Lipid nanoparticle, targeted lipid nanoparticle, preparation method and application

Abstract

The invention discloses a lipid nanoparticle, a targeted lipid nanoparticle, and a preparation method and application thereof. The lipid nanoparticle has a core-shell structure, the core is formed by PLGA nanoparticles containing drug molecules, and the shell is formed by raw materials including phospholipid, cholesterol and a modification material; wherein the PLGA nanoparticles containing the drug molecules are formed by raw materials comprising PLGA and the drug molecules; the mass ratio of the PLGA to the phospholipid is (1-5): 1; the mass ratio of the phospholipid to the cholesterol is (3-10): 1, and the mass ratio of the phospholipid to the modification material is (3-10): 1; the drug molecules are selected from one or more of insoluble drugs, water-soluble drugs, genes and proteins; and the modification material is PEG-DSPE (Polyethylene Glycol Distearoyl The hardness of the lipid nanoparticles can be regulated and controlled, and cellular uptake can be improved by regulating and controlling the hardness; the targeted lipid nanoparticles can better improve cellular uptake, promote intestinal epithelial cell absorption and improve oral curative effect.

Description

Technical Field

[0001] The present invention relates to lipid nanoparticles, targeted lipid nanoparticles, preparation methods and applications, and in particular to orally administrable lipid nanoparticles, targeted lipid nanoparticles, preparation methods and applications. Background Art

[0002] Currently, oral administration is the most commonly used form of drug delivery. However, oral administration faces limitations due to the physiological barriers of the gastrointestinal tract, as well as physical barriers such as the solubility and permeability of the drug itself, resulting in unsatisfactory oral administration of many drugs. Oral administration of nanoparticles can improve drug solubility and membrane permeability, thereby enhancing oral absorption of drugs to a certain extent.

[0003] PLGA has good biocompatibility and biodegradability and has been approved by the U.S. Food and Drug Administration (FDA) and the European Medicines Agency (EMA) for use in drug delivery systems. Research on oral drug delivery systems based on PLGA nanoparticles is relatively extensive and can encapsulate a variety of drugs, including hydrophilic and hydrophobic drugs.

[0004] CN107126425A discloses a tanshinone IIA PEG-PLGA-PEG nanoparticle, wherein the carrier material used is a PEG-PLGA-PEG triblock copolymer compound, the drug entrapped is tanshinone IIA, and the nanoparticles have a particle size range of 100 to 400 nm.

[0005] Liposomes are closed vesicles with a double-layer membrane structure composed of amphiphilic molecules such as phospholipids. They are widely used in drug delivery, cosmetics, vaccines, and other fields. Liposomes have been widely used due to their high biocompatibility.

[0006] CN102697721A discloses a salidroside block copolymer lipid nanoparticle preparation, which is mainly made of phospholipids and / or cholesterol and a block copolymer. The block copolymer is selected from PLGA and its derivatives.

[0007] CN103768012A discloses a method for preparing a curcumin lipid nanoparticle suspension or nanoparticles. One part by weight of curcumin and 5 to 20 parts by weight of an amphoteric biodegradable polymer are dissolved in an organic solvent to form a lipid phase solution; a surfactant is dissolved in water to form an aqueous phase solution; the lipid phase solution and the aqueous phase solution are mixed through a microchannel to form particles, and the organic solvent is removed to obtain a curcumin lipid nanoparticle suspension.

[0008] CN112220932A discloses a VCAM-1 monoclonal antibody-tanshinone IIA nanostructured lipid carrier. The VCAM-1 monoclonal antibody-tanshinone IIA lipid nanoparticles are made from 10 parts by mass of tanshinone IIA, 84-89 parts by mass of glycerol monostearate, 0-5 parts by mass of polyethylene glycol monostearate, 0.5 parts by mass of monoamino-terminated polyethylene glycol stearate, and 0.5 parts by mass of VCAM-1 monoclonal antibody.

[0009] So far, there has been no report on combining PLGA, liposomes, and targeted modification materials (or modification materials) to construct nanoparticles that target transporters, improve cellular uptake efficiency, promote intestinal epithelial cell absorption, and have controllable hardness. Summary of the Invention

[0010] One object of the present invention is to provide a lipid nanoparticle whose hardness can be regulated to improve cellular uptake. Another object of the present invention is to provide a targeted lipid nanoparticle that can better improve cellular uptake, promote intestinal epithelial cell absorption, and enhance oral efficacy. Another object of the present invention is to provide a method for preparing the above-mentioned lipid nanoparticles. Another object of the present invention is to provide a method for preparing the above-mentioned targeted lipid nanoparticles. Another object of the present invention is to provide the use of the above-mentioned targeted lipid nanoparticles in the preparation of a drug for treating myocardial ischemia.

[0011] The present invention adopts the following technical solutions to achieve the above-mentioned purpose.

[0012] In one aspect, the present invention provides a lipid nanoparticle having a core-shell structure, wherein the core is formed by PLGA nanoparticles containing drug molecules, and the shell is formed by raw materials including phospholipids, cholesterol and a modifying material;

[0013] Wherein, the PLGA nanoparticles containing drug molecules are formed from raw materials including PLGA and drug molecules;

[0014] The mass ratio of PLGA to phospholipid is 1 to 5:1; the mass ratio of phospholipid to cholesterol is 3 to 10:1; and the mass ratio of phospholipid to modification material is 3 to 10:1.

[0015] Wherein, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins;

[0016] Wherein, the modification material is PEG-DSPE.

[0017] On the other hand, the present invention provides a targeted lipid nanoparticle having a core-shell structure, wherein the core is formed by PLGA nanoparticles containing drug molecules, and the shell is formed by raw materials including phospholipids, cholesterol and a targeting modification material;

[0018] Wherein, the PLGA nanoparticles containing drug molecules are formed from raw materials including PLGA and drug molecules;

[0019] The mass ratio of PLGA to phospholipid is 1 to 5:1; the mass ratio of phospholipid to cholesterol is 3 to 10:1; and the mass ratio of phospholipid to targeting modification material is 3 to 10:1.

[0020] Wherein, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins;

[0021] Wherein, the targeting modification material is selected from one or more of fructose-PEG-DSPE, cholic acid-PEG-DSPE, mannose-PEG-DSPE and valine-valine-PEG-DSPE.

[0022] In another aspect, the present invention further provides a targeted lipid nanoparticle having a core-shell structure, wherein the core is a PLGA nanoparticle and the shell is formed from raw materials including phospholipids, cholesterol, a targeting modification material and a drug molecule;

[0023] The mass ratio of PLGA to phospholipid is 1 to 5:1; the mass ratio of phospholipid to cholesterol is 3 to 10:1; and the mass ratio of phospholipid to targeting modification material is 3 to 10:1.

