Lipid body encapsulating fat-soluble nutrient ingredient, and preparation method therefor and use thereof

Abstract

The present invention relates to the fields of pharmaceutical techniques / health care products, particularly to a lipid body preparation encapsulating a fat-soluble nutritional ingredient, and a preparation method therefor and the use thereof. The lipid body encapsulating a fat-soluble nutritional ingredient constructed in the present invention can improve the solubility, stability and bioavailability of the fat-soluble nutritional ingredient, and the effect thereof is better than that of free EPA. Moreover, compared with some lipid bodies constructed with inappropriate parameters, the lipid body provided in the present invention is more effective in improving the stability and bioavailability, and has good encapsulation effects.

Description

Fat bodies encapsulating fat-soluble nutritional ingredients and preparation method and application thereof

[0001] The present application claims priority to the Chinese patent application No. 202411391042.2, filed on September 30, 2024, and entitled "Fat bodies encapsulating fat-soluble nutritional ingredients and preparation method and application thereof", the content of which is incorporated herein by reference in its entirety. TECHNICAL FIELD

[0002] The present application relates to the field of pharmaceutical technology, in particular to fat bodies encapsulating fat-soluble nutritional ingredients and preparation method and application thereof. BACKGROUND

[0003] Fat-soluble nutritional ingredients refer to those nutritional ingredients that are not soluble or have very low solubility (solubility is generally less than 0.1 mg / ml) in water, which play a variety of important roles in the body, such as Omega-3 / 6 / 9 unsaturated fatty acids and other functional neutral lipids, fat-soluble vitamins A / D / E / K, steroid hormones estrogen / androgen, fat-soluble extracts lutein, ginsenosides, puerarin, berberine, etc.

[0004] Fat-soluble nutritional ingredients are easily soluble in neutral lipids, but have poor water solubility, often poor oral absorption, low bioavailability and poor stability in biological systems. In order to improve the solubility and bioavailability of fat-soluble ingredients, some strategies are often needed to improve, such as chemical modification of drug structure or the use of nanocarriers for delivery.

[0005] In order to improve the solubility, effectiveness and safety of fat-soluble nutritional ingredients, many delivery carriers are applied to encapsulate fat-soluble nutritional ingredients, but these methods also have some disadvantages, such as: 1) liposomes, liposomes are prepared by simple mixing, extrusion, oscillation and ultrasonication, etc. However, liposomes have a hydrophilic core, only the hydrophobic environment between their phospholipid bilayers can be used for fat-soluble nutritional ingredients, but their encapsulation capacity is low. In addition, liposomes carrying fat-soluble nutritional ingredients result in poor membrane stability, which is prone to fusion, rupture or leakage. 2) Solid lipid nanoparticles, solid lipid nanoparticles are prepared by melting method, solvent method, supercritical fluid method, etc. High temperature or organic solvent is needed in the preparation process, which may cause the crystallization or separation of fat-soluble nutritional ingredients, thereby affecting their stability. 3) Lipid microspheres, micelles and other polymer nanocarriers, lipid microspheres are prepared by emulsion-solvent evaporation method, co-precipitation method, solution polymerization method, etc. However, certain materials may not be stable enough in the body environment, and are easily affected by body fluids or enzymes to lose activity, and even may cause safety problems.

[0006] The new type of nanoparticle fat body has a hydrophobic core of neutral lipids, and is a nanosphere wrapped by a monomolecular phospholipid film, which is similar to naturally occurring lipid droplets and lipoprotein structures, can efficiently dissolve and encapsulate hydrophobic small molecule compounds, and has good biocompatibility due to the natural existence of the components of the fat body in the body. In addition, the preparation method is simple and efficient. If the fat body is applied to load fat-soluble nutritional ingredients, it is expected to solve the problems of poor water solubility, poor stability, poor oral absorption and low bioavailability of fat-soluble nutritional ingredients, and help to improve the physiological function of the body and maintain the health of the body, and has a wide application prospect in the fields of food, cosmetics or medical treatment. SUMMARY

[0007] Therefore, the technical problem to be solved by the present application is to provide a fat body preparation encapsulating fat-soluble nutritional ingredients and a preparation method and application thereof, which has good solubility, bioavailability and stability, and can load targeting molecules.

[0008] The fat body preparation encapsulating fat-soluble nutritional ingredients provided by the present application comprises: a monomolecular phospholipid film and a fat-soluble nutritional ingredient and a neutral lipid wrapped inside the monomolecular phospholipid film.

[0009] In the present application, the fat-soluble nutritional ingredients include at least one of unsaturated fatty acids, fat-soluble vitamins, steroid hormones and / or fat-soluble extracts.

[0010] The unsaturated fatty acids include monounsaturated fatty acids and polyunsaturated fatty acids, and as a feasible case, the unsaturated fatty acids include at least one of Omega-3, Omega-6, Omega-9 unsaturated fatty acids; further, the unsaturated fatty acids include but are not limited to at least one of oleic acid, linoleic acid, linolenic acid, eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), docosahexaenoic acid (DHA), fish oil, snakegourd seed oil, soybean oil, castor oil, and corn oil.

[0011] The fat-soluble vitamin is a kind of vitamin that is easily soluble in organic solvents but difficult to dissolve in water, and as a feasible case, the fat-soluble vitamin includes but is not limited to at least one of vitamin A, vitamin D, vitamin E or vitamin K.

[0012] The steroid hormone, also known as steroid hormone, is a lipid hormone synthesized from cholesterol. As a feasible case, the steroid hormone includes but is not limited to at least one of androgen, estrogen, progestogen, anti-estrogen, anti-progestogen or non-steroidal estrogen.

[0013] The liposoluble extract in the present application refers to a compound that can be extracted by using an organic solvent. As a feasible case, the liposoluble extract includes, but is not limited to, at least one of berberine, berberine, coenzyme Q10, retinol, ginsenoside, lutein, quercetin, anthocyanin, lycopene, panax notoginseng saponin, puerarin, chlorophyll, carotenoid, polyphenol or phytosterol.

[0014] In the present application, the neutral lipid is selected from one or more of glyceryl trioleate, fish oil, castor oil, corn oil, glyceryl trioctanoate, retinol ester, wax ester, sterol ester, stanols ester, sunflower oil, soybean oil, peanut oil, clove oil, simethicone, cinnamon oil, tea oil, liquid paraffin, star anise oil, mixed fatty acid glyceride (stearin), hydrogenated vegetable oil, refined olive oil, refined olive oil, retinol ester and fat-soluble vitamins.

