CN103006560B - Hyaluronic acid oligosaccharide encased paclitaxel liposome and preparation method thereof - Google Patents

Abstract

The invention relates to a preparation method of paclitaxel liposomes wrapped in hyaluronic acid oligosaccharides, comprising the following steps: weighing phospholipids and paclitaxel to prepare a phospholipid-paclitaxel honeycomb film; hydrating the prepared film; Pre-incubation of sugar and ethyldimethylaminopropylcarbodiimide in acetate buffer; add the hydrated suspension to oligosaccharide-acetate buffer and incubate; the obtained The mixture was separated to prepare paclitaxel liposomes coated with oligosaccharides. The paclitaxel liposome encapsulated by the oligosaccharide has uniform size, high encapsulation efficiency and stable property, and effectively improves the solubility of paclitaxel in water. In addition, for tumor cells with high expression of CD44 (such as breast cancer, etc.), the complex has strong targeting (hyaluronic acid specifically binds to CD44), can effectively increase the concentration of paclitaxel in the tumor site, and reduce the dosage of paclitaxel , while reducing the toxic effect of the drug on the human body and enhancing its bioavailability.

Description

Paclitaxel liposome encapsulated by hyaluronic acid oligosaccharide and preparation method thereof

technical field

The invention relates to a liposome, in particular to a hyaluronic acid oligosaccharide-wrapped paclitaxel liposome and a preparation method thereof.

Background technique

Tumor treatment is an unsolved problem today and a major challenge that the medical community has been facing. Drug therapy is an important means of tumor treatment. Although traditional anti-tumor drugs have achieved certain curative effects, their allergic reactions, inhibition of bone marrow hematopoietic system, hair loss and other toxic and side effects greatly limit their clinical application and become the mainstay of clinical treatment. One of the puzzles. How to improve the curative effect of tumor treatment is an urgent problem to be solved in modern medicine. In response to this shortcoming, targeted therapy has become a research hotspot in tumor therapy, and the use of nanometer as a targeting carrier has become the focus of research by medical scholars.

The so-called targeted drug therapy is to make the drug target the tumor site, store a relatively high concentration in the local area, prolong the time of the drug, and improve the lethality of the tumor, but has little effect on normal tissue cells. At present, the drugs used for tumor targeted therapy include chemotherapy drugs (such as slow-release chemotherapy drugs, liposome chemotherapy drugs), chemical ablation drugs (such as absolute ethanol, glacial acetic acid, hydrochloric acid, sulfuric acid and other protein coagulants), genes and Molecular targeted drugs, traditional Chinese medicine, etc. Drugs can be administered through multiple routes, such as percutaneous puncture administration, intraoperative administration, local tumor administration under endoscopy or laparoscopy, vascular interventional administration, drug percutaneous ultrasound conduction therapy, intracavitary drug infusion, etc. At the same time, new treatment methods such as cryochemotherapy, thermochemotherapy, radiochemotherapy and synchronous chemotherapy have been derived, which greatly relieve the pain of patients and significantly improve the curative effect.

Paclitaxel (PTX) is a commonly used broad-spectrum antineoplastic drug, which is extracted from Taxus genus plants. It has a unique anticancer mechanism and belongs to a new type of anti-microtubule drug. The protein is stable, inhibits cell mitosis, and has a certain effect on various advanced cancers such as ovarian cancer, breast cancer, and lung cancer. However, due to its extremely low solubility in water, it cannot be absorbed when taken orally. Currently, PTX injection in clinical use uses polyoxyethylene castor oil and absolute ethanol as solubilizing carriers, but it causes various toxic and side effects when used, among which allergic reactions are the most serious , coupled with its lack of targeting, the dosage is relatively large, and its toxic and side effects are further increased, so that its application is severely limited.

In view of the poor water solubility and lack of targeting of PTX, in the prior art, PTX is made into liposomes. Liposome is a new type of nano-drug carrier, which has the advantages of high efficiency and stability in drug delivery. It is a molecularly ordered assembly spontaneously formed by phospholipids in water by hydrophobic association. It is a multilayered vesicle structure, hydrophobic Sexual drugs are embedded in the hydrophobic bilayer of liposomes through hydrophobic interaction. Using liposomes as carriers can not only reduce the hypersensitivity of the body, but also increase the solubility of hydrophobic drugs in water and reduce the dosage of drugs. In recent years, great progress has been made in the connection of nanoparticles with targeting molecules for tumor-targeted therapy.