[0024] Wherein, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins;

[0025] Wherein, the targeting modification material is selected from one or more of fructose-PEG-DSPE, cholic acid-PEG-DSPE, mannose-PEG-DSPE and valine-valine-PEG-DSPE.

[0026] According to the lipid nanoparticles of the present invention, preferably, the molecular weight of PLGA is 10KDa to 100KDa; the drug molecule is a poorly soluble drug, and the poorly soluble drug is selected from one or more of paclitaxel, tanshinone IIA, curcumin and quercetin; the phospholipid is selected from one or more of distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, lecithin and dipalmitoylphosphatidylcholine.

[0027] According to the targeted lipid nanoparticles of the present invention, preferably, the drug molecule is tanshinone IIA or a salt thereof.

[0028] In another aspect, the present invention further provides a method for preparing the lipid nanoparticles as described above, comprising the following steps:

[0029] (1) mixing the drug molecules and PLGA in an organic solvent to obtain an organic phase;

[0030] (2) forming an aqueous phase containing a stabilizer;

[0031] (3) adding the organic phase to the aqueous phase, mixing by oscillation or ultrasound, then removing the organic solvent and separating to obtain PLGA nanoparticles containing drug molecules, and then resuspending them in PBS to obtain an aqueous solution of PLGA nanoparticles containing drug molecules;

[0032] (4) mixing phospholipids, cholesterol, and a modifying material in an organic solvent, and then removing the organic solvent to obtain a film;

[0033] (5) adding an aqueous solution of PLGA nanoparticles containing drug molecules to the film, oscillating or ultrasonicating, and then filtering to obtain lipid nanoparticles;

[0034] Among them, step (1) and step (2) are performed in no particular order.

[0035] In another aspect, the present invention further provides a method for preparing the targeted lipid nanoparticles as described above, comprising the following steps:

[0036] 1) mixing the drug molecule and PLGA in an organic solvent to obtain an organic phase;

[0037] 2) forming an aqueous phase containing a stabilizer;

[0038] 3) adding the organic phase to the aqueous phase, mixing by oscillation or ultrasound, then removing the organic solvent and separating to obtain PLGA nanoparticles containing drug molecules, which are then resuspended in PBS to obtain an aqueous solution of PLGA nanoparticles containing drug molecules;

[0039] 4) mixing phospholipids, cholesterol, and targeted modification materials in an organic solvent, and then removing the organic solvent to obtain a film;

[0040] 5) adding an aqueous solution of PLGA nanoparticles containing drug molecules to the film, oscillating or sonicating, and then filtering to obtain targeted lipid nanoparticles;

[0041] There is no particular order for step 1) and step 2).

[0042] In yet another aspect, the present invention provides a method for preparing the targeted lipid nanoparticles as described above, comprising the following steps:

[0043] 1′) dissolving PLGA in an organic solvent to obtain an organic phase;

[0044] 2') forming an aqueous phase containing a stabilizer;

[0045] 3') adding the organic phase to the aqueous phase, mixing by oscillation or ultrasound, then removing the organic solvent, separating to obtain PLGA nanoparticles, and then resuspending in PBS to obtain a PLGA nanoparticle aqueous solution;

[0046] 4′) mixing phospholipids, cholesterol, targeting modification materials and drug molecules in an organic solvent, and then removing the organic solvent to obtain a film;

[0047] 5′) adding the PLGA nanoparticle aqueous solution to the film, shaking or sonicating, and then filtering to obtain targeted lipid nanoparticles;

[0048] There is no particular order between step 1') and step 2').

[0049] According to the preparation method of the present invention, preferably:

[0050] Dissolving a stabilizer in water to form an aqueous phase; wherein the stabilizer is selected from polyvinyl alcohol and / or Pluronic F127;

[0051] The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform and dichloromethane.

[0052] In the final aspect, the present invention also provides a use of the targeted lipid nanoparticles as described above in the preparation of a drug for preventing myocardial ischemia.

[0053] The hardness of the lipid nanoparticles of the present invention can be regulated, and the hardness can be regulated to improve cellular uptake. Compared with non-targeted lipid nanoparticles, the targeted lipid nanoparticles of the present invention can better improve cellular uptake, promote intestinal epithelial cell absorption of lipid nanoparticles, and improve transmembrane transport efficiency. According to the preferred technical solution of the present invention, the double-modified targeted lipid nanoparticles obtained by using sodium tanshinone IIA sulfonate as the drug molecule and mannose-PEG2K-DSPE and cholic acid-PEG2K-DSPE as targeting modification materials have a good anti-myocardial ischemia effect. The preparation method of the present invention is stable and can obtain lipid nanoparticles with a particle size of 130 to 160 nm, and can obtain targeted lipid nanoparticles with stable performance. BRIEF DESCRIPTION OF THE DRAWINGS

[0054] Figure 1 The cellular uptake of different lipid nanoparticles and targeted lipid nanoparticles by Caco-2 cells (Mean±SD, n=3).

[0055] Figure 2 These are cardiac ultrasound images of the different drug-administered groups.

[0056] Figure 3 The results show the effects of different drug groups on HE staining of mouse myocardial tissue. DETAILED DESCRIPTION

[0057] The present invention will be further described below with reference to specific embodiments, but the protection scope of the present invention is not limited thereto.

[0058] <Lipid Nanoparticles>

[0059] The present invention provides a lipid nanoparticle having a core-shell structure. The core is formed from PLGA nanoparticles containing drug molecules. The shell is formed from raw materials including phospholipids, cholesterol, and a modifying material. According to one embodiment of the present invention, the shell is composed of phospholipids, cholesterol, and a modifying material. The modifying material is PEG-DSPE.

[0060] The drug-molecule-containing PLGA nanoparticles of the present invention are formed from raw materials including PLGA and drug molecules. According to one embodiment of the present invention, the drug-molecule-containing PLGA nanoparticles are formed from PLGA and drug molecules.

[0061] In the present invention, PLGA is poly(lactic-co-glycolic acid), a linear copolymer formed by ring-opening polymerization of lactide and glycolide. Lactide is a cyclic dimer of lactic acid (LA) containing methyl side chains (-CH3); glycolide is a cyclic dimer of glycolic acid (GA) without side chains (only -H).

[0062] In the present invention, when forming PLGA, the molar ratio of lactide to glycolide is 25:75 to 80:20, preferably 75:25.

[0063] In the present invention, the molecular weight of PLGA can be 10KDa to 100KDa, preferably 15KDa to 50KDa, and more preferably 15KDa to 30KDa. The present invention has found through extensive research and experiments that such PLGA is more conducive to regulating the hardness of the obtained lipid nanoparticles to improve cellular uptake.

[0064] In the present invention, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins. In certain embodiments, the drug molecule is a poorly soluble drug. According to one embodiment of the present invention, the poorly soluble drug is selected from one or more of paclitaxel, tanshinone IIA, curcumin and quercetin. The drug molecule may include its corresponding salt. According to a specific embodiment of the present invention, the drug molecule is tanshinone IIA or a salt thereof (e.g., sodium tanshinone IIA sulfonate). The CAS number of tanshinone IIA is 568-72-9.