[0015] In the present application, the neutral lipid further includes an antioxidant selected from at least one of vitamin E, carotenoid, ubiquinone, guaiac wood resin, gallic acid, butylated hydroxyanisole. In some embodiments, the mass fraction of the antioxidant in the neutral lipid is 1% to 10%. For example, the mass fraction of the antioxidant in the neutral lipid is 1% to 5%, or 5% to 10%, or 3% to 7%, or 4 to 6%, or 4.8% to 5.2%, or 4.5% to 5.5%. As preferred, the mass fraction of the antioxidant in the neutral lipid is 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9% or 10%.

[0016] In the present application, the membrane material of the monomolecular phospholipid film includes one or more of phospholipids or functional polar lipids.

[0017] In some embodiments, the phospholipid is selected from one or more of egg yolk lecithin, glycocholic acid, soybean phospholipid, trimethyl-2,3-dioleoylpropyl ammonium bromide, dioleoyl phosphatidyl ethanolamine, distearoyl phosphatidyl choline, egg yolk lecithin, dipalmitoyl phosphatidyl choline, dipalmitoyl phosphatidic acid, distearoyl phosphatidyl glycerol sodium, dimyristoyl phosphatidyl choline, 1-stearoyl-hydrolyzed phosphatidyl choline, 1,2-dimyristoyl-rac-glycerol-3-methoxypolyethylene glycol 2000, phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidylinositol, phosphatidyl serine, phosphatidyl glycerol, phosphatidic acid, cardiolipin and sphingomyelin.

[0018] In some embodiments, the functional polar lipid is selected from one or more of cholesterol, polyethylene glycol modified sterol, biotin modified sterol, amino acid modified sterol, polypeptide modified sterol, polysaccharide modified sterol, nucleic acid modified sterol, polyethylene glycol modified phospholipid, biotin modified phospholipid, amino acid modified phospholipid, polypeptide modified phospholipid, polysaccharide modified phospholipid and nucleic acid modified phospholipid.

[0019] In some embodiments, the monomolecular phospholipid film further comprises a cationic lipid selected from one or more of DOTAP, DODAP, DODMA, DOTMA, D-Lin-MC3-DMA.

[0020] Further, the preparation of the present application further comprises a targeting molecule which targets an organ, tissue or cell.

[0021] Further, the preparation of the present application further comprises a targeting molecule which targets an organ, tissue or cell.

[0022] In the present application, the organ, tissue or cell is from a human or animal body;

[0023] The organ is an endocrine organ, a digestive organ, a circulatory organ, a urinary organ, a reproductive organ, a motor organ, a nervous system and a sensory organ. The endocrine organ includes thyroid, pancreas. The digestive organ includes stomach, liver, gallbladder, spleen, pancreas, small intestine, large intestine. The respiratory organ includes lung. The circulatory organ includes heart, blood vessel. The urinary organ includes kidney, ureter, bladder. The reproductive organ includes uterus, ovary. The motor organ includes muscle, bone. The nervous system includes brain, cerebellum. The sensory organ includes skin, eye, ear.

[0024] The tissue or cell is from a tumor of a human or animal body. The tumor includes lung cancer, kidney cancer, laryngeal cancer, liver cancer, muscle tissue cancer, blood tumor, bone cancer, brain cancer, breast cancer, neck cancer, oral or nasal mucosa cancer, bladder cancer, central nervous system cancer, cervical cancer, head and neck cancer, colon cancer, endometrial cancer, cancer of the external genitalia, esophageal cancer, gallbladder cancer, gastrointestinal cancer, urogenital cancer, head cancer, ovarian cancer, pancreatic cancer, prostate cancer, skin cancer, spleen cancer, small intestine cancer, large intestine cancer, stomach cancer, melanoma, testicular cancer and / or thyroid cancer.

[0025] The targeting molecule can be embedded in the monomolecular phospholipid film, can be combined with the phospholipid on the monomolecular phospholipid film through the avidin-biotin system, can be connected with a substance specifically targeting the phospholipid to be combined on the monomolecular phospholipid film, or can be a combination of any two or more of the above.

[0026] In the present application, the targeting molecule is at least one of LTA-P33, ApoE, BCMA antibody, Nrp-B, Trf-B, LDLR-B, ErbB2-B, CXCR4-B, GRP78-B or Soma-B.

[0027] In some embodiments, the targeting molecule is linked to biotin, and streptavidin-labeled phospholipid molecules are added during the preparation of the fat body. Alternatively, the targeting molecule is linked to streptavidin, and biotin-labeled phospholipid molecules are added during the preparation of the fat body.

[0028] In some embodiments, the targeting molecule is linked to a peptide segment that targets and recognizes a single-molecule phospholipid membrane, and the peptide segment comprises at least one of AAMB, ALDI, CYB5R3-N, LDAMP1, HSD17B13-N28, MDT-28-P, MLDS-P, DHS-3-P, HSD17B11-N28, PspA-H1, Vipp1-H1, Snf7-H1, Chmp1B-H1, PB, PE, and PF. The targeting molecule and the peptide segment that targets and recognizes a single-molecule phospholipid membrane can be linked via a linker or directly linked without a linker, and the present application does not limit the same. The linker is a cleavable linker or a self-cleavable linker. The amino acid sequence of the cleavable linker is LEAGCKNFFPRSFTSCGSLE, and the self-cleavable linker is P2A, T2A, or E2A.

[0029] In some embodiments, the fat-soluble nutritional ingredient is eicosapentaenoic acid ethyl ester.

[0030] In this embodiment, the single-molecule phospholipid membrane comprises at least one of egg yolk lecithin, glycocholic acid, trimethyl-2,3-dioleoyloxypropyl ammonium bromide, cholesterol, trimethyl-2,3-dioleoyloxypropyl ammonium bromide, and / or phosphatidyl ethanolamine-polyethylene glycol 2000; and the neutral fat is triolein and / or soybean oil.

[0031] Specifically, the single-molecule phospholipid membrane comprises egg yolk lecithin, and the eicosapentaenoic acid ethyl ester and triolein are wrapped inside the single-molecule phospholipid membrane.

[0032] Alternatively, the single-molecule phospholipid membrane comprises egg yolk lecithin and glycocholic acid, and the eicosapentaenoic acid ethyl ester is wrapped inside the single-molecule phospholipid membrane.

