Liposome drug delivery system, its preparation method and use

Abstract

The application discloses a liposome drug delivery system, which comprises (a) a first phospholipid as a main membrane material, (b) a second phospholipid as an auxiliary membrane material, and (c) a stabilizer. The application also discloses a composition for dissolving adipose tissue prepared by using the drug delivery system, a preparation method and application of the composition. Finally, the application discloses a medicine / cosmetic product prepared by using the composition. The liposome drug delivery system, the cosmetic product or the medicine disclosed by the application has a good fat-reducing effect, can achieve a good sustained-release effect, improves the selectivity to fat cells, achieves the effect of dissolving adipose tissue, has a small side effect, and has a good market application prospect.

Description

Technical Field

[0001] This invention relates to the field of medical aesthetic products or pharmaceuticals, specifically to a liposome drug delivery system and a fat-dissolving composition prepared by encapsulating active ingredients for dissolving fat cells in the liposome drug delivery system. Background Technology

[0002] With the improvement of people's living standards, the incidence of obesity is increasing, and it has been listed by the World Health Organization as one of the diseases that seriously endanger human health. At the same time, obesity also affects the image of those who pursue a high-quality life and value appearance. Among obese individuals, the accumulation of localized fat in the body has the most significant impact on health and beauty, such as a double chin and abdomen. Therefore, developing products with localized fat reduction functions has great economic value.

[0003] Currently, Kybella is the only product globally approved by the U.S. Food and Drug Administration (FDA) in 2015 for use in adults with moderate to severe double chins. In my country, no topical lipolysis injections have been approved by the National Medical Products Administration. Kybella's main active ingredient is sodium deoxycholate, which exerts its pharmacological effect by causing extensive necrosis of fat cells through cell membrane lysis. However, because deoxycholic acid is not specific, it can damage not only fat cells but also other normal cells, causing inflammation at the injection site. Furthermore, Kybella's weakly alkaline pH significantly increases the incidence of severe pain and swelling after injection.

[0004] Lipobean, a phosphatidylcholine injection developed by AMI Pharm for the treatment of hepatic encephalopathy, was launched in South Korea in December 2010. Its main active ingredient is phosphatidylcholine, with a small amount of sodium deoxycholate as a solubilizer. The company also initiated clinical trials to evaluate the lipolysis effect of Lipobean. While the lipolysis clinical evaluation has been ongoing since December 2010, no related reports have been disclosed. Simultaneously, Lipobean has also been used off-label for local lipolysis, but according to clinical and market feedback, the lipolysis effect and side effects of Lipobean are highly controversial. The pharmacological mechanism by which the drug exerts its lipolysis effect is also controversial. Some studies suggest that sodium deoxycholate is the active ingredient responsible for Lipobean's lipolysis pharmacological effect. In addition, other non-compliant fat-dissolving injections are circulating in the medical aesthetics market. These products have complex ingredients, mostly compound preparations, with main components such as L-carnitine, deoxycholic acid, phosphatidylcholine, insulin-like growth factor-1, lipase, saponifying agents, triamcinolone acetonide, and hyaluronic acid. These compound products are mostly cosmetics, and in clinical use, they are often injected locally into the subcutaneous fat layer. According to clinical and market feedback, these non-compliant products are accompanied by adverse reactions during clinical use. The main complications include: redness, swelling, infection and ulceration at the injection site, pigmentation, depigmentation, lumps, local depression, local swelling, allergic reactions, parotid fistula, parotitis, and kidney damage, most of which are irreversible. Summary of the Invention

[0005] This invention provides a medical aesthetic product or pharmaceutical composition for weight loss, its preparation method, and its uses.

[0006] This invention provides a liposome drug delivery system comprising:

[0007] (a) The first phospholipid as the main membrane material

[0008] (b) The second phospholipid, as an auxiliary membrane material, and

[0009] (c) Stabilizers.

[0010] The first phospholipid is selected from neutral phospholipids.

[0011] The first phospholipid is selected from at least one of dioleoylphosphatidylcholine, distearylphosphatidylcholine, hydrogenated soybean lecithin, egg yolk lecithin, soybean lecithin, distearylphosphatidylethanolamine, and dioleoylphosphatidylethanolamine; preferably, the first phospholipid is dioleoylphosphatidylcholine.

[0012] The second phospholipid is selected from at least one of negatively charged phospholipids, positively charged phospholipids, and neutral phospholipids.

[0013] The positively charged phospholipid is selected from at least one of (2-dioleoylhydroxypropyl)trimethylammonium chloride and 1,2-dioleoyl-3-dimethylamino-propane; preferably, the positively charged phospholipid is selected from 1,2-dioleoyl-3-dimethylamino-propane.

[0014] Wherein, the second phospholipid is a polymer-modified phospholipid; preferably, the polymer-modified phospholipid is a polyethylene glycol-modified phospholipid; preferably, the polyethylene glycol-modified phospholipid is distearate-phosphatidylethanolamine-polyethylene glycol 2000.

[0015] The negatively charged phospholipid is selected from at least one of distearylphosphatidylglycerol, dioleoylphosphatidylglycerol, phosphatidylserine, dioleoylphosphatidylserine, and distearylphosphatidylserine; preferably, the negatively charged phospholipid is selected from at least one of distearylphosphatidylglycerol, dioleoylphosphatidylglycerol, and phosphatidylserine.

[0016] Wherein, the mass ratio of the first phospholipid to the second phospholipid is 1-25:1-10; preferably, the mass ratio of the first phospholipid to the second phospholipid is 10-18:1-8.

[0017] The aforementioned liposome delivery system is prepared using raw materials containing a first phospholipid and a second phospholipid in the following ratio: the mass ratio of the first phospholipid to the second phospholipid is (3-10):1; preferably, it is prepared using raw materials containing a first phospholipid and a second phospholipid in the following ratio: the mass ratio of the first phospholipid to the second phospholipid is 14:4, 15:4, 15:3, 15:2 or 90:19.

[0018] The stabilizer is selected from steroids; preferably, the steroid is selected from at least one of cholesterol, cholesterol hexasuccinate, ergosterol, and lanosterol; preferably, the steroid is cholesterol.

