Liposome with daucosterol or daucosterol analogue as membrane material as well as preparation method and application of liposome

Abstract

The invention belongs to the technical field of medicines, and particularly relates to a liposome taking daucosterol or an analogue thereof as a membrane material as well as a preparation method and application of the liposome. The daucosterol lipidosome prepared by the invention is small in particle size, relatively good in cell entry ability and good in blood long-circulation effect and drug delivery ability, and meanwhile, the daucosterol lipidosome carrying different drugs is good in stability and dispersity and high in drug encapsulation efficiency, is beneficial to effective delivery and release of the drugs, can be used for safely and efficiently treating diseases such as tumors and the like, and has a good application prospect. Wide application prospects are realized.

Description

Technical Field

[0001] The invention belongs to the field of medical technology, and in particular relates to a liposome using carotene or its analogues as membrane material, and a preparation method and application thereof. Background Art

[0002] Liposomes are artificial vesicular structures composed of monolayer or multilayer membranes with closed compartments. Most typically, liposome membranes are composed of lipid bilayers, but they can also be composed of other amphiphilic compounds, including other types of amphiphiles, polymers, and peptides. Liposomes form spontaneously when lipids are dispersed in an aqueous environment under appropriate conditions. Liposomes have molecular characteristics of mammalian cell membranes and are non-toxic, non-antigenic, and biodegradable.

[0003] Liposomes have become an important tool for delivering drugs in the pharmaceutical industry. Lipophilic or amphiphilic drugs and compounds can be incorporated into the liposome membrane, and hydrophilic drugs and compounds can also be encapsulated in the hydrophilic core of the liposome. Liposomes can change the in vivo distribution of encapsulated drugs, so that the drugs are mainly accumulated in the target tissues, thereby improving the therapeutic index of the drugs, reducing the therapeutic dose of the drugs and reducing the toxicity of the drugs. When the drug is present in the lipid bilayer, the drug is easily released suddenly, thereby significantly reducing its blood circulation time and targeted distribution. Therefore, for amphiphilic drugs or water-soluble drugs, it can be considered to encapsulate the drugs in the inner aqueous phase of the liposomes by active drug loading. In addition, since liposomes can adsorb opsonins in plasma, liposomes often passively concentrate in organs such as the liver and spleen, thereby reducing their blood circulation time and targeted drug delivery capabilities. In order to increase the blood circulation time of liposomes, PEGylated phospholipids are usually added to liposomes. However, the addition of PEGylated phospholipids will significantly reduce the cellular entry ability of liposomes and reduce the drug delivery ability within cells. In addition, the introduction of PEGylated phospholipids activates complement, resulting in accelerated blood clearance of liposomes after multiple administrations.

[0004] Therefore, in order to increase the stability of drugs, prolong their circulation time in the body, and treat diseases safely and effectively, it is crucial to find a liposome with strong intracellular ability and good drug delivery capability. Summary of the invention

[0005] In view of the problems of the prior art, the present invention provides a liposome using carotene or its analogues as membrane material, and a preparation method and application thereof.

[0006] Use of carotene or its analogues as membrane material in preparing liposomes.

[0007] The present invention also provides a blank liposome with carotene or its analogues as membrane material, wherein the blank liposome has a membrane, and the membrane is made of the following raw materials in parts by weight:

[0008] 80-120 parts of lipid substances,

[0009] 15-40 parts of carotene or its analogues.

[0010] Preferably, the raw materials further include at least one of cholesterol and distearoylphosphatidylethanolamine-polyethylene glycol 2000.

