A nanoparticulate lipid body encapsulating a hydrophobic small molecule drug, and methods of making and using the same

Abstract

A nanoparticle liposome encapsulating a hydrophobic small molecule drug, its preparation method, and its applications are disclosed. The nanoparticle liposome comprises a monomolecular phospholipid membrane and a neutral lipid containing the hydrophobic small molecule drug encapsulated within the monomolecular phospholipid membrane. The nanoparticle liposome has a neutral lipid containing the hydrophobic small molecule drug as a hydrophobic core, and the nanospheres encapsulated by the monomolecular phospholipid membrane can efficiently dissolve and encapsulate hydrophobic small molecule compounds. Furthermore, the liposome encapsulated with the hydrophobic small molecule drug exhibits bioactivity, significantly killing cancer cells with a superior killing effect compared to free drugs. Additionally, its preparation method is simple and efficient. Therefore, liposomes are a promising delivery platform for hydrophobic small molecule drugs and can be widely used in the treatment of diseases such as cancer, infectious diseases, and metabolic disorders.

Description

Nanoparticle fat body encapsulating hydrophobic small molecule drugs and its preparation method and application

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on November 14, 2023, with application number 202311512646.3, and invention name “A nanoparticle fat body encapsulating hydrophobic small molecule drugs, its preparation method and application”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention relates to the field of pharmaceutical technology, and in particular to a nanoparticle fat body encapsulating a hydrophobic small molecule drug, and a preparation method and application thereof. Background Art

[0003] Small molecule drugs are an important means of treating various diseases. Among them, hydrophobic small molecule drugs refer to small molecule compounds that are insoluble or have very low solubility in water. Due to their hydrophobicity (i.e., not easily soluble in water or other polar solvents), these drugs often have low solubility and bioavailability in biological systems. Many common drugs are hydrophobic small molecule drugs, such as antibiotics, anticancer drugs, lipid-lowering drugs, etc. In addition, 90% of the small molecule drugs under development are hydrophobic small molecule compounds. Due to their poor water solubility, their absorption and bioavailability are seriously affected, and they can also cause varying degrees of toxic side effects, which seriously limit their clinical application. Therefore, improving the solubility of hydrophobic small molecule drugs is an important challenge in drug research and development.

[0004] To improve the solubility and bioavailability of hydrophobic small molecule drugs, the following methods are often used: 1) Drug modification: chemically modifying the structure of hydrophobic small molecule drugs, such as adding polar functional groups; 2) cosolvents: adding organic solvents and surfactants such as ethanol, Tween 80, and polyoxyethylated castor oil to increase the solubility of hydrophobic drugs; 3) Drug delivery: utilizing lipid nanocarriers such as liposomes, polymer nanoparticles, and nanomicelles to encapsulate and deliver hydrophobic small molecule drugs. However, drug modification often results in loss of small molecule drug activity. Surfactants such as ethanol and Tween 80, used as excipients, often cause toxic side effects such as hemolysis and allergic reactions.

[0005] In addition, although there are currently a variety of delivery vehicles for encapsulating and delivering hydrophobic small molecule compounds, there are also some disadvantages, such as: 1) Liposomes. Liposomes are prepared by simple mixing, oscillation, and ultrasound methods. However, liposomes have poor stability and are prone to fusion, rupture, or drug leakage, which will affect their distribution in the body and drug release. 2) Lipid microspheres. Lipid microspheres are prepared by emulsification-solvent evaporation, coprecipitation, solution polymerization, and other methods. However, these methods may require the use of organic solvents, and there may be problems with solvent residues. In addition, the preparation process of lipid microspheres will affect the stability of the drug. 3) Solid lipid nanoparticles: Solid lipid nanoparticles are prepared by melting, solvent, supercritical fluid, and other methods. High temperatures or organic solvents are required during the preparation process, which may cause the drug to crystallize or separate, thereby affecting its stability.

[0006] Summary of the Invention

[0007] In view of this, the technical problem to be solved by the present invention is to provide a nanoparticle fat body encapsulating hydrophobic small molecule drugs and its preparation method and application. The prepared nanoparticle fat body encapsulating drugs can efficiently dissolve and encapsulate hydrophobic small molecule compounds.

[0008] To achieve the above-mentioned purpose, one of the inventive objects of the present invention is to provide a nanoparticle fat body encapsulating a hydrophobic small molecule drug, comprising a monomolecular phospholipid membrane and a neutral lipid containing a hydrophobic small molecule drug encapsulated inside the monomolecular phospholipid membrane.

[0009] The schematic structural diagram of the nanoparticle fat body carrying a hydrophobic small molecule drug provided by the present invention is shown in FIG1 .

[0010] As can be seen from Figure 1, the nanoparticle fat body provided by the present invention that encapsulates hydrophobic small molecule drugs has a hydrophobic core formed by neutral lipids (shown in the light gray part in Figure 1), which can effectively encapsulate one or more hydrophobic small molecule drugs (shown as hydrophobic small molecule drug A and hydrophobic small molecule drug B in Figure 1), improve the solubility and bioavailability of hydrophobic small molecule drugs, and the outer layer of neutral lipids is wrapped with a single molecule phospholipid membrane to form nanoparticles. It is similar to the structure of naturally occurring lipid droplets and lipoproteins and can efficiently dissolve and encapsulate hydrophobic small molecule compounds. In addition, the components of the fat body exist naturally in the body and have good biocompatibility. Therefore, the above-mentioned fat body is a promising delivery platform for hydrophobic small molecule drugs and can be widely used in the treatment of diseases such as cancer, infectious diseases and metabolic diseases.

[0011] The nanoparticle fat body loaded with hydrophobic small molecule drugs provided by the present invention is suitable for most hydrophobic small molecule drugs, including but not limited to one or more of paclitaxel, magnolol, camptothecin, auristatin derivative MMAE, etc.

[0012] The nanoparticle fat body containing hydrophobic small molecule drugs provided by the present invention can contain one hydrophobic small molecule drug or can contain multiple hydrophobic small molecule drugs at the same time, such as paclitaxel-camptothecin.

[0013] Optionally, the neutral lipid is selected from one or more of fish oil, corn oil, tricaprylin, and retinol ester.

[0014] In some specific embodiments of the present invention, the neutral lipid is fish oil or corn oil.