[0019] Cholesterol accounts for 0% to 30% of the total mass of the primary phospholipid, secondary phospholipid, and stabilizer.

[0020] The mass ratio of the first phospholipid to the second phospholipid to the stabilizer is 1-25:1-10:0-10; preferably, the mass ratio of the first phospholipid to the second phospholipid to the stabilizer is 10-18:1-8:1-5.

[0021] The aforementioned liposome drug delivery system is prepared using raw materials containing a first phospholipid, a second phospholipid, and a stabilizer in the following ratio: the mass ratio of the first phospholipid to the second phospholipid to the stabilizer is 15:

[0022] (1.5~5):(2~2.5); Preferably, it is prepared from raw materials containing a first phospholipid, a second phospholipid, and a stabilizer in the following ratio, wherein the mass ratio of the first phospholipid: the second phospholipid: the stabilizer is 14:4:2, 15:4:2, 15:3:2, 15:2:2 or 90:19:12.

[0023] The aforementioned liposome drug delivery system was prepared using raw materials containing the following components in the indicated weight ratios:

[0024] Dioleoylphosphatidylcholine 140-450 parts, second phospholipid 40-150 parts, cholesterol 20-60 parts;

[0025] The second phospholipid is selected from dioleoylphosphatidylglycerol, 1,2-dioleoyl-3-dimethylamino-propane, distearylphosphatidylglycerol, distearylphosphatidylethanolamine-polyethylene glycol 2000 or phosphatidylserine;

[0026] Preferably, the liposome drug delivery system is prepared using raw materials containing the following components in the indicated weight ratios:

[0027] 450 parts of dioleoylphosphatidylcholine, 75-117 parts of 1,2-dioleoyl-3-dimethylamino-propane, and 60 parts of cholesterol;

[0028] Dioleoylphosphatidylcholine 450 parts, phosphatidylserine 45-72 parts, cholesterol 60 parts;

[0029] Dioleoylphosphatidylcholine 450 parts, dioleoylphosphatidylglycerol 90-150 parts, cholesterol 60 parts;

[0030] Dioleoylphosphatidylcholine 450 parts, distearate phosphatidylglycerol 90-150 parts, cholesterol 60 parts; or

[0031] 450 parts of dioleoylphosphatidylcholine, 63-108 parts of distearate phosphatidylethanolamine-polyethylene glycol 2000, and 60 parts of cholesterol;

[0032] Preferably, the liposome drug delivery system is prepared using raw materials containing the following components in the indicated weight ratios:

[0033] Dioleoylphosphatidylcholine 140 parts, dioleoylphosphatidylglycerol 40 parts, cholesterol 20 parts;

[0034] 140 parts of dioleoylphosphatidylcholine, 40 parts of 1,2-dioleoyl-3-dimethylamino-propane, and 20 parts of cholesterol;

[0035] 450 parts of dioleoylphosphatidylcholine, 120 parts of distearylphosphatidylglycerol, and 60 parts of cholesterol;

[0036] 450 parts of dioleoylphosphatidylcholine, 96 parts of 1,2-dioleoyl-3-dimethylamino-propane, and 60 parts of cholesterol;

[0037] Dioleoylphosphatidylcholine 450 parts, dioleoylphosphatidylglycerol 120 parts, cholesterol 60 parts;

[0038] Dioleoylphosphatidylcholine 450 parts, distearate phosphatidylethanolamine-polyethylene glycol 2000 90 parts, cholesterol 60 parts; or,

[0039] Dioleoylphosphatidylcholine 450 parts, phosphatidylserine 60 parts, cholesterol 60 parts.

[0040] It also includes a buffer solution; preferably, the buffer solution is selected from at least one of sodium chloride buffer, PBS buffer, HEPES buffer, Tris buffer, and His buffer; preferably, the buffer solution is selected from at least one of 0.9% w / w sodium chloride buffer and PBS buffer.

[0041] The buffer solution is an isotonic buffer solution.

[0042] The pH of the buffer solution is 5 to 8.

[0043] The aforementioned liposome drug delivery system is prepared using raw materials containing a buffer solution and total phospholipids in the following proportions: the volume-to-mass ratio of the buffer solution to total phospholipids is 1-5:1-30, ml / mg; preferably, it is prepared using raw materials containing a buffer solution and total phospholipids in the following proportions: the volume-to-mass ratio of the buffer solution to total phospholipids is 1:9, 3:51, 1:19, 6:109, or 1:18, ml / mg.

[0044] The liposomes have an average particle size of 50–500 nanometers; preferably, the liposomes have an average particle size of 50–200 nanometers; preferably, the liposomes have an average particle size of 100–200 nanometers; preferably, the liposomes have an average particle size of 150–200 nanometers.

[0045] The liposomes are positively charged, negatively charged, or neutral; preferably, the liposomes are positively charged.

[0046] The liposomes described therein are monocompartmental liposomes, large monocompartmental liposomes, multicompartmental liposomes, liposome nanoparticles, or multiporous liposomes.

[0047] The present invention also provides the use of the aforementioned liposome drug delivery system in the preparation of pharmaceuticals or medical aesthetic products for treating adipose cell-related diseases.

[0048] The present invention also provides a composition for dissolving fat, which is prepared by encapsulating the active ingredient for dissolving fat cells in the aforementioned liposome delivery system.

[0049] Wherein, the active ingredient for dissolving fat cells is bile acid; preferably, the bile acid is selected from at least one of the following: deoxycholic acid, cholic acid, lithocholic acid, chenodeoxycholic acid, porphyrin deoxycholic acid, trihydroxycosanoic acid, ursodeoxycholic acid, taurocholic acid, glycocholic acid, or physiologically tolerable salts of the above compounds; preferably, the bile acid is selected from deoxycholic acid and / or its salts; preferably, the bile acid is selected from deoxycholic acid and / or sodium deoxycholate.

[0050] The mass ratio of the first phospholipid, the second phospholipid, and the stabilizer to the active ingredient is 1-10:1-10.

