutes to obtain a milky white uniform W/0 type agent. The solvent was removed by rotary evaporation under reduced pressure of 40 Torr under nitrogen to give a pale-yellow gum, which was added to a mixture of <RTIgt; The microporous membrane was used for filtration and sterilization, and 100 pieces of lentinan liposome injection were obtained by dispensing.

 Evaluation of the fat shield 1:

Take 0.2 ml of liposome 1 in a 10 ml conical centrifuge tube, add 0.2 ml protamine (10 mg/ml), shake well, let stand for 3 min, add 0.05 mol/L sodium hydroxide solution to 5 ml Centrifuge at 3000 r/min for 30 min at room temperature and take 2 ml. To the clear solution, add 0. 2% sulphonate solution (prepared from 0.2 g of anthrone plus 100 ml of concentrated sulfuric acid) 4 ml, mix by shaking, heat in a boiling water bath for 6 min, and cool in an ice water bath. 2min, room temperature for 10 min, measure the absorbance; take the amount of lentinan reference substance dried to constant weight by phosphorus pentoxide, accurately weighed, add a small amount

0.5 mol / L sodium hydroxide solution was ground and dissolved, diluted with water to make a solution containing 20 g per 1 ml, measured by the same method, calculated, that is, the amount of lentinan was not encapsulated. The encapsulation rate is calculated by the following formula:

 Encapsulation efficiency (%) = (1 - unencapsulated amount of lentinan / total amount of lentinan) χ ΐοο%

 The encapsulation efficiency of the lentinan was determined to be 82.8%.

 (2) Particle size and microscopic morphology

The particle size range of liposome 1 was determined to be 200 to 400 nm and the average particle size was 316 nm using a laser particle size analyzer (US Backmman DELSA 44 OS laser particle size analyzer) (Fig. 1). Observed by transmission electron microscopy (FEI Tecnai G ² 12 transmission electron microscope), the appearance of liposome 1 was spherical and the structure was clear (Fig. 2). Example 2

 Raw material prescription for liposome 2:

制备： 称取 8.0 g大豆磷脂、 1.0 g胆固醇、 0.2 g聚乙二醇 2000和 0.2 g维生素 E, 加氯仿 900 ml, 搅拌使溶解， 得油相混 合物。 称取 0.1 g注射级香菇多糖和 0.9 g去氧胆酸钠， 加入注射 用水 300 ml, 并加入 0.1 mol/L氢氧化钠液适壹至香菇多糖溶解， 用 0.1 mol/L盐酸调节 pH值为 7.0~8.0， 得水相混合物。 水水浴 冷却下， 将水相混合物緩緩加入油相混合物中， 然后间歇超声乳化 约 5min， 得乳白色均一的 W/0型乳剂。 在 40 和氮气保护下减压 旋转蒸发除去溶剂得淡黄色浓稠胶状物， 加入 2%甘露醇的磷酸緩 冲液（ρΗ值 6.0 ~ 6.5 ) 200 ml, 40Ό孵化 1 h, 得到脂质体 2的 溶液。

Preparation: Weigh 8.0 g of soybean phospholipid, 1.0 g of cholesterol, 0.2 g of polyethylene glycol 2000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0.1 g of injection-grade lentinan and 0.9 g of sodium deoxycholate, add 300 ml of water for injection, add 0.1 mol/L sodium hydroxide solution to dissolve the lentinan, and adjust the pH with 0.1 mol/L hydrochloric acid. From 7.0 to 8.0, a mixture of aqueous phases is obtained. Water bath The aqueous phase mixture was slowly added to the oil phase mixture under cooling, and then intermittently emulsified for about 5 minutes to obtain a milky white uniform W/0 type emulsion. The solvent was removed by rotary evaporation under reduced pressure of 40 and nitrogen to give a pale yellow thick gel, which was added to a 2% mannitol phosphate buffer (pH 6.0 6.0 ~ 6.5) 200 ml, 40 Ό for 1 h to obtain liposomes. 2 solution.

 The obtained solution was filtered and sterilized by a microporous membrane, and packed into 100 pieces, pre-frozen for 2 hours at -50 Torr, gradually warmed to -6 Torr for about 8 hours, and gradually heated to 30 for 12 hours to obtain a pale yellow mass. , that is, the powder injection form of liposome 2.