For example, Chinese invention patent application number 200610137900.6 (publication number 101176719A), the invention provides a paclitaxel docetaxel liposome combination drug and its preparation method, the drug provided can reduce the drug dosage by nearly one time, and the curative effect can be increased by 15% Above, the toxic and side effects of medicine are greatly reduced.

Another example is the Chinese invention patent application number 200710010448.1, which provides a paclitaxel liposome and its preparation method, which solves the water-insoluble defect of paclitaxel, and paclitaxel heat-sensitive long-circulation liposome not only has the low toxicity of paclitaxel common liposome preparations , high stability, and reduced toxic and side effects. It also has the long circulation of long-circulation liposomes and the heat sensitivity of heat-sensitive liposomes, which can prolong the circulation time of paclitaxel in the body and improve the targeting of tumors, thereby Improve its anti-tumor effect.

However, there are no reports about the preparation of PTX nanomaterials from hyaluronan oligosaccharides (oHA) at home and abroad. As a target molecule, oHA targets CD44 on the surface of tumor cells to effectively kill tumor cells without affecting normal cells. Compared with ordinary liposomes, it has the characteristics of strong stability and high encapsulation efficiency, and is an ideal antitumor drug. Therefore, if the potential of oHA-PTX-Lipid to target tumors is developed, it will be beneficial to tumor treatment have a huge impact.

Contents of the invention

In view of the disadvantages of poor water solubility and lack of targeting of PTX in the prior art, the object of the present invention is to provide a hyaluronic acid oligosaccharide-coated paclitaxel liposome (oHA-PTX-Lipid) and a preparation method thereof. oHA-PTX-Lipid can effectively increase the amount of PTX dissolved in water, target the CD44 receptor on the surface of tumor cells through oHA, effectively increase its concentration in the tumor site, reduce the dosage of PTX, and enhance its bioavailability.

According to another aspect of the present invention, there is provided a preparation method of hyaluronic acid oligosaccharide-coated paclitaxel liposomes, that is, at first by film hydration method, phospholipids are used to prepare drug-loaded liposomes, and then, it is mixed with oHA Incubate overnight, fully dialyze with double-distilled water, and homogenize with a high-pressure homogenizer to obtain paclitaxel liposome oHA-PTX-Lipid coated with hyaluronic acid oligosaccharides. This method is simple and easy to operate, and the obtained oHA-PTX- Lipid has uniform size, high encapsulation efficiency and stable properties.

Specifically, the present invention provides a method for preparing paclitaxel liposomes encapsulated by hyaluronic acid oligosaccharides, comprising the following steps:

Step 1, weighing phospholipids and paclitaxel to prepare a phospholipid-paclitaxel honeycomb film, wherein the mass ratio of phospholipids to paclitaxel is (20-50) : 1;

Step 2, hydrating the prepared film so that the mass percent concentration of phospholipids is 1% to 5%;

Step 3, pre-incubating hyaluronic acid oligosaccharides and ethyldimethylaminopropylcarbodiimide in acetate buffer solution with a pH value of 3~6 at 35~50°C to obtain hyaluronic acid oligosaccharides Sugar-acetate buffer;

Step 4, adding the suspension obtained in step 2 to the hyaluronic acid oligosaccharide-acetate buffer solution obtained in step 3, and incubating at 35-50°C;

Step 5, separating the mixture obtained in step 4 to prepare paclitaxel liposomes coated with hyaluronic acid oligosaccharides;

Among them, the mass ratio of phospholipids, paclitaxel, hyaluronic acid oligosaccharides and ethyldimethylaminopropyl carbodiimide is (20~50) : 1 : (0.25~1) : (2.5~10), transparent The mass-volume ratio of oligosaccharides to acetate buffer is (2~8) g : 1 mL.

Wherein, the method that can adopt described honeycomb thin film is as: phospholipid and paclitaxel are dissolved in solvent, and preferably dissolve in volatile solvent, then solvent volatilization makes the paclitaxel liposome that hyaluronic acid oligosaccharide encapsulates .

The volatile solvent can be selected from at least one of chloroform, ethanol, dichloromethane, ethyl acetate, acetone, tert-butyl ether, n-heptane, diethyl ether, methanol, toluene, benzene and the like.

Preferably, the volatile solvent is a mixture of chloroform and methanol, and the volume ratio of the two is 65 : 35.