[0011] Preferably, the lipid substance is at least one of natural phospholipids, semi-synthetic phospholipids, and fully synthetic phospholipids; the natural phospholipids are at least one of natural phosphatidylcholine, sphingomyelin, glycerophospholipids, soybean phosphatidylcholine, egg yolk phosphatidylcholine, and cephalin; the semi-synthetic phospholipids or fully synthetic phospholipids are at least one of phosphatidylcholine phospholipids, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine phospholipids, phosphatidylglycerol, dicetyl phosphate, PEG-modified phospholipids, and succinic acid cholesterol ester;

[0012] The phosphatidylcholine phospholipid is at least one of hydrogenated soybean lecithin, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, dilauroylphosphatidylcholine, dioleoylphosphatidylcholine, phosphatidylcholine, monopalmitoylphosphatidylcholine and glycerylphosphatidylcholine;

[0013] And / or, the phospholipid of the phosphatidylethanolamine class is at least one of 1-palmitoyl-2-oleoylphosphatidylethanolamine, dilauroylphosphatidylethanolamine, dierucoylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, distearoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine and dimyristoylphosphatidylethanolamine.

[0014] Preferably, its preparation method comprises the following steps:

[0015] Step 1: dissolving the raw material in an organic solvent to obtain solution A;

[0016] Step 2: Evaporating and removing the organic solvent in solution A to form a film;

[0017] Step 3: adding a protective agent to the film to hydrate and remove the film, and obtaining a liposome aqueous solution by dispersion and filtration;

[0018] Step 4: freeze-dry the liposome aqueous solution to obtain.

[0019] Preferably, in step 1, the organic solvent is selected from at least one of methanol, ethanol, chloroform and dichloromethane;

[0020] And / or, in step 2, the evaporation temperature is 40-50°C;

[0021] And / or, in step 3, the protective agent is selected from at least one of trehalose, glucose, sucrose, lactose, mannitol, and ammonium sulfate;

[0022] And / or, in step 4, the freeze-drying time is 48-72 hours.

[0023] The present invention also provides a method for preparing the blank liposome, comprising the following steps:

[0024] Step 1: dissolving the raw material in an organic solvent to obtain solution A;

[0025] Step 2: Evaporating and removing the organic solvent in solution A to form a film;

[0026] Step 3: adding a protective agent to the film to hydrate and remove the film, and obtaining a liposome aqueous solution by dispersion and filtration;

[0027] Step 4: freeze-dry the liposome aqueous solution to obtain.

[0028] The present invention also provides the use of the blank liposome as a drug carrier.

[0029] Preferably, the method for using the blank liposomes for drug delivery comprises the following steps: forming a film with the raw materials and active ingredients for preparing the blank liposomes, and performing dispersion filtration and freeze drying to obtain the film;

[0030] Alternatively, the blank liposome is reacted with the active ingredient and freeze-dried to obtain the liposome.

[0031] The present invention also provides a pharmaceutical composition, which is prepared by using the blank liposome as a drug carrier, adding active ingredients and pharmaceutically acceptable excipients; the active ingredient is selected from at least one of paclitaxel, docetaxel, adriamycin and irinotecan.

[0032] The present invention is the first to prepare liposomes using daucosterol as a membrane material. The daucosterol liposomes prepared by the present invention have a small particle size, good cell-entering ability, good blood long circulation effect and drug delivery ability, and at the same time, the daucosterol liposomes carrying different drugs have good stability and dispersibility, high drug encapsulation rate, are conducive to the effective delivery and release of drugs, can safely and efficiently treat diseases such as tumors, and have broad application prospects.

[0033] Obviously, according to the above contents of the present invention, in accordance with common technical knowledge and customary means in the art, without departing from the above basic technical ideas of the present invention, other various forms of modification, replacement or change may be made.

[0034] The above contents of the present invention are further described in detail below through specific implementation methods in the form of embodiments. However, this should not be understood as the scope of the above subject matter of the present invention being limited to the following examples. All technologies realized based on the above contents of the present invention belong to the scope of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0035] Figure 1 The drug-time curves of blank cholesterol liposomes and blank carotene liposomes were injected into the tail vein of rats (n=5).

[0036] Figure 2 The therapeutic effect of carotene doxorubicin liposomes on 4T1 breast cancer.

[0037] Figure 3 This is the therapeutic effect of carotene doxorubicin liposomes on RM-1 prostate cancer. DETAILED DESCRIPTION

[0038] In the following examples and experimental examples, reagents and raw materials not specifically described are all commercially available products.