 After the obtained powder injection was reconstituted, the encapsulation efficiency of liposome 2 was determined to be 84.3%, and the average particle diameter was 318 nm. The appearance of liposome 2 was spherical after observation by transmission electron microscopy. Clear structure. Example 3

 Prescription of the raw material of liposome 3:

制备：称取 6.1 g大豆磷脂、 0.8 g胆固醇、 0.2 g聚乙二醇 1000 和 0.2 g维生素 E, 加氯仿 900 ml, 搅拌使溶解， 得油相混合物。 称取 0.1 g注射级香菇多糖和 0.5 g去氧胆酸钠， 加入注射用水 300 ml, 并加入 0.1mol/L氢氧化钠液适量至香菇多糖溶解， 用 0.1 mol/L盐酸调节 pH值为 7.0~8.0,得水相混合物。水水浴冷却下， 将水相混合物緩緩加入油相混合物中，然后间歇超声乳化约 6min， 得乳白色均一的 WO型乳剂。 在 35 和氮气保护下减压旋转蒸发 除去溶剂得淡黄色浓稠胶状物，加入 pH值 6.0~ 6.5磷酸盐緩冲溶 液 200 ml， 40 孵化 1.5 h， 得到脂质体溶液。 如实施例 1所述方 法测得香菇多糖包封率为 82.2% , 平均粒径为 302 nm。 经透射电 镜观察， 可见脂质体 3的外观呈球状， 结构清晰。 实施例 4

Preparation: Weigh 6.1 g of soybean phospholipid, 0.8 g of cholesterol, 0.2 g of polyethylene glycol 1000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0.1 g of injection-grade lentinan and 0.5 g of sodium deoxycholate, add 300 ml of water for injection, and add 0.1 mol/L sodium hydroxide solution to dissolve the lentinan, and adjust the pH to 7.0 with 0.1 mol/L hydrochloric acid. ~8.0, a mixture of aqueous phases. The aqueous phase mixture was slowly added to the oil phase mixture under cooling with a water bath, and then intermittently emulsified for about 6 minutes to obtain a milky white uniform WO type emulsion. The solvent was removed by rotary evaporation under reduced pressure of nitrogen and nitrogen to give a pale-yellow thick gel, which was added to a pH 6.0-6.5 phosphate buffer solution, 200 ml, and 40 incubated for 1.5 h to obtain a liposome solution. As described in Example 1 The encapsulation efficiency of lentinan was 82.2% and the average particle size was 302 nm. Observed by transmission electron microscopy, it can be seen that the appearance of liposome 3 is spherical and the structure is clear. Example 4

 Prescription of the raw material of liposome 4:

制备： 称取 5.2 g大豆磷脂、 0.7 g胆固醇、 0.2 g聚乙二醇 2000和 0.2 g维生素 E， 加氯仿 900 ml, 搅拌使溶解， 得油相混 合物。 称取 0.1 g注射级香菇多糖和 0.7 g去氧胆酸钠， 加入注射 用水 300 ml, 并加入 0.1 mol/L氢氧化钠液适量至香菇多糖溶解， 用 0.1 mol/L盐酸调节 pH值为 7.0 ~ 8.0, 得水相混合物。 冰水浴 冷却下， 将水相混合物緩緩加入油相混合物中， 然后间歇超声乳化 约 6min， 得乳白色均一的 W/0型乳剂。 在 和氪气保护下减压 旋转蒸发除去溶剂得淡黄色浓稠胶状物， 加入 2%甘露醇水溶液 200 ml, 45 孵化 1.5 h， 用微孔滤膜过滤， 得到脂质体 4的溶液。 如实施例 2的方法制备脂质体 2的粉针剂。将其复溶后，按实施例 1的方法测得香菇多糖包封率为 75.1%， 平均粒径为 297 nm。 经 透射电镜观察， 可见脂质体 4的外观呈球状， 结构清晰。 实施例 5

Preparation: Weigh 5.2 g of soybean phospholipid, 0.7 g of cholesterol, 0.2 g of polyethylene glycol 2000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0.1 g of injection-grade lentinan and 0.7 g of sodium deoxycholate, add 300 ml of water for injection, and add 0.1 mol/L sodium hydroxide solution to dissolve the lentinan, and adjust the pH to 7.0 with 0.1 mol/L hydrochloric acid. ~ 8.0, a mixture of aqueous phases. Under ice cooling, the aqueous phase mixture was slowly added to the oil phase mixture, and then intermittently emulsified for about 6 minutes to obtain a milky white uniform W/0 type emulsion. The solvent was removed by rotary evaporation under reduced pressure under a helium atmosphere to give a pale yellow thick gel, which was then taken to a mixture of 2% mannitol aqueous solution, 200 ml, 45 incubated for 1.5 h, and filtered through a microporous membrane to obtain a solution of liposome 4. A powder injection of liposome 2 was prepared as in Example 2. After reconstitution, the encapsulation efficiency of the lentinan was 75.1% and the average particle diameter was 297 nm as measured by the method of Example 1. Observed by transmission electron microscopy, the appearance of liposome 4 was spherical and the structure was clear. Example 5