[0065] In the present invention, the phospholipid is selected from one or more of distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, lecithin and dipalmitoylphosphatidylcholine, preferably lecithin, more preferably soybean lecithin.

[0066] In the present invention, PEG-DSPE is a biomaterial composed of polyethylene glycol (PEG) and distearoylphosphatidylethanolamine (DSPE), wherein the molecular weight of PEG is 1000-5000 Da, preferably 2000-2500 Da.

[0067] In the present invention, the mass ratio of PLGA to phospholipid can be 1 to 5:1, preferably 1 to 3:1, more preferably 1 to 2:1. The mass ratio of PLGA to drug molecule is 6 to 12:1, preferably 7 to 11:1, more preferably 7 to 10:1, and even more preferably 8 to 9:1.

[0068] In the present invention, the mass ratio of phospholipid to cholesterol is 3 to 10:1, preferably 4 to 8:1, and more preferably 5 to 6:1.

[0069] In the present invention, the mass ratio of phospholipid to modifying material is 3 to 10:1, preferably 5 to 10:1.

[0070] This is conducive to obtaining nanoparticles with a particle size of 130 to 160 nm, preferably 135 to 155 nm, and is conducive to regulating hardness, thereby improving cellular uptake.

[0071] <Targeted lipid nanoparticles>

[0072] Targeted lipid nanoparticles A

[0073] Targeted lipid nanoparticles A have a core-shell structure. The core is formed from PLGA nanoparticles containing drug molecules. The shell is formed from raw materials including phospholipids, cholesterol, and a targeting modification material. According to one embodiment of the present invention, the shell is composed of phospholipids, cholesterol, and a targeting modification material.

[0074] In the present invention, the PLGA nanoparticles containing drug molecules are formed from raw materials including PLGA and drug molecules. According to one embodiment of the present invention, the PLGA nanoparticles containing drug molecules are formed from PLGA and drug molecules.

[0075] In the present invention, PLGA is poly(lactic-co-glycolic acid), a linear copolymer formed by ring-opening polymerization of lactide and glycolide. Lactide is a cyclic dimer of lactic acid (LA) containing methyl side chains (-CH3); glycolide is a cyclic dimer of glycolic acid (GA) without side chains (only -H).

[0076] In the present invention, when forming PLGA, the molar ratio of lactide to glycolide is 25:75 to 80:20, preferably 75:25.

[0077] In the present invention, the molecular weight of PLGA can be 5KDa to 100KDa, preferably 10KDa to 50KDa, and more preferably 15KDa to 30KDa. The present invention has found through extensive research and experiments that such PLGA is more conducive to increasing the hardness of the resulting lipid nanoparticles and can improve cellular uptake by regulating the hardness.

[0078] In the present invention, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins. In certain embodiments, the drug molecule is a poorly soluble drug. According to one embodiment of the present invention, the poorly soluble drug is selected from one or more of paclitaxel, tanshinone IIA, curcumin and quercetin. The drug molecule includes a corresponding salt. According to a specific embodiment of the present invention, the drug molecule is tanshinone IIA or a salt thereof (e.g., sodium tanshinone IIA sulfonate). The CAS number of tanshinone IIA is 568-72-9.

[0079] In the present invention, the phospholipid is selected from one or more of distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, lecithin and dipalmitoylphosphatidylcholine, preferably lecithin, more preferably soybean lecithin.

[0080] In the present invention, the targeting modification material is preferably selected from one or more of fructose-PEG-DSPE, cholic acid-PEG-DSPE, and mannose-PEG-DSPE. In certain embodiments, the targeting modification material is fructose-PEG-DSPE. In other embodiments, the targeting modification material is a mixture of cholic acid-PEG-DSPE and mannose-PEG-DSPE, with the mass ratio of the two being 1:1.

[0081] In the present invention, fructose-PEG-DSPE is a biomaterial composed of fructose, polyethylene glycol (PEG) and distearoylphosphatidylethanolamine (DSPE), wherein the molecular weight of PEG is 1000-5000 Da, preferably 2000-2500 Da.

[0082] Cholic acid-PEG-DSPE is a biomaterial composed of cholic acid, polyethylene glycol (PEG), and distearoylphosphatidylethanolamine (DSPE). The molecular weight of PEG is 1000-5000 Da, preferably 2000-2500 Da.

[0083] Mannose-PEG-DSPE is a biomaterial composed of mannose, polyethylene glycol (PEG), and distearoylphosphatidylethanolamine (DSPE), wherein the molecular weight of PEG is 1000-5000 Da, preferably 2000-2500 Da.

[0084] In the present invention, the mass ratio of PLGA to phospholipid can be 1 to 5:1, preferably 1 to 3:1, more preferably 1 to 2:1. The mass ratio of PLGA to drug molecule is 6 to 12:1, preferably 7 to 11:1, more preferably 7 to 10:1, and even more preferably 8 to 9:1.

[0085] In the present invention, the mass ratio of phospholipid to cholesterol is 3 to 10:1, preferably 4 to 8:1, and more preferably 5 to 6:1.

[0086] In the present invention, the mass ratio of phospholipid to targeting modification material is 3 to 10:1, preferably 5 to 10:1.

[0087] This is conducive to obtaining nanoparticles with a particle size of 130 to 160 nm, preferably nanoparticles with a particle size of 135 to 155 nm, which is conducive to better improving cellular uptake and promoting intestinal epithelial cell absorption of lipid nanoparticles.

[0088] Targeted lipid nanoparticles B

[0089] Targeted lipid nanoparticles B have a core-shell structure. The core is formed from PLGA nanoparticles. The PLGA nanoparticles do not contain drug molecules. The shell is formed from raw materials including phospholipids, cholesterol, a targeting modification material, and drug molecules. According to one embodiment of the present invention, the shell is composed of phospholipids, cholesterol, a targeting modification material, and drug molecules.

[0090] According to one embodiment of the present invention, the PLGA nanoparticles consist of PLGA.

[0091] PLGA, phospholipids, drug molecules, and targeting modification materials are as described above and are not described in detail here. According to a specific embodiment of the present invention, the drug molecule is sodium tanshinone IIA sulfonate.

[0092] In the present invention, the targeting modification material is preferably selected from one or more of fructose-PEG-DSPE, cholic acid-PEG-DSPE, and mannose-PEG-DSPE. According to a specific embodiment of the present invention, the targeting modification material is a mixture of cholic acid-PEG-DSPE and mannose-PEG-DSPE, with a mass ratio of 1:1.

[0093] In the present invention, the mass ratio of PLGA to phospholipid may be 1 to 5:1, preferably 1 to 3:1, and more preferably 1 to 2:1.