[0033] Alternatively, the single-molecule phospholipid membrane comprises egg yolk lecithin and glycocholic acid, and the eicosapentaenoic acid ethyl ester and triolein are wrapped inside the single-molecule phospholipid membrane.

[0034] Alternatively, the single-molecule phospholipid membrane comprises egg yolk lecithin, and the eicosapentaenoic acid ethyl ester, vitamin E, and soybean oil are wrapped inside the single-molecule phospholipid membrane.

[0035] Alternatively, the single-molecule phospholipid membrane comprises egg yolk lecithin and glycocholic acid, and the eicosapentaenoic acid ethyl ester, vitamin E, and soybean oil are wrapped inside the single-molecule phospholipid membrane.

[0036] Alternatively, the monomolecular phospholipid film comprises egg yolk phosphatidylcholine, DOTAP, cholesterol, and DSPE-PEG2000, and encapsulated inside the monomolecular phospholipid film are eicosapentaenoic acid ethyl ester, vitamin E, and soybean oil;

[0037] Alternatively, the monomolecular phospholipid film comprises egg yolk phosphatidylcholine, DOTAP, cholesterol, DSPE-PEG2000, and glycocholic acid, and encapsulated inside the monomolecular phospholipid film are eicosapentaenoic acid ethyl ester, vitamin E, and soybean oil.

[0038] More specifically, the volume ratio of the eicosapentaenoic acid ethyl ester to the neutral lipid is (1-5):(1-5). In some embodiments, the volume ratio of the eicosapentaenoic acid ethyl ester to the neutral lipid is (1-5): 1, or (1-5):2, or (1-5):3, or (1-5):4, or (1-5):5, or 1:(1-5), or 2:(1-5), or 3:(1-5), or 3:(1-5), or 4:(1-5), or 5:(1-5). In some embodiments, the volume ratio of the eicosapentaenoic acid ethyl ester to the neutral lipid is 1:1, 2:1, 3:1, 4:1, 5:1, 1:2, 3:2, 5:2, 1:3, 2:3, 4:3, 5:3, 1:4, 3:4, 5:4, 1:5, 2:5, 3:5, or 4:5.

[0039] More specifically, the mass ratio of the egg yolk phosphatidylcholine to the glycocholic acid is (10-40):(2-40). In some embodiments, the volume ratio of the eicosapentaenoic acid ethyl ester to the neutral lipid is (10-40):(10-40) or (10-40):(2-8). For example, the volume ratio of the eicosapentaenoic acid ethyl ester to the neutral lipid is 10:10, 20:10, 30:10, 40:10, 10:20, 30:20, 10:30, 20:30, 40:30, 10:40, or 30:40, or 10:2, 10:3, 10:4, 10:5, 10:6, 10:7, 10:8, 20:2, 20:3, 20:4, 20:5, 20:6, 20:7, 20:8, 30:2, 30:3, 30:4, 30:5, 30:6, 30:7, 30:8, 40:2, 40:3, 40:4, 40:5, 40:6, 40:7, or 40:8.

[0040] More specifically, the volume ratio of the egg yolk phosphatidylcholine, DOTAP, cholesterol, and DSPE-PEG2000 is 300:(30-300):(30-150):(50-300). In some embodiments, the volume ratio of the egg yolk phosphatidylcholine, DOTAP, cholesterol, and DSPE-PEG2000 is 300:(33-267):(30-120):(75-300). For example, the volume ratio of the egg yolk phosphatidylcholine, DOTAP, cholesterol, and DSPE-PEG2000 is 300:33:(30-120):(75-300), or 300:90:(30-120):(75-300), or 300:267:(30-120):(75-300), or 300:(33-267):30:(75-300), or 300:(33-267):79.5:(75-300), or 300:(33-267):120:(75-300), or 300:(33-267):(30-120):75, or 300:(33-267):(30-120):199.5, or 300:(33-267):(30-120):300, or 300:33:30:75, or 300:90:79.5:199.5, or 300:267:120:300.

[0041] More specifically, the volume ratio of the egg yolk phosphatidylcholine, DOTAP, cholesterol, DSPE-PEG2000, and glycocholic acid is 300:(30-300):(30-150):(50-300):(1-10). In some embodiments, the volume ratio of the egg yolk phosphatidylcholine, DOTAP, cholesterol, DSPE-PEG2000, and glycocholic acid is 300:(30-40):(20-25):(50-100):(4-5), and preferably, the volume ratio of the egg yolk phosphatidylcholine, DOTAP, cholesterol, DSPE-PEG2000, and glycocholic acid is 300:33:22.5:75:4.5.

[0042] In other embodiments, the fat-soluble nutritional ingredient is vitamin A palmitate, and the monomolecular phospholipid membrane comprises DOPC, and the triolein and vitamin A palmitate are encapsulated inside the monomolecular phospholipid membrane.

[0043] In this embodiment, the volume ratio of the glyceryl trioleate and vitamin A palmitate is (1-10): 1. For example, the volume ratio of the glyceryl trioleate and vitamin A palmitate is (1-8): 1, or (1-6): 1, or (2-5): 1, or (3-5): 1, or 1:1, 2:1, 3:1, 4:1, 5:1, 6:1, 7:1, 8:1, 9:1, 10:1.

[0044] In other embodiments, the fat-soluble nutrient is lutein, the single-molecule phospholipid membrane comprises egg yolk lecithin, and the neutral fat is at least one of soybean oil, castor oil, or glyceryl tricaprylate.

[0045] In this embodiment, the neutral fat is soybean oil, castor oil, glyceryl tricaprylate, or a mixture of soybean oil, castor oil, and glyceryl tricaprylate.

[0046] In this embodiment, the mass ratio of lutein to egg yolk lecithin is (1-10):(1-10); preferably (1-5):(1-5). For example, the mass ratio of lutein to egg yolk lecithin is (1-5): 1, or (1-5): 2, or (1-5): 3, or (1-5): 4, or (1-5): 5, or 1:(1-5), or 2:(1-5), or 3:(1-5), or 3:(1-5), or 4:(1-5), or 5:(1-5). For example, the mass ratio of lutein to egg yolk lecithin is 1:1, 2:1, 3:1, 4:1, 5:1, 1:2, 3:2, 5:2, 1:3, 2:3, 4:3, 5:3, 1:4, 3:4, 5:4, 1:5, 2:5, 3:5, or 4:5.