[0051] The phospholipid is prepared using raw materials containing the following weight ratio components, wherein the total mass ratio of the first phospholipid, the second phospholipid, and the stabilizer to the active ingredient is (3-10):1; preferably, it is prepared using raw materials containing the following weight ratio components, wherein the total mass ratio of the first phospholipid, the second phospholipid, and the stabilizer to the active ingredient is (3.5-4.5):1; preferably, it is prepared using raw materials containing the following weight ratio components, wherein the total mass ratio of the first phospholipid, the second phospholipid, and the stabilizer to the active ingredient is (3.7-4.4):1; more preferably, it is prepared using raw materials containing the following weight ratio components, wherein the total mass ratio of the first phospholipid, the second phospholipid, and the stabilizer to the active ingredient is 4:1, 21:5, 121:30, or 19:5.

[0052] The buffer solution is prepared using raw materials containing a buffer solution and an active ingredient in the following ratio: the volume-to-mass ratio of the buffer solution to the active ingredient is 1-5:1-10, ml / mg. Preferably, the buffer solution is prepared using raw materials containing a buffer solution and an active ingredient in the following ratio: the volume-to-mass ratio of the buffer solution to the active ingredient is 2:5 or 1:5.

[0053] In some embodiments, the fat-dissolving composition provided by the present invention is liposomes with an average particle size of 50-500 nanometers; preferably, the average particle size of the liposomes is 50-200 nanometers; preferably, the average particle size of the liposomes is 100-200 nanometers; preferably, the average particle size of the liposomes is 100-150 nanometers.

[0054] The present invention also provides a method for preparing the aforementioned liposome drug delivery system or the aforementioned composition, characterized in that the method employs solvent injection, reverse evaporation, thin film dispersion, microfluidics, active drug loading, or ammonium sulfate gradient method.

[0055] The present invention also provides a method for preparing the aforementioned liposome drug delivery system or the aforementioned composition, characterized by comprising the following steps:

[0056] a. Take the active ingredient, the first phospholipid, the second phospholipid, and the stabilizer according to the specified ratio, and disperse them in an organic solvent to obtain a mixed solution; or, take the first phospholipid, the second phospholipid, and the stabilizer according to the specified ratio, and disperse them in an organic solvent to obtain a mixed solution;

[0057] b. Inject the mixed solution obtained in step a into the buffer solution to obtain a suspension;

[0058] c. Collect the liposomes from the suspension in step b.

[0059] Wherein, the organic solvent in step a is selected from ethanol and / or diethyl ether; preferably, the organic solvent in step a is ethanol.

[0060] In step a, the mixed solution is heated to 20-80°C; preferably, in step a, the mixed solution is heated to 65°C.

[0061] In step b, the buffer solution is heated to 20–80°C; preferably, step b is heated to 65°C.

[0062] In step b, the mixture is stirred for 0.5 to 2 hours after injection; preferably, the mixture is stirred for 1 hour after injection.

[0063] In step c, the obtained suspension is granulated and then the organic solvent is removed to obtain liposomes; preferably, step c granulates the obtained suspension using a 0.05-0.6 μm extruder; preferably, step c granulates the obtained suspension using a 0.45 μm or 0.22 μm extruder.

[0064] The present invention also provides a method for preparing the aforementioned liposome drug delivery system or the aforementioned composition, characterized by comprising the following steps:

[0065] I. Take the active ingredient, the first phospholipid, the second phospholipid, and the stabilizer according to the specified ratio, add an organic solvent and a first buffer solution, and disperse to form an emulsion; or, take the first phospholipid, the second phospholipid, and the stabilizer according to the specified ratio, add an organic solvent and a first buffer solution, and disperse to form an emulsion;

[0066] II. Remove the organic solvent from the emulsion in step I, add the second part of the buffer solution, and obtain a suspension;

[0067] III. Collect the liposomes from the suspension in step II.

[0068] Wherein, the organic solvent in step I is chloroform and / or diethyl ether; preferably, the organic solvent in step I is chloroform.

[0069] In step I, an emulsion is formed by ultrasonic treatment; preferably, the ultrasonic time is 1 to 40 minutes; preferably, the ultrasonic time is 10 minutes; preferably, the ultrasonic temperature is 10 to 30°C; preferably, the ultrasonic temperature is below 20°C.

[0070] The emulsion formed in step I is a W / O type emulsion.

[0071] In step II, the organic solvent is removed by vacuum evaporation.

[0072] Wherein, the volume ratio of the first part of the buffer solution to the second part of the buffer solution is 1-5:1-10; preferably, the volume ratio of the first part of the buffer solution to the second part of the buffer solution is 1:4.

[0073] In step III, the obtained suspension is granulated to obtain liposomes; preferably, in step c, the obtained suspension is granulated using a 0.05-0.6 μm extruder; preferably, in step c, the obtained suspension is granulated using a 0.45 μm or 0.22 μm extruder.

[0074] The present invention also provides the use of the aforementioned liposome delivery system or the aforementioned composition in the preparation of fat reduction drugs or medical aesthetic products; preferably, the fat reduction drugs or medical aesthetic products are local fat reduction drugs or medical aesthetic products.

[0075] The drug or medical aesthetic product lyses the cell membrane of adipocytes and / or promotes adipocyte apoptosis; preferably, the drug or medical aesthetic product is taken up by adipocytes.

[0076] Finally, the present invention provides a weight-loss drug or medical aesthetic product containing the aforementioned composition, as well as acceptable excipients or auxiliary ingredients in the drug or medical aesthetic product.

[0077] The excipients or auxiliary ingredients are selected from at least one of vitamin C, benzoic acid, and lidocaine.

[0078] The drug or medical aesthetic product mentioned is an injectable.

[0079] The drug or medical aesthetic product is a topical drug delivery preparation for adipose tissue.

[0080] Among them, (a) is the first phospholipid, which is the main membrane material and its main function is to form the backbone of the liposome bilayer.

[0081] Among them, (b) the second phospholipid, serving as both an auxiliary membrane material and a liposome membrane material, primarily functions to enhance the selectivity of adipocytes, thereby promoting the uptake of liposomes by adipocytes and / or reducing the uptake of liposomes by macrophages. This allows the active ingredients to exert their effects more effectively within adipocytes, while simultaneously reducing the impact on non-adipocytes (such as vascular cells), improving therapeutic efficacy, and reducing adverse reactions.

[0082] Among them, (c) liposome stabilizers mainly play a role in regulating the fluidity of liposome membrane structure and stabilizing liposome membrane structure.

