A lung-selective lipid nanoparticle and its preparation method and application

Abstract

The present invention belongs to the field of nanopharmaceutical technology and discloses a lung-selective lipid nanoparticle, its preparation method, and application. The preparation method comprises: dissolving four phospholipids in anhydrous ethanol to obtain a phospholipid ethanol solution; then uniformly adding anhydrous ethanol dissolved in an anticancer drug to obtain a phospholipid ethanol solution containing the anticancer drug; uniformly adding pure water to the phospholipid ethanol solution containing the anticancer drug under stirring, followed by dialysis for 6 hours to obtain the lipid nanoparticles of the present invention. The lipid nanoparticles prepared by the preparation method of the present invention have a lipophilic shell composed of a single layer of phospholipids, a small particle size and a narrow distribution, and can encapsulate hydrophobic drugs, thereby increasing drug stability. They are lung-selective and, after varying the molar percentage of 2,3-dioleoyloxypropyl-1-trimethylammonium bromide, can increase the accumulation of the anticancer drug in the lungs while reducing its accumulation in the liver. The lipid nanoparticles can be used to prepare drugs for treating lung-related diseases.

Description

Technical Field

[0001] The present invention relates to the technical field of nanopharmaceuticals, and in particular to a lipid nanoparticle with lung selectivity, a preparation method and an application thereof. Background Art

[0002] Globally, hundreds of millions of people suffer from chronic respiratory diseases, with lung disease (such as lung cancer, asthma, and infections) being one of the most common causes of death. Lung disease is also the organ most commonly affected by climate change, smoking, pollution, and genetic factors. While substantial progress has been made in the long-term management of most lung diseases, knowledge of their etiology, early identification, basic diagnostic procedures including biomarkers, and, particularly, disease-modifying treatments for acute exacerbations remains in its infancy. Therefore, the diagnosis and treatment of lung diseases present a formidable challenge. Due to the unique physiological environment of the lung, drug delivery within the lung is becoming increasingly important. Traditional drug therapies (oral or parenteral) suffer from poor selectivity and bioavailability, limiting their effectiveness in treating lung diseases and leading to associated toxicity and numerous systemic side effects. Furthermore, while pulmonary drug delivery is an effective route for treating lung diseases, rapid drug elimination due to pulmonary defense mechanisms complicates inhaled therapy. Nanosystems can reduce therapeutic doses and side effects, improve patient compliance, avoid clearance by alveolar macrophages, protect against drug degradation, and provide controlled and targeted drug release. Therefore, the use of nanocarriers to achieve pulmonary drug delivery is the best therapeutic strategy to overcome the above problems.

[0003] There are many types of nanosystems, such as lipid nanoparticles (LNPs), polymer nanoparticles, and metal nanoparticles. Compared with traditional nanosystems and chemotherapy, LNPs have many advantages. They have good biocompatibility, controlled sustained release of anti-tumor drugs, and low toxicity. They have become a promising carrier for delivering various therapeutic agents throughout the pharmaceutical industry. The application of LNPs has also expanded to other fields such as medical imaging, cosmetics, nutrition, agriculture, and other innovative fields such as nanoreactors. As the most representative lipid nanosystem, its particle size is 50-1000nm and it has a spherical structure. As an important component of the COVID-19 mRNA vaccine, LNPs play a key role in effectively protecting and transporting mRNA to cells. The encapsulation of anti-cancer drugs in LNPs has also been deeply studied. LNPs have gradually become the focus of research. Because LNPs have the ability to control the location and time of drug delivery in the body, they can be used to provide treatment for various diseases.

[0004] As one of the most advanced non-viral synthetic nanoparticles, LNPs have been shown to deliver drugs to the liver or spleen after intravenous administration, either through passive targeting or phagocytosis by macrophages as foreign bodies. To date, much of the clinical interest has focused on the treatment of liver-related diseases such as hepatitis, liver parasites, and liver tumors. Therefore, there is an urgent need to optimize multiple aspects of drug delivery systems to develop efficient drug delivery systems for lung delivery, thereby improving the therapeutic index and fully realizing the potential of drugs for the treatment of lung-related diseases. Summary of the Invention

[0005] The present invention aims to provide lung-selective lipid nanoparticles, their preparation method, and applications. These lipid nanoparticles, composed of a monolayer of phospholipids forming a lipophilic shell, have a small, narrowly distributed particle size, enabling them to encapsulate hydrophobic drugs and enhance their stability. Furthermore, they exhibit lung selectivity, and by varying the molar percentage of 2,3-dioleoyloxypropyl-1-trimethylamine bromide, they can enhance the accumulation of anticancer drugs in the lungs while reducing their accumulation in the liver. These particles can be used in the preparation of drugs for treating lung-related diseases.