 Formulation of liposome 5

聚乙二醇 2000 0. 3 g

Polyethylene glycol 2000 0. 3 g

 Sodium deoxycholate 1. 0 g Preparation: Weigh 10. O g soybean phospholipid, 1.5 g cholesterol and 0.3 g polyethylene glycol 2000, add chloroform 900 ml, stir to dissolve, and obtain an oil phase mixture. Weigh 0. 2 g of Astragalus polysaccharide and 1.0 g of sodium deoxycholate, add 300 ml of water for injection, stir until dissolved to obtain an aqueous mixture. Under ice cooling, the aqueous phase mixture was slowly added to the oil phase mixture, and then intermittently emulsified for about 6 minutes to obtain a milky white uniform W/0 type emulsion. The solvent was removed by rotary evaporation under reduced pressure of 40 and nitrogen to give a pale yellow thick gel, and then water (200 ml of water for injection, and incubated for 1.5 h in 40 ,, filtered and sterilized by a microporous membrane, and packed into 100 pieces, ie, Astragalus polysaccharide liposome injection was obtained. The encapsulation efficiency of the polysaccharide was 75.2% and the average particle size was 293 nm. Observation by transmission electron microscopy showed that the appearance of liposome 5 was spherical and the structure was clear.

 Example 6

 Formulation of liposome 6

制备： 称取 10. 0 g大豆磷脂、 1. 5 g胆固醇和 0. 7g聚乙二醇 2000, 加氯仿 900 ml , 搅拌使溶解， 得油相混合物。 称取 0. 2 g 黄芪多糖和 0. 9g去氧胆酸钠， 加注射用水 300 ml , 搅拌至溶解， 得水相混合物。冰水浴冷却下，将水相混合物緩緩加入油相混合物 中， 然后间歇超声乳化约 6 min， 得乳白色均一的 W/0型乳剂。 在 40 和氮气保护下减压旋转蒸发除去溶剂得淡黄色浓稠胶状物，加 入含 2%甘露醇的磷酸盐緩冲溶液（pH值 6. 5 ) 200 ml , 40Ό孵化 1. 5 h， 得到脂质体 6溶液。

Preparation: Weigh 10.0 g of soybean phospholipid, 1.5 g of cholesterol and 0.7 g of polyethylene glycol 2000, add chloroform 900 ml, stir to dissolve, and obtain an oil phase mixture. Weigh 0. 2 g of Astragalus polysaccharide and 0.9 g of sodium deoxycholate, add 300 ml of water for injection, stir until dissolved to obtain an aqueous mixture. The aqueous phase mixture was slowly added to the oil phase mixture under cooling in an ice water bath, and then intermittently emulsified for about 6 minutes to obtain a milky white uniform W/0 type emulsion. 5小时,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,,, A liposome 6 solution was obtained.

A powder injection of liposome 6 was obtained as described in Example 2. After the reconstitution, the encapsulation efficiency of the astragalus polysaccharide was determined to be 84.5 %, and the average particle diameter was 308 nm. Observed by transmission electron microscopy, the appearance of liposome 6 was spherical and the structure was clear. Example 7

 Raw material prescription for Litosan 7

制备：称取 5.0 g大豆磷脂、 0.6 g胆固醇、 0.2 g聚乙二醇 2000 和 0.2 g维生素 E， 加氯仿 900 ml, 搅拌使溶解， 得油相混合物。 称取 0.1 g注射级香菇多糖和 0.4 g吐温 80，加入注射用水 300 ml, 并加入 0.1 mol/L氢氧化钠液适量至香菇多糖溶解， 用 0.1 mol/L 盐酸调节 pH值为 7.0~8.0， 得水相混合物。 水水浴冷却下， 将水 相混合物緩緩加入油相混合物中， 然后循环高压乳化约 6 min， 得 乳白色均一的 W/0型乳剂。 在 40Ό和氮气保护下减压旋转蒸发除 去溶剂得淡黄色浓稠胶状物， 加入 pH 值 6.0 ~ 6.5磷酸盐緩沖溶 液 200 ml, 40TC孵化 1.5 h， 得到脂质体 7的溶液。 按实施例 1所 述方法测得香菇多糖包封率为 80.2%， 平均粒径为 293 nm。 经透 射电镜观察， 可见脂质体 7的外观呈球状， 结构清晰。

Preparation: Weigh 5.0 g of soybean phospholipid, 0.6 g of cholesterol, 0.2 g of polyethylene glycol 2000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0.1 g of injection-grade lentinan and 0.4 g of Tween 80, add 300 ml of water for injection, and add 0.1 mol/L sodium hydroxide solution to dissolve the lentinan, and adjust the pH to 7.0-8.0 with 0.1 mol/L hydrochloric acid. , get the water phase mixture. Under the cooling of the water bath, the aqueous phase mixture was slowly added to the oil phase mixture, and then emulsified by high pressure for about 6 minutes to obtain a milky white uniform W/0 type emulsion. The solvent was removed by rotary evaporation under reduced pressure of 40 Torr under nitrogen to give a pale-yellow thick gel, which was added to a pH 6.0-6.5 phosphate buffer solution 200 ml, and 40 TC for 1.5 h to obtain a liposome 7 solution. According to the method described in Example 1, the encapsulation efficiency of Lentinus edodes polysaccharide was 80.2%, and the average particle diameter was 293 nm. Observed by transmission electron microscopy, it can be seen that the appearance of liposome 7 is spherical and the structure is clear.