[0039] Example 1 Preparation of Carotene Blank Liposomes

[0040] 100 mg of lecithin PC-98T and 20 mg of carotene were added to 100 mL of anhydrous ethanol / chloroform (volume ratio of 2:1), stirred at room temperature to form a clear solution, and the organic solvent was removed by rotary evaporation in a constant temperature water bath at 40°C to form a film, and 10 mL of 5% trehalose aqueous solution was added to remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a water solution of carotene blank liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 48 hours, and nitrogen was introduced and sealed to obtain carotene blank liposomes. After testing, the particle size of the liposome was 124 nm.

[0041] Example 2 Preparation of Carotene Blank Liposomes

[0042] 100 mg of egg yolk lecithin E80 and 15 mg of carotene were added to 200 mL of methanol / chloroform mixture (V / V is 2:1), stirred at room temperature to form a clear solution, and the organic solvent was removed by rotary evaporation in a constant temperature water bath at 40°C to form a film, and 10 mL of distilled water was added to hydrate and remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a water solution of carotene blank liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 48 hours, and nitrogen was introduced and sealed to obtain carotene blank liposomes. After testing, the particle size of the liposome was 123 nm.

[0043] Example 3 Preparation of Carotene Blank Liposomes

[0044] 100 mg of egg yolk lecithin E80 and 15 mg of carotene were added to 200 mL of methanol / chloroform mixture (V / V is 2:1), stirred at room temperature to form a clear solution, and in a constant temperature water bath at 40°C, the organic solvent was removed by rotary evaporation to form a film, and 10 mL of 5% glucose aqueous solution was added to hydrate and remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a water solution of carotene blank liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 48 hours, nitrogen was introduced, and sealed to obtain carotene blank liposomes. After testing, the particle size of the liposomes was 114 nm.

[0045] Example 4 Preparation of Carotene Blank Liposomes

[0046] 120 mg of soybean lecithin S100 and 40 mg of carotene were added to 200 mL of methanol / chloroform mixture (V / V is 2:1), stirred at room temperature to form a clear solution, and in a constant temperature water bath at 50°C, the organic solvent was removed by rotary evaporation to form a film, and 10 mL of 10% sucrose aqueous solution was added to hydrate and remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a water solution of carotene blank liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 72 hours, nitrogen was introduced, and sealed to obtain carotene blank liposomes. After testing, the particle size of the liposomes was 107 nm.

[0047] Example 5 Preparation of Carotene Blank Liposomes

[0048] 90 mg of soybean lecithin S100, 20 mg of carotene, 10 mg of cholesterol, and 10 mg of mPEG2000-DSPE were added to 200 mL of a methanol / chloroform mixture (V / V of 2:1), stirred at room temperature to form a clear solution, and in a constant temperature water bath at 50°C, the organic solvent was removed by rotary evaporation to form a film, and 10 mL of a 10% sucrose aqueous solution was added to hydrate and remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a water solution of carotene blank liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 72 hours, and nitrogen was introduced and sealed to obtain carotene blank liposomes. After testing, the particle size of the liposome was 102 nm.

[0049] Example 6 Preparation of paclitaxel liposomes containing daunoside

[0050] 80 mg of egg yolk lecithin, 20 mg of carotene, and 10 mg of paclitaxel were added to 200 mL of a methanol / chloroform mixture (V / V of 2:1), stirred at room temperature to form a clear solution, and in a constant temperature water bath at 50°C, the organic solvent was removed by rotary evaporation to form a film, and 10 mL of an 8% trehalose aqueous solution was added to hydrate and remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a carotene paclitaxel liposome aqueous solution, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 72 hours, and nitrogen gas was introduced as a protective gas, and sealed to obtain carotene paclitaxel liposomes. After testing, the particle size of the liposome was 125 nm, and the paclitaxel encapsulation rate was ≥90%.