[0006] To solve the above problems, the present invention provides a method for preparing lung-selective lipid nanoparticles, comprising the following steps:

[0007] 1) dissolving four phospholipids in anhydrous ethanol and mixing them uniformly to obtain a phospholipid ethanol solution; the four phospholipids are 1,2-dioleoylglycerol-3-phosphoethanolamine, cholesterol, 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol, and 2,3-dioleoyloxypropyl-1-trimethylammonium bromide;

[0008] 2) adding anhydrous ethanol containing the anticancer drug dropwise to the phospholipid ethanol solution at a uniform rate, stirring for 5 minutes to uniformly mix the solution, thereby obtaining a phospholipid ethanol solution containing the anticancer drug; then, adding pure water dropwise to the phospholipid ethanol solution containing the anticancer drug at a uniform rate while stirring; transferring the solution to a dialysis bag; and dialyzing the solution against pure water for 6 hours while stirring to remove unencapsulated anticancer drug, thereby obtaining lung-selective lipid nanoparticles.

[0009] Furthermore, in step 1), the molar ratio of 1,2-dioleoylglycerol-3-phosphoethanolamine to cholesterol, 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol, and 2,3-dioleoyloxypropyl-1-trimethylamine bromide is 69:40:4:X, where X is 12.6-48.4.

[0010] Furthermore, the anticancer drug in step 2) is docetaxel, and the mass ratio of the anticancer drug to the total amount of the four phospholipids is 1:4.

[0011] Furthermore, the volume ratio of the phospholipid ethanol solution to pure water in step 2) is 7:3.

[0012] Furthermore, the drug concentration of the anhydrous ethanol containing the anticancer drug in step 2) is 6.75 mg / mL.

[0013] Furthermore, the molecular weight cut-off of the dialysis bag in step 2) is 14,000.

[0014] The present invention also provides a lipid nanoparticle with lung selectivity prepared by the above preparation method.

[0015] Furthermore, the hydrated particle size of the lung-selective lipid nanoparticles is between 140-170 nm.

[0016] The present invention also provides a use of the above-mentioned lung-selective lipid nanoparticles in the preparation of drugs for treating lung-related diseases.

[0017] Compared with the prior art, the present invention has the following beneficial effects:

[0018] 1. The lipid nanoparticles of the present invention can change the charge size of the nanoparticles of the same component by fixing the molar ratio of the three lipids in the components and changing the molar percentage of 2,3-dioleoyloxypropyl-1-trimethylammonium bromide, thereby improving the passive tissue targeting ability of the lipid nanoparticles and making it easier for drugs to enter cells.

[0019] 2. The lipid nanoparticles of the present invention can prevent inter-particle aggregation, reduce the binding of particles to plasma proteins in the body, prolong the blood circulation time of drugs, and improve the bioavailability of drugs in lipid nanocarriers.

[0020] 3. The lipid nanoparticles of the present invention have lung selectivity, which can allow more anticancer drugs to accumulate in the lungs while reducing their accumulation in the liver.

[0021] 4. The lipid nanoparticles of the present invention have higher biocompatibility than other nanocarriers, can control drug release and targeting, are biodegradable, and have low immunogenicity. Furthermore, they are simple and convenient to prepare, and their surface is easily modified, allowing for rapid large-scale production. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1Statistical graphs of the hydrated particle size of the empty 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs) provided in Example 1-1, the empty 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles (10% DOTMA-LNPs, 30% DOTMA-LNPs) provided in Example 1-2, the docetaxel-loaded 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs / DTX) provided in Example 2-1, the docetaxel-loaded 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) provided in Example 2-2, the empty 50% DOTMA lipid nanoparticles (50% DOTMA-LNPs) provided in Comparative Example 1-1, and the docetaxel-loaded 50% DOTMA lipid nanoparticles (50% DOTMA-LNPs / DTX) provided in Comparative Example 1-2.