[0015] In some specific embodiments of the present invention, the neutral lipid is a mixture of fish oil and tricaprylin. Optionally, the volume ratio of the fish oil to tricaprylin is (1-5):(1-2), more preferably (1-5):1, specifically 5:1, 4:1, 3:1, 2:1, 1:1, etc.

[0016] In some specific embodiments of the present invention, the neutral lipid is a mixture of fish oil and retinol ester. Optionally, the volume ratio of the fish oil to retinol ester is (1-5): (1-2), specifically 5:1, 4:1, 3:1, 2:1, 1:1, 1:2, etc.

[0017] In some specific embodiments of the present invention, the neutral lipid is a mixture of tricaprylin and corn oil. Optionally, the volume ratio of tricaprylin to corn oil is (1-5):(1-2), preferably (1-5):1, specifically 5:1, 4:1, 3:1, 2:1, 1:1, etc.

[0018] In some specific embodiments of the present invention, the neutral lipid is a mixture of fish oil, tricaprylin, and retinol esters. Optionally, the volume ratio of the fish oil, tricaprylin, and retinol esters is (2-4):(1-3):(1-2), specifically 4:1:1, 3:2:1, 3:1:2, 2:3:1, 2:2:2, etc.

[0019] Optionally, the phospholipid membrane is selected from one or more of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine, phosphatidylcholine, glycerophosphatidic acid, phosphatidylinositol, phosphatidylethanolamine, and polyethylene glycol-modified phosphatidylethanolamine.

[0020] Optionally, the phospholipid membrane is selected from any one of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine, glycerophosphatidic acid, phosphatidylinositol, phosphatidylethanolamine, polyethylene glycol-modified phosphatidylethanolamine and a mixture of phosphatidylcholine.

[0021] The molar content of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine, glycerophosphatidic acid, phosphatidylinositol, phosphatidylethanolamine or polyethylene glycol-modified phosphatidylethanolamine in the mixture is 5% to 10%.

[0022] Optionally, the phospholipid membrane is selected from glycerophosphatidic acid and phosphatidylcholine, and the molar ratio of glycerophosphatidic acid to phosphatidylcholine is (5-10): (90-95).

[0023] Optionally, the phospholipid membrane is selected from phosphatidylinositol and phosphatidylcholine, and the molar ratio of phosphatidylinositol to phosphatidylcholine is (5-10): (90-95).

[0024] Optionally, the phospholipid membrane is selected from phosphatidylethanolamine and phosphatidylcholine, and the molar ratio of phosphatidylethanolamine to phosphatidylcholine is (5-10): (90-95).

[0025] Optionally, the phospholipid membrane is selected from polyethylene glycol-modified phosphatidylethanolamine and phosphatidylcholine, and the molar ratio of the polyethylene glycol-modified phosphatidylethanolamine to phosphatidylcholine is (5-10): (90-95).

[0026] The nanoparticle fat body containing hydrophobic small molecule drugs provided by the present invention has a uniform spherical structure.

[0027] Experimental results show that when encapsulating the hydrophobic small molecule drug paclitaxel, fish oil and tricaprylin as neutral lipids, especially at a 1:1 volume ratio, can achieve high drug encapsulation concentrations, reaching 230 μg / mL. The prepared paclitaxel-encapsulated nanoparticle fat bodies have a particle size of approximately 130 nm. Using fish oil and retinol esters as neutral lipids, the encapsulation concentration is approximately 400 μg / mL, with a particle size of approximately 200 nm. In particular, a 1:2 volume ratio achieves a higher drug encapsulation concentration of 600 μg / mL, and the prepared paclitaxel-encapsulated nanoparticle fat bodies have a particle size of approximately 800 nm. Using a mixture of fish oil, tricaprylin, and retinol esters as neutral lipids, the paclitaxel encapsulation concentration is approximately 600 μg / mL, with a particle size of approximately 200 nm.

[0028] Experimental results indicate that the choice of phospholipid also influences drug entrapment concentration and particle size. For example, when PE or PEG-PE are added to phospholipids, the paclitaxel content in the paclitaxel liposomes increases, with loading rates exceeding 500 μg / mL. The addition of PI, PE, and PEG-PE increases the average particle size of the paclitaxel liposomes to approximately 200-300 nm. The addition of 10% PE significantly increases the concentration of the paclitaxel liposomes, reaching an OD600 of 120.

[0029] When encapsulating magnolol, the present invention selected triolein as the neutral lipid and 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution as the phospholipid. The encapsulation concentration of magnolol was 17 μg / mL, and the particle size was approximately 114.2 nm.

[0030] When camptothecin is entrapped, the present invention uses tricaprylin or a mixture of fish oil and tricaprylin as the neutral lipid, optionally with a volume ratio of 1:1 between the fish oil and tricaprylin. The phospholipid used is a 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution. The camptothecin entrapment concentration is 4 mg / L, and the particle size is approximately 120 nm.

[0031] When encapsulating auristatin derivatives with MMAE, the present invention employed triolein or corn oil as the neutral lipid, and a 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution as the phospholipid. When corn oil was used as the neutral lipid, the MMAE encapsulation concentration was 100 μg / mL, while when tricaprylin was used as the neutral lipid, the MMAE encapsulation concentration was 40 μg / mL. The resulting MMAE-encapsulated nanoparticles had a particle size of approximately 140 nm.

[0032] When co-encapsulating paclitaxel and camptothecin, the present invention employed a mixed neutral lipid of fish oil and tricaprylin, optionally in a 1:1 volume ratio, and a phospholipid solution of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine. Test results demonstrated that the prepared nanoparticle fat bodies achieved an average particle size of approximately 115.5 nm, with an encapsulation concentration of 65 mg / L for paclitaxel and 4 mg / L for camptothecin.

[0033] The experimental results show that after the hydrophobic small molecule drugs are made into nanoparticle fat bodies using the above-mentioned neutral lipids, the hydrophobic small molecule drugs that originally had very low solubility in the solvent can appear in a uniform milky white solution state, indicating that their solubility is greatly improved.

[0034] The second object of the present invention is to provide a method for preparing the above-mentioned nanoparticle fat body containing hydrophobic small molecule drugs, comprising the following steps:

[0035] S1) mixing a hydrophobic small molecule drug solution and a neutral lipid, and removing the solvent to obtain a neutral lipid containing the hydrophobic small molecule drug;

[0036] S2) mixing the neutral lipid and phospholipid containing the hydrophobic small molecule drug to obtain a lipid mixture, and separating the mixture to obtain nanoparticle fat bodies encapsulating the hydrophobic small molecule drug.