Preferably, in step 1, the phospholipids are selected from the group consisting of dioleoyl phosphatidyl choline, dioleoyl phosphatidyl choline (DOPC), dioleoyl phosphatidyl olamine, dioleoyl phosphatidyl olamine, dioleoyl phosphatidyl olamine, At least one of phosphatidylglycerol, dialkanoylphosphatidylamino acid, and the like.

Preferably, the di-C ₁₀ -C ₂₀ alkanoyl phosphatidylcholine is di-C 10-C 20 alkanoyl phosphatidyl choline, such as di-C ₁₂ alkanoyl phosphatidyl choline (DLPC), di-C ₁₄ alkanoyl phosphatidyl choline ( DMPC), di-C ₁₆ alkanoylphosphatidylcholine (DPPC), di-C ₁₈ alkanoylphosphatidylcholine (DSPC), etc.

Preferably, the dialkanoylphosphatidylethanolamine is dialkanoylphosphatidylethanolamine.

Preferably, the di-C ₁₀ -C ₂₀ alkanoyl phosphatidylethanolamine, such as di-C ₁₂ alkanoyl phosphatidylethanolamine (also known as dilauroyl phosphatidylethanolamine, DLPE), di-C ₁₄ Alkanoylphosphatidylethanolamine (DMPE), diC ₁₈ alkanoylphosphatidylethanolamine (DSPE), etc.

Preferably, the dioleoylphosphatidylethanolamine is dioleoylphosphatidylethanolamine (DOPE).

Preferably, the dialkanoylphosphatidylglycerol is di-C ₁₀ -C ₂₀ alkanoylphosphatidylglycerol, such as di-C ₁₂ alkanoylphosphatidylglycerol (also known as dilauroylphosphatidylglycerol, DLPG).

Preferably, the dialkanoylphosphatidyl amino acid is dialkanoylphosphatidylserine.

Preferably, the dialkanoylphosphatidylserine is diC ₁₀ -C ₂₀ alkanoylphosphatidylserine, such as diC ₁₈ alkanoylphosphatidylserine (DSPS).

Preferably, the phospholipid is a mixture of DLPE and DLPG, wherein the mass ratio of DLPE and DLPG is (9˜1) : 1.

Preferably, in step 2, hydration is performed for at least 2 hours.

Preferably, in step 2, the film is spin hydrated at 45° C. for at least 2 h.

Preferably, in step 3, the pH of the acetate buffer is 4.5.

Preferably, the pre-incubation in step 3 is at least 1~3h, and the incubation in step 4 is at least 4~12h.

Preferably, the pre-incubation in step 3 is at least 2 hours, and the incubation in step 4 is at least 8 hours.

Preferably, the pre-incubation temperature in step 3 is 37°C, and the incubation temperature in step 4 is 37°C.

Preferably, in step 5, the mixture obtained in step 4 is dialyzed, homogenized, and dried to prepare paclitaxel liposomes coated with hyaluronic acid oligosaccharides.

Preferably, the dialysis specifically includes: transferring the mixture obtained in step 4 to a dialysis bag with a molecular weight cut-off of 10,000, and fully dialysis with double distilled water for 24 hours.

Preferably, the homogenization specifically includes: using a high-pressure homogenizer for homogenization, the pressure is 15,000-20,000 PSI, and the number of cycles is 5-10 times.

The present invention also provides a hyaluronic acid oligosaccharide-coated paclitaxel liposome prepared by the above preparation method.

Preferably, the content of paclitaxel in the paclitaxel liposome encapsulated by hyaluronic acid oligosaccharides is 2-5 wt%, and the content of hyaluronic acid oligosaccharides is 0.8-1.2 wt%.

Preferably, the content of the hyaluronic acid oligosaccharides is 1 wt%.

The paclitaxel liposomes encapsulated by hyaluronic acid oligosaccharides provided by the invention have uniform size, high encapsulation efficiency, stable properties, and can be stored for a long time to overcome the shortcomings of PTX such as poor water solubility and lack of targeting, and can be used for breast cancer and ovarian cancer , targeted therapy for malignant tumors with high expression of CD44 such as head and neck cancer, can effectively improve the bioavailability of PTX. Experimental research results show that the oHA-PTX-Lipid prepared by the present invention exhibits good anti-tumor effects both in vitro and in vivo, and compared with traditional PTX, its anti-tumor effect is significantly improved.

Detailed ways

The technical solutions of the present invention will be further described below through specific examples, so as to better understand the present invention.