[0051] Example 7 Preparation of carotene docetaxel liposomes

[0052] 90 mg of egg yolk lecithin E80, 30 mg of carotene, and 10 mg of docetaxel were added to 200 mL of a methanol / chloroform mixture (V / V of 2:1), stirred at room temperature to form a clear solution, and in a constant temperature water bath at 50°C, the organic solvent was removed by rotary evaporation to form a film, and 10 mL of an 8% trehalose aqueous solution was added to hydrate and remove the film, and a high-pressure homogenizer was used to homogenize the liposome particles to 0.1-0.3 microns, and the liposomes were passed through a 0.22 micron microporous filter membrane to obtain an aqueous solution of carotene docetaxel liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 72 hours, and nitrogen gas was introduced as a protective gas, and sealed to obtain carotene docetaxel liposomes. After testing, the particle size of the liposomes was 116 nm, and the encapsulation rate was ≥90%.

[0053] Example 8 Preparation of carotene adriamycin liposomes

[0054] 100mg egg yolk lecithin, 30mg carotene, 10mg cholesterol, added to 200mL of dichloromethane: ethanol 1: 1 (volume ratio) mixed solvent, stirred at room temperature to form a clear solution, and the organic solvent was removed by rotary evaporator at 50℃ to form a film. 10mL of 8% ammonium sulfate aqueous solution was added, ultrasonicated until the blank liposome particles were 0.1-0.3 microns, passed through a 0.22 micron microporous filter membrane, and the carotene blank liposome solution was obtained. Then, 0.15M (0.15mol / L) glucose solution was used as the eluent, and the carotene blank liposome solution was replaced by a gel column to remove the external aqueous phase. Then, 20mg of doxorubicin hydrochloride was added, and the solution was kept warm in a 37℃ water bath for 30min to obtain a carotene doxorubicin liposome aqueous solution, and then the solution was divided into a vial. Place it in a freeze dryer and freeze-dry for 72 hours, pass a protective gas (argon), and seal it to obtain carotene doxorubicin liposomes. The results showed that the particle size of the liposome was 128 nm and the doxorubicin encapsulation efficiency was ≥ 95%.

[0055] Example 9 Preparation of carotene irinotecan liposomes

[0056] 90g soybean lecithin S100, 25mg carotene, added to 200mL chloroform: methanol = 2: 1 (volume ratio) mixed solvent, stirred at 50℃ to form a clear solution, and evaporated to remove the organic solvent using a film evaporator at 35℃ to form a film. Add 10mL 6% ammonium sulfate aqueous solution, ultrasonicate until the blank liposome particles are 0.1-0.3 microns, pass through a 0.22 micron microporous filter membrane to obtain a carotene blank liposome solution, and then use phosphate buffered saline (PBS) as an eluent to replace the external aqueous phase through a gel column. Add 10mg irinotecan hydrochloride and keep warm in a 60℃ water bath for 30min to obtain a carotene irinotecan liposome aqueous solution. Then divide the solution into vials, put them into a freeze dryer for freeze drying for 72 hours, pass protective gas (argon), seal, and obtain carotene irinotecan liposomes. The results of the test showed that the particle size of the liposome was 122 nm, and the irinotecan encapsulation rate was ≥ 95%.

[0057] The differences between the preparation methods of Examples 1-9 are summarized as follows (Table 1). As shown in Table 1, the carotenoid liposomes prepared by the present invention have a small particle size, good cell-entry ability, and a high drug encapsulation rate.

[0058] Table 1

[0059]

[0060]

[0061] The following is the preparation method of the control sample.

[0062] Comparative Example 1 Preparation of Paclitaxel Ordinary Liposomes

[0063] 80 mg of egg yolk lecithin, 20 mg of cholesterol and 10 mg of paclitaxel were added to 200 mL of a methanol / chloroform mixture (V / V of 2:1), stirred at room temperature to form a clear solution, and the organic solvent was removed by rotary evaporation in a constant temperature water bath at 50°C to form a film, and 10 mL of an 8% trehalose aqueous solution was added to hydrate and remove the film, and ultrasonicated until the liposome particles were 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain an aqueous solution of paclitaxel common liposomes, and then the solution was divided into vials, placed in a freeze dryer for freeze drying for 72 hours, and nitrogen as a protective gas was introduced, and the vials were sealed to obtain paclitaxel common liposomes.