[0047] In this embodiment, the mass-volume ratio of lutein to neutral fat is (0.1-10) mg:(0.1-10) mL. For example, the mass-volume ratio of lutein to neutral fat is (0.5-5) mg:(0.5-5) mL, preferably, the mass-volume ratio of lutein to neutral fat is (0.8-1.2) mg:(0.8-1.2) mL, for example, the mass-volume ratio of lutein to neutral fat is 0.8 mg:1.2 mL, 0.8 mg:1.0 mL, 0.8 mg:0.8 mL, 1.0 mg:1.2 mL, 1.0 mg:1.0 mL, 1.0 mg:0.8 mL, 1.2 mg:1.2 mL, 1.2 mg:1.0 mL, or 1.2 mg:0.8 mL.

[0048] In this embodiment, the volume ratio of the soybean oil, castor oil and trioctanoin in the mixture of the soybean oil, castor oil and trioctanoin is (1-10):(1-10):(1-10). Preferably, the volume ratio of the soybean oil, castor oil and trioctanoin is (1-5):(1-5):(1-5), for example, the volume ratio of the soybean oil, castor oil and trioctanoin is (1-3):(1-3):(1-3), or (1-2):(1-2):(1-2). For example, the volume ratio of the soybean oil, castor oil and trioctanoin is (1-2):(1-2):1, or (1-2):(1-2):2, or (1-2):1:(1-2), or (1-2):2:(1-2), or 1:(1-2):(1-2), or 2:(1-2):(1-2), preferably, the volume ratio of the soybean oil, castor oil and trioctanoin is 1:1:1.

[0049] In other embodiments, the fat-soluble nutritional ingredient is ginsenoside, the single-molecule phospholipid membrane comprises egg yolk lecithin, and the neutral fat is at least one of soybean oil, fish oil or trioctanoin.

[0050] In this embodiment, the neutral fat is soybean oil, castor oil, trioctanoin, or a mixture of soybean oil, castor oil and trioctanoin.

[0051] In this embodiment, the mass ratio of ginsenoside to egg yolk lecithin is (1-50):(1-10); preferably (10-50):(2-10). For example, the mass ratio of ginsenoside to egg yolk lecithin is (10-25):(2-5), or (10-15):(2-3). Preferably, the mass ratio of ginsenoside to egg yolk lecithin is (10-15):2 or (10-15):3, for example, the mass ratio of ginsenoside to egg yolk lecithin is 10:2, 11:2, 12:2, 13:2, 14:2, 15:2, 10:3, 11:3, 12:3, 13:3, 14:3 or 15:3.

[0052] In this embodiment, the mass-volume ratio of ginsenoside to neutral fat is (0.1-10) mg:(0.1-5) mL. For example, the mass-volume ratio of ginsenoside to neutral fat is (1-10) mg:(0.1-1) mL, preferably, the mass-volume ratio of ginsenoside to neutral fat is (1-5) mg:(0.1-0.5) mL, for example, the mass-volume ratio of ginsenoside to neutral fat is 5 mg:0.5 mL, 4 mg:0.4 mL, 3 mg:0.3 mL, 2 mg:0.2 mL, 1.0 mg:0.1 mL.

[0053] In this embodiment, the volume ratio of the soybean oil, castor oil and tricaprylin in the mixture is (1-10):(1-10):(1-10). As a preference, the volume ratio of the soybean oil, castor oil and tricaprylin is (1-5):(1-5):(1-5), for example, the volume ratio of the soybean oil, castor oil and tricaprylin is (1-3):(1-3):(1-3), or (1-2):(1-2):(1-2). For example, the volume ratio of the soybean oil, castor oil and tricaprylin is (1-2):(1-2):1, or (1-2):(1-2):2, or (1-2):1:(1-2), or (1-2):2:(1-2), or 1:(1-2):(1-2), or 2:(1-2):(1-2), as a preference, the volume ratio of the soybean oil, castor oil and tricaprylin is 1:1:1.

[0054] In other embodiments, the fat-soluble nutritional ingredient is berberine, the monomolecular phospholipid membrane comprises egg yolk lecithin and / or polyoxyethylene ether castor oil, and the neutral fat is at least one of castor oil, fish oil or medium-chain triglyceride.

[0055] In this embodiment, the neutral fat is castor oil, fish oil or medium-chain triglyceride. The monomolecular phospholipid membrane is a mixture of egg yolk lecithin and polyoxyethylene ether castor oil.

[0056] In this embodiment, the mass ratio of berberine to castor oil is (0.001-0.01):(1-10); preferably (0.003-0.005):(2-8). For example, the mass ratio of berberine to castor oil is 0.004:4. In this embodiment, the mass ratio of berberine to fish oil is (0.001-0.01):(1-10); preferably (0.003-0.005):(2-8). For example, the mass ratio of berberine to fish oil is 0.004:4. In this embodiment, the mass ratio of berberine to medium-chain triglyceride is (0.001-0.01):(1-10); preferably (0.003-0.005):(2-8). For example, the mass ratio of berberine to medium-chain triglyceride is 0.004:4.

[0057] In this embodiment, the mass ratio of the egg yolk lecithin and polyoxyethylene ether castor oil in the mixture is (1-10):(1-10). As a preference, the mass ratio of the egg yolk lecithin and polyoxyethylene ether castor oil is (2-8):(2-8), for example, the mass ratio of the egg yolk lecithin and polyoxyethylene ether castor oil is (3-5):(3-5), as a preference, the mass ratio of the egg yolk lecithin and polyoxyethylene ether castor oil is 4:4.

[0058] Further, the application also provides a preparation method of the preparation as described above, comprising:

[0059] After mixing the fat-soluble nutritional ingredient and the neutral fat, the fat-soluble nutritional ingredient and the neutral fat are mixed with the medium and the membrane material of the monomolecular phospholipid membrane, and after ultrasonic treatment and centrifugation, the lower milky white solution is collected, and after vortexing, the fat body preparation loaded with the fat-soluble nutritional ingredient is obtained.

[0060] Or the preparation method of the preparation as described above, comprising: after mixing the fat-soluble nutritional ingredient and the neutral fat, the fat-soluble nutritional ingredient and the neutral fat are mixed with the medium and the membrane material of the monomolecular phospholipid membrane, and after repeated vortexing, the fat body preparation loaded with the fat-soluble nutritional ingredient is obtained; the medium is water or a buffer solution.

[0061] In the application, the buffer solution is a PBS buffer solution, a HEPES buffer solution, a sucrose solution, a NaCl solution, a KCl solution, or a MgCl2 solution.