Example 1

Weigh 820mg DLPE, 380mg DLPG and 40mg PTX, add 20ml organic solvent to dissolve in a 250ml round bottom flask, mix well at 50°C for 10-20min, vacuum-dry for 4h, freeze-dry at -80°C for 2h Water hydration, 45°C, 150r/min, at least 2h; at the same time, dissolve 10mg hyaluronic acid oligosaccharides (oHA) and 200mg ethyldimethylaminepropylcarbodiimide (EDAC) in 5ml acetate buffer solution, 37°C, 150 r/min, pre-incubated for 2 hours; add the suspension after hydration to the buffer after pre-incubation, and incubate overnight at 37°C, 150 r/min; then dialyze with double distilled water for 24 hours; Homogenize under high pressure, with a pressure of about 15,000, and cycle 10 times; filter with a 0.22 μm filter in an ultra-clean bench; sub-package, freeze overnight at -80°C, and drain into powder to prepare paclitaxel liposomes wrapped in hyaluronic acid oligosaccharides ( oHA-PTX-Lipid).

The average particle size of the oHA-PTX-Lipid prepared in this example is about 120nm, with uniform size and stable properties. The content of PTX measured by HPLC method is 3wt%, and the content of oHA measured by carbazole method is 1wt%. It acted on breast tumor cell MDA-MB-231, and its killing effect on breast tumor cells was detected. The results showed that its killing effect was enhanced by 2 times compared with the same concentration of simple PTX-commercially available drug Taxol, indicating that this The oHA-PTX-Lipid prepared in the example achieves the effect of targeting and killing tumor cells, reduces the dosage of PTX, and effectively improves the bioavailability of PTX.

Example 2

Weigh 1440.3mgDLPE, 175.0mgDLPG and 50.0mgPTX, add 15ml of organic solvent to dissolve in a 250ml round bottom flask, mix well at 50°C for 10-20min, vacuum dry for 2h, freeze-dry at -80°C for 2h, freeze-dry overnight, then distill with 35ml Water hydration, 45°C, 150 r/min, at least 2h; at the same time, 14mg oHA and 240mg EDAC were dissolved in 7ml acetate buffer, 37°C, 150r/min, pre-incubated for 2h; The turbid solution was added to the buffer after pre-incubation, incubated overnight at 37°C, 150 r/min; dialyzed in double distilled water for 24 hours; homogenized under high pressure, with a pressure of about 15,000, circulated 10 times, filtered through a 0.22 μm filter in an ultra-clean bench, and separated frozen overnight at -80°C, and dried into a powder to obtain paclitaxel liposomes (oHA-PTX-Lipid) encapsulated by hyaluronic acid oligosaccharides.

The average particle size of the oHA-PTX-Lipid prepared in this example is about 120nm, with uniform size and stable properties. The content of PTX measured by HPLC method is 5wt%, and the content of oHA measured by carbazole method is 1wt%. It acts on breast tumor cell MDA-MB-231, and detects its killing effect on breast tumor cell. The results show that: compared with the same concentration of simple PTX-the commercially available drug Taxol, its killing effect is enhanced by 2 times, indicating that the oHA-PTX-Lipid prepared in this example has achieved the effect of targeting and killing tumor cells, reducing the use of PTX dose, effectively improving the bioavailability of PTX.

Example 3

Weigh 600.0mg DLPE, 600.0mg DLPG and 40.0mg PTX, add 15ml organic solvent to dissolve in a 250ml round bottom flask, mix at 50°C for 10-20min, vacuum-dry for 2h, freeze-dry at -80°C for 2h, freeze-dry overnight, use Hydrate with 35ml double distilled water, 45℃, 150 r/min, at least 2h; at the same time, dissolve 14mg oHA and 240mg EDAC in 7ml acetate buffer, 37℃, 150r/min, pre-incubate for 2h; After completion, the suspension was added to the pre-incubation buffer, incubated overnight at 37°C, 150 r/min; double-distilled water dialysis for 24 hours, high-pressure homogenization, pressure about 15,000, 10 cycles, ultra-clean bench 0.22μm filter Filtrate, subpackage, freeze at -80°C overnight, and drain into powder to prepare paclitaxel liposomes encapsulated by hyaluronic acid oligosaccharides.