 Formulation of liposome 8

吐温 80 0.4 g 制备： 称取 6.1 g大豆磷脂、 1.5 g胆固醇、 0.2 g聚乙二醇 2000和 0.2 g维生素 E， 加氯仿 900 ml, 搅拌使溶解， 得油相混 合物。 称取 0.1 g注射级香菇多糖和 0.4 g吐温 80， 加入注射用 水 300 ml, 并加入 0.1 mol/L氢氧化钠液适量至香菇多糖溶解， 用 0.1 mol/L盐酸调节 pH值为 7.0 ~ 8.0, 得水相混合物。 水水浴 冷却下， 将水相混合物緩緩加入油^混合物中， 然后用间歇超声乳 化约 6min, 得乳白色均一的 W/0型乳剂。 在 401和氮气保护下减 压旋转蒸发除去溶剂得淡黄色浓稠胶状物， 加入 2%含甘露醇的磷 酸盐緩冲液（pH值 6.0 ~ 6.5 ) 200 ml, 40Ό孵化 1.5 h, 得到脂 质体 8的溶液。 采用如实施例 2的方法制备脂质体 8的粉针剂。将 其复溶后，按实施例 1的方法测得香菇多糖包封率为 82.1%， 平均 粒径为 305 nm。 经透射电镜观察， 可见 ^质体 8的外观呈球状， 结构清晰。 实施例 9

Tween 80 0.4 g Preparation: Weigh 6.1 g of soybean phospholipid, 1.5 g of cholesterol, 0.2 g of polyethylene glycol 2000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0.1 g of injection-grade lentinan and 0.4 g of Tween 80, add 300 ml of water for injection, and add 0.1 mol/L sodium hydroxide solution to dissolve the lentinan, and adjust the pH to 7.0-8.0 with 0.1 mol/L hydrochloric acid. , get the water phase mixture. The aqueous phase mixture was slowly added to the oil mixture under cooling with a water bath, and then intermittently emulsified for about 6 minutes to obtain a milky white uniform W/0 type emulsion. The solvent was removed by rotary evaporation under reduced pressure of 401 under a nitrogen atmosphere to give a pale yellow thick gum, which was added to a 2% mannitol-containing phosphate buffer (pH 6.0-6.5), 200 ml, and incubated for 1.5 h at 40 Torr to obtain a lipid. A solution of plastid 8. A powder injection of liposome 8 was prepared as in Example 2. After reconstitution, the encapsulation efficiency of the lentinan was 82.1% and the average particle diameter was 305 nm as measured by the method of Example 1. Observed by transmission electron microscopy, it can be seen that the appearance of the plastid 8 is spherical and the structure is clear. Example 9

 Prescription of the raw material of liposome 9:

制备：称取 7.0g大豆磷月旨、 0.8 g胆固醇、 0.5 g聚乙二醇 1000 和 0.2 g维生素 E, 加氯仿 900 ml, 搅拌使溶解， 得油相混合物。 称取 0.1 g注射级香菇多糖和 0.6 g去氧胆酸钠， 加入注射用水 300 ml, 并加入 0. lmol/L氢氧化钠液适量至香菇多糖溶解， 用 0.1 mol/L盐酸调节 pH值为 7.0~8.0，得水相混合物。水水浴冷却下， 将水相混合物緩緩加入油相混合物中，然后间歇超声乳化约 6min， 得乳白色均一的 W/0型乳剂。 在 35Ό和氮气保护下减压旋转蒸发 除去溶剂得淡黄色浓稠^状物，加入 pH值 6.0-6.5磷酸盐緩冲溶 液 200 ml, 40X孵化 1.5 h， 得到脂质体溶液。 如实施例 1所述方 法测得香菇多糖包封率为 75.0%， 平均粒径为 312 nm。 经透射电 镜观察， 可见脂质体 9的外观呈球状， 结构清晰。 实施例 10

Preparation: Weigh 7.0 g of soybean phosphorus, 0.8 g of cholesterol, 0.5 g of polyethylene glycol 1000 and 0.2 g of vitamin E, add 900 ml of chloroform, stir to dissolve, and obtain an oil phase mixture. Weigh 0.1 g of injection-grade lentinan and 0.6 g of sodium deoxycholate, add 300 ml of water for injection, and add 0.1 ml of sodium hydroxide solution to dissolve the lentinan, and adjust the pH with 0.1 mol/L hydrochloric acid. From 7.0 to 8.0, a mixture of aqueous phases is obtained. The water bath is cooled, The aqueous phase mixture was slowly added to the oil phase mixture and then intermittently emulsified for about 6 minutes to obtain a milky white uniform W/0 type emulsion. The solvent was removed by rotary evaporation under reduced pressure of 35 Torr and nitrogen to give a pale-yellow thick substance, and then added to a pH 6.0-6.5 phosphate buffer solution 200 ml, and 40X incubation for 1.5 h to obtain a liposome solution. The encapsulation efficiency of lentinan was 75.0% and the average particle size was 312 nm as described in Example 1. Observed by transmission electron microscopy, it can be seen that the appearance of liposome 9 is spherical and the structure is clear. Example 10

 The vegetal liposome liposome (liposome 9) prepared as described above was used as an example to examine the immunomodulatory effects and antitumor effects of the polysaccharide liposome of the present invention. The above therapeutic effects are determined by measuring the activity of macrophages, natural killer cells (NK cells) and lymphocytes, and the tumor suppressing rate.