[0083] Total phospholipids refer to the total mass of primary and secondary phospholipids.

[0084] According to some specific embodiments of the present invention, the present invention provides a deoxycholic acid liposome prepared from the following components: deoxycholic acid, phospholipids, cholesterol, and an external aqueous buffer.

[0085] According to some specific embodiments of the present invention, the present invention provides a deoxycholic acid liposome, the formulation of which is: deoxycholic acid and / or sodium deoxycholate, phosphatidylcholine or modified phosphatidylcholine, cholesterol, phosphatidylserine, vitamin C, benzoic acid, and lidocaine.

[0086] The fat reduction medical aesthetic products or pharmaceutical compositions provided by this invention have a good fat reduction effect. By optimizing and preparing liposomes containing active ingredients, a good sustained-release effect can be achieved, while improving the selectivity for fat cells and achieving the effect of dissolving fat tissue. With few side effects, it has good market application prospects.

[0087] This invention provides a novel fat-dissolving or fat-reducing injection, using a substance with fat-dissolving or fat-reducing effects as the active ingredient, such as deoxycholic acid (sodium). Microcarrier formulation technology, such as liposome technology, is used to prepare a microcarrier formulation containing the active ingredient, such as deoxycholic acid liposomes. By optimizing the formulation and dosage form of the drug-containing microcarrier, the selectivity for killing adipocytes is improved, and the method of killing cells is changed from extracellular to intracellular, ultimately reducing the fat-dissolving side effects of the active ingredient.

[0088] The present invention provides a novel local lipolysis injection that can efficiently dissolve adipose tissue, reduce fat accumulation, and at the same time reduce damage to other non-fat cells (such as vascular cells, muscle cells, and adipose precursor cells). The therapeutic effect and adverse reactions are far superior to lipolysis injections on the market.

[0089] Meaning of abbreviations:

[0090] PBS: Phosphate Buffer

[0091] HEPES: 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid buffer

[0092] DOPC: Dioleoylphosphatidylcholine, or also known as dioleoyllecithin

[0093] DSPC: Distearylphosphatidylcholine

[0094] HSPC: Hydrogenated Soy Lecithin

[0095] EPC: Natural Phospholipids, Egg Yolk Lecithin

[0096] DSPG: Distearylphosphatidylglycerol

[0097] DODMA: 1,2-Dioleol-3-dimethylamino-propane

[0098] DOPG: Dioleoylphosphatidylglycerol

[0099] DSPE-PEG2000: Distearylphosphatidylethanolamine-polyethylene glycol 2000

[0100] PS: Phosphatidylserine

[0101] DOPS: Dioleoylphosphatidylserine

[0102] DSPS: Distearylphosphatidylserine

[0103] DOTAP: (2-Dioleoylhydroxypropyl)trimethylammonium chloride

[0104] DSPE: Distearylphosphatidylethanolamine

[0105] DOPE: Dioleoylphosphatidylethanolamine

[0106] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0107] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following examples. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached image description:

[0108] Figure 1a, 1b, 1c: Describe and evaluate the killing effects of high, medium, and low concentrations of DODMA liposomes (DC-LIP(H), DC-LIP(M), DC-LIP(L)), SDC (sodium deoxycholate solution), and LIP (unloaded blank liposomes) on mature adipocytes, HUVECs (human umbilical vein endothelial cells), and 3T3-L1 (mouse embryonic fibroblasts) at dosages of 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml, and 0.0625 mg / ml, respectively.

[0109] Figures 2a, 2b, 2c This study describes and evaluates the killing effects of high, medium, and low concentrations of PS liposomes (DC-LIP(H), DC-LIP(M), DC-LIP(L)), SDC (sodium deoxycholate solution), and LIP (unloaded blank liposomes) on mature adipocytes, HUVECs (human umbilical vein endothelial cells), and 3T3-L1 (mouse embryonic fibroblasts) at dosages of 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml, and 0.0625 mg / ml, respectively.

[0110] Figures 3a, 3b, and 3c This study describes and evaluates the killing effects of high, medium, and low concentrations of DOPG liposomes (DC-LIP(H), DC-LIP(M), DC-LIP(L)), SDC (sodium deoxycholate solution), and LIP (unloaded blank liposomes) on mature adipocytes, HUVECs (human umbilical vein endothelial cells), and 3T3-L1 (mouse embryonic fibroblasts) at dosages of 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml, and 0.0625 mg / ml, respectively.

[0111] Figures 4a, 4b, and 4c This study describes and evaluates the killing effects of high, medium, and low concentrations of DSPG liposomes (DC-LIP(H), DC-LIP(M), DC-LIP(L)), SDC (sodium deoxycholate solution), and LIP (unloaded blank liposomes) on mature adipocytes, HUVECs (human umbilical vein endothelial cells), and 3T3-L1 (mouse embryonic fibroblasts) at dosages of 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml, and 0.0625 mg / ml, respectively.

[0112] Figures 5a, 5b, and 5cThis study describes and evaluates the killing effects of three concentrations of DSPE-PEG2000 liposomes (DC-LIP(H), DC-LIP(M), DC-LIP(L)), SDC (sodium deoxycholate solution), and LIP (unloaded blank liposomes) at dosages of 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml, and 0.0625 mg / ml on mature adipocytes, HUVECs (human umbilical vein endothelial cells), and 3T3-L1 (mouse embryonic fibroblasts).

[0113] Figures 6a, 6b, 6c, 6d, 6e , representing the cellular uptake rates of DOPC+DODMA prescription blank liposomes, DOPC+PS prescription blank liposomes, DOPC+DOPG prescription blank liposomes, DOPC+DSPG prescription blank liposomes, and DOPC+DSPE-PEG2000 prescription blank liposomes on 3T3-L1 cells, mature adipocytes, and HUVECs, respectively.

[0114] Figures 7a and 7b , respectively, represent the skin pathological structures at the injection sites of Kybella and DC-LIP (PEG).