The average particle size of the oHA-PTX-Lipid prepared in this example is about 120nm, with uniform size and stable properties. The content of PTX calculated by HPLC method is 2wt%, and the content of oHA measured by carbazole method is 1wt%. It acts on breast tumor cell MDA-MB-231, and detects its killing effect on breast tumor cell. The results show that: compared with the same concentration of simple PTX-the commercially available drug Taxol, its killing effect is enhanced by 2 times, indicating that the oHA-PTX-Lipid prepared in this example has achieved the effect of targeting and killing tumor cells, reducing the use of PTX dose, effectively improving the bioavailability of PTX.

Example 4

Weigh 800.0mg DLPE, 600.0mg DLPG and 40.0mg PTX, add 15ml of organic solvent to dissolve in a 250ml round bottom flask, mix well at 50°C for 10-20min, vacuum dry for 2h, freeze-dry at -80°C for 2h, freeze-dry overnight, use Hydrate with 35ml double distilled water, 45℃, 150 r/min, at least 2h; at the same time, dissolve 16mg oHA and 280mg EDAC in 8ml acetate buffer, 37℃, 150r/min, pre-incubate for 2h; After completion, the suspension was added to the pre-incubation buffer, incubated overnight at 37°C, 150 r/min; double-distilled water dialysis for 24 hours, high-pressure homogenization, pressure about 15,000, 10 cycles, ultra-clean bench 0.22μm filter Filtrate, subpackage, freeze at -80°C overnight, and drain into powder to prepare paclitaxel liposomes encapsulated by hyaluronic acid oligosaccharides.

Example 5

Weigh 900.0mg DLPE, 600.0mg DLPG and 40.0mg PTX, add 15ml organic solvent to dissolve in a 250ml round bottom flask, mix at 50°C for 10-20min, vacuum-dry for 2h, freeze-dry at -80°C for 2h, freeze-dry overnight, use Hydrate with 35ml double distilled water, 45℃, 150 r/min, at least 2h; at the same time, dissolve 20mg oHA and 300mg EDAC in 10ml acetate buffer, 37℃, 150r/min, pre-incubate for 2h; After completion, the suspension was added to the pre-incubation buffer, incubated overnight at 37°C, 150 r/min; double-distilled water dialysis for 24 hours, high-pressure homogenization, pressure about 15,000, 10 cycles, ultra-clean bench 0.22μm filter Filtrate, subpackage, freeze at -80°C overnight, and drain into powder to prepare paclitaxel liposomes encapsulated by hyaluronic acid oligosaccharides.

The oHA-PTX-Lipid prepared in Examples 4-5 is uniform in size and stable in properties, and the experimental results show that Examples 4-5 act on breast tumor cells MDA-MB-231 and can also achieve targeted killing of tumor cells. Effect, reducing the dosage of PTX, effectively improving the bioavailability of PTX.

Liposome is a new type of nano-drug carrier. It has the advantages of high efficiency and stability in drug delivery. It is a molecularly ordered assembly spontaneously formed by phospholipids in water by hydrophobic association. It is a multilayered vesicle structure, hydrophobic Sexual drugs are embedded in the hydrophobic bilayer of liposomes through hydrophobic interaction. Using liposomes as carriers can not only reduce the hypersensitivity of the body, but also increase the solubility of hydrophobic drugs in water and reduce the dosage of drugs. The liposome in the oHA-PTX-Lipid of the present invention can enhance the dispersion of PTX in water, and the oHA connected to the liposome can target and bind to the CD44 receptor on the cell surface. CD44 receptor is a transmembrane glycoprotein receptor widely distributed on the cell surface, belonging to the adhesion factor family, mediating cell and extracellular matrix adhesion, lymphocyte homing and many other physiological and pathological processes. Studies have found that CD44 receptor is highly expressed on many tumor cells, and its main ligand is hyaluronic acid (HA). oHA is a degradation product of HA, which has a similar uronic acid structure to HA, but the number of disaccharide units in oHA is less, and oHA also has the ability to specifically bind to CD44. However, compared with HA, oHA has the advantages of biological functions such as low metabolic clearance rate and removing the sugar coat on the surface of tumor cells. At the same time, oHA has higher solubility than HA, and is superior to HA in technical production as a target molecule. Based on the fact that CD44 on the surface of tumor cells can bind to oHA, oHA can become a targeting ligand for treating tumor cells with high expression of CD44. The use of oHA to carry chemotherapy drugs can reduce the tumor size in animal models of CD44 high expression tumors, increase the drug concentration at the tumor site, and achieve the effect of targeted therapy for tumors. In recent years, great progress has been made in the connection of nanoparticles with targeting molecules for tumor-targeted therapy. The oHA on the surface of the liposome of the present invention can directly target the CD44 receptor on the surface of tumor cells, remove the HA sugar coat on the surface of tumor cells, increase the concentration of PTX at the tumor site, enhance its killing activity and bioavailability, and open up a new path for tumor targeting therapy new way. At present, there are no reports on the preparation of PTX nanomaterials from oHA at home and abroad.