[0037] Optionally, the solvent of the hydrophobic small molecule drug solution is selected from anhydrous ethanol, chloroform, methanol, acetone and other solvents that can dissolve hydrophobic small molecule drugs.

[0038] Optionally, the concentration of the hydrophobic small molecule drug solution is 0-40 mg / mL. In some specific embodiments, the concentration of the hydrophobic small molecule drug solution is 20 mg / mL.

[0039] The concentration of the hydrophobic small molecule drug solution can also be adjusted according to the actual situation of the drug.

[0040] Optionally, the ratio of the hydrophobic small molecule drug solution and the neutral lipid can be adjusted according to actual conditions. Taking paclitaxel as an example, the ratio of paclitaxel to neutral lipid can be 1:1-1:5.

[0041] Optionally, the method for removing the solvent in step S1) is drying with high-purity nitrogen.

[0042] First, the phospholipid solution is placed in a centrifuge tube, and the solvent is blown dry with high-purity nitrogen; then the neutral lipid containing the hydrophobic small molecule drug is added to achieve mixing of the two.

[0043] Optionally, the step S2) is specifically as follows:

[0044] A) vortexing the neutral lipid and phospholipid containing the hydrophobic small molecule drug in a buffer solution to obtain a milky white lipid mixture;

[0045] B) Repeatedly centrifuging and vortexing the lipid mixture to separate and obtain nanoparticle fat bodies encapsulating hydrophobic small molecule drugs.

[0046] The buffer includes but is not limited to phosphate buffer, HEPES buffer, sucrose solution, NaCl solution, KCl solution, MgCl2 solution and the like.

[0047] Optionally, the vortexing time in the above step A) is 3 to 7 minutes, and the vortexing speed is 3000 to 4000 rpm.

[0048] Optionally, the milky white lipid mixture is purified by repeatedly centrifuging and vortexing the lipid mixture. The number of repetitions can be 1 to 3 times.

[0049] Optionally, the obtained milky white lipid mixture is subjected to a first centrifugation. Optionally, the first centrifugation speed is 800-1200g for 3-7 minutes. After centrifugation, the liquid phase system presents two layers. The lower milky white solution is collected by extraction and vortexed to obtain a milky white lipid mixture 2.

[0050] Optionally, the lipid mixture 2 is centrifuged for a second time. Optionally, the speed of the second centrifugation is 18,000 to 22,000 g for 3 to 7 minutes. After centrifugation, the precipitated components at the bottom of the centrifuge tube are removed and vortexed to obtain a milky white lipid mixture 3.

[0051] Optionally, the lipid mixture 3 is centrifuged for a third time. Optionally, the third centrifugation speed is 800-1200g for 3 to 7 minutes. After centrifugation, the liquid phase system presents two layers. The lower milky white solution is collected by extraction and vortexed to obtain a pure milky white lipid mixture.

[0052] The third object of the present invention is to provide a use of the above-mentioned nanoparticle fat body encapsulating hydrophobic small molecule drugs in the preparation of drugs for preventing, alleviating or treating tumors, infectious diseases and / or metabolic diseases.

[0053] The fourth object of the present invention is to provide an application of the above-mentioned nanoparticle fat body encapsulating hydrophobic small molecule drugs in preventing, alleviating or treating tumors, infectious diseases and / or metabolic diseases.

[0054] The results of cell tests show that the nanoparticle fat bodies loaded with hydrophobic small molecule drugs prepared by the present invention have biological activity and can significantly kill cancer cells, with a killing effect better than that of free drugs.

[0055] Optionally, the cancer cells are one or more of human embryonic kidney epithelial cells HEK293, human breast cancer cells MCF7, and human breast cancer cells MDA-MB-231.

[0056] Optionally, the tumor is one or more of renal cancer, breast cancer, and lung cancer.

[0057] Optionally, the infectious disease is one or more of influenza and tuberculosis.

[0058] Optionally, the metabolic disease is one or more of diabetes, obesity, and cardiovascular disease.

[0059] The fifth object of the present invention is to provide a drug for alleviating or treating tumors, infectious diseases and / or metabolic diseases, comprising the above-mentioned nanoparticle fat body encapsulating hydrophobic small molecule drugs.

[0060] Compared to the prior art, the present invention provides a nanoparticle fat body that encapsulates a hydrophobic small molecule drug, comprising a monomolecular phospholipid membrane and a neutral lipid containing the hydrophobic small molecule drug encapsulated within the monomolecular phospholipid membrane. The novel nanoparticle fat body provided by the present invention has a neutral lipid containing the hydrophobic small molecule drug as a hydrophobic core, and the nanospheres encapsulated by the monomolecular phospholipid membrane can efficiently dissolve and encapsulate hydrophobic small molecule compounds. In addition, the fat body encapsulating the hydrophobic small molecule drug has biological activity and can significantly kill cancer cells, and the killing effect is better than that of free drugs. In addition, the preparation method is simple and efficient. Therefore, the fat body is a promising delivery platform for hydrophobic small molecule drugs and can be widely used to treat diseases such as cancer, infectious diseases, and metabolic diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0061] FIG1 is a schematic structural diagram of a nanoparticle fat body loaded with a hydrophobic small molecule drug provided by the present invention;

[0062] FIG2 is a test diagram of the fat body carrying the hydrophobic small molecule compound paclitaxel prepared in 1.1 of Example 1;

[0063] FIG3 is a test diagram of the fat body carrying the hydrophobic small molecule compound paclitaxel prepared in 1.2 of Example 1;

[0064] FIG4 is a test diagram of the fat body carrying the hydrophobic small molecule compound paclitaxel prepared in 1.3 of Example 1;

[0065] FIG5 is a test diagram of the fat body carrying the hydrophobic small molecule compound paclitaxel prepared in 1.4 of Example 1;

[0066] FIG6 is a test graph of the fat body carrying the hydrophobic small molecule compound paclitaxel prepared in 1.5 of Example 1;

[0067] FIG7 is a test diagram of the fat body carrying the hydrophobic small molecule compound magnolol prepared in Example 2;

[0068] FIG8 is a side view of the fat body carrying the hydrophobic small molecule compound camptothecin prepared in Example 3;

[0069] FIG9 is a test graph of the hydrophobic small molecule auristatin derivative MMAE prepared in Example 4;

[0070] FIG10 is a test image of the fat body prepared in Example 5 carrying the hydrophobic small molecule compounds paclitaxel and camptothecin;

[0071] FIG11 is a graph comparing the anti-tumor properties of paclitaxel fat bodies and free paclitaxel;

[0072] FIG12 is a graph comparing the survival rates of magnolol fat bodies and free magnolol;

[0073] FIG13 is a graph comparing the survival rates of MMAE fat bodies and free MMAE. DETAILED DESCRIPTION

[0074] To further illustrate the present invention, the nanoparticle fat body carrying hydrophobic small molecule drugs, its preparation method and application provided by the present invention are described in detail below with reference to the examples.