[0094] In the present invention, the mass ratio of phospholipid to cholesterol is 3 to 10:1, preferably 4 to 8:1, and more preferably 5 to 6:1.

[0095] In the present invention, the mass ratio of phospholipid to targeting modification material is 3 to 10:1, preferably 5 to 10:1. The mass ratio of drug molecule to PLGA is 1:6 to 12, preferably 1:7 to 11, and more preferably 1:8 to 10.

[0096] This is conducive to obtaining nanoparticles with a particle size of 130 to 160 nm, preferably nanoparticles with a particle size of 135 to 155 nm, which is conducive to better improving cellular uptake and promoting intestinal epithelial cell absorption of lipid nanoparticles.

[0097] <Preparation method>

[0098] Preparation method of lipid nanoparticles

[0099] The present invention provides a method for preparing lipid nanoparticles as described above, comprising the following steps: (1) forming an organic phase containing PLGA and drug molecules; (2) forming an aqueous phase; (3) forming PLGA nanoparticles containing drug molecules; (4) forming a film free of drug molecules; and (5) forming lipid nanoparticles. Steps (1) and (2) may be performed in any order. A detailed description is provided below.

[0100] In step (1), the drug molecule and PLGA are mixed in an organic solvent to obtain an organic phase. The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform, and dichloromethane, preferably dichloromethane or chloroform, more preferably dichloromethane. The mass volume ratio of PLGA to the organic solvent can be 15 to 30 mg / mL, preferably 20 to 30 mg / mL, and more preferably 25 to 28 mg / mL.

[0101] In step (2), an aqueous phase containing a stabilizer is formed. Specifically, the stabilizer is dissolved in water to obtain an aqueous phase. The stabilizer is selected from polyvinyl alcohol and / or Pluronic F127. The stabilizer is preferably polyvinyl alcohol (PVA) having a molecular weight of 5 to 200 kDa, preferably 30 to 100 kDa, and more preferably 30 to 70 kDa. The concentration of the stabilizer in the aqueous phase can be 1.5 to 3.5 mg / mL, preferably 2 to 3 mg / mL, and more preferably 2 to 2.5 mg / mL.

[0102] In step (3), the organic phase is added to the aqueous phase, mixed by oscillation or ultrasound, and then the organic solvent is removed and separated to obtain PLGA nanoparticles containing drug molecules. The PLGA nanoparticles are then resuspended in PBS to obtain an aqueous solution of PLGA nanoparticles containing drug molecules. The volume ratio of the organic phase to the aqueous phase is 1:8 to 15, preferably 1:9 to 13, and more preferably 1:9 to 11.

[0103] The organic phase can be added dropwise to the aqueous phase. Preferably, ultrasonic mixing is employed, with ultrasonication continued for 8 to 15 minutes, preferably 10 to 15 minutes. After ultrasonication, stirring is continued for 8 to 12 hours. The organic solvent can then be removed by rotary evaporation. Separation can be performed by centrifugation, with a centrifugal speed of 8,000 to 15,000 r / min, preferably 10,000 to 15,000 r / min, and more preferably 12,000 to 13,000 r / min. In the present invention, PBS is an abbreviation for phosphate buffered saline, which is known in the art.

[0104] In step (4), the phospholipid, cholesterol, and the modifying material are mixed in an organic solvent, and then the organic solvent is removed to obtain a film. The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform, and dichloromethane. Preferably, the organic solvent is a mixed solvent of chloroform and methanol, and the volume ratio of the two is 3 to 4:1. According to one embodiment of the present invention, the phospholipid, cholesterol, and the modifying material are sequentially added to the organic solvent and mixed. After uniform mixing, the organic solvent is removed using a rotary evaporator to obtain a film.

[0105] In step (5), an aqueous solution of PLGA nanoparticles containing drug molecules is added to the film, shaken or sonicated, and then filtered to obtain lipid nanoparticles. Preferably, the method is ultrasonication. Specifically, water bath ultrasonication is performed for 5 to 7 minutes, followed by probe ultrasonication for 10 to 15 minutes (ultrasonic power is 300 to 350 W, and the parameters are cycled such that ultrasonication is performed for 2 to 3 seconds followed by a 2 to 3 second pause), and the resulting solution is then filtered using a filter membrane with a diameter of less than 0.5 μm, preferably a 0.22 μm filter membrane, to obtain lipid nanoparticles.

[0106] Preparation method of targeted lipid nanoparticles A

[0107] The preparation method of targeted lipid nanoparticles A comprises the following steps: 1) forming an organic phase containing PLGA and drug molecules; 2) forming an aqueous phase; 3) forming an aqueous solution of PLGA nanoparticles containing drug molecules; 4) forming a film free of drug molecules; and 5) forming the targeted lipid nanoparticles. Steps 1) and 2) can be performed in any order. A detailed description is provided below.

[0108] Formation of an organic phase containing PLGA and drug molecules

[0109] In step 1), the drug molecule and PLGA are mixed in an organic solvent to obtain an organic phase. The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform, and dichloromethane, preferably dichloromethane or chloroform, and more preferably dichloromethane. The mass volume ratio of PLGA to the organic solvent can be 15 to 30 mg / mL, preferably 20 to 30 mg / mL, and more preferably 25 to 28 mg / mL.

[0110] Forming an aqueous phase

[0111] In step 2), an aqueous phase containing a stabilizer is formed. Specifically, the stabilizer is dissolved in water to form an aqueous phase. The stabilizer is selected from polyvinyl alcohol and / or Pluronic F127. The stabilizer is preferably polyvinyl alcohol (PVA) having a molecular weight of 5 to 200 kDa, preferably 30 to 100 kDa, and more preferably 30 to 70 kDa. The concentration of the stabilizer in the aqueous phase may be 1.5 to 3.5 mg / mL, preferably 2 to 3 mg / mL, and more preferably 2 to 2.5 mg / mL.

[0112] Formation of PLGA nanoparticle aqueous solution containing drug molecules

[0113] In step 3), the organic phase is added to the aqueous phase and mixed by oscillation or ultrasound. The organic solvent is then removed and separated to obtain drug-containing PLGA nanoparticles, which are then resuspended in PBS to obtain an aqueous solution of drug-containing PLGA nanoparticles. The volume ratio of the organic phase to the aqueous phase is 1:8-15, preferably 1:9-13, and more preferably 1:9-11. The organic phase can be added dropwise to the aqueous phase. Preferably, ultrasonic mixing is performed, with sonication continued for 8-15 minutes, preferably 10-15 minutes. After sonication, stirring is continued for 8-12 hours. The organic solvent can then be removed by rotary evaporation. Separation can be performed by centrifugation at a speed of 8,000-15,000 r / min, preferably 10,000-15,000 r / min, and more preferably 12,000-13,000 r / min. The resulting PLGA nanoparticles are resuspended in PBS to obtain an aqueous solution of drug-containing PLGA nanoparticles.