 Mouse tumor model preparation:

Take healthy and mature Kunming male mice, weighing 20~24g (provided by Experimental Animal Center, Tongji Medical College, Huazhong University of Science and Technology). The mice were inoculated with H ₂₂ liver cancer cells (given by the teaching and research section of Wuhan University Medical College) and passaged, and at least three generations of passage replication were performed in the laboratory. The ascites containing tumor cells was taken from the abdominal cavity of the passaged mice, diluted with RPMI1640 medium (Gibco) to a final concentration of 2× ^{10 7} /ml, and the dilution was subcutaneously injected from the right axilla of the mice to inoculate the tumors at a dose of 0.2 ml. / mice only. The average tumor weight of the model mice was greater than 0.4 g, and the tumor weight ranged from 1.13 to 3. lg.

 Experimental grouping and administration:

 According to the test drugs and the doses administered by the mice, they were divided into a tumor model group, a cyclophosphamide group, a lentinan and a cyclophosphamide combined administration group, and a combination of lentinan liposome and cyclophosphamide. The specific groupings are listed in Table 1 below. Table 1 The test drugs are divided into

Experimental group and doses of test drug preparation concentration and mode of administration

Tumor model group 5% glucose injection; 0. lmi/iog intraperitoneal injection of cyclophosphamide group (30 mg / kg) 3mg / ml cyclophosphamide; 0. lml / 10g intraperitoneal injection of lentinan + cyclophosphamide 6mg/ml cyclophosphamide + 200 g/ml lentinan; each

(1.0mg/kg + 30 mg/kg) 0.05ml/10g intraperitoneal injection Lentinus edodes liposome + cyclophosphamide 6mg / ml cyclophosphamide + 200μ ₈ / ιη1 lentinan liposome; ( 1. Omg / kg + 30 mg / kg) 0.05ml / 10g each intraperitoneal injection of liver macrophages assay phagocytic activity, administration of the active splenic lymphocyte transformation and NK cell activity assay used to assay, animals hepatoma cells were seeded the next day were randomly divided into the four groups H ₂₂ intraperitoneally administered continuously for 15 days. The mode of administration used in the inhibition of tumor inhibition rate was that the animals were randomly divided into the above four groups on the fourth day of inoculation of H ₂₂ hepatoma cells, and the abdominal cavity was continuously administered for 15 days.

 (1) Determination of macrophage activity

 After 24 hours of the last administration, the mice were sacrificed. Preparation of hepatocyte suspension: The mice were soaked in 75% alcohol for a while, the mouse liver was aseptically removed, washed 3 times with D-Hank, s solution, weighed, and then the liver of the same group of animals was mixed according to weight and volume 1 : 1 ratio was added to RPMI1640 medium (Gibco) to grout, and the final concentration of the cells was diluted to 2 X lO mK

 The hepatocyte suspension obtained above was added to each well of a 96-well microtiter plate, and cultured in a carbon dioxide incubator for 24 hours. The cells were washed three times with RPMI1640 medium (Gibco) to obtain adherent monolayer macrophages. Then, add 0.072% neutral red to each well.

(Solarbio Company) physiological saline solution ΙΟμ Ι, cultured for 4 hours, the supernatant was discarded, each well was added acetic acid - ethanol _(1: 1) 150μ1 ο 96-well plate placed in a shaker shaken for 10 minutes to completely dissolve the crystals, is set The absorbance of each well was measured at a wavelength of 492 nm on a microplate reader (ZS-3 type, Beijing Xinfeng Electric Technology Co., Ltd.), and macrophage phagocytic activity (0D value) was calculated. The experimental results were analyzed using SPSS statistical software. All data is expressed in ±s. Inter-group comparisons were performed using the t test. The results are shown in Table 2 below. Table 2 Effect of test drugs on liver phagocytic activity (^ soil s) Group 0D values

 Tumor model group 0.501 ± 0.119 cyclophosphamide group (30 mg/kg) 0.558 ± 0.128 lentinan + cyclophosphamide (1.0 mg/kg + 30 mg/kg) 0.559 ± 0.176 lentinan liposome + cyclophosphamide (1.0 mg /kg + 30