[0023] Figure 2 Statistical graphs of potential changes for the empty 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs) provided in Example 1-1, the empty 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles (10% DOTMA-LNPs, 30% DOTMA-LNPs) provided in Example 1-2, the docetaxel-loaded 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs / DTX) provided in Example 2-1, the docetaxel-loaded 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) provided in Example 2-2, the empty 50% DOTMA lipid nanoparticles (50% DOTMA-LNPs) provided in Comparative Example 1-1, and the docetaxel-loaded 50% DOTMA lipid nanoparticles (50% DOTMA-LNPs / DTX) provided in Comparative Example 1-2.

[0024] Figure 3 Statistical graphs of the encapsulation efficiency of 0% DOTMA lipid nanoparticles loaded with docetaxel (0% DOTMA-LNPs / DTX) provided in Example 2-1, 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) provided in Example 2-2, and 50% DOTMA lipid nanoparticles loaded with docetaxel (50% DOTMA-LNPs / DTX) provided in Comparative Example 1-2.

[0025] Figure 4This is a transmission electron micrograph of the empty 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs) provided in Example 1-1.

[0026] Figure 5 The in vitro cumulative release curves of docetaxel at room temperature are shown for docetaxel, 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) provided in Example 2-2, and 50% DOTMA lipid nanoparticles loaded with docetaxel (50% DOTMA-LNPs / DTX) provided in Comparative Example 1-2.

[0027] Figure 6 The drug distribution results of docetaxel, 0% DOTMA lipid nanoparticles loaded with docetaxel (0% DOTMA-LNPs / DTX) provided in Example 2-1, 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) provided in Example 2-2, and 50% DOTMA lipid nanoparticles loaded with docetaxel (50% DOTMA-LNPs / DTX) provided in Comparative Example 1-2 in major organs (heart, liver, spleen, lung, and kidney) in the body 6 hours after tail vein injection.

[0028] Figure 7 This is a schematic diagram of the structure of a lung-selective lipid nanoparticle prepared by the present invention. DETAILED DESCRIPTION

[0029] In order to further understand the present invention, preferred embodiments of the present invention are described in detail below in conjunction with examples. However, it should be understood that these descriptions are only for further illustrating the features and advantages of the present invention, rather than for limiting the claims of the present invention.

[0030] All raw materials of the present invention are not particularly limited in their sources and can be purchased from the market or prepared according to conventional methods well known to those skilled in the art.

[0031] Example 1-1

[0032] Preparation of unloaded 0% DOTMA lipid nanoparticles:

[0033] 2.57 mg (69 equivalents) 1,2-dioleoylglycerol-3-phosphoethanolamine (DOPE), 0.77 mg (40 equivalents) cholesterol (CHOL), 0.50 mg (4 equivalents) 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol (DMG-PEG), and 0 mg (0 equivalents) 2,3-dioleoyloxypropyl-1-trimethylamine bromide (DOTMA) were dissolved in anhydrous ethanol and mixed evenly to obtain a phospholipid ethanol solution; pure water was added dropwise at a uniform rate under stirring conditions and stirred for 5 minutes, and the solution was transferred to a dialysis bag (MW = 14000) and dialyzed against pure water under stirring for 6 hours to obtain an empty 0% DOTMA lipid nanoparticle solution (0% DOTMA-LNPs).

[0034] The particle size of the empty 0% DOTMA lipid nanoparticles was tested and the results were as follows: Figure 1 As shown in FIG, the particle size of the unloaded 0% DOTMA lipid nanoparticles is between 140-170 nm; Figure 2 As shown, the potential of the empty 0% DOTMA lipid nanoparticles is close to neutral, indicating that the empty 0% DOTMA lipid nanoparticles are successfully constructed.

[0035] The empty 0% DOTMA lipid nanoparticle solution was dropped onto the copper mesh, and the excess liquid was absorbed by filter paper. The mesh was stained with 1% phosphotungstic acid solution, and then washed with clean water for 3 times after staining for 1 to 2 minutes. The excess liquid was absorbed by filter paper, and the mesh was dried and then observed using a transmission electron microscope (TEM, JEM-1230, Japan). The transmission electron microscope image of the empty 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs) is shown in FIG. Figure 4 shown.

[0036] Example 1-2

[0037] Preparation of unloaded 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles:

[0038] (1) 2.32 mg (69 equivalents) of 1,2-dioleoylglycerol-3-phosphoethanolamine (DOPE), 0.70 mg (40 equivalents) of cholesterol (CHOL), 0.45 mg (4 equivalents) of 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol (DMG-PEG), and 0.37 mg (12.6 equivalents) of 2,3-dioleoyloxypropyl-1-trimethylamine bromide (DOTMA) were dissolved in anhydrous ethanol and mixed evenly to obtain a phospholipid ethanol solution; the remaining steps were the same as those in Example 1-1 (except for transmission observation), and an empty 10% DOTMA lipid nanoparticle solution (10% DOTMA-LNPs) was obtained.