[0115] Figure 8 A schematic diagram of drug administration in an evaluation test of local nonspecific skin damage in New Zealand white rabbits. Detailed Implementation

[0116] Example 1 (Drug-loaded liposomes DOPC+DOPG)

[0117] Measure 20 ml of isotonic sodium chloride into an Erlenmeyer flask, add a magnetic stir bar, and place it on a heat-collecting stirrer. Heat to 65°C to prepare solution A. Weigh out the prescribed amounts of deoxycholic acid (50 mg), dioleoylphosphatidylcholine (DOPC) (140 mg), dioleoylphosphatidylglycerol (DOPG) (40 mg), and cholesterol (20 mg). Add 2 ml of anhydrous ethanol and sonicate for 5 min to dissolve. Set the temperature to 65°C to prepare solution B. Slowly inject solution B into solution A using a 5 ml syringe and continue stirring for 1 h. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders, and then remove the organic solvent using a tangential flow ultrafiltration device to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 180.0 nm, and the Zeta potential was -61.0 mV.

[0118] Example 2 (Drug-loaded liposomes DOPC+DODMA)

[0119] Measure 20 ml of isotonic sodium chloride into an Erlenmeyer flask, add a magnetic stir bar, and place it on a heat-collecting stirrer. Heat to 65°C to prepare solution A. Weigh out the prescribed amounts of deoxycholic acid (50 mg), dioleoylphosphatidylcholine (DOPC) (140 mg), 1,2-dioleoyl-3-dimethylaminopropane (DODMA) (40 mg), and cholesterol (20 mg). Add 2 ml of anhydrous ethanol and sonicate for 5 min to dissolve. Set the temperature to 65°C to prepare solution B. Slowly inject solution B into solution A using a 5 ml syringe and continue stirring for 1 h. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders, and then remove the organic solvent using a tangential flow ultrafiltration device to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 172.3 nm, and the Zeta potential was 37.8 mV.

[0120] Example 3 (Drug-loaded liposomes DOPC+DODMA)

[0121] Weigh out the prescribed amounts of 150 mg deoxycholic acid, 450 mg dioleoylphosphatidylcholine (DOPC), 95 mg 1,2-dioleoyl-3-dimethylaminopropane (DODMA), and 60 mg cholesterol. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 133.4 nm, and the Zeta potential was 26.7 mV.

[0122] Example 4 (Drug-loaded liposomes DOPC+PS)

[0123] Weigh out the prescribed amounts of 150 mg deoxycholic acid, 450 mg dioleoylphosphatidylcholine (DOPC), 60 mg phosphatidylserine (PS), and 60 mg cholesterol. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 122.7 nm, and the Zeta potential was -43.0 mV.

[0124] Example 5 (Drug-loaded liposomes DOPC+DOPG)

[0125] Weigh out the prescribed amounts of 150 mg deoxycholic acid, 450 mg dioleoylphosphatidylcholine (DOPC), 120 mg dioleoylphosphatidylglycerol (DOPG), and 60 mg cholesterol. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 126.7 nm, and the Zeta potential was -51.6 mV.

[0126] Example 6 (Drug-loaded liposomes DOPC+DSPG)

[0127] Weigh out the prescribed amounts of 150 mg deoxycholic acid, 450 mg dioleoylphosphatidylcholine (DOPC), 120 mg distearate phosphatidylglycerol (DSPG), and 60 mg cholesterol. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 130.9 nm, and the Zeta potential was -51.6 mV.

[0128] Example 7 (Drug-loaded liposomes DOPC + DSPE-PEG2000)

[0129] Weigh out the prescribed amounts of 150 mg deoxycholic acid, 450 mg dioleoylphosphatidylcholine (DOPC), 90 mg distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), and 60 mg cholesterol. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Disperse the mixture in a W / O emulsion by sonication at below 20°C for 10 min. Remove the organic solvent by vacuum evaporation in a rotary evaporator. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue vacuum evaporation for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 130.8 nm, and the Zeta potential was -35.6 mV.

[0130] Example 8 (Liposomes with fluorescent blank DOPC+DOPG)

[0131] Weigh out the prescribed amounts of 450 mg dioleoylphosphatidylcholine (DOPC), 120 mg dioleoylphosphatidylglycerol (DOPG), 60 mg cholesterol, and 1.5 mg Dil fluorescent dye. Add 18 ml of chloroform and 6 ml of phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml of phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 179.8 nm, and the Zeta potential was -63.3 mV.

[0132] Example 9 (Liposomes with fluorescent blank DOPC+DSPG)

[0133] Weigh out the prescribed amounts of 450 mg dioleoylphosphatidylcholine (DOPC), 120 mg distearate phosphatidylglycerol (DSPG), 60 mg cholesterol, and 1.5 mg Dil fluorescent dye. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 186.7 nm, and the Zeta potential was -65.1 mV.

[0134] Example 10 (Fluorescent blank liposome DOPC+PS)

[0135] Weigh out the prescribed amounts of 450 mg dioleoylphosphatidylcholine (DOPC), 60 mg phosphatidylserine (PS), 60 mg cholesterol, and 1.5 mg Dil fluorescent dye. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 193.7 nm, and the Zeta potential was -51.9 mV.

[0136] Example 11 (Fluorescent blank liposomes DOPC + DSPE-PEG2000)

[0137] Measure 30 ml of isotonic sodium chloride into an Erlenmeyer flask, add a magnetic stir bar, and place it on a heat-collecting stirrer. Heat to 65°C to prepare solution A. Weigh out the prescribed amounts of 450 mg of dioleoylphosphatidylcholine (DOPC), 90 mg of distearate phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000), 60 mg of cholesterol, and 1.5 mg of Dil fluorescent dye. Add 3 ml of anhydrous ethanol and sonicate for 5 min to dissolve. Set the temperature to 65°C to prepare solution B. Slowly inject solution A into solution B using a 5 ml syringe and continue stirring for 1 h. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders, and then remove the organic solvent using a tangential flow ultrafiltration device to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 183.9 nm, and the Zeta potential was -59.7 mV.