1.234 ±1.05**## mg/kg ) Note: **P<0.01 compared with cyclophosphamide group; ##P<0.01 compared with lentinan + cyclophosphamide group. The results of the experiment showed that: the cyclophosphamide group had a small difference in macrophage activity compared with the tumor model group, indicating that cyclophosphamide had little effect on the phagocytic activity of hepatic macrophages of the tumor-bearing sputum; lentinan + cyclophosphamide group Compared with the cyclophosphamide group, the phagocytic activity of macrophages was very small, indicating that the effect of lentinan on the phagocytic activity of hepatic macrophages in tumor-bearing mice was also small. Comparison of lentinan liposome + cyclophosphamide group and tumor model group Compared with cyclophosphamide group, the phagocytic activity of macrophages was significantly increased (P<0.01), indicating that lentinan liposome can significantly enhance the phagocytic activity of liver macrophages in tumor-bearing mice; lentinan liposome+loop Compared with the same dose of lentinan + cyclophosphamide group, the phagocytic activity of macrophages in tumor-bearing mice was significantly increased (P<0.01). It was also confirmed that the lentinan polysaccharide encapsulated as a liposome can significantly increase the activity of liver macrophages.

 (2) Natural killer cell test

After 24 hours of the last administration, the mice were sacrificed. Preparation of spleen cell suspension: The mice were soaked in 75% alcohol for a while, the mouse spleens were aseptically removed, washed 3 times with D-Hank, s solution, weighed, and then the spleens of the same group of animals were mixed according to weight and volume. The ratio of 1:1 was added to RPMI1640 medium (Gibco) for slurrying, and the final concentration of the cells was diluted to 2x10 ⁶ /ml.

Adding effector cells (spleen cells prepared as above) 100 μ 1 , RPMI 1640 medium (Gibco) 100 μ 1 in the well of 96-well microtiter plate; pupil plus target cells (Κ562 cells (Basic Medical College, Tongji Medical College, Huazhong University of Science and Technology) The experimental platform center provides)) 100 μl, RPMI1640 medium (Gibco) 100 μl, Ε+Τ plus effector cells 100 μl, target cells 100 μl. Each has three duplicate holes. Place in a 37-inch carbon dioxide incubator for 24 h. Add 4 mg/ml of MTT (sigma) 20 μl 4 h before termination of culture. Place in 371 C, 5% CO ₂ incubator for 4 h, then remove the supernatant, each well. 150 μl DMS0 was added. The 96-well plate was shaken in a shaker for 10 minutes until the crystals were completely dissolved. The absorbance of each well was measured at 492 nm using DMS0 as a blank solvent. Calculation of results: N cell killing rate = [1 -(D _E+T -D _B ) /D _T ] 100%. The experimental results were analyzed using SPSS statistical software. All data were expressed as ± s. The t-test was used to compare between groups. The results are shown in Table 3 below. Effect of test drug on 杀κ cell killing rate (j ± s) Group killing rate (%) Tumor model group 65.95 soil 17.05 Cyclophosphamide group (30 mg/kg) 48.97 士12.87 lentinan + cyclophosphamide (1. Omg/kg + 30

 56.19 ±23.25

 Mg/kg ) Lentinus edodes liposome + cyclophosphamide ( 1. Omg/kg +

 91.02 soil 9.156*#

 30 mg/kg)

 Note: *P<0.05 compared with the cyclophosphamide group; #P<0.05 compared with the lentinan + cyclophosphamide group. The results of this experiment showed that the killing rate of NK cells was significantly lower in the cyclophosphamide group than in the tumor model group (P < 0.05), indicating that cyclophosphamide inhibited the killing rate of NK cells in tumor-bearing mice; lentinan + cyclophosphamide Compared with the tumor model group, the NK cell killing rate was lower, but the NK cell killing rate was increased compared with the cyclophosphamide group, but it was not statistically significant, indicating that the combination of lentinan and cyclophosphamide partially eliminated the ring. The toxic side effects of phosphoramide on tumor-bearing mouse NK cells; the lentinan liposome + cyclophosphamide group compared with the tumor model group, and the NK cell killing rate were significantly increased compared with the cyclophosphamide group (P < 0.05). , indicating that the combination of lentinan liposome and cyclophosphamide can not only eliminate the toxic side effects of cyclophosphamide on NK cells of tumor-bearing mice, but also significantly improve the killing rate of spleen NK cells in tumor-bearing mice; Compared with the same dose of lentinan + cyclophosphamide group, the NK cell killing rate was significantly increased in the plastid + cyclophosphamide group (P < 0.05), indicating that the lentinan liposome and Compared mushroom polysaccharide can significantly enhance the tumor-bearing mice NK cell killing rate.