[0075] The oil-water partition coefficient (logP) is a common indicator used to measure the hydrophobicity of a drug. The logP value refers to the logarithm of the ratio of the partition coefficients of a substance in n-octane (oil) and water, reflecting the distribution of the substance between the oil and water phases. The larger the logP value, the more lipophilic and hydrophobic the substance is. To demonstrate that fat bodies are a platform for encapsulating and delivering hydrophobic small molecule drugs, the present invention selected several hydrophobic small molecule compounds with logP values ​​greater than 1.5 (shown in Table 1) for encapsulation and corresponding biological activity verification, such as paclitaxel, honokiol, camptothecin, and the auristatin derivative MMAE (Monomethyl auristatin E).

[0076] Table 1 LogP values ​​of hydrophobic small molecule drugs entrapped in fat bodies

[0077] Example 1 Construction of Fatsomes Carrying the Hydrophobic Small Molecule Compound Paclitaxel (Paclitaxel Fatsomes)

[0078] 1.1. Changing the neutral lipid conditions, namely the ratio of fish oil to tricaprylin (8:0 TAG), to prepare fat bodies carrying the hydrophobic small molecule compound paclitaxel

[0079] 1.1.1) Prepare neutral lipids according to the following six volume ratios: fish oil / tricaprylin = 1 / 0, 5 / 1, 4 / 1, 3 / 1, 2 / 1, and 1 / 1;

[0080] 1.1.2) Dissolve paclitaxel (PTX) in anhydrous ethanol to a concentration of 20 mg / mL;

[0081] 1.1.3) Take 200 μL of the neutral lipid prepared in different volume ratios above and add 200 μL of paclitaxel solution (PTX). Drain the ethanol with high-purity nitrogen to obtain neutral lipid containing PTX.

[0082] 1.1.4) Add 80 μL of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (containing 2 mg DOPC) to a microcentrifuge tube and dry the solvent with high-purity nitrogen.

[0083] 1.1.5) 100 μL of PBS and 5 μL of the neutral lipid containing PTX prepared in step 3) were added to a microcentrifuge tube, and the mixture was vortexed for 4 min (vortexed for 10 s, rested for 10 s) (in actual application, vortexed for 3-7 min, vortexed at 3000-4000 rpm, and 4000 rpm was used in this example) to obtain a milky white lipid mixture 1. The lipid mixture 1 was centrifuged at 1000 g for 5 min (in actual application, centrifuged at 800-1200 g for 3-7 min). After centrifugation, the liquid phase system showed two layers. The lower milky white solution was collected by extraction and vortexed to obtain a milky white lipid mixture 2.

[0084] 1.1.6) The lipid mixture 2 obtained in step 5) is centrifuged at 20,000 g for 5 minutes (in practice, 18,000-22,000 g for 3-7 minutes is acceptable). After centrifugation, the precipitate at the bottom of the microcentrifuge tube is removed and vortexed to obtain a milky white lipid mixture 3.

[0085] 1.1.7) The lipid mixture 3 obtained in step 6) is centrifuged at 1000 g for 5 minutes (in practice, 800-1200 g for 3-7 minutes is acceptable). After centrifugation, the liquid phase system exhibits two stratified layers. The lower milky white solution is collected by extraction and vortexed to obtain a milky white lipid mixture 4, which is the final fat body carrying the hydrophobic small molecule compound paclitaxel (paclitaxel fat body).

[0086] Paclitaxel and the paclitaxel fat body prepared above were dissolved in PBS respectively. The dissolution diagrams are shown in Figure 2A, where A is the morphology of paclitaxel in PBS and fat body in PBS. Paclitaxel has a very low solubility in PBS due to its hydrophobicity and appears turbid, while the paclitaxel encapsulated in the fat body appears as a uniform milky white solution.

[0087] The prepared paclitaxel fat bodies were further observed using a laser confocal microscope, and the paclitaxel fat bodies showed a uniform spherical structure (see Figure 2B, where a is the result of paclitaxel fat bodies stained red with the neutral lipid-specific dye LipidTox Red, and b is the DIC result of paclitaxel fat bodies, and the scale bar is 2 μm).

[0088] Paclitaxel fat bodies were dissolved in methanol (chromatographic grade) and the PTX content in the paclitaxel fat bodies was measured by HPLC. The detection conditions were: an Agilent Zorbax SB-C18 column, a mobile phase of methanol:acetonitrile:water = 4:3:3 (volume ratio), a column temperature of 25°C, a detection wavelength of 227 nm, and a flow rate of 1 mL / min. Paclitaxel fat bodies prepared at a 1:1 volume ratio of fish oil to tricaprylin showed the highest PTX content, reaching 230 μg / mL. At other ratios, the PTX content in the paclitaxel fat bodies was relatively low (see Figure 2C, where C shows the PTX content in the paclitaxel fat bodies measured by HPLC). The average particle size of the paclitaxel fat bodies prepared with different neutral lipid volume ratios was similar, approximately 130 nm (see Figure 2D, where D shows the average particle size of the paclitaxel fat bodies measured by dynamic light scattering particle size analysis). When the volume ratio of fish oil to tricaprylin was 1:0, the concentration of the prepared paclitaxel fat body was the highest, with an OD600 of approximately 45. Under other ratio conditions, the concentration of the paclitaxel fat body was not much different, with an OD600 of approximately 30 (see E in Figure 2, where E is the concentration of the paclitaxel fat body measured by a multifunctional microplate reader, expressed as OD600).

[0089] 1.2. Changing the neutral lipid conditions, i.e., the ratio of fish oil to retinol ester, to prepare fat bodies carrying the hydrophobic small molecule compound paclitaxel

[0090] 1.2.1) Neutral lipids were prepared according to the following 7 volume ratios: fish oil / retinol ester = 1 / 0, 5 / 1, 4 / 1, 3 / 1, 2 / 1, 1 / 1, 1 / 2.