[0138] Example 12 (Fluorescent blank liposomes DOPC+DODMA)

[0139] Weigh out the prescribed amounts of 450 mg dioleoylphosphatidylcholine (DOPC), 95 mg 1,2-dioleoyl-3-dimethylamino-propane (DODMA), 60 mg cholesterol, and 1.5 mg Dil fluorescent dye. Add 18 ml chloroform and 6 ml phosphate-buffered saline (PBS). Sonicate at below 20°C for 10 min to disperse into a W / O emulsion. Place the emulsion in a rotary evaporator and evaporate under reduced pressure to remove the organic solvent. Once the solution in the flask reaches a colloidal state, add 24 ml phosphate buffer, shake, and continue evaporating under reduced pressure for 1 h to form a suspension. Granulate the resulting suspension using 0.45 μm and 0.22 μm extruders to obtain deoxycholic acid liposomes. The particle size of the deoxycholic acid liposomes was measured to be 192.8 nm, and the Zeta potential was 36.4 mV.

[0140] Example 1: Evaluation of the cell-killing effects of the medical aesthetic products or pharmaceutical compositions of the present invention on different cells.

[0141] 1 Experimental Methods

[0142] 1.1 Preparation of drug-loaded liposomes

[0143] Three concentrations of DODMA liposomes (high, medium, and low) were prepared according to the method in Example 3, with the difference being that the prescribed amounts of DODMA were 117 mg, 96 mg, and 75 mg, respectively.

[0144] Three concentrations of PS liposomes, namely high, medium and low, were prepared according to the method of Example 4, with the difference being that the PS mass in the formulation was 72 mg, 60 mg and 45 mg, respectively.

[0145] Three concentrations of DOPG liposomes (high, medium, and low) were prepared according to the method in Example 5, with the difference being that the DOPG mass in the formulation was 150 mg, 120 mg, and 90 mg, respectively.

[0146] Three concentrations of DSPG liposomes (high, medium, and low) were prepared according to the method in Example 6, with the difference being that the mass of DSPG in the formulation was 150 mg, 120 mg, and 90 mg, respectively.

[0147] Three concentrations of DSPE-PEG2000 liposomes (high, medium, and low) were prepared according to the method in Example 7, with the difference being that the mass of DSPE-PEG2000 in the formulation was 108 mg, 90 mg, and 63 mg, respectively.

[0148] Blank liposomes were prepared according to Examples 3 to 7, except that the active ingredient deoxycholic acid was not added during the preparation process.

[0149] To prepare sodium deoxycholate solution, weigh 100 mg of sodium deoxycholate and dissolve it completely in 10 mL of phosphate buffer to obtain sodium deoxycholate solution.

[0150] 1.2 Evaluation of cell killing effect

[0151] 3T3-L1 (mouse embryonic fibroblasts) were pre-seeded into 96-well plates and induced to differentiate into mature adipocytes for subsequent cell evaluation. 3T3-L1 (mouse embryonic fibroblasts) and HUVEC (human umbilical vein endothelial cells) cells in logarithmic growth phase were added to 10,000 cells per well of 96-well plates (100 μL per well) and cultured in a CO2 incubator for 24 h. Cells were then evaluated. Each drug-loaded liposome and sodium deoxycholate solution was diluted with complete culture medium to concentrations of deoxycholic acid of 0.5 mg / ml, 0.25 mg / ml, 0.125 mg / ml, and 0.0625 mg / ml, respectively. Blank liposomes were diluted to the same proportions as the corresponding drug-loaded liposomes to obtain blank liposome solutions. The drug-loaded liposomes containing different concentrations of deoxycholic acid, sodium deoxycholate solution, and blank liposome solution were added to wells of mature adipocytes, 3T3-L1 cells in logarithmic growth phase, and HUVECs (n=2), 100 μL per well. After drug administration, the cells were cultured in a CO2 incubator for 24 h. 100 μL of supernatant was aspirated from each well, and 50 μL of cell viability assay reagent (CellCounting-Lite 3D Luminescent Cell Viability Assay) was added to each well. The cells were shaken at room temperature for 5 min until the cell clusters were completely lysed. After incubation for 30 min, the cells were transferred to opaque 96-well plates, 50 μL per well, and the fluorescence intensity (FI) was detected using a fluorescence microplate reader.

[0152] Cell viability is calculated using the following formula based on fluorescence intensity:

[0153] Cell viability (%) = (FI) 样品 -FI 空白 ) / (FI 对照 –FI 空白 )×100%

[0154] FI 空白 : Indicates the average fluorescence intensity value after no cells were present and no sample treatment was performed;

[0155] FI 对照 : Indicates the presence of cells, but the average fluorescence intensity value without sample treatment.

[0156] The differences in cell killing effects were statistically analyzed using Graphpad software based on cell survival rates.

[0157] 2 Experimental Results

[0158] 2.1 By Figure 1 (a-c) shows that when the concentration of the active ingredient deoxycholic acid exceeds 0.25 mg / mL, there is no significant difference in the killing ability of DOPC+DODMA and SDC against adipocytes; however, for non-adipocytes (HUVEC and 3T3-L1), the cell killing ability of DOPC+DODMA is significantly weaker than that of the SDC group.

[0159] 2.2 As shown in Figures 2(a-c), when the concentration of the active ingredient deoxycholic acid exceeds 0.25 mg / mL, there is no significant difference in the killing ability of DOPC+PS formulation and SDC against adipocytes; however, for non-adipocytes (3T3-L1), the cell killing ability of DOPC+PS formulation is significantly weaker than that of SDC group.

[0160] 2.3 As shown in Figures 3(a-c), when the concentration of the active ingredient deoxycholic acid exceeds 0.25 mg / mL, there is no significant difference in the cytotoxic ability of the DOPC+DOPG formulation and SDC against adipocytes; however, for non-adipocytes (3T3-L1), the cytotoxic ability of the DOPC+DOPG formulation is significantly weaker than that of the SDC group, and for non-adipocytes (HUVEC), the cytotoxic ability of the DOPC+DOPG formulation is also weaker than that of the SDC group.

[0161] 2.4 As shown in Figures 4(a-c), when the concentration of the active ingredient deoxycholic acid exceeds 0.25 mg / mL, there is no significant difference in the cell-killing ability of the DOPC+DSPG formulation and SDC against adipocytes; however, for non-adipocytes (3T3-L1), the cell-killing ability of the DOPC+DSPG formulation is significantly weaker than that of the SDC group, and for non-adipocytes (HUVEC), the cell-killing ability of the DOPC+DSPG formulation is also weaker than that of the SDC group.