[0062] In the membrane material, not only phospholipids are contained, and each component in the membrane material is dissolved in an organic solvent and then mixed, and the organic solvent is then volatilized. The organic solvent is a mixture of methanol and at least one of the following solvents: anhydrous ethanol, chloroform, benzene, toluene, xylene, butanol, isopropyl alcohol, diethyl ether, acetone, cyclohexanone, methyl isobutyl ketone, ethyl acetate, butyl acetate, cyclohexanone, or petroleum ether, and preferably a mixture of methanol and chloroform.

[0063] In the application, the parameters of the ultrasonic treatment include: power: 100-300 W, water temperature: 20-40℃, 1-10 min, and ultrasonic treatment for 1-5 times. For example, the power of the ultrasonic treatment is 200-300 W, preferably 200-250 W, and specifically 200 W, 210 W, 211 W, 212 W, 213 W, 214 W, 215 W, 216 W, 217 W, 218 W, 219 W, 220 W, 230 W, 240 W, or 250 W. The water temperature of the ultrasonic treatment is 20-30℃, and preferably 20℃, 21℃, 22℃, 23℃, 24℃, 25℃, 26℃, 27℃, 28℃, 29℃, or 30℃. The time of the ultrasonic treatment is 1-5 min, and preferably 1 min, 2 min, 3 min, 4 min, or 5 min. The number of times of the ultrasonic treatment is 1 time, 2 times, 3 times, 4 times, or 5 times. In a specific embodiment, the parameters of the ultrasonic treatment include: power: 216 W, water temperature: 25℃, 3 min, and ultrasonic treatment for three times.

[0064] In the application, the repeated vortexing and centrifugation includes: after mixing, vortexing to obtain a mixture 1, collecting the lower solution after centrifugation of the mixture 1, and vortexing again to obtain a mixture 2; removing the precipitate after centrifugation of the mixture 2, and vortexing again to obtain a mixture 3; collecting the lower solution after centrifugation of the mixture 3, and vortexing again to obtain a mixture 4 containing the fat body.

[0065] Specifically: after mixing, vortex to obtain mixture 1.

[0066] In this step, the vortex parameters include: 3000-4000 rpm vortex for 3-7 min, work for 1-10 s, stop for 1-10 s. As a preferred, the vortex speed is 3000, 3200, 3400, 3500, 3600, 3700, 3800, 3900 or 4000 rpm. As a preferred, the vortex time is 3 min, 4 min, 5 min, 6 min or 7 min. As a preferred, the vortex works for 1 s and stops for 1 s, or works for 2 s and stops for 2 s, or works for 2 s and stops for 2 s, or works for 3 s and stops for 3 s, or works for 4 s and stops for 4 s, or works for 5 s and stops for 5 s, or works for 6 s and stops for 6 s, or works for 7 s and stops for 7 s, or works for 8 s and stops for 8 s, or works for 9 s and stops for 9 s, or works for 10 s and stops for 10 s, or works for 5 s and stops for 6 s, or works for 5 s and stops for 7 s, or works for 5 s and stops for 8 s, or works for 5 s and stops for 9 s, or works for 5 s and stops for 10 s, or works for 6 s and stops for 7 s, or works for 6 s and stops for 8 s, or works for 6 s and stops for 9 s, or works for 6 s and stops for 10 s, or works for 7 s and stops for 8 s, or works for 7 s and stops for 9 s, or works for 7 s and stops for 10 s, or works for 8 s and stops for 9 s, or works for 8 s and stops for 10 s, or works for 9 s and stops for 10 s. In some specific embodiments, the vortex parameters in this step include 4000 rpm, vortex for 10 s and stop for 10 s, vortex for 4 min. Or the vortex parameters in this step include 4000 rpm, vortex for 10 s and stop for 5 s, vortex for 3 min.

[0067] After the lower solution of the mixture 1 is collected by centrifugation, vortex again to obtain mixture 2.

[0068] In this step, the vortex parameters include 1000-4000 rpm, and the centrifugation parameters include: 800-1200 g centrifugation at room temperature for 3-7 min. As a preferred, the centrifugation speed is 800 g, 900 g, 1000 g, 1100 g or 1200 g, and the centrifugation time is 3 min, 4 min, 5 min, 6 min or 7 min. In some embodiments, the centrifugation conditions include 1000 g centrifugation at room temperature for 5 min.

[0069] After the precipitate of the mixture 2 is removed by centrifugation, vortex again to obtain mixture 3;

[0070] In this step, the parameters of vortexing include 1000-4000 rpm, and the parameters of centrifugation include 18000-22000 g centrifugation for 3-7 min in practical application. Preferably, the speed of centrifugation is 18000 g, 19000 g, 20000 g, 21000 g or 22000 g, and the time of centrifugation is 3 min, 4 min, 5 min, 6 min or 7 min. In some embodiments, the centrifugation condition includes 20000 g centrifugation for 5 min at room temperature.

[0071] The mixture 3 is collected by centrifugation to obtain a lower solution, and vortexing again to obtain a mixture 4 containing the fat body. In this step, the parameters of vortexing include 1000-4000 rpm, and the parameters of centrifugation include 800-1200 g centrifugation for 3-7 min in practical application. Preferably, the speed of centrifugation is 800 g, 900 g, 1000 g, 1100 g or 1200 g, and the time of centrifugation is 3 min, 4 min, 5 min, 6 min or 7 min. In some embodiments, the centrifugation condition includes 1000 g centrifugation for 5 min at room temperature.

[0072] Further, the present application also provides the use of the preparation as described above or the preparation prepared by the method as described above in the preparation of health food, cosmetics and / or drugs.

[0073] The health food provided by the present application includes the preparation as described above or the preparation prepared by the method as described above.

[0074] The health food provided by the present application also includes a health food acceptable auxiliary material. The health food acceptable auxiliary material refers to the health food acceptable auxiliary material and its use regulation in the health food record product, and the present application does not limit this.

[0075] The cosmetics provided by the present application include the preparation as described above or the preparation prepared by the method as described above.