[0064] Comparative Example 2 Preparation of Docetaxel Ordinary Liposomes

[0065] 90 mg of egg yolk lecithin E80, 30 mg of cholesterol, and 10 mg of docetaxel were added to 200 mL of methanol / chloroform mixture (V / V is 2:1), stirred at room temperature to form a clear solution, and in a constant temperature water bath at 50°C, the organic solvent was removed by rotary evaporation to form a film, and 10 mL of 8% trehalose aqueous solution was added to hydrate and remove the film, and the high-pressure homogenizer was used to homogenize the liposome particles to 0.1-0.3 microns, and the 0.22 micron microporous filter membrane was passed to obtain an aqueous solution of docetaxel ordinary liposomes. The solution was then divided into vials, placed in a freeze dryer for freeze drying for 72 hours, and nitrogen protective gas was introduced and sealed to obtain docetaxel ordinary liposomes.

[0066] Comparative Example 3 Preparation of conventional doxorubicin liposomes

[0067] 100mg egg yolk lecithin and 30mg cholesterol were added to 200mL of a mixed solvent of dichloromethane: ethanol 1:1 (volume ratio), stirred at room temperature to form a clear solution, and the organic solvent was removed by a rotary evaporator at 50°C to form a film. 10mL of 8% ammonium sulfate aqueous solution was added, and the blank liposome particles were ultrasonically filtered to 0.1-0.3 microns, and passed through a 0.22 micron microporous filter membrane to obtain a blank liposome solution, and then 0.15mol / L glucose solution was used as an eluent to replace the external aqueous phase through a gel column. Then 20mg of doxorubicin hydrochloride was added, and the mixture was kept warm in a 37°C water bath for 30min to obtain an aqueous solution of doxorubicin common liposomes. The solution was then divided into vials, placed in a freeze dryer for freeze drying for 72 hours, and a protective gas (argon) was introduced, and the vials were sealed to obtain doxorubicin common liposomes.

[0068] The technical solution of the present invention is further illustrated by experiments below.

[0069] Experimental Example 1 Study on the Stability of Carotene Liposomes

[0070] According to Example 6, Example 7, Comparative Example 1 and Comparative Example 2, beta-carotene paclitaxel liposomes, beta-carotene docetaxel liposomes, paclitaxel ordinary liposomes and docetaxel ordinary liposomes were prepared respectively.

[0071] 1. Experimental Methods

[0072] The prepared liposomes were placed at room temperature for 6 hours, and each liposome was observed with the naked eye to see whether drug crystals were precipitated.

[0073] 2. Experimental Results

[0074] The results are shown in Table 2. After standing at room temperature for 6 hours, no drug was precipitated in the paclitaxel liposomes (Example 6) and the docetaxel liposomes (Example 7), while drug precipitation occurred in the paclitaxel common liposomes (Comparative Example 1) and the docetaxel common liposomes (Comparative Example 2). This indicates that the drug has good stability and dispersibility in the paclitaxel liposomes loaded with the drug prepared by the present invention.

[0075] Table 2

[0076] Example 6 Example 7 Comparative Example 1 Comparative Example 2 Drug precipitation no no yes yes

[0077] Experimental Example 2 Pharmacokinetics of Carotene Liposomes

[0078] In this experimental example, the cholesterol blank liposome (Cho-Lip) encapsulating the fluorescent dye DID was prepared according to Comparative Example 1, except that paclitaxel was replaced by DiD;

[0079] Dau-Lip liposomes containing fluorescent dye DID were prepared according to Example 6, except that paclitaxel was replaced by DiD.