[0039] (2) 1.83 mg (69 equivalents) of 1,2-dioleoylglycerol-3-phosphoethanolamine (DOPE), 0.54 mg (40 equivalents) of cholesterol (CHOL), 0.36 mg (4 equivalents) of 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol (DMG-PEG), and 1.11 mg (48.4 equivalents) of 2,3-dioleoyloxypropyl-1-trimethylamine bromide (DOTMA) were dissolved in anhydrous ethanol and mixed evenly to obtain a phospholipid ethanol solution; the remaining steps were the same as those in Example 1-1 (except for transmission observation), and an empty 30% DOTMA lipid nanoparticle solution (30% DOTMA-LNPs) was obtained.

[0040] The particle sizes of the empty 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles were tested and the results were as follows: Figure 1 As shown in FIG, the particle size of the unloaded 10% DOTMA lipid nanoparticles and the 30% DOTMA lipid nanoparticles is between 140 and 170 nm; Figure 2 As shown, the empty 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles were positively charged, indicating that the empty 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles were successfully constructed.

[0041] Example 2-1

[0042] Preparation of 0% DOTMA lipid nanoparticles loaded with docetaxel:

[0043] After obtaining the phospholipid ethanol solution in Example 1-1, 0.96 mg of docetaxel (DTX) dissolved in anhydrous ethanol was added dropwise at a constant rate and stirred for 5 minutes to uniformly mix the solution. This yielded a phospholipid ethanol solution containing docetaxel. The remaining steps were the same as in Example 1-1. After dialysis, 0% DOTMA lipid nanoparticles loaded with docetaxel (0% DOTMA-LNPs / DTX) were obtained.

[0044] The schematic diagram of the structure of 0% DOTMA lipid nanoparticles loaded with docetaxel in this example is shown in FIG. Figure 7 As shown by Figure 7 It can be seen that the 0% DOTMA lipid nanoparticle loaded with docetaxel is a spherical nanoparticle with a single-layer phospholipid shell on the outside, and the hydrophobic drug docetaxel is wrapped inside the nanoparticle.

[0045] The particle size of 0% DOTMA lipid nanoparticles loaded with docetaxel was tested and the results were as follows: Figure 1 As shown in FIG, the particle size of the 0% DOTMA lipid nanoparticles loaded with docetaxel is between 140 and 170 nm; Figure 2As shown, the potential of the 0% DOTMA lipid nanoparticles loaded with docetaxel is neutral, indicating that the 0% DOTMA lipid nanoparticles loaded with docetaxel are successfully constructed.

[0046] Example 2-2

[0047] Preparation of docetaxel-loaded 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles:

[0048] After obtaining the phospholipid ethanol solution in steps (1) and (2) of Example 1-2, 0.96 mg of docetaxel (DTX) dissolved in anhydrous ethanol was added dropwise at a uniform rate, and the solution was stirred for 5 minutes to mix uniformly, thereby obtaining a phospholipid ethanol solution containing docetaxel. The remaining steps were the same as those of Example 1-2. After dialysis, 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel (10% DOTMA-LNPs / DTX and 30% DOTMA-LNPs / DTX) were obtained.

[0049] The particle sizes of 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel were tested, and the results were as follows: Figure 1 As shown in FIG, the particle size of the 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel is between 140 and 170 nm; Figure 2 As shown, the 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel were positively charged, indicating that the 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel were successfully constructed.

[0050] Comparative Example 1-1

[0051] Preparation of unloaded 50% DOTMA lipid nanoparticles:

[0052] 1.32 mg (69 equivalents) of 1,2-dioleoylglycerol-3-phosphoethanolamine (DOPE), 0.39 mg (40 equivalents) of cholesterol (CHOL), 0.26 mg (4 equivalents) of 1,2-dimyristoyl-sn-glycero-3-methoxypolyethylene glycol (DMG-PEG), and 1.87 mg (113 equivalents) of 2,3-dioleoyloxypropyl-1-trimethylamine bromide (DOTMA) were dissolved in anhydrous ethanol and mixed evenly to obtain a phospholipid ethanol solution; the remaining steps were the same as in Example 1-1 (except for transmission observation) to obtain an empty 50% DOTMA lipid nanoparticle solution (50% DOTMA-LNPs).