[0091] The subsequent steps are the same as the preparation method in 1.1 above.

[0092] Laser confocal microscopy observation of the resulting paclitaxel-containing fat bodies revealed that paclitaxel-containing fat bodies could be prepared by varying the neutral lipid content, using fish oil and retinol esters. The paclitaxel-containing fat bodies exhibited a uniform, spherical structure (Figure 3A, where a is the result of paclitaxel-containing fat bodies stained green with the neutral lipid-specific dye LipidTox Green, and b is the DIC result of paclitaxel-containing fat bodies; scale bar is 1 μm). HPLC analysis of the PTX content in the paclitaxel-containing fat bodies revealed the highest PTX content, reaching 600 μg / mL, when the volume ratio of fish oil to retinol ester was 1:2. The average particle size was also the largest, at approximately 800 nm. Under other ratio conditions, the PTX content in the paclitaxel fat body was relatively low, approximately 400 μg / mL (Figure 3B, PTX content in paclitaxel fat body measured by HPLC), and the average particle size was similar, approximately 200 nm (see Figure 3B and C, where C is the average particle size of the paclitaxel fat body measured by dynamic light scattering particle size analyzer). The concentration of the paclitaxel fat body prepared under different neutral lipid ratio conditions was similar, with an OD600 value of approximately 60 (see Figure 3D, where D is the concentration of the paclitaxel fat body measured by a multifunctional microplate reader, expressed as OD600).

[0093] 1.3. Changing the neutral lipid conditions, that is, the ratio of corn oil to tricaprylin (8:0 TAG) to prepare fat bodies carrying the hydrophobic small molecule compound paclitaxel

[0094] 1.3.1) Neutral lipids were prepared according to the following five volume ratios: tricaprylin / corn oil = 1 / 0, 5 / 1, 4 / 1, 3 / 1, and 2 / 1.

[0095] The subsequent steps are the same as the preparation method in 1.1 above.

[0096] The obtained paclitaxel fat bodies were observed under laser confocal microscopy. Paclitaxel-loaded fat bodies were also prepared using corn oil and tricaprylin, varying the neutral lipid content. The paclitaxel fat bodies exhibited uniform, spherical structures (Figure 4A, a, shows paclitaxel fat bodies stained red with the neutral lipid-specific dye LipidTox Red; b, shows DIC results of paclitaxel fat bodies; scale bar: 2 μm). The average particle size of the paclitaxel fat bodies prepared with different neutral lipid volume ratios was similar, approximately 150 nm (Figure 4B, B, shows the average particle size of the paclitaxel fat bodies measured by dynamic light scattering particle size analysis). The highest concentration of paclitaxel fat bodies was achieved when the volume ratio of tricaprylin to corn oil was 3:1, with an OD600 of approximately 80. The lowest concentration of paclitaxel fat bodies was achieved when the volume ratio of tricaprylin to corn oil was 2:1, with an OD600 of approximately 20. Under other ratio conditions, the concentration of paclitaxel fat bodies was not much different, and OD600 was about 40 (C in Figure 4, C is the concentration of paclitaxel fat bodies measured by a multifunctional microplate reader, expressed as OD600).

[0097] 1.4. Changing the neutral lipid conditions, i.e., the ratio of fish oil, tricaprylin, and retinol esters, to prepare fat bodies carrying the hydrophobic small molecule compound paclitaxel

[0098] Neutral lipids were prepared according to the following 6 volume ratios: fish oil / tricaprylin / retinol ester = 6 / 0 / 0, 4 / 1 / 1, 3 / 2 / 1, 3 / 1 / 2, 2 / 3 / 1, and 2 / 2 / 2.

[0099] The subsequent steps are the same as the preparation method in 1.1 above.

[0100] The obtained paclitaxel fat bodies were observed under laser confocal microscopy. The experimental results showed that by changing the neutral lipid conditions, using fish oil, tricaprylin, and retinol esters, paclitaxel-carrying fat bodies could be prepared. The paclitaxel fat bodies exhibited a uniform spherical structure (see Figure 5A, where a is the result of paclitaxel fat bodies stained red with the neutral lipid-specific dye LipidTox Red, and b is the result of DIC of paclitaxel fat bodies; scale bar is 2 μm). HPLC measurement of the PTX content in the paclitaxel fat bodies showed similar PTX content, reaching 600 μg / mL, under conditions of varying ratios of fish oil, tricaprylin, and retinol ester (Figure 5B, B shows the PTX content in the paclitaxel fat bodies measured by HPLC). The average particle size was similar, approximately 200 nm (see Figure 5C, C shows the average particle size of the paclitaxel fat bodies measured by dynamic light scattering particle size analysis).

[0101] 1.5. Modifying phospholipid conditions to prepare fat bodies carrying the hydrophobic small molecule compound paclitaxel

[0102] The neutral lipid conditions remained unchanged, namely fish oil and tricaprylin (1 / 1, volume ratio), and the types and proportions of phospholipids were changed to prepare paclitaxel fat bodies. The following are the proportions and types of phospholipids used:

[0103] Phosphatidyl cholines (PC), phosphatidic acid (PA), phosphatidylinositols (PI), phosphatidyl ethanolamines (PE), and polyethylene glycol-modified phosphatidyl ethanolamines (PEG-PE)

[0104] 1.5.1) Change the type and ratio of phospholipids. Prepare phospholipid solutions in chloroform at a concentration of 32 mM. Prepare the types and ratios of phospholipids for the preparation of paclitaxel liposomes according to the following scheme.

[0105] Table 2 Changes in the types and ratios of phospholipids used in the preparation of paclitaxel fat bodies

[0106] 1.5.2) Fish oil and tricaprylin (1 / 1, volume ratio) were used to prepare a neutral lipid containing PTX.

[0107] 1.5.3) Add 80 μL of the prepared phospholipids to each microcentrifuge tube and dry the solvent with high-purity nitrogen.

[0108] 1.5.4) Add 100 μL of PBS and 5 μL of the neutral lipid containing PTX prepared in step 2) to a microcentrifuge tube.

[0109] The subsequent steps are the same as the preparation method in 1.1 above.