[0076] The cosmetic provided by the present application further comprises a cosmetic base. The cosmetic provided by the present application comprises a cleaning cosmetic, a care cosmetic, and a beauty / cosmetic cosmetic. The skin-applicable cleaning cosmetic comprises facial cleanser, makeup remover, cleansing cream, facial mask, floral water, itch powder, body powder, or bath liquid. The skin-applicable care cosmetic comprises skin cream, skin lotion, or skin water. The skin-applicable beauty / cosmetic cosmetic comprises powder, rouge, eye shadow, eyeliner, eyebrow pencil, perfume, or cologne. The hair-applicable cleaning cosmetic comprises shampoo, hair cream, or shaving cream. The hair-applicable care cosmetic comprises hair conditioner, hair cream, hair oil / wax, or hair pomade. The hair-applicable beauty / cosmetic cosmetic comprises hair styling mousse / hair gel, hair dye, hair relaxer, mascara, hair tonic, or hair removal agent. The nail-applicable cleaning cosmetic comprises nail polish remover. The nail-applicable care cosmetic comprises nail cream, nail hardener. The nail-applicable beauty / cosmetic cosmetic comprises nail polish. The lip-applicable cleaning cosmetic comprises lip makeup remover. The lip-applicable care cosmetic comprises lip balm. The lip-applicable beauty / cosmetic cosmetic comprises lipstick, lip gloss, or lip liner.

[0077] The medicine provided by the present application comprises the preparation as described above or the preparation prepared by the method as described above. The medicine further comprises pharmaceutically acceptable excipients.

[0078] In the present application, the pharmaceutically acceptable excipients are selected according to the needs of the dosage form. The dosage form of the medicine or vaccine provided by the present application is oral preparation, inhalant, or injection. Alternatively, the medicine or vaccine is oral preparation, for example, tablet, pill, oral liquid, capsule, syrup, drop pill, or granule. In some embodiments provided by the present application, the capsule is hard capsule or soft capsule. In some embodiments provided by the present application, the tablet is oral tablet or oral cavity tablet.

[0079] The oral tablet refers to tablet for oral administration. The medicine in most of such tablets is absorbed through the gastrointestinal tract to exert effect. The medicine in some tablets exerts effect locally in the gastrointestinal tract. In some embodiments provided by the present application, the oral tablet is ordinary compressed tablet, dispersible tablet, effervescent tablet, chewable tablet, coated tablet, or controlled-release tablet. The medicine is inhalant, which is alternatively inhalation aerosol, inhalation powder aerosol, and liquid preparation for use in a nebulizer. The medicine is injection, for example, injection liquid or injection powder.

[0080] This invention provides a fat body formulation encapsulating fat-soluble nutrients, its preparation method, and its applications. The fat bodies constructed by this invention encapsulate fat-soluble nutrients, which can improve the solubility, stability, and bioavailability of the fat-soluble nutrients, and the effect is superior to that of free EPA. Moreover, compared with fat bodies constructed under some inappropriate parameters, the fat bodies provided by this invention can more effectively improve stability and bioavailability, and have a good encapsulation effect. Attached Figure Description

[0081] Figure 1 shows the preparation of eicosapentaenoic acid ethyl ester fat bodies under the first group of lipid conditions, wherein:

[0082] Photograph A shows the ethyl eicosapentaenoic acid (EPA) fat bodies prepared under the first lipid conditions;

[0083] B indicates the particle size of EPA fat bodies prepared under the first group of lipid conditions;

[0084] C represents the PDI of EPA fat bodies prepared under the first lipid conditions;

[0085] D shows the HPLC peak diagram of EPA fat bodies prepared under the first lipid conditions;

[0086] E represents the concentration of EPA in EPA fat bodies prepared under the first lipid conditions;

[0087] F indicates the raw material utilization rate of EPA in EPA fat bodies prepared under the first group of lipid conditions;

[0088] G represents EPA fat bodies prepared under the first lipid conditions;

[0089] H shows the morphology and structure of EPA fat bodies prepared under the first lipid conditions;

[0090] Figure 2 shows the preparation of eicosapentaenoic acid ethyl ester fat bodies under the second group of lipid conditions, wherein:

[0091] Photograph A shows ethyl eicosapentaenoic acid (EPA) fat bodies prepared under the second lipid conditions;

[0092] B indicates the particle size of EPA fat bodies prepared under the second group of lipid conditions;

[0093] C represents the PDI of EPA fat bodies prepared under the second lipid condition;

[0094] D indicates the concentration of EPA in EPA-containing fat bodies prepared under the second lipid conditions;

[0095] E indicates the raw material utilization rate of EPA in EPA fat bodies prepared under the second lipid conditions;

[0096] F shows the TLC lipid analysis of EPA in the EPA fatty body prepared under the third set of lipid conditions;

[0097] Figure 3 shows the preparation of eicosapentaenoic acid ethyl ester fatty body under the third set of lipid conditions, wherein:

[0098] A shows the photograph of eicosapentaenoic acid ethyl ester fatty body (EPA fatty body) prepared under the third set of lipid conditions;

[0099] B shows the particle size of EPA fatty body prepared under the third set of lipid conditions;

[0100] C shows the PDI of EPA fatty body prepared under the third set of lipid conditions;

[0101] D shows the concentration of EPA in the EPA fatty body prepared under the third set of lipid conditions;

[0102] E shows the raw material utilization rate of EPA in the EPA fatty body prepared under the third set of lipid conditions;

[0103] F shows the TLC lipid analysis of EPA in the EPA fatty body prepared under the third set of lipid conditions;

[0104] Figure 4 shows the preparation of eicosapentaenoic acid ethyl ester fatty body under the fourth set of lipid conditions, wherein:

[0105] A shows the photograph of eicosapentaenoic acid ethyl ester fatty body (EPA fatty body) prepared under the fourth set of lipid conditions;

[0106] B shows the particle size of EPA fatty body prepared under the fourth set of lipid conditions;

[0107] C shows the PDI of EPA fatty body prepared under the fourth set of lipid conditions;

[0108] D shows the concentration of EPA in the EPA fatty body prepared under the fourth set of lipid conditions;

[0109] Figure 5 shows the preparation of eicosapentaenoic acid ethyl ester fatty body under the fifth set of lipid conditions, wherein:

[0110] A shows the photograph of eicosapentaenoic acid ethyl ester fatty body (EPA fatty body) prepared under the fifth set of lipid conditions;

[0111] B shows the concentration of EPA in the EPA fatty body prepared under the fifth set of lipid conditions;

[0112] C shows the change in particle size of EPA fatty body prepared under the fifth set of lipid conditions after being placed for different weeks;

[0113] D shows the change in PDI of EPA fatty body prepared under the fifth set of lipid conditions after being placed for different weeks;