[0053] The particle size of the empty 50% DOTMA lipid nanoparticle solution was tested and the results were as follows: Figure 1As shown in FIG, the particle size of the 50% DOTMA lipid nanoparticles is between 140 and 170 nm; Figure 2 As shown in FIG, the potential of the 50% DOTMA lipid nanoparticles is positively charged, indicating that the unloaded 50% DOTMA lipid nanoparticles are successfully constructed.

[0054] Comparative Example 1-2

[0055] Preparation of 50% DOTMA lipid nanoparticles loaded with docetaxel:

[0056] After obtaining the phospholipid ethanol solution in Comparative Example 1-1, 0.96 mg of docetaxel (DTX) dissolved in anhydrous ethanol was added dropwise at a uniform rate and stirred for 5 minutes to uniformly mix the solution. This yielded a phospholipid ethanol solution containing docetaxel. The remaining steps were the same as those in Comparative Example 1-1. After dialysis, 50% DOTMA lipid nanoparticles loaded with docetaxel (50% DOTMA-LNPs / DTX) were obtained.

[0057] The particle size of 50% DOTMA lipid nanoparticles loaded with docetaxel was tested and the results were as follows: Figure 1 As shown in FIG, the particle size of the 50% DOTMA lipid nanoparticles loaded with docetaxel is between 140 and 170 nm; Figure 2 As shown in FIG, the potential of the 50% DOTMA lipid nanoparticles loaded with docetaxel is positively charged, indicating that the 50% DOTMA lipid nanoparticles loaded with docetaxel are successfully constructed.

[0058] Test Example 1

[0059] In vitro drug release of docetaxel and DOTMA lipid nanoparticles loaded with different molar percentages of docetaxel at different time points:

[0060] Using docetaxel, 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) loaded with docetaxel obtained in Example 2-2, and 50% DOTMA lipid nanoparticles (50% DOTMA-LNPs / DTX) loaded with docetaxel obtained in Comparative Example 1-2, these four solutions were divided into three equal parts and allowed to stand at room temperature. At 0 h, 0.05 h, 0.25 h, 0.5 h, 1 h, 2 h, 4 h, 6 h, 12 h, and 24 h, 0.2 mL of supernatant was taken from each solution. After each 0.2 mL of supernatant was taken, the same volume of fresh PBS solution was added to keep the volume of the entire system unchanged. The supernatant was then added to 3 mL of methanol solution, and the DTX content in the solution was detected by high performance liquid chromatography (HPLC) to calculate the cumulative release of DTX.

[0061] The DTX cumulative release curve is shown in Figure 5 As shown, according to Figure 5 It can be found that the drug release of 10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX, and 50% DOTMA-LNPs / DTX is not much different, but less drug is released than free DTX at the same time. This shows that the DOTMA content in DOTMA-LNPs / DTX and the charge size of the carrier have no significant effect on drug release, but it has advantages over free drugs, and can prolong the drug release time to a certain extent and increase the stability of the drug in vitro.

[0062] Example 3

[0063] Distribution of docetaxel and DOTMA lipid nanoparticles loaded with different molar percentages of docetaxel in different major organs:

[0064] ICR mice were randomly divided into 5 groups, 3 mice in each group, and each group was injected with docetaxel, 0% DOTMA lipid nanoparticles loaded with docetaxel (0% DOTMA-LNPs / DTX) obtained in Example 2-1, 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles loaded with docetaxel (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) obtained in Example 2-2, and 50% DOTMA lipid nanoparticles loaded with docetaxel (50% DOTMA-LNPs / DTX) obtained in Comparative Example 1-2 at a dose of 5 mg DTX / kg body weight via tail vein. 6 h after administration, mice were anesthetized with intraperitoneal injection of chloral hydrate, followed by perfusion with normal saline, and major organs such as the heart, liver, spleen, lungs, and kidneys were removed. The organs were weighed and homogenized using a homogenizer. Then, 5 mL of a mixed solution (chloroform: methanol = 4:1, v / v) was added to the homogenized tissue fluid, and the mixture was crushed and ultrasonicated for 10 min. The mixture was centrifuged at 8000 rpm / min for 10 min. The chloroform solution at the bottom layer was removed and dried, and 3 mL of methanol solution was added to re-dissolve the protein to denature and precipitate it. The mixture was centrifuged at 8000 rpm / min for 10 min. The drug content in the supernatant of each organ was measured by HPLC. The percentage of the drug detected per gram of tissue to the total amount of drug injected was calculated as follows: Figure 6 shown.