[0110] Laser confocal microscopy observation of the resulting paclitaxel-containing fat bodies revealed that varying the phospholipid profile allowed the preparation of paclitaxel-containing fat bodies. However, the ratio and type of phospholipids affected the morphology and paclitaxel content of the fat bodies. While PA-containing fat bodies exhibited relatively heterogeneous morphology, fat bodies containing other phospholipids exhibited uniform, spherical structures (Figure 6, Panels A to C; Panels A to C show the results of paclitaxel-containing fat bodies stained red with the neutral lipid-specific dye LipidTox Red, and Panels B show the results of DIC of paclitaxel-containing fat bodies. Scale bar: 2 μm). HPLC analysis of the PTX content in paclitaxel liposomes prepared using different phospholipids revealed low PTX content of approximately 300 μg / mL in paclitaxel liposomes prepared with PA and PI. However, the addition of PE and PEG-PE significantly increased the PTX content, reaching over 500 μg / mL (see Figure 6, Panel D, which shows the PTX content in paclitaxel liposomes measured by HPLC). The addition of PI, PE, and PEG-PE increased the average particle size of the paclitaxel liposomes to approximately 200-300 nm (see Figure 6, Panel E, which shows the average particle size of the paclitaxel liposomes measured by dynamic light scattering particle size analysis). The addition of 10% PE significantly increased the concentration of the paclitaxel liposomes, reaching an OD600 of 120. The OD600 concentration of paclitaxel liposomes prepared using the remaining phospholipids was approximately 30 (see Figure 6, Panel F, which shows the OD600 concentration of the paclitaxel liposomes measured by a multifunctional microplate reader).

[0111] Example 2 Construction of a Fat Body Carrying the Hydrophobic Small Molecule Compound Magnolol (Magnolol Fat Body)

[0112] According to the chemical structure of magnolol and the encapsulation conditions of the paclitaxel fat body in Example 1, the lipid conditions selected in this example are: the phospholipid is 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (DOPC), the neutral lipid is triolein (18:1TAG), the paclitaxel in Example 1 is replaced by magnolol, and the other operating methods remain unchanged to prepare fat bodies encapsulated with magnolol (denoted as magnolol fat bodies).

[0113] Due to its hydrophobicity, the solubility of magnolol in PBS is very low, presenting a turbid state, while the magnolol encapsulated in the fat body presents a uniform milky white solution state (see A in Figure 7, A is a morphological diagram of magnolol in PBS and the fat body in PBS). The prepared magnolol fat body is further observed by laser microscopy, and the magnolol fat body presents a uniform spherical structure (see B in Figure 7, a is the result of magnolol fat body dyed red by the neutral lipid-specific dye LipidTox Red, and b is the DIC result of magnolol fat body, and the scale is 5 microns). The average particle size of the magnolol fat body is 114.2nm (see C in Figure 7, C is a diagram of the average particle size of the magnolol fat body measured by a dynamic light scattering particle size analyzer). The magnolol fat body is dissolved in methanol (chromatographic grade) to prepare a sample, and the content of magnolol in the paclitaxel fat body is measured by HPLC. The detection conditions are as follows: the chromatographic column is Agilent Zorbax SB-C18, the sample volume is 10 μL, the mobile phase is methanol: water = 7:3 (volume ratio), the flow rate is 1.0 mL / min, the detection wavelength is UV = 254 nm, and the content of magnolol in magnolol fat body is about 17 μg / mL (see D in Figure 7, D is a graph showing the content of magnolol in magnolol fat body measured by high performance liquid chromatography).

[0114] Example 3 Construction of Fat Bodies Carrying the Hydrophobic Small Molecule Compound Camptothecin (Camptothecin Fat Bodies)

[0115] Based on the chemical structure of camptothecin and the encapsulation conditions of the paclitaxel fat body in Example 1, the lipid conditions selected in this example are: the phospholipids are 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (DOPC) and cholesterol (Cholesterol, Cho), the neutral lipid is tricaprylin (8:0TAG) or a mixed neutral lipid of fish oil and tricaprylin (1:1 volume ratio), the paclitaxel in Example 1 is replaced with camptothecin, and the remaining operating procedures remain unchanged to prepare fat bodies encapsulating camptothecin (denoted as camptothecin fat bodies).

[0116] Due to its hydrophobicity, camptothecin has a very low solubility in PBS, resulting in a turbid state. However, camptothecin encapsulated in fat bodies appears as a uniform milky white solution (see Figure 8A, where A shows the morphology of camptothecin in PBS and fat bodies in PBS). Further observation of the prepared camptothecin fat bodies using a laser confocal microscope revealed that the camptothecin fat bodies exhibited a uniform spherical structure. The average particle size of camptothecin fat bodies prepared using different phospholipid and neutral lipid conditions was similar, approximately 120 nm (see Figure 8B, where ac in B shows the results of camptothecin fat bodies stained red with the neutral lipid-specific dye LipidTox Red, and df shows the average particle size of camptothecin fat bodies measured using a dynamic light scattering particle size analyzer. The scale bar is 2 microns). The camptothecin content in the camptothecin fat body was measured by HPLC. Compared with the mixed neutral lipid of fish oil and tricaprylin, only the camptothecin fat body prepared with tricaprylin had a relatively high camptothecin content of about 4 mg / L (C, C in Figure 8 are graphs showing the camptothecin content in the camptothecin fat body measured by HPLC).

[0117] Example 4 Construction of Fat Body Carrying Hydrophobic Small Molecule Compound Auristatin Derivative MMAE (Monomethyl auristatin E) (MMAE Fat Body)

[0118] Based on the chemical structure of MMAE and the encapsulation conditions of the paclitaxel fat body in Example 1, the lipid conditions selected in this example are: the phospholipid is 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (DOPC), and the neutral lipid is triolein (18:1TAG) or corn oil. The paclitaxel used in Example 1 is replaced with MMAE, and the remaining operating procedures remain unchanged to prepare fat bodies encapsulated with MMAE (denoted as MMAE fat bodies).