[0114] Formation of a film free of drug molecules

[0115] In step 4), the phospholipid, cholesterol, and targeting modification material are mixed in an organic solvent, and the organic solvent is then removed to obtain a thin film. The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform, and dichloromethane. Preferably, the organic solvent is a mixed solvent of chloroform and methanol in a volume ratio of 3 to 4:1. According to one embodiment of the present invention, the phospholipid, cholesterol, and targeting modification material are sequentially added to the organic solvent and mixed. After uniform mixing, the organic solvent is removed using a rotary evaporator to obtain a thin film.

[0116] Formation of targeted lipid nanoparticles A

[0117] In step 5), an aqueous solution of PLGA nanoparticles containing drug molecules is added to the film, shaken or sonicated, and then filtered to obtain targeted lipid nanoparticles. Preferably, ultrasound is used. Specifically, water bath ultrasound is first performed for 5 to 7 minutes, followed by probe ultrasound for 10 to 15 minutes (ultrasound power is 300 to 350 W, and the parameters are cycled such that ultrasound is continued for 2 to 3 seconds and then stopped for 2 to 3 seconds). The resulting solution is then filtered using a filter membrane with a diameter of less than 0.5 μm, preferably a 0.22 μm filter membrane, to obtain lipid nanoparticles A.

[0118] Preparation method of targeted lipid nanoparticles B

[0119] The preparation method of targeted lipid nanoparticles B comprises the following steps: 1) forming an organic phase free of drug molecules; 2) forming an aqueous phase; 3) forming an aqueous solution of PLGA nanoparticles; 4) forming a film containing drug molecules; and 5) obtaining the targeted lipid nanoparticles. Steps 1) and 2) can be performed in any order. A detailed description is provided below.

[0120] Formation of an organic phase free of drug molecules

[0121] In step 1'), PLGA is dissolved in an organic solvent to obtain an organic phase. The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform, and dichloromethane, preferably dichloromethane or chloroform, and more preferably dichloromethane. The mass volume ratio of PLGA to organic solvent can be 15 to 30 mg / mL, preferably 20 to 30 mg / mL, and more preferably 25 to 28 mg / mL.

[0122] Forming an aqueous phase

[0123] In step 2'), an aqueous phase containing a stabilizer is formed. Specifically, the stabilizer is dissolved in water to form an aqueous phase. The stabilizer is selected from polyvinyl alcohol and / or Pluronic F127. The stabilizer is preferably polyvinyl alcohol (PVA) having a molecular weight of 5 to 200 kDa, preferably 30 to 100 kDa, and more preferably 30 to 70 kDa. The concentration of the stabilizer in the aqueous phase can be 1.5 to 3.5 mg / mL, preferably 2 to 3 mg / mL, and more preferably 2 to 2.5 mg / mL.

[0124] Formation of PLGA nanoparticle aqueous solution

[0125] In step 3'), the organic phase is added to the aqueous phase and mixed by oscillation or ultrasound. The organic solvent is then removed and the precipitate is separated to obtain PLGA nanoparticles (without drug molecules). The precipitate is then resuspended in PBS to obtain a PLGA nanoparticle aqueous solution. The volume ratio of the organic phase to the aqueous phase is 1:8-15, preferably 1:9-13, and more preferably 1:9-11. The organic phase can be added dropwise to the aqueous phase. Preferably, ultrasonic mixing is performed, and the ultrasonication is continued for 8-15 minutes, preferably 10-15 minutes. After the ultrasonication, stirring is continued for 8-12 hours. The organic solvent can then be removed by rotary evaporation. The separation method can be centrifugation, and the centrifugal speed can be 8000-15000 r / min, preferably 10000-15000 r / min, and more preferably 12000-13000 r / min. The obtained PLGA nanoparticles are resuspended in PBS to obtain a PLGA nanoparticle aqueous solution.

[0126] Formation of a thin film containing drug molecules

[0127] In step 4'), phospholipids, cholesterol, targeting modification materials and drug molecules are mixed in an organic solvent, and then the organic solvent is removed to obtain a thin film.

[0128] The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform, and dichloromethane. Preferably, the organic solvent is a mixed solvent of chloroform and methanol, with a volume ratio of 3 to 4:1. According to one embodiment of the present invention, phospholipids, cholesterol, targeting modification materials, and drug molecules are sequentially added to the organic solvent and mixed. After uniform mixing, the organic solvent is removed using a rotary evaporator to obtain a film.

[0129] Formation of targeted lipid nanoparticles B

[0130] In step 5'), an aqueous solution of PLGA nanoparticles is added to the film, shaken or sonicated, and then filtered to obtain targeted lipid nanoparticles. Ultrasound is preferred. Specifically, water bath sonication is performed for 5 to 7 minutes, followed by probe sonication for 10 to 15 minutes (ultrasound power of 300 to 350 W, with a parameter cycle of sonicating for 2 to 3 seconds and then stopping for 2 to 3 seconds). The resulting solution is then filtered through a filter membrane with a diameter of less than 0.5 μm, preferably a 0.22 μm filter membrane, to obtain targeted lipid nanoparticles B.

[0131] <Application>

[0132] The present invention also provides the use of the targeted lipid nanoparticles as described above in the preparation of a drug for treating myocardial ischemia. The drug molecule used in the targeted lipid nanoparticles is tanshinone IIA or a salt thereof (e.g., sodium tanshinone IIA sulfonate). The targeted lipid nanoparticles of the present invention can better improve cellular uptake. Moreover, compared with non-targeted lipid nanoparticles, the targeted lipid nanoparticles can improve the intestinal epithelial cell absorption efficiency of the lipid nanoparticles. At the same time, by regulating the hardness of the nanoparticles, the intestinal epithelial cell uptake of the nanoparticles can also be significantly regulated. Constructing nanoparticles with controllable hardness and receptor targeting can improve transmembrane transport efficiency. The targeted lipid nanoparticles of the present invention have a good anti-myocardial ischemia effect.

[0133] <Test Method>

[0134] Particle size determination: The particle size of lipid nanoparticles in the solution was measured using a laser particle size analyzer.

[0135] Determination of Young's modulus: Atomic force microscope was used for testing.

[0136] The sources of some of the raw materials in the following preparation examples and embodiments are introduced as follows:

[0137] PLGA (polylactic-co-glycolic acid) with molecular weights of 15 KDa, 30 KDa, and 100 KDa, respectively; when forming PLGA, the molar ratio of lactide to glycolide is 75:25; sourced from Jinan Daigang Biotechnology Co., Ltd. Tanshinone IIA: sourced from Shanghai Yuanye Biotechnology Co., Ltd. PEG2K-DSPE: wherein the molecular weight of PEG is 2000 Da; sourced from Xi'an Ruixi Biotechnology Co., Ltd. Fructose-PEG2K-DSPE: wherein the molecular weight of PEG is 2000 Da; sourced from Chongqing Yusi Pharmaceutical Technology Co., Ltd. Mannose-PEG2K-DSPE, cholic acid-PEG2K-DSPE: wherein the molecular weight of PEG is 2000 Da; sourced from Xi'an Ruixi Biotechnology Co., Ltd. Caco-2 cells: sourced from Nanjing KeyGen Biotechnology Co., Ltd.