[0065] according to Figure 6 It can be seen that DOTMA-LNPs / DTX is widely distributed in the liver, spleen and lungs, but with the change of the molar percentage of DOTMA, docetaxel can accumulate more in the lungs, while reducing its accumulation in the liver. When the molar percentage of DOTMA accounts for 10%-30% of the total phospholipids, the prepared DOTMA-LNPs / DTX has better lung selectivity.

[0066] The encapsulation efficiency of the 0% DOTMA lipid nanoparticles (0% DOTMA-LNPs / DTX) loaded with anticancer drugs provided in Example 2-1, the 10% DOTMA lipid nanoparticles and 30% DOTMA lipid nanoparticles (10% DOTMA-LNPs / DTX, 30% DOTMA-LNPs / DTX) loaded with docetaxel obtained in Example 2-2, and the 50% DOTMA lipid nanoparticles (50% DOTMA-LNPs / DTX) loaded with docetaxel obtained in Comparative Example 1-2 were measured by HPLC. The encapsulation efficiency statistics are as follows: Figure 3 shown.

[0067] according to Figure 3 It can be seen that DOTMA lipid nanoparticles loaded with different molar percentages of docetaxel have a higher encapsulation efficiency, indicating that DOTMA lipid nanoparticles loaded with docetaxel can reduce the number of dosing times. In addition, the encapsulation efficiency of each group is not much different, which ensures that the dosage during treatment is relatively consistent and avoids the impact of different drug dosages.

[0068] The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention. It is obvious to those skilled in the art that the present invention is not limited to the details of the above exemplary embodiments, and that the present invention can be implemented in other specific forms without departing from the spirit or basic features of the present invention. Therefore, no matter from which point of view, the embodiments should be regarded as exemplary and non-restrictive. The scope of the present invention is defined by the appended claims rather than the above description, and it is intended that all changes that fall within the meaning and scope of the equivalent elements of the claims are included in the present invention. Although embodiments of the present invention have been shown and described, it is understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of the present invention, and the scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A method for preparing lung-selective lipid nanoparticles, characterized in that: The following steps are involved: 1) dissolving four phospholipids in anhydrous ethanol and mixing them uniformly to obtain a phospholipid ethanol solution; the four phospholipids are 1,2-dioleoylglycerol-3-phosphoethanolamine, cholesterol, 1,2-dimyristoyl-sn-glycerol-3-methoxypolyethylene glycol, and 2,3-dioleoyloxypropyl-1-trimethylamine bromide; the molar ratio of 1,2-dioleoylglycerol-3-phosphoethanolamine to cholesterol, 1,2-dimyristoyl-sn-glycerol-3-methoxypolyethylene glycol, and 2,3-dioleoyloxypropyl-1-trimethylamine bromide is 69:40:4:X, where X is 12.6-48.4; 2) adding anhydrous ethanol containing an anticancer drug dropwise to the phospholipid ethanol solution at a uniform rate, and stirring for 5 minutes to uniformly mix the solution to obtain a phospholipid ethanol solution containing the anticancer drug; then, adding pure water dropwise to the phospholipid ethanol solution containing the anticancer drug at a uniform rate while stirring; transferring the solution to a dialysis bag, and dialyzing against pure water with stirring for 6 hours to remove unencapsulated anticancer drug, thereby obtaining lung-selective lipid nanoparticles; the anticancer drug is docetaxel.

2. The preparation method according to claim 1, characterized in that In step 2), the mass ratio of the anticancer drug to the total amount of the four phospholipids is 1:

4.

3. The preparation method according to claim 1, characterized in that In step 2), the volume ratio of the phospholipid ethanol solution to pure water is 7:

3.

4. The preparation method according to claim 1, characterized in that The drug concentration of the anhydrous ethanol containing the anticancer drug in step 2) is 6.75 mg / mL.

5. The preparation method according to claim 1, characterized in that The molecular weight cut-off of the dialysis bag in step 2) is 14,000.

6. A lung-selective lipid nanoparticle prepared according to the preparation method according to any one of claims 1 to 5.

7. The lung-selective lipid nanoparticle according to claim 6, characterized in that The hydrated particle size of the lung-selective lipid nanoparticles is between 140 and 170 nm.

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