[0080] 1. Experimental Methods

[0081] Ten SD rats weighing 180-220 g were randomly divided into two groups, with 5 rats in each group. They were given cholesterol blank liposomes encapsulating the fluorescent dye DID and carotene blank liposomes encapsulating the fluorescent dye DID, respectively, and injected through the rat tail vein at a dose of 150 μg / kg. Blood was collected from the ophthalmic venous plexus at 5 min, 30 min, 1 h, 2 h, 4 h, 8 h, and 30 h after administration, and then the DID fluorescence intensity in the blood samples was detected by fluorescence photometry. The fluorescence value at 5 minutes was taken as the highest fluorescence point, i.e. 100%, and the remaining time points were used as the basis for calculating the remaining DID content in the body.

[0082] 2. Experimental Results

[0083] from Figure 1 It can be seen that after intravenous injection, the clearance rate of the carotenoid liposomes is significantly lower than that of ordinary liposomes, indicating that the carotenoid liposomes prepared by the present invention have a good long blood circulation effect.

[0084] Experimental Example 3 Antitumor Effect of Carotene Liposomes

[0085] According to Example 8 and Comparative Example 3, glycoside doxorubicin liposomes and doxorubicin ordinary liposomes were prepared respectively.

[0086] 1. Experimental Methods

[0087] Cell culture: Mouse breast cancer cell line 4T1, mouse prostate cancer cell line RM-1.

[0088] Animal culture:

[0089] (1) Forty female BALB / c mice (6-8 weeks old) were purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0090] Take 2-4×10 7 4T1 cells in the logarithmic growth phase were slowly injected into the right axillary subcutaneous tissue of 18-20 g mice using a 1 mL syringe. 100 μL was injected into each mouse and the tumor growth was observed until the tumor volume reached about 100 mm. 3 The animals were randomly divided into 4 groups for drug administration, with 10 animals in each group, and the tumor volume was measured every two days.

[0091] The above 4 groups were the saline group: injected with saline;

[0092] Adriamycin solution group: The mice were injected into the tail vein with a dose of 5 mg adriamycin / kg, once every two days for a total of 4 times.

[0093] Adriamycin common liposome group (Comparative Example 3): 5 mg adriamycin / kg was injected into the rat tail vein once every two days for a total of 4 times.

[0094] Carotene doxorubicin liposome group (Example 8 group): 5 mg doxorubicin / kg was injected into the rat tail vein once every two days for a total of 4 times.

[0095] (2) Forty male C57BL / 6 mice (6-8 weeks old) were purchased from Chengdu Dashuo Experimental Animal Co., Ltd.

[0096] Take 2-4×10 7 RM-1 cells in the logarithmic growth phase were slowly injected into the right axillary subcutaneous tissue of 18-20 g mice using a 1 mL syringe. 100 μL was injected into each mouse and the tumor mass was observed to grow until the tumor mass reached a volume of about 80 mm. 3 The animals were randomly divided into 4 groups for drug administration, with 10 animals in each group, and the tumor volume was measured every two days.

[0097] The above 4 groups were the saline group: injected with saline;

[0098] Adriamycin solution group: The mice were injected into the tail vein with a dose of 6 mg adriamycin / kg, once every other day for a total of 5 times.

[0099] Adriamycin common liposome group (Comparative Example 3): 6 mg of adriamycin / kg was injected into the rat tail vein once every other day for a total of 5 times.

[0100] Carotene doxorubicin liposome group (Example 8 group): 6 mg doxorubicin / kg was injected into the rat tail vein once every other day for a total of 5 times.

[0101] 2. Experimental Results

[0102] like Figure 2 , 3 As shown, compared with the conventional doxorubicin liposome group (Comparative Example 3), the carotene glycoside doxorubicin liposome group (Example 8) can significantly inhibit the increase of tumor volume and has a stronger anti-tumor effect. The results show that the drug-loaded carotene liposome has a better drug delivery ability and can effectively treat breast cancer, prostate cancer and other tumors.

[0103] In summary, the carotenoid liposomes prepared by the present invention have small particle size, good cell entry ability, good blood long circulation effect and drug delivery ability, and the carotenoid liposomes loaded with different drugs have good stability and dispersibility, high drug encapsulation rate, which is conducive to the effective delivery and release of drugs, can safely and efficiently treat diseases such as tumors, and have broad application prospects.