[0138] Preparation Example 1 - Preparation of PLGA Nanoparticles Containing Drug Molecules

[0139] 40 mg of PLGA (PLGA molecular weight: 15 KDa) and 5 mg of Tanshinone IIA were sequentially added to 1.5 mL of dichloromethane and dissolved to obtain an organic phase.

[0140] PVA (polyvinyl alcohol, molecular weight 30-70 kDa) was dissolved in water to obtain an aqueous phase containing PVA. The concentration of PVA in the aqueous phase was 2 mg / mL.

[0141] The organic phase was added to 15 mL of the PVA-containing aqueous phase under ice-cooling and ultrasonication (ultrasonic power of 300 W, with a 1-second on / 1-second off cycle). Ultrasonication was continued for 10 minutes and then stirred overnight to remove the organic solvent. The mixture was then centrifuged at 12,000 rpm, the precipitate collected, and washed with deionized water three times to remove excess PVA and unencapsulated tanshinone IIA, yielding drug-containing PLGA nanoparticles. The mixture was then resuspended in PBS to obtain a drug-containing PLGA nanoparticle aqueous solution.

[0142] Preparation Example 2-Preparation of PLGA Nanoparticles Containing Drug Molecules

[0143] The only difference from Preparation Example 1 is that the molecular weight of the PLGA used is 100 KDa.

[0144] Preparation Example 3 - Preparation of PLGA Nanoparticles Containing Drug Molecules

[0145] The only difference from Preparation Example 1 is that the molecular weight of the PLGA used is 30 KDa.

[0146] Example 1 - Preparation of lipid nanoparticles

[0147] 20 mg of soybean lecithin (denoted as PC), 4 mg of cholesterol, and 2 mg of PEG2K-DSPE were sequentially added to a chloroform-methanol mixed solvent (the volume ratio of chloroform to methanol was 3:1) and mixed. After mixing evenly, the organic solvent was removed using a rotary evaporator to form a uniform film.

[0148] The film was added with the drug-containing aqueous solution of PLGA nanoparticles prepared in Preparation Example 1, such that the mass ratio of PLGA in the drug-containing aqueous solution of PLGA nanoparticles to the mass ratio of phospholipids in the film was 1:1. Water bath sonication was performed for 5 minutes, followed by probe sonication for 10 minutes (ultrasonic power of 300 W, with a 2-second sonication and 2-second rest cycle). The resulting solution was filtered through a 0.22 μm filter membrane to obtain the final lipid nanoparticles, designated PLGA15K-LP (1:1).

[0149] Example 2 - Preparation of lipid nanoparticles

[0150] The only difference from Example 1 is that the mass ratio of PLGA in the aqueous solution of PLGA nanoparticles containing drug molecules to the mass ratio of phospholipids in the film is 2:1. The obtained lipid nanoparticles are designated as PLGA15K-LP (2:1).

[0151] Example 3 - Preparation of lipid nanoparticles

[0152] The only difference from Example 1 is that the mass ratio of PLGA in the aqueous solution of PLGA nanoparticles containing drug molecules to the mass ratio of phospholipids in the film is 5:1. The obtained lipid nanoparticles are designated as PLGA15K-LP (5:1).

[0153] Example 4 - Preparation of lipid nanoparticles

[0154] The only difference from Example 1 was that the drug-containing PLGA nanoparticle aqueous solution used was from Preparation Example 2, and the mass ratio of PLGA in the drug-containing PLGA nanoparticle aqueous solution to the phospholipid in the film was 5:1. The resulting lipid nanoparticles were designated PLGA100K-LP (5:1).

[0155] Table 1 Particle size and Young's modulus of different lipid nanoparticles (Mean ± SD, n = 3)

[0156]

[0157] As can be seen from the table, the present invention can obtain lipid nanoparticles with a particle size of 135 to 155 nm. As can be seen from the polydispersity index (PDI), the particle size distribution of the obtained lipid nanoparticles is relatively uniform.

[0158] As can be seen from the results of Examples 1 to 3, as the amount of PLGA increases, the Young's modulus of the lipid nanoparticles first increases and then decreases. By comparing Example 3 and Example 4, it can be seen that as the molecular weight of PLGA increases, the Young's modulus increases. This shows that the present invention can produce lipid nanoparticles with controllable hardness.

[0159] Example 5 - Preparation of lipid nanoparticles

[0160] The only difference from Example 1 is that the aqueous solution of PLGA nanoparticles containing drug molecules used is from Preparation Example 3. The obtained lipid nanoparticles are designated as PLGA30K-LP.

[0161] Example 6 - Preparation of targeted lipid nanoparticles

[0162] 20 mg of phospholipid (soy lecithin), 4 mg of cholesterol, and 2 mg of fructose-PEG2K-DSPE were sequentially added to a chloroform-methanol mixed solvent (the volume ratio of chloroform to methanol was 3:1) and mixed. After mixing evenly, the organic solvent was removed using a rotary evaporator to form a uniform film.

[0163] The PLGA nanoparticle aqueous solution containing drug molecules prepared in Preparation Example 3 was added to the film (so that the mass ratio of PLGA to phospholipid was 1:1), and water bath ultrasound was performed for 5 minutes, and probe ultrasound was continued for 10 minutes (ultrasonic power was 300 W, and the parameter cycle was set to ultrasound for 2 seconds and then stop for 2 seconds). The resulting solution was filtered with a 0.22 μm filter membrane to obtain fructose-modified targeted lipid nanoparticles, which were recorded as FD-PLGA30K-LP.

[0164] Preparation Example 4-Preparation of PLGA Nanoparticles

[0165] 40 mg of PLGA (PLGA molecular weight: 30 KDa) was added to 1.5 mL of dichloromethane and dissolved to obtain an organic phase.

[0166] PVA (polyvinyl alcohol, molecular weight 30-70 kDa) was dissolved in water to obtain an aqueous phase containing PVA. The concentration of PVA in the aqueous phase was 2 mg / mL.

[0167] The organic phase was added to 15 mL of the PVA-containing aqueous phase under ultrasonic conditions (ultrasonic power of 300 W, with a 1-second on / 1-second off cycle) in an ice bath. Ultrasonication was continued for 10 minutes, followed by stirring overnight to remove the organic solvent. The mixture was then centrifuged at 12,000 rpm, the precipitate collected, and washed with deionized water three times to remove excess PVA, yielding PLGA nanoparticles (drug-free). The PLGA nanoparticles were then resuspended in PBS to obtain an aqueous solution.