 (3) Lymphocyte proliferation activity test

A spleen cell suspension was prepared as described above. A spleen cell suspension of 100 μl was added to each well of a 96-well culture plate. Control wells and test wells were set for each test drug group. PHA-P (sigma) 100 μl (12.5 μg / ml) was added to the test well, and RPMI1640 (Gibco) ΙΟΟμ Ι was added to the control well. Incubate in a 37V, 5% CO ₂ incubator for 24h. The subsequent steps are as described in part (2) above. Calculation of results: Conversion rate = (0D test hole - 0D vs. _3⁄4 hole) / 0D vs. _3⁄4 hole > 100%. The test results are based on SPSS statistical software. Line analysis. All data is expressed in ±s. Comparisons were made between groups using the t test. The results are shown in Table 4 below. Table 4 Effect of test drug on proliferation rate of spleen lymphocytes (± s) Group proliferation rate (%) Tumor model group 29.58 ± 18.63 Cyclophosphamide group (30 mg/kg) 14.95 ± 10.90 Lentinus edodes + cyclophosphamide ( 1. Omg/kg + 30 mg/kg ) 28.58 ± 12.35 Lentinus edodes liposome + cyclophosphamide ( 1. Omg/kg + 30

 45.00 + 22.76*# mg/kg )

 Note: *P<0.05 compared with the cyclophosphamide group; #P<0.05 compared with the lentinan + cyclophosphamide group. The results of the experiment showed that the proliferation rate of spleen lymphocytes was significantly lower in the cyclophosphamide group than in the tumor model group (P < 0.05), indicating that cyclophosphamide inhibited the proliferation rate of spleen lymphocytes in tumor-bearing mice; Compared with the tumor model group, the phosphoric acid amide group had a small difference in lymphocyte proliferation rate, but the lymphocyte proliferation rate increased compared with the cyclophosphamide group, indicating that the combination of lentinan and cyclophosphamide can eliminate cyclophosphamide. The toxic side effects of spleen lymphocytes in tumor mice; the lentinan liposome + cyclophosphamide group compared with the tumor model group, and the lymphocyte proliferation rate was significantly increased compared with the cyclophosphamide group (Ρ < 0.05), indicating lentinan When combined with cyclophosphamide, liposome can not only eliminate the toxic side effects of cyclophosphamide on lymphocytes of tumor-bearing mice, but also significantly increase the proliferation rate of spleen lymphocytes in tumor-bearing mice; lentinan liposome + ring Compared with the same dose of lentinan + cyclophosphamide group, the lymphocyte proliferation rate was significantly increased in the phosphoramide group (P < 0.05), indicating that the scent Lentinan polysaccharide liposomes significantly increased as compared to tumor-bearing mouse spleen lymphocyte proliferation.

 (4) Determination of tumor inhibition rate

After 24 hours of the last administration, the mice were sacrificed. Mouse tumors were isolated, tumors were weighed, and tumor inhibition rates were calculated. The results are shown in Table 2 below. The experimental results were analyzed by SPSS statistical software. All data is represented by f soil s. Inter-group comparisons were performed using the t test. The experimental results are shown in Table 5 below. Table 5 Comparison of tumor inhibition rates of test drugs (士s) Tumor weight (g) tumor inhibition rate (%)

 Tumor model group 1. 88 ± 1. 40 - cyclophosphamide group (30 mg/kg) 0. 69 soil 0. 29 61. 40 soil 16. 48

 Lentinus edodes polysaccharide + cyclophosphamide

 0. 54士 0. 28 69. 89 soil

 ( 1. Omg/kg + 30 mg/kg )

 Lentinus edodes liposome + cyclophosphamide

 0. 46土 0. 20 75. 22 soil 10· 75*

 ( 1. Omg/kg + 30 mg/kg )

 Compared with cyclophosphamide, *P<0.05. According to the "Guidelines for the Pharmacodynamics of Antitumor Drugs", the evaluation criteria were: 40% inhibition rate was ineffective, tumor inhibition rate was > 40% and statistically processed P < 0 05 (compared to the tumor model group) is an effective evaluation criterion.

 The experimental results showed that: the inhibition rate of lentinan and cyclophosphamide group was higher than that of cyclophosphamide group, indicating that lentinan can enhance the inhibitory effect of cyclophosphamide, but the difference was not statistically significant; Compared with the cyclophosphamide group, the tumor inhibition rate was significantly increased in the plastid + cyclophosphamide group (P < 0.05), indicating that the lentinan liposome can significantly enhance the tumor suppressive effect of cyclophosphamide. This proves that the lentinan liposome has superior antitumor activity over lentinan.