[0162] 2.5 As shown in Figures 5(a-c), when the concentration of the active ingredient deoxycholic acid exceeds 0.25 mg / mL, there is no significant difference in the cell-killing ability of the DOPC+DSPE-PEG200 formulation and SDC against adipocytes; however, for non-adipocytes (3T3-L1), the cell-killing ability of the DOPC+DSPE-PEG200 formulation is significantly weaker than that of the SDC group, and for non-adipocytes (HUVEC), the cell-killing ability of the DOPC+DSPE-PEG200 formulation is also weaker than that of the SDC group.

[0163] The results showed that the deoxycholic acid liposome preparation prepared in this application has significant in vitro killing ability against adipocytes, but its killing ability against 3T3-L1 or HUVEC non-adipocytes is very weak, indicating that the deoxycholic acid liposome preparation prepared in this application has good adipocyte killing specificity.

[0164] Example 2: Evaluation of the uptake capacity of different cells of the medical aesthetic products or pharmaceutical compositions of the present invention.

[0165] 1 Experimental Methods

[0166] 1.1 Preparation of blank liposomes with different fluorescent formulations

[0167] Blank liposomes with different formulations containing Dil fluorescence were prepared in accordance with Examples 8, 9, 10, 11 and 12.

[0168] 1.2 Comparison of in vitro cellular uptake.

[0169] 3T3-L1 (mouse embryonic fibroblasts) were pre-seeded into 96-well plates and induced to differentiate into mature adipocytes for subsequent cell evaluation. HUVECs (human umbilical vein endothelial cells) and 3T3-L1 (mouse embryonic fibroblasts) in logarithmic growth phase were added to 10,000 cells per well of 96-well plates, with a volume of 100 μL per well. The plates were incubated in a CO2 incubator, and after 24 hours of cell culture and cell adhesion, drug was administered. Blank liposomes were diluted with complete culture medium to achieve the same DOPC concentration as the 0.5 mg / mL deoxycholic acid-loaded liposomes in Example 1. The diluted blank liposomes were then added to wells containing mature adipocytes, 3T3-L1 cells, and HUVECs (n=6), with 100 μL added to each well. The cells were cultured in a CO2 incubator for 24 h. The liquid in the wells was aspirated, and the plates were washed twice with PBS. 100 μL of 0.1% Triton sulfate was added to each well, and the plates were shaken at room temperature for 5 min. The cells were then transferred to opaque 96-well plates (50 μL per well), and the fluorescence intensity (FI) was measured using a fluorescence microplate reader (maximum excitation wavelength 540 nm, maximum emission wavelength 590 nm). A separate 96-well plate was set up for each cell type for cell mass calculation. After 48 h of cell culture, mature adipocytes, 3T3-L1 cells, and HUVECs were digested with trypsin and collected from each well. The average cell count per well was determined using a cell counter. Cell uptake capacity was calculated using the following formula:

[0170] Cellular uptake = (FI) 样本 —FI 空白 ) / cells per well × 10 4

[0171] FI 空白 : Indicates the average fluorescence intensity value of cells present but without the addition of blank liposomes.

[0172] Cell uptake results were calculated based on average fluorescence intensity, and the differences in cellular uptake capacity of liposomes were statistically analyzed using Graphpad software.

[0173] 2 Experimental Results

[0174] 2.1 As shown in Figure 6a, when the concentration of DOPC phospholipid in the DOPC+DODMA prescription liposome is 1.5 mg / mL, the cellular uptake capacity of adipocytes for the DOPC+DODMA prescription liposome is significantly higher than that of non-adipocytes (3T3-L1 and HUVEC).

[0175] 2.2 As shown in Figure 6b, when the concentration of DOPC phospholipids in the DOPC+PS formulation liposome is 1.5...

[0176] At a concentration of mg / mL, adipocytes showed significantly higher cellular uptake of DOPC+PS prescription liposomes than non-adipocytes (3T3-L1 and HUVEC).

[0177] 2.3 As shown in Figure 6c, when the concentration of DOPC phospholipid in the DOPC+DOPG formulation liposome is 1.5 mg / mL, the cellular uptake capacity of adipocytes for the DOPC+DOPG formulation liposome is significantly higher than that of non-adipocytes (3T3-L1 and HUVEC).

[0178] 2.4 As shown in Figure 6d, when the concentration of DOPC phospholipid in the DOPC+DSPG prescription liposome is 1.5 mg / mL, the cellular uptake capacity of adipocytes for the DOPC+DSPG prescription liposome is significantly higher than that of non-adipocytes (3T3-L1 and HUVEC).

[0179] 2.5 As shown in Figure 6e, when the concentration of DOPC phospholipid in the DOPC+DSPE-PEG200 formulation liposome is 1.5 mg / mL, the cellular uptake capacity of adipocytes of the DOPC+DSPE-PEG200 formulation liposome is significantly higher than that of non-adipocytes (3T3-L1 and HUVEC).

[0180] The results showed that the different blank liposomes prepared in this application had higher uptake capacity in adipocytes than in other non-adipocytes.

[0181] Example 3: Evaluation of the effects of the medical aesthetic products or pharmaceutical compositions of the present invention on local nonspecific skin damage.

[0182] 1 Experimental Methods

[0183] 1.1 Preparation of liposomes

[0184] Referring to Example 7, drug-loaded liposomes of the DOPC+DSPE-PEG2000 formulation, DC-LIP(PEG), were prepared.

[0185] 1.2 Evaluation of local nonspecific skin lesions

[0186] The experiment used healthy adult New Zealand white rabbits weighing 2.0-2.5 kg. A self-controlled, left-right comparison method was used to evaluate the local skin damage caused by DC-LIP (PEG) and a similar marketed product (Kybella). Three 1cm × 1cm areas were divided on each side of the spine of each animal, as shown in the attached diagram. Figure 8 The protocol shown involved subcutaneous administration of 2 mg per area, administered once daily for 3 consecutive days. After completing the administration, skin tissue from each injection site was collected for HE staining to evaluate the histopathological lesions at the injection site.