Claims

1. Use of carotene or its analogs as membrane materials in the preparation of liposomes.

2. A blank liposome with carotene or its analogues as membrane material, characterized in that: The blank liposome has a membrane, and the membrane is made of the following raw materials in parts by weight: 80-120 parts of lipid substances, 15-40 parts of carotene or its analogues.

3. The blank liposome according to claim 2, characterized in that: The raw materials also include at least one of cholesterol and distearoylphosphatidylethanolamine-polyethylene glycol 2000.

4. The blank liposome according to claim 2, characterized in that: The lipid substance is at least one of natural phospholipids, semi-synthetic phospholipids, and fully synthetic phospholipids; the natural phospholipids are at least one of natural lecithin, sphingomyelin, glycerophospholipids, soybean lecithin, egg yolk lecithin, and cephalin; the semi-synthetic phospholipids or fully synthetic phospholipids are at least one of phospholipids of phosphatidylcholine, phosphatidylserine, phosphatidylinositol, phosphatidylethanolamine, phosphatidylglycerol, dicetyl phosphate, PEG-modified phospholipids, and succinic acid cholesterol ester; The phosphatidylcholine phospholipid is at least one of hydrogenated soybean lecithin, dipalmitoylphosphatidylcholine, distearoylphosphatidylcholine, dimyristoylphosphatidylcholine, dilauroylphosphatidylcholine, dioleoylphosphatidylcholine, phosphatidylcholine, monopalmitoylphosphatidylcholine and glycerylphosphatidylcholine; And / or, the phospholipid of the phosphatidylethanolamine class is at least one of 1-palmitoyl-2-oleoylphosphatidylethanolamine, dilauroylphosphatidylethanolamine, dierucoylphosphatidylethanolamine, dioleoylphosphatidylethanolamine, distearoylphosphatidylethanolamine, dipalmitoylphosphatidylethanolamine and dimyristoylphosphatidylethanolamine.

5. The blank liposome according to any one of claims 2 to 4, characterized in that: Its preparation method comprises the following steps: Step 1: dissolving the raw material in an organic solvent to obtain solution A; Step 2: Evaporating and removing the organic solvent in solution A to form a film; Step 3: adding a protective agent to the film to hydrate and remove the film, and obtaining a liposome aqueous solution by dispersion and filtration; Step 4: freeze-dry the liposome aqueous solution to obtain.

6. The blank liposome according to claim 5, characterized in that: In step 1, the organic solvent is selected from at least one of methanol, ethanol, chloroform and dichloromethane; And / or, in step 2, the evaporation temperature is 40-50°C; And / or, in step 3, the protective agent is selected from at least one of trehalose, glucose, sucrose, lactose, mannitol, and ammonium sulfate; And / or, in step 4, the freeze-drying time is 48-72 hours.

7. The method for preparing blank liposomes according to any one of claims 2 to 6, characterized in that: The steps include: Step 1: dissolving the raw material in an organic solvent to obtain solution A; Step 2: Evaporating and removing the organic solvent in solution A to form a film; Step 3: adding a protective agent to the film to hydrate and remove the film, and obtaining a liposome aqueous solution by dispersion and filtration; Step 4: freeze-dry the liposome aqueous solution to obtain.

8. Use of blank liposomes according to any one of claims 2 to 6 as drug carriers.

9. The use according to claim 8, characterized in that The method for using blank liposomes for drug loading comprises the following steps: forming a film with raw materials and active ingredients for preparing blank liposomes, and performing dispersion filtration and freeze drying to obtain the film; Alternatively, the blank liposome is reacted with the active ingredient and freeze-dried to obtain the liposome.

10. A pharmaceutical composition, characterized in that The invention is prepared by using the blank liposome described in any one of claims 2 to 6 as a drug carrier, adding active ingredients and pharmaceutically acceptable excipients; the active ingredient is selected from at least one of paclitaxel, docetaxel, adriamycin and irinotecan.

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