 o

Claims

Rights request A polysaccharide liposome characterized in that the raw material for preparing the lipid body comprises a polysaccharide, a phospholipid, a cholesterol, a surfactant, and polyethylene glycol.  The liposome according to claim 1, characterized in that the polysaccharide is lentinan, sassafras polysaccharide, tremella polysaccharide, trehalose, ginseng polysaccharide, ganoderma lucidum polysaccharide, porcine polysaccharide, or a mixture thereof.  The liposome according to claim 1, characterized in that the polysaccharide is lentinan or sassafras polysaccharide.  The liposome according to any one of claims 1 to 3, characterized in that the phospholipid is soybean phospholipid, lecithin, hydrogenated soybean phospholipid, hydrogenated lecithin, phosphatidylethanolamine, distearoylphosphatidylcholine, poly Ethylene glycol derivatized phospholipids, or mixtures thereof.  The liposome according to any one of claims 1 to 3, characterized in that the surfactant is deoxycholate, poloxamer, Tween 80, or a mixture thereof.  6. Liposomes according to claim 5, characterized in that the surfactant is sodium deoxycholate or potassium deoxycholate.  The liposome according to any one of claims 1 to 3, wherein the polyethylene glycol is selected from the group consisting of polyethylene glycol 800-5000, phospholipidized polyethylene glycol derivatives, and mixtures thereof.  8. The liposome of claim 7, characterized in that the polyethylene glycol is polyethylene glycol 1000-2400.  9. The liposome of claim 8, characterized in that the polyethylene glycol is polyethylene glycol 2000.  The liposome according to any one of claims 1 to 3, characterized in that the raw material further comprises an antioxidant, and/or a lyoprotectant.  The liposome according to any one of claims 1 to 3, characterized in that the raw material comprises 1 part of polysaccharide, 30-100 parts of phospholipid, 2-20 parts of cholesterol, 3- 12 parts of surfactant, polyethylene glycol 1-8 servings.  The liposome according to claim 11, characterized in that the raw material has a weight ratio of 1 part of polysaccharide, 40-90 parts of phospholipids, and 5-15 parts of cholesterol. The liposome according to claim 11, wherein the raw material has a weight ratio of 1 part of polysaccharide, 50-70 parts of phospholipid, and 5-15 parts of cholesterol. 14. The liposome of claim 12, wherein the antioxidant is from 1 to 8 parts.  The liposome according to any one of claims 1 to 3, characterized in that the liposome has an entrapment ratio of 65% or more.  The lipopolysaccharide according to any one of claims 1 to 3, wherein the liposome has an encapsulation ratio of 70% or more.  The liposome according to any one of claims 1 to 3, wherein the liposome has an encapsulation efficiency of 80% or more.  The liposome according to any one of claims 1 to 3, wherein the liposome has a particle diameter of 1 μηη or less.  The liposome according to any one of claims 1 to 3, characterized in that the liposome has a particle diameter of from 100 to 500 nm.  The liposome according to any one of claims 1 to 3, characterized in that the liposome has a particle diameter of 140 300 nm.  The liposome according to any one of claims 1 to 3, characterized in that the liposome is present in the form of an injection or a powder injection.  22. A composition comprising a liposome according to any one of claims 1-21.  23. The composition of claim 22, characterized in that said composition further comprises a pharmaceutically acceptable carrier.  24. The composition of claim 22 or 23, characterized in that the composition further comprises one or more anti-tumor drugs or immunomodulatory drug active ingredients.  25. Composition according to claim 22 or 23, characterized in that the composition is present in the form of an injection or a powder injection.  26. A method of preparing the liposome of claims 1-21, comprising the steps of: a. dissolving a feedstock comprising phospholipids, cholesterol and polyethylene glycol in an organic solvent to provide an oil phase mixture;  b. dissolving the raw material comprising the polysaccharide and the surfactant in water or an aqueous solution to obtain an aqueous phase mixture;  c injecting the aqueous phase mixture into the oil phase mixture and emulsification;  d. The solvent is removed by evaporation under constant pressure under reduced pressure in a protective gas;  e. Add water or aqueous solution and incubate in a water bath. 27. The method of claim 26, wherein the method further comprises the steps of: f. The solution obtained in step e is filtered and then freeze-dried.  28. Process according to claim 26 or 27, characterized in that the organic solvent of step a is chloroform or chloroform-methanol.  29. Process according to claim 25 or 26, characterized in that the emulsification treatment of step c is an intermittent sonication or a cyclic high pressure emulsification treatment.  30. The method of claim 26 or 27, characterized in that the water bath incubation of step e is carried out at 35-501C.  31. The method of claim 30, characterized in that the water bath incubation of step e is carried out at 40.  32. The method of claim 26 or 27, wherein the step b is inconsistent with the pH of the e-step reaction environment.  33. The method of claim 26 or 27, wherein the step b is performed in opposition to the acidity and alkalinity of the e-step reaction environment.  34. The method of claim 33, wherein the step of incubating in the water bath in the step e is 1-2 h.  35. The method of claim 26 or 27, wherein said starting material of said step a further comprises an antioxidant.  36. The method of claim 26 or 27, wherein said aqueous solution of step e comprises a lyoprotectant.  37. Use of a liposome according to any one of claims 1 to 21 for the manufacture of a medicament for anti-tumor or for regulating immunity of the body.  38. Use of a composition according to any one of claims 22 to 25 for the manufacture of a medicament for anti-tumor or for regulating immune function.  39. A method of treating a tumor or modulating immune function comprising administering to a subject a liposome according to any one of claims 1-21.  40. A method of treating a tumor or modulating immune function, comprising administering to a subject a composition according to any one of claims 22-25.