[0168] Example 7 - Preparation of targeted lipid nanoparticles

[0169] 20 mg of phospholipid (soy lecithin), 4 mg of cholesterol, 2 mg of mannose-PEG2K-DSPE, 2 mg of cholic acid-PEG2K-DSPE, and (2 mg) of tanshinone IIA sodium sulfonate were sequentially added to a chloroform-methanol mixed solvent (the volume ratio of chloroform to methanol was 3:1) and mixed. After mixing evenly, the organic solvent was removed using a rotary evaporator to form a uniform film.

[0170] The film was added with the drug-free aqueous solution of PLGA nanoparticles prepared in Preparation Example 4 (so that the mass ratio of PLGA to phospholipid was 1:1). The film was sonicated in a water bath for 5 minutes and then probe-ultrasounded for 10 minutes (ultrasound power of 300 W, with a 2-second sonication cycle and a 2-second rest cycle). The resulting solution was filtered through a 0.22 μm filter membrane to obtain mannose- and bile acid-coated targeted lipid nanoparticles, designated MD-PLGA-LP.

[0171] Comparative Preparation Example 1-Preparation of LP Liposomes

[0172] 20 mg of soy lecithin, 4 mg of cholesterol, 2 mg of PEG2K-DSPE, and 2.5 mg of tanshinone IIA were sequentially added to a chloroform-methanol mixture (chloroform:methanol volume ratio of 3:1) and mixed. After thorough mixing, the organic solvent was removed using a rotary evaporator to form a uniform film. PBS solution was added to the film, and water bath sonication was performed for 5 minutes, followed by probe sonication for 10 minutes (ultrasonic power of 300 W, with a 2-second on / off cycle). The resulting solution was filtered through a 0.22 μm filter to obtain drug-loaded liposomes, designated as LP.

[0173] Comparative Preparation Example 2-Preparation of LP Liposomes

[0174] 20 mg of soy lecithin, 4 mg of cholesterol, and 2 mg of PEG2K-DSPE were sequentially added to a chloroform-methanol mixture (chloroform:methanol volume ratio of 3:1) and mixed. After thorough mixing, the organic solvent was removed using a rotary evaporator to form a uniform film. A PBS solution of 2.5 mg of sodium tanshinone IIA sulfonate was added to the film. The film was sonicated in a water bath for 5 minutes, followed by a probe sonication cycle for 10 minutes (ultrasonic power of 300 W, with a 2-second interval and a 2-second interval). The resulting solution was filtered through a 0.22 μm filter membrane to obtain drug-loaded liposomes, designated as LP.

[0175] Application test case

[0176] 1. Uptake Efficiency

[0177] 1.1 Experimental methods

[0178] Caco-2 cells were plated at 2×10 5 Cells were seeded into 12-well plates at 100 μL / well and cultured for 48 hours. The culture medium was discarded, and Caco-2 cells were incubated with different fluorescently labeled lipid nanoparticles for 2 hours, with triplicate wells per group. The solution was then discarded, and the cells were washed three times with PBS. The cells were digested with 0.25% trypsin, harvested, washed twice with PBS, and resuspended in 500 μL of PBS. The cells were then filtered through a 200-mesh nylon cloth and quantified using flow cytometry.

[0179] The lipid nanoparticles used were those of Example 1, Example 2, Example 3, Example 4, and Example 5, respectively. The liposomes of Preparation Example 1 were compared, and the targeted lipid nanoparticles (FD-PLGA30K-LP) of Example 6 were also tested. The relationship between lipid nanoparticles of different hardness and cellular uptake was investigated. The results are shown in FIG. Figure 1 A shown and Figure 1 As shown in B. Figure 1 In B, “*” indicates P < 0.05, “***” indicates P < 0.001, and “****” indicates P < 0.0001.

[0180] 1.2 Experimental Results

[0181] Depend on Figure 1 As shown in Figure 1, the lipid nanoparticle group (PLGA30K-LP) improved the cellular uptake of liposome LP compared to the LP group (derived from Comparative Preparation Example 1). Compared to the non-targeted lipid nanoparticles (PLGA30K-LP), the fructose-modified targeted lipid nanoparticles (FD-PLGA30K-LP) significantly improved the intestinal epithelial cell absorption of lipid nanoparticles.

[0182] Depend on Figure 1 B shows that the uptake of lipid nanoparticles varies with the mass ratio of PLGA to phospholipid PC. The hardest-working PLGA15K-LP (5:1) (PLGA to PC mass ratio of 5:1; Young's modulus of 173.67 MPa) exhibits the lowest uptake, while the harder PLGA15K-LP (1:1) and PLGA15K-LP (2:1) (Young's moduli of 310.67 MPa and 348.33 MPa, respectively) exhibit higher uptake. These results indicate that the uptake of lipid nanoparticles increases with increasing hardness. Therefore, adjusting the hardness of lipid nanoparticles can improve their cellular uptake.

[0183] 2. In vivo anti-myocardial ischemia efficacy

[0184] 2.1 Experimental methods

[0185] Coronary artery ligation was used to establish the model. Using ICR mice as the model, the mice were anesthetized (sodium pentobarbital, intraperitoneal injection), fixed in the supine position with tape, and assisted breathing was performed using a small animal ventilator; a small horizontal skin incision was made between the 3rd and 4th intercostal spaces at the left sternal border, and the heart was exposed after separating the underlying muscles; under an in vitro microscope, the left anterior descending artery was ligated with 8-0 silk suture; the reduction in myocardial activity on the anterior wall of the left ventricle indicated that the ligation was successful. The sham-operated group (n=8) used the same surgical method except for the ligation. 3 days after the operation, the surviving ICR mice were randomly divided into 7 groups (n=7): sham-operated group (Sham group), model group (Model group), sodium tanshinone IIA sulfonate group (STS group), sodium tanshinone IIA sulfonate liposome group (i.e., LP group, LP derived from Comparative Preparation Example 2), MD-PLGA-LP group (derived from Example 7) (20 mg·kg by oral gavage) -1 ·d -1 ) and administered intragastrically for three consecutive days. Cardiac function was then assessed using echocardiography. Simultaneously, myocardial tissue from the ischemic and marginal ischemic zones of the post-MI heart failure group was obtained and HE staining was performed to examine the effect of improving cardiac pathology.

[0186] Echocardiography was used to determine the phenotype of heart failure after myocardial infarction and the effects of each preparation group on cardiac function. Figure 2 The results of HE staining of myocardial tissue are shown in Figure 3 shown.

[0187] 2.2 Experimental Results

[0188] Depend on Figure 2 Compared with the Sham group, the ventricular cavity of the Model group mice was enlarged and the ventricular wall was thinned. Compared with the Model group, the ventricular cavity of the STS group, LP group, and MD-PLGA-LP group mice was reduced to varying degrees, and the ventricular wall was thickened to varying degrees. The ventricular remodeling effect of the LP group and MD-PLGA-LP group was better than that of the STS group. Compared with the LP group, the ventricular cavity of the MD-PLGA-LP group was more significantly reduced, the ventricular wall was thickened, and the ventricular remodeling effect was better.