[0119] Due to its hydrophobicity, MMAE has a very low solubility in PBS and presents a turbid state, while the MMAE encapsulated in the fat body presents a uniform milky white solution state (see Figure 9 A, A is the morphology of MMAE in PBS and fat body in PBS). The prepared MMAE fat body was further observed by optical microscopy. The MMAE fat body prepared by corn oil or triolein all presented a uniform spherical structure (see Figure 9 B, B a is the DIC result of MMAE fat body prepared by corn oil, b is the DIC result of MMAE fat body prepared by triolein, and the scale is 5 microns). In addition, the average particle size of the MMAE fat body prepared by corn oil or triolein is similar, about 140nm, and the polymer dispersibility index (PDI) is also less than 0.2 (see Figure 9 C, C is the average particle size of MMAE fat body measured by dynamic light scattering particle size analyzer). 10 μL of MMAE fat was added to 190 μL of methanol (1% TCA), vortexed for 10 seconds, and centrifuged at 15,000 rpm for 3 minutes. 150 μL of the supernatant was added to a built-in sample tube and the MMAE content in the MMAE fat was measured by HPLC. The detection conditions were: an Agilent Zorbax SB-C18 column, a 10 μL sample volume, a mobile phase of 20% acetonitrile (1% TFA)-80% water (1% TFA) at 0 min, 80% acetonitrile (1% TFA)-20% water (1% TFA) at 5 min, and 20% acetonitrile (1% TFA)-80% water (1% TFA) at 7 min, at a flow rate of 1.0 mL / min, and detection at a UV wavelength of 210 nm. Consistent with the peak time of the MMAE standard, the MMAE signal of MMAE prepared from corn oil or triolein appeared at the same peak time. The MMAE content in the MMAE fat body prepared from corn oil was 100 μg / mL, and the MMAE content in the MMAE fat body prepared from triolein was 40 μg / mL (D in Figure 9, D is a graph showing the MMAE content in the MMAE fat body measured by high performance liquid chromatography).

[0120] Example 5 Construction of Fatsomes Carrying Both the Hydrophobic Small Molecule Compounds Paclitaxel and Camptothecin (Paclitaxel-Camptothecin Fatsomes)

[0121] Lipid conditions: The phospholipid was 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (DOPC) or egg phosphatidylcholine (Egg PC), the neutral lipid was a mixed neutral lipid of fish oil and tricaprylin (1:1 volume ratio), the paclitaxel used in Example 1 was replaced with a mixture of paclitaxel and camptothecin (1:1 mass ratio), and the rest of the operation method remained unchanged to prepare fat bodies carrying the hydrophobic small molecule compounds paclitaxel and camptothecin (paclitaxel-camptothecin fat bodies).

[0122] The prepared paclitaxel-camptothecin fat bodies were observed under a laser confocal microscope. The paclitaxel-camptothecin fat bodies exhibited a uniform spherical structure with an average particle size of 115.5 nm (see Figure 10A, where a represents the paclitaxel-camptothecin fat bodies stained red with the neutral lipid-specific dye LipidTox Red, and b represents the average particle size of the paclitaxel-camptothecin fat bodies measured using a dynamic light scattering particle size analyzer; the scale bar is 2 μm). Due to different detection conditions for camptothecin and paclitaxel, the camptothecin and paclitaxel contents in the paclitaxel-camptothecin fat bodies were measured by HPLC according to the following method. The HPLC detection conditions for camptothecin were: an Agilent Zorbax SB-C18 column, a mobile phase of methanol:acetonitrile:water = 3:3:4 (volume ratio), a column temperature of 35°C, a detection wavelength of 366 nm, and a flow rate of 0.8 mL / min. The detection conditions for paclitaxel were: an Agilent Zorbax SB-C18 column, a mobile phase of methanol:acetonitrile:water (4:3:3 by volume), a column temperature of 25°C, a detection wavelength of 227 nm, and a flow rate of 1 mL / min. When using the HPLC detection conditions for camptothecin, the elution time of camptothecin in the paclitaxel-camptothecin liposomes was consistent with that of the camptothecin standard, at 3.4 minutes, indicating that the paclitaxel-camptothecin liposomes had successfully entrapped camptothecin (see arrow B in Figure 10 , where a is the HPLC profile of the camptothecin standard and b is the HPLC profile of camptothecin in the paclitaxel-camptothecin liposomes). Similarly, when using HPLC detection conditions for paclitaxel, the peak elution time of paclitaxel in the paclitaxel-camptothecin fat body was consistent with that of the paclitaxel standard at 5.6 min, indicating that the paclitaxel-camptothecin fat body successfully entrapped paclitaxel (see arrow C in Figure 10 , where a is the HPLC graph of the paclitaxel standard and b is the HPLC graph of paclitaxel in the paclitaxel-camptothecin fat body). The camptothecin content in the paclitaxel-camptothecin fat body was approximately 4 mg / L, and the paclitaxel content was approximately 65 mg / L (see D in Figure 10 , where D is a graph showing the paclitaxel and camptothecin contents in the paclitaxel-camptothecin fat body prepared under Egg PC and DOPC conditions). These results demonstrate that the paclitaxel-camptothecin fat body successfully entrapped both paclitaxel and camptothecin.

[0123] Example 6: Fat bodies carrying hydrophobic small molecule compounds have biological activity and significantly kill cancer cells, with a killing effect superior to that of free drugs.

[0124] 6.1. Paclitaxel fat bodies significantly kill cancer cells, with a killing effect superior to free paclitaxel

[0125] The constructed paclitaxel fat bodies were incubated with human embryonic kidney epithelial HEK293 cells for 60 minutes, then washed twice with physiological saline. High-sensitivity laser scanning confocal microscopy was used to observe whether the paclitaxel fat bodies could enter the cells. The paclitaxel fat bodies were labeled with fluorescently labeled TAG (shown as green dots). The results showed that the paclitaxel fat bodies could be taken up by the cells and significantly killed the HEK293 cells with increasing paclitaxel content (see Figure 11A, where a and b show the entry of paclitaxel fat bodies into cells observed using high-sensitivity laser scanning confocal microscopy, and c shows the cell survival rate after treatment with different concentrations of paclitaxel fat bodies). Similarly, when the constructed paclitaxel fat bodies were incubated with human breast cancer MCF7 cells for 60 minutes, the paclitaxel fat bodies were taken up by the MCF7 cells and significantly killed the MCF7 cells as the paclitaxel content increased (see Figure 11B, where a and b in B show the entry of paclitaxel fat bodies into cells observed using a highly sensitive laser scanning confocal microscope, and c shows the cell survival rate after treatment with different concentrations of paclitaxel fat bodies). Furthermore, the cancer cell killing effects of the paclitaxel fat bodies were compared with those of unencapsulated free paclitaxel. The paclitaxel fat bodies prepared in 4 and 5 of Example 1 were selected to treat human breast cancer cells MDA-MB-231, respectively. The results showed that compared with free, unencapsulated paclitaxel drugs, the paclitaxel fat bodies had a better killing effect on cancer cells, especially when the concentration of paclitaxel was 5 μg / mL, the killing effect of the paclitaxel fat bodies was significantly higher than that of free paclitaxel (see Figure 11C and D, C is the survival rate graph of the paclitaxel fat bodies prepared in 4 of Example 1, D is the survival rate graph of the paclitaxel fat bodies prepared in 5 of Example 1, * indicates p < 0.05, ** indicates p < 0.01).