[0114] Figure 6 shows the preparation of ethyl eicosapentaenoate fat bodies under the sixth set of lipid conditions, wherein:

[0115] A shows the photos of ethyl eicosapentaenoate fat bodies (EPA fat bodies) prepared under the sixth set of lipid conditions;

[0116] B shows the concentration of EPA in the EPA fat bodies prepared under the sixth set of lipid conditions;

[0117] C shows the change in particle size of the EPA fat bodies prepared under the sixth set of lipid conditions after being placed for different weeks;

[0118] D shows the change in PDI of the EPA fat bodies prepared under the sixth set of lipid conditions after being placed for different weeks;

[0119] Figure 7 shows that the bioavailability of ethyl eicosapentaenoate fat bodies is significantly improved, wherein:

[0120] A shows that the following samples were selected for gavage to mice: corn oil dissolved EPA (fat bodies not encapsulated, EPA group), sample 1-1 EPA fat bodies of the fifth set (1-1 group), sample 4-1 EPA fat bodies of the fifth set (4-1 group), sample P1-1 EPA fat bodies of the sixth set (P1-1 group), sample P3-1 EPA fat bodies of the sixth set (P3-1 group), and the concentration of EPA in the plasma of each mouse was detected by ELISA kit after gavage for 2 hours;

[0121] B shows the increase rate of the concentration of EPA in the plasma of mice in the ethyl eicosapentaenoate fat body treatment group relative to the EPA group without fat body encapsulation;

[0122] Figure 8 shows the preparation of vitamin A palmitate fat bodies, wherein:

[0123] A shows the optical microscope photos of the prepared vitamin A palmitate fat bodies (VA fat bodies) (a-c), the scale is 5 microns, and the electron microscope photo of the prepared VA fat bodies (d), the scale is 200 nanometers;

[0124] B shows the particle size and PDI of the prepared VA fat bodies;

[0125] C shows the HPLC peak chart of the prepared VA fat bodies;

[0126] Figure 9 shows the preparation of lutein fat bodies, wherein:

[0127] A shows the photos of the prepared lutein-containing fat bodies (Lu fat bodies);

[0128] B shows the change in particle size of the prepared Lu fat bodies after being placed for different days;

[0129] C shows PDI change of Lu fat bodies prepared after different days of placement;

[0130] D shows HPLC peak profile of Lu fat bodies prepared;

[0131] E shows concentration of lutein in Lu fat bodies prepared;

[0132] F shows optical microscope photo of Lu fat bodies prepared, scale bar is 5 microns;

[0133] Figure 10 shows ginsenoside fat bodies prepared, wherein:

[0134] A shows photo of ginsenoside-containing fat bodies (Gi fat bodies) prepared;

[0135] B shows particle size of Gi fat bodies prepared;

[0136] C shows PDI of Gi fat bodies prepared;

[0137] D shows HPLC peak profile of Gi fat bodies prepared;

[0138] E shows concentration of ginsenosides in Gi fat bodies prepared;

[0139] F shows optical microscope photo of Gi fat bodies prepared, scale bar is 2 microns;

[0140] Figure 11 shows berberine fat bodies prepared, wherein:

[0141] A shows photo of berberine-containing fat bodies (Be fat bodies) prepared;

[0142] B shows particle size of Be fat bodies prepared;

[0143] C shows PDI of Be fat bodies prepared;

[0144] D shows HPLC peak profile of Be fat bodies prepared;

[0145] E shows concentration of berberine in Be fat bodies prepared;

[0146] F shows optical microscope photo of Be fat bodies prepared, scale bar is 5 microns. DETAILED DESCRIPTION

[0147] The present application provides a fat body preparation for encapsulating fat-soluble nutritional ingredients, its preparation method and application. Those skilled in the art can refer to the content herein, and appropriately improve the process parameters to achieve. It is particularly pointed out that all similar substitutions and changes are obvious to those skilled in the art, and they are considered to be included in the present application. The method and application of the present application have been described by preferred embodiments, and relevant personnel can obviously modify or appropriately change and combine the method and application herein without departing from the content, spirit and scope of the present application, to realize and apply the present application technology.

[0148] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. For the purposes of the present application, the phrase "art" shall mean the collective term of art and science, including the techniques and methodologies described herein. With respect to the definitions and terms used in the art, the skilled artisan can refer to Current Protocols in Molecular Biology (Ausubel). The abbreviations for amino acid residues are the standard three letter and / or one letter codes used in the art to designate one of the 20 commonly occurring L-amino acids.

[0149] In this application, the term "and / or", describing the association relationship of the associated objects, means that there can be three kinds of relationships, for example, A and / or B, which can represent: the case of A alone, the case of A and B existing at the same time, and the case of B alone. Wherein A, B can be singular or plural.

[0150] The terms "comprise", "contain" and "have" are used interchangeably in this document, and are intended to mean the inclusivity of the scheme, meaning that the scheme can have other elements in addition to the listed elements. It should also be understood that the use of "comprise", "contain" and "have" in this document also provides a "consisting of" scheme.

[0151] In this application, "at least one" means one or more, and "multiple" means two or more. "At least one of the following" or similar expressions means any combination of these items, including any combination of single item or multiple items.

[0152] The term "drug" herein refers to a preparation which exists in a form effective for the biological activity of the active ingredient contained therein, and does not contain additional components which have unacceptable toxicity to the subject to which the pharmaceutical composition is administered.

[0153] The term "treatment" refers to a surgical or therapeutic treatment whose purpose is to prevent, slow down (reduce) or halt an unwanted physiological change or pathology, such as cancer and tumors, in a subject. Beneficial or desired results include, but are not limited to, alleviation of symptoms, diminishment of extent of disease, stabilized (i.e., not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, and remission (whether partial or total), whether detectable or undetectable. A subject in need of treatment includes a subject who has a disorder or disease, a subject who is predisposed to a disorder or disease, or a subject who is intended to prevent a disorder or disease. When referring to terms such as slow down, reduce, diminish, palliate, alleviate, etc., the meaning also includes elimination, disappearance, non-occurrence, etc.

[0154] The term "subject" herein refers to an organism that receives treatment for a particular disease or disorder as described herein. Illustratively, a "subject" includes a mammal, such as a human, a primate (e.g., a monkey) or a non-primate mammal, that receives treatment for a disease or disorder.