[0187] 2 Experimental Results

[0188] Figure 7a shows that the stratified squamous epithelium of the skin in the injection area of ​​Kybella, a similar product, is thinner, the epidermis is degenerated and necrotic, and the cell nuclei are dissolved and disappeared; the collagen fibers in the dermis are degenerated and necrotic, the cell nuclei are condensed and disintegrated, eosinophilicity is increased, and more hair follicles and sebaceous glands and other appendages are degenerated and necrotic.

[0189] Figure 7b shows that the epidermal structure of the skin tissue in the injection area of ​​the DC-LIP (PEG) group is intact and clear, the keratinized layer is obvious, the stratified squamous epithelium is relatively thin, and the cells are arranged neatly and tightly. The collagen fibers in the dermis are interwoven and relatively dense, the cytoplasm is red-stained and uniform, and no obvious pathological changes are observed.

[0190] The results show that the deoxycholic acid liposomes prepared in this application do not cause significant damage to non-adipocytes and tissues of the skin, and have better local application specificity and safety.

Claims

1. A composition for dissolving fats, characterized in that, It was prepared by encapsulating active ingredients that dissolve adipocytes using a liposome delivery system; The active ingredient for dissolving fat cells is bile acid; The bile acids mentioned are selected from deoxycholic acid and / or sodium deoxycholate; The liposome delivery system includes: (a) The first phospholipid as the main membrane material, and (b) The second phospholipid as an auxiliary membrane material; The liposome delivery system also includes a stabilizer; The stabilizer is selected from steroids; The steroid in question is cholesterol; The total mass ratio of the first phospholipid, the second phospholipid, and the stabilizer to the active ingredient is 4:

1. The first phospholipid is selected from dioleoylphosphatidylcholine; The second phospholipid is a polymer-modified phospholipid; The polymer-modified phospholipid is a polyethylene glycol-modified phospholipid; The polyethylene glycol-modified phospholipid is distearate phosphatidylethanolamine-polyethylene glycol 2000; The composition includes 450 parts by weight of dioleoylphosphatidylcholine, 90 parts by weight of distearate phosphatidylethanolamine-polyethylene glycol 2000, and 60 parts by weight of cholesterol.

2. The composition according to claim 1, characterized in that, The liposome drug delivery system is prepared using raw materials containing a first phospholipid and a second phospholipid in the following ratio: the mass ratio of the first phospholipid to the second phospholipid is 15:

3.

3. The composition according to claim 1, characterized in that, The liposome drug delivery system is prepared using raw materials containing a first phospholipid, a second phospholipid, and a stabilizer in the following ratio: the mass ratio of the first phospholipid to the second phospholipid to the stabilizer is 15:3:

2.

4. The composition according to claim 1, characterized in that, The liposome drug delivery system also includes a buffer solution.

5. The composition according to claim 4, characterized in that, The buffer solution is selected from PBS buffer.

6. The composition according to claim 4, characterized in that, The buffer solution is an isotonic buffer solution.

7. The composition according to claim 4, characterized in that, The pH of the buffer solution is 5-8.

8. The composition according to claim 4, characterized in that, The liposome drug delivery system is prepared using raw materials containing a buffer solution and total phospholipids in the following ratio: the volume-to-mass ratio of the buffer solution to total phospholipids is 1:18, ml / mg.

9. The composition according to claim 1, characterized in that, The liposomes in the liposome drug delivery system are negatively charged.

10. The composition according to claim 1, characterized in that, The liposomes in the liposome drug delivery system are single-compartment liposomes.

11. The composition according to claim 4, characterized in that, The volume-to-mass ratio of the buffer solution to the active ingredient is 1:

5.

12. The composition according to claim 1, characterized in that, The average particle size of the liposomes is 100-200 nanometers.

13. The composition according to claim 12, characterized in that, The average particle size of the liposomes is 100-150 nanometers.

14. A method for preparing the composition according to any one of claims 1 to 13, characterized in that, Includes the following steps: I. Take the active ingredient, first phospholipid, second phospholipid and stabilizer according to the formula, add organic solvent and first part of buffer solution, and disperse to form an emulsion; II. Remove the organic solvent from the emulsion in step I, add the second part of the buffer solution, and obtain a suspension; III. Collect the liposomes from the suspension in step II to obtain the product; The organic solvent mentioned in step I is chloroform and / or diethyl ether; Step I involves ultrasonic treatment to form an emulsion; The volume ratio of the first buffer portion to the second buffer portion is 1:

4.

15. The preparation method according to claim 14, characterized in that, The organic solvent mentioned in step I is chloroform.

16. The preparation method according to claim 14, characterized in that, The ultrasound time is 1 to 40 minutes.

17. The preparation method according to claim 14, characterized in that, The ultrasound time was 10 minutes.

18. The preparation method according to claim 14, characterized in that, The ultrasonic temperature is 10~30℃.

19. The preparation method according to claim 14, characterized in that, The ultrasonic temperature is below 20℃.

20. The preparation method according to claim 14, characterized in that, The emulsion formed in step I is a W / O type emulsion.

21. The preparation method according to claim 14, characterized in that, Step II involves removing the organic solvent by vacuum evaporation.

22. The preparation method according to claim 14, characterized in that, Step III involves granulating the obtained suspension to obtain liposomes.

23. The preparation method according to claim 14, characterized in that, Step III involves granulating the obtained suspension using a 0.05~0.6μm extruder.

24. The preparation method according to claim 14, characterized in that, Step III involves granulating the obtained suspension using 0.45μm and 0.22μm extruders.

25. Use of the composition according to any one of claims 1 to 13 in the preparation of a fat-reducing drug or a medical aesthetic product.

26. The use as described in claim 25, characterized in that, The fat-reducing drugs or cosmetic products mentioned are local fat-reducing drugs or cosmetic products.

27. Weight loss drugs or cosmetic products, characterized by: The composition contains any one of claims 1 to 13, and an excipient or auxiliary ingredient acceptable in pharmaceuticals or medical aesthetic products; The excipients or auxiliary ingredients are selected from at least one of vitamin C, benzoic acid, and lidocaine.

28. The drug or medical aesthetic product according to claim 27, characterized in that, The drug or cosmetic product mentioned is an injectable.

29. The drug or medical aesthetic product according to claim 28, characterized in that, The drug or medical aesthetic product is a topical administration preparation for adipose tissue.

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