Moreover, oHA in the oHA-PTX-Lipid of the present invention acts as a target molecule to target CD44 on the surface of tumor cells to effectively kill tumor cells without affecting normal cells. Compared with ordinary liposomes, it has the characteristics of strong stability and high encapsulation efficiency, and is an ideal antitumor drug. The oHA-PTX-Lipid of the present invention has the potential to enter clinical application as a targeted antitumor drug, and can produce a great breakthrough in the aspect of tumor treatment.

The specific embodiments of the present invention have been described in detail above, but they are only examples, and the present invention is not limited to the specific embodiments described above. For those skilled in the art, any equivalent modifications and substitutions to the present invention are also within the scope of the present invention. Therefore, equivalent changes and modifications made without departing from the spirit and scope of the present invention shall fall within the scope of the present invention.

Claims (10)

1. a preparation method for the Paclitaxel liposome of hyaluronic acid oligosaccharide parcel, is characterized in that, comprises the following steps:

Step 1, takes phospholipid and paclitaxel, prepares the cellular thin film of phospholipid-paclitaxel;

Step 2, by the thin film aquation making, making the mass percentage concentration of phospholipid is 1% ~ 5%, obtains suspension;

Step 3,35 ~ 50 DEG C of preincubates in the acetate buffer that is 3 ~ 6 at pH value by hyaluronic acid oligosaccharide and ethyl dimethyl amine propyl carbodiimide diimine, obtain oligosaccharide-acetate buffer;

Step 4, the suspension that step 2 is obtained joins in hyaluronic acid oligosaccharide-acetate buffer that step 3 obtains, and hatches for 35 ~ 50 DEG C;

Step 5, the mixture separation that step 4 is obtained, makes the Paclitaxel liposome that hyaluronic acid oligosaccharide wraps up;

Wherein, the mass ratio of phospholipid, paclitaxel, hyaluronic acid oligosaccharide and ethyl dimethyl amine propyl carbodiimide diimine is (20 ~ 50) :1 :(0.25 ~ 1) :(2.5 ~ 10), the mass volume ratio of hyaluronic acid oligosaccharide and acetate buffer is (2 ~ 8) g :1mL.

2. preparation method according to claim 1, is characterized in that, in step 1, phospholipid and paclitaxel is dissolved in solvent, and then solvent evaporates makes the cellular thin film of phospholipid-paclitaxel.

3. preparation method according to claim 1; it is characterized in that; in step 1, described phospholipid is selected from least one in dioxane acyl phospholipids phatidylcholine, dioleyl phosphatidyl choline, dioxane acyl phospholipids acyl hydramine, dioleoyl phosphatidyl hydramine, dioxane acyl phospholipids acyl glycerol and dioxane acyl phospholipids amic acid.

4. preparation method according to claim 1, it is characterized in that, described phospholipid is the mixture of two lauroyl PHOSPHATIDYL ETHANOLAMINE and PE, and wherein, the mass ratio of two lauroyl PHOSPHATIDYL ETHANOLAMINE and PE is (9 ~ 1) :1.

5. preparation method according to claim 1, is characterized in that, in step 3, the pH value of described acetate buffer is 4.5.

6. preparation method according to claim 1, is characterized in that, preincubate at least 1 ~ 3h in step 3 is hatched in step 4 as at least 4 ~ 12h.

7. preparation method according to claim 1, is characterized in that, in step 3, preincubate temperature is 37 DEG C, and in step 4, incubation temperature is 37 DEG C.

8. preparation method according to claim 1, is characterized in that, in step 5, and the mixture dialysis that step 4 is obtained, homogenizing, dry, make the Paclitaxel liposome that oligosaccharide wraps up.

9. the Paclitaxel liposome of the hyaluronic acid oligosaccharide parcel that according to claim 1 prepared by preparation method.

10. the Paclitaxel liposome of hyaluronic acid oligosaccharide according to claim 9 parcel, is characterized in that, shown in the Paclitaxel liposome of hyaluronic acid oligosaccharide parcel the content of paclitaxel be 2 ~ 5wt%, hyaluronic acid oligosaccharide content is 0.8 ~ 1.2 wt%.