[0189] Depend on Figure 3 It can be seen that the cardiomyocytes of the blank control group (normal saline group, i.e., Sham group) mice were normal in morphology, with no cell edema, no inflammatory cell infiltration in the intercellular matrix, and uniform myocardial fiber staining. Compared with the blank control group, the cardiomyocytes of the model group mice were irregular in morphology, with enlarged nuclei and darker colors, disordered myocardial fiber arrangement, and even myocardial fiber rupture and widened intercellular spaces. Compared with the model group, myocardial damage in different drug administration groups was alleviated to varying degrees, with more normal cardiomyocyte morphology, more regular myocardial fiber distribution, and reduced intercellular spaces and inflammatory cell infiltration to varying degrees. Among them, the effect of the MD-PLGA-LP group was better. It can better improve myocardial ischemia.

[0190] The present invention is not limited to the above-mentioned embodiments. Any modification, improvement, or substitution that can be conceived by those skilled in the art without departing from the essential content of the present invention shall fall within the scope of the present invention.

Claims

1. A lipid nanoparticle, characterized in that It has a core-shell structure, wherein the core is formed by PLGA nanoparticles containing drug molecules, and the shell is formed by raw materials including phospholipids, cholesterol and modification materials; Wherein, the PLGA nanoparticles containing drug molecules are formed from raw materials including PLGA and drug molecules; The mass ratio of PLGA to phospholipid is 1 to 5:1; the mass ratio of phospholipid to cholesterol is 3 to 10:1; and the mass ratio of phospholipid to modification material is 3 to 10:

1. Wherein, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins; Wherein, the modification material is PEG-DSPE.

2. A targeted lipid nanoparticle, characterized in that: It has a core-shell structure, wherein the core is formed by PLGA nanoparticles containing drug molecules, and the shell is formed by raw materials including phospholipids, cholesterol and targeting modification materials; Wherein, the PLGA nanoparticles containing drug molecules are formed from raw materials including PLGA and drug molecules; The mass ratio of PLGA to phospholipid is 1 to 5:1; the mass ratio of phospholipid to cholesterol is 3 to 10:1; and the mass ratio of phospholipid to targeting modification material is 3 to 10:

1. Wherein, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins; Wherein, the targeting modification material is selected from one or more of fructose-PEG-DSPE, cholic acid-PEG-DSPE, mannose-PEG-DSPE and valine-valine-PEG-DSPE.

3. A targeted lipid nanoparticle, characterized in that: It has a core-shell structure, with the core being PLGA nanoparticles and the shell being formed from raw materials including phospholipids, cholesterol, targeting modification materials and drug molecules; The mass ratio of PLGA to phospholipid is 1 to 5:1; the mass ratio of phospholipid to cholesterol is 3 to 10:1; and the mass ratio of phospholipid to targeting modification material is 3 to 10:

1. Wherein, the drug molecule is selected from one or more of poorly soluble drugs, water-soluble drugs, genes and proteins; Wherein, the targeting modification material is selected from one or more of fructose-PEG-DSPE, cholic acid-PEG-DSPE, mannose-PEG-DSPE and valine-valine-PEG-DSPE.

4. The lipid nanoparticle according to claim 1, wherein The molecular weight of PLGA is 10KDa to 100KDa; the drug molecule is a poorly soluble drug, and the poorly soluble drug is selected from one or more of paclitaxel, tanshinone IIA, curcumin and quercetin; the phospholipid is selected from one or more of distearoylphosphatidylcholine, dioleoylphosphatidylethanolamine, lecithin and dipalmitoylphosphatidylcholine.

5. The targeted lipid nanoparticle according to claim 2 or 3, characterized in that The drug molecule is tanshinone IIA or a salt thereof.

6. The method for preparing lipid nanoparticles according to claim 1, wherein The following steps are involved: (1) mixing the drug molecules and PLGA in an organic solvent to obtain an organic phase; (2) forming an aqueous phase containing a stabilizer; (3) adding the organic phase to the aqueous phase, mixing by oscillation or ultrasound, then removing the organic solvent and separating to obtain PLGA nanoparticles containing drug molecules, and then resuspending them in PBS to obtain an aqueous solution of PLGA nanoparticles containing drug molecules; (4) mixing phospholipids, cholesterol, and a modifying material in an organic solvent, and then removing the organic solvent to obtain a film; (5) adding an aqueous solution of PLGA nanoparticles containing drug molecules to the film, oscillating or ultrasonicating, and then filtering to obtain lipid nanoparticles; Among them, step (1) and step (2) are performed in no particular order.

7. The method for preparing targeted lipid nanoparticles according to claim 2, wherein The following steps are involved: 1) mixing the drug molecule and PLGA in an organic solvent to obtain an organic phase; 2) forming an aqueous phase containing a stabilizer; 3) adding the organic phase to the aqueous phase, mixing by oscillation or ultrasound, then removing the organic solvent and separating to obtain PLGA nanoparticles containing drug molecules, which are then resuspended in PBS to obtain an aqueous solution of PLGA nanoparticles containing drug molecules; 4) mixing phospholipids, cholesterol, and targeted modification materials in an organic solvent, and then removing the organic solvent to obtain a film; 5) adding an aqueous solution of PLGA nanoparticles containing drug molecules to the film, oscillating or sonicating, and then filtering to obtain targeted lipid nanoparticles; There is no particular order for step 1) and step 2).

8. The method for preparing targeted lipid nanoparticles according to claim 3, wherein The following steps are involved: 1′) dissolving PLGA in an organic solvent to obtain an organic phase; 2') forming an aqueous phase containing a stabilizer; 3') adding the organic phase to the aqueous phase, mixing by oscillation or ultrasound, then removing the organic solvent, separating to obtain PLGA nanoparticles, and then resuspending in PBS to obtain a PLGA nanoparticle aqueous solution; 4′) mixing phospholipids, cholesterol, targeting modification materials and drug molecules in an organic solvent, and then removing the organic solvent to obtain a film; 5′) adding the PLGA nanoparticle aqueous solution to the film, shaking or sonicating, and then filtering to obtain targeted lipid nanoparticles; There is no particular order between step 1') and step 2').

9. The preparation method according to any one of claims 6 to 8, characterized in that: Dissolving a stabilizer in water to form an aqueous phase; wherein the stabilizer is selected from polyvinyl alcohol and / or Pluronic F127; The organic solvent is selected from one or more of ethyl acetate, methanol, ethanol, acetone, chloroform and dichloromethane.

10. Use of the targeted lipid nanoparticles according to claim 5 in preparing a drug for resisting myocardial ischemia.

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