[0126] Cell viability assay: HEK293, MCF7, or MDA-MB-231 cells to be treated were plated at 1000 cells / well in a 96-well plate. After overnight attachment, the cells were treated with the indicated drug concentrations and incubated for 72 hours. Subsequently, the original culture medium was replaced with culture medium containing 10% CCK8. After incubation for 1 hour, the absorbance at 450 nm was read using a microplate reader. Cell viability was calculated using the following formula: Cell viability = (Ae-Ab) / (Ac-Ab)*100%

[0127] Wherein, Ae represents the absorbance value of the well containing cells after drug treatment;

[0128] Ac represents the absorbance value of the wells containing cells without drug treatment;

[0129] Ab represents the absorbance value of blank wells containing only culture medium and CCK8 reagent, which is used to correct background noise.

[0130] 6.2. Magnolia officinalis fat bodies significantly kill cancer cells, and the killing effect is better than free magnolia officinalis

[0131] The experimental method was the same as in 6.1, comparing the cancer cell killing effects of magnolol fat bodies and unencapsulated free magnolol. The magnolol fat bodies prepared in Example 2 were used to treat human breast cancer MCF7 cells. The results showed that compared with free, unencapsulated magnolol, magnolol fat bodies were more effective in killing cancer cells. In particular, when the concentration of magnolol was 10 μg / mL, the killing effect of magnolol fat bodies was significantly higher than that of free magnolol (see Figure 12, which is a comparison of the survival rates of magnolol fat bodies and free magnolol, * indicates p < 0.05).

[0132] 6.3. MMAE fat bodies significantly kill cancer cells, with a killing effect superior to free MMAE

[0133] The experimental method was the same as in 6.1, comparing the cancer cell killing effects of MMAE fat bodies and unencapsulated free MMAE. The MMAE fat bodies prepared in Example 4 were used to treat mouse breast cancer 4T1 cells. The results showed that MMAE fat bodies were more effective in killing cancer cells than free, unencapsulated MMAE drugs. In particular, when the MMAE concentration was 500 nM, the killing effect of MMAE fat bodies was significantly higher than that of free MMAE (see Figure 13, which is a comparison of the survival rates of MMAE fat bodies and free MMAE, *** indicates p < 0.001).

[0134] The above embodiments are only intended to help understand the method and core concept of the present invention. It should be noted that, without departing from the principles of the present invention, a number of improvements and modifications may be made to the present invention by those skilled in the art, and such improvements and modifications also fall within the scope of protection of the claims of the present invention.

Claims

1. A nanoparticle fat body encapsulating a hydrophobic small molecule drug, characterized in that: It includes a monomolecular phospholipid membrane and a neutral lipid containing a hydrophobic small molecule drug encapsulated inside the monomolecular phospholipid membrane.

2. The nanoparticle fat body containing hydrophobic small molecule drugs according to claim 1, characterized in that: The hydrophobic small molecule drug is selected from one or more of paclitaxel, magnolol, camptothecin, and auristatin derivative MMAE.

3. The nanoparticle fat body loaded with hydrophobic small molecule drugs according to claim 1, characterized in that: The neutral lipid is selected from one or more of fish oil, corn oil, tricaprylin, and retinol ester.

4. The nanoparticle fat body loaded with hydrophobic small molecule drugs according to claim 3, characterized in that: The neutral lipid is selected from a mixture of fish oil and tricaprylin, or a mixture of fish oil and retinol ester, or a mixture of tricaprylin and corn oil, or a mixture of fish oil, tricaprylin and retinol ester; The volume ratio of the fish oil to tricaprylin is (1-5):(1-2); The volume ratio of the fish oil to the retinol ester is (1-5):(1-2); The volume ratio of tricaprylin to corn oil is (1-5):(1-2); The volume ratio of the fish oil, tricaprylin and retinol ester is (2-4): (1-3): (1-2).

5. The nanoparticle fat body loaded with hydrophobic small molecule drugs according to claim 1, characterized in that: The phospholipid membrane is selected from one or more of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine, phosphatidylcholine, glycerophosphatidic acid, phosphatidylinositol, phosphatidylethanolamine, and polyethylene glycol-modified phosphatidylethanolamine.

6. The nanoparticle fat body loaded with hydrophobic small molecule drugs according to claim 5, characterized in that: The phospholipid membrane is selected from any one of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine, glycerophosphatidic acid, phosphatidylinositol, phosphatidylethanolamine, polyethylene glycol-modified phosphatidylethanolamine and a mixture of phosphatidylcholine; The molar content of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine, glycerophosphatidic acid, phosphatidylinositol, phosphatidylethanolamine or polyethylene glycol-modified phosphatidylethanolamine in the mixture is 5% to 10%.

7. The method for preparing the nanoparticle fat body containing hydrophobic small molecule drugs according to any one of claims 1 to 6, comprising the following steps: S1) mixing a hydrophobic small molecule drug solution and a neutral lipid, and removing the solvent to obtain a neutral lipid containing the hydrophobic small molecule drug; S2) mixing the neutral lipid and phospholipid containing the hydrophobic small molecule drug to obtain a lipid mixture, and separating to obtain nanoparticle fat bodies encapsulating the hydrophobic small molecule drug.

8. The preparation method according to claim 7, characterized in that: The step S2) is specifically as follows: A) vortexing the neutral lipid and phospholipid containing the hydrophobic small molecule drug in a buffer to obtain a milky white lipid mixture; B) Repeatedly centrifuging and vortexing the lipid mixture to separate the nanoparticle fat bodies containing the hydrophobic small molecule drug.

9. Use of the nanoparticle fat body containing hydrophobic small molecule drugs according to any one of claims 1 to 6 in the preparation of drugs for preventing, alleviating or treating tumors, infectious diseases and / or metabolic diseases.

10. A drug for preventing, alleviating or treating tumors, infectious diseases and / or metabolic diseases, characterized in that: The invention comprises the nanoparticle fat body containing the hydrophobic small molecule drug as claimed in any one of claims 1 to 6.

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