[0155] The term "subject" herein refers to an organism that receives treatment for a particular disease or disorder as described herein. Illustratively, a "subject" includes a mammal, such as a human, a primate (e.g., a monkey) or a non-primate mammal, that receives treatment for a disease or disorder.

[0156] The term "effective amount" refers to that amount of a therapeutic agent, alone or in combination with another therapeutic agent, which is effective in preventing or alleviating a disease condition or the progression of the disease when administered to a cell, tissue, or subject. An "effective amount" also refers to the amount of a compound which is sufficient to effect a reduction in a symptom, e.g., to treat, cure, prevent, or alleviate an associated medical condition, or to increase the rate of treatment, cure, prevention, or alleviation of such conditions. When the active ingredient is administered individually to an individual, the therapeutically effective dose refers to that ingredient alone. When a combination is used, the therapeutically effective dose refers to the combined amounts of the active ingredients that produce the therapeutic effect, whether administered in combination, consecutively, or simultaneously.

[0157] The term "IC50" herein refers to the half maximal inhibitory concentration of an antagonist being measured. It is understood that a certain drug induces 50% of tumor cell death at a certain concentration, and this concentration is called the 50% inhibitory concentration, i.e., the concentration corresponding to the ratio of dead cells to the total number of cells equal to 50%. The IC50 value can be used to measure the ability of a drug to induce death, i.e., the stronger the inducing ability, the lower the value.

[0158] The term "logP" value herein refers to the logarithmic value of the partition coefficient ratio of a certain substance in n-octane (oil) and water, reflecting the distribution of the substance in oil and water phases. The larger the logP value, the more lipophilic and the stronger the hydrophobicity of the substance. The logP values of these fat-soluble nutrients are from the ChemSpider website: https: / / www.chemspider.com / Default.aspx. The solubility values of these fat-soluble nutrients in water are from the DrugBank website: https: / / go.drugbank.com / drugs / DB08887.

[0159] In this paper, the particle size refers to the diameter of the particles or molecules of an object, and the average particle size refers to the diameter of a hypothetical particle group composed of uniform spherical particles, compared with an actual particle group composed of particles of different sizes and shapes, if the total length of the particle size of the two is the same, the diameter of the spherical particle is called the average particle size of the actual particle group. In the examples, the average particle size is the linear average diameter, which measures the diameter or length of each object in linear scale, and then the average of these measurements is obtained.

[0160] In this paper, the polydispersity index (PDI) is also known as molecular weight distribution coefficient, non-uniformity index, dispersity, etc. PDI is mainly used to describe the uniformity of particle or molecular size distribution. The larger the pdi, the wider the molecular weight distribution; the smaller the pdi, the more uniform the molecular weight distribution.

[0161] In this paper, the raw material utilization rate refers to the percentage of the total consumption of raw materials consumed in the preparation of the fat body containing fat-soluble nutrients, and the calculation formula is: raw material utilization rate = the amount of raw materials contained in the product / the total consumption of raw materials for producing the product * 100%

[0162] It should be understood that in various embodiments of the present application, the size of the sequence number of the above-mentioned processes does not mean the order of execution, and part or all of the steps can be executed in parallel or in sequence, and the execution order of each process should be determined according to its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0163] The test materials used in the present application are all ordinary commercially available products, which can be purchased in the market.

[0164] The fragment names and sequences involved in this paper are as follows:

[0165] Table 1 Name and amino acid sequence of peptide segment targeting and recognizing single-molecule phospholipid membrane

[0166] Table 2 Name and amino acid sequence of targeting molecule and targeted protein

[0167] Table 3 logP values of fat body encapsulated fat-soluble nutritional ingredients

[0168] The application is further illustrated below with reference to examples.

[0169] Example 1: Construction of fat body containing eicosapentaenoic acid ethyl ester (EPA fat body)

[0170] Eicosapentaenoic acid ethyl ester (EPA) is the ethyl ester form of eicosapentaenoic acid, belongs to unsaturated fatty acid ester, belongs to Omega-3 fatty acid. It has many important physiological functions and effects in the human body: 1. Improving cardiovascular health, 2. Anti-inflammatory effect, 3. Improving mood and cognitive function. In order to improve the solubility, stability and bioavailability of EPA, we prepared a fat body containing eicosapentaenoic acid ethyl ester (EPA fat body). The solubility of EPA in water is only 0.1 μg / mL, while the solubility of EPA in EPA fat body can reach 40 mg / ml, which is 400,000 times that of its aqueous phase. In addition, the stability of EPA fat body is good, and it significantly increases the bioavailability of EPA. The specific experimental procedures and results are as follows:

[0171] I. Construction of EPA fat body

[0172] 1) Neutral lipid condition: triolein (TAG) or soybean oil.

[0173] 2) Polar lipid condition: chloroform was used to prepare egg yolk phosphatidylcholine (Egg PC) at a concentration of 25 mg / ml, glycocholic acid (GA) at a concentration of 5 mg / ml and 25 mg / ml, trimethyl-2,3-dioleoylpropyl ammonium bromide (DOTAP) at a concentration of 25 mg / ml, cholesterol at a concentration of 12.5 mg / ml, and phosphatidylethanolamine-polyethylene glycol 2000 (DSPE-PEG2000) at a concentration of 8.96 mg / ml.

[0174] 3) Preparation of eicosapentaenoic acid ethyl ester containing 5% vitamin E (mass ratio), denoted as EPA-E. Eicosapentaenoic acid ethyl ester without vitamin E is denoted as EPA, and vitamin E is prepared with anhydrous ethanol at a concentration of 200 mg / ml.

[0175] 4) The phospholipid in step 2) was added into a 1.5ml microcentrifuge tube and dried by blowing high purity nitrogen (see Table 4 for the formulation).

[0176] 5) 100μl ddH2O and 5μl neutral lipids containing EPA (see Table 2 for the formulation) were added into the dried microcentrifuge tube in step 4), and ultrasonicated for 5 times (power: 216W, water temperature: 25°C, 3min) to obtain a milky white lipid mixture 1. The lipid mixture 1 was centrifuged at 1,000g for 5min. After centrifugation, the liquid phase system showed two layers of stratification, and the lower milky white solution was collected by suction, vortexed to obtain a milky white lipid mixture 2, which was the final ethyl eicosapentaenoate-carrying fat body (EPA fat body).

[0177] Table 4 Ethyl eicosapentaenoate fat body of different lipid formulations

[0178] II. Characteristic detection of EPA fat body in each group

[0179] The EPA liposomes prepared under the first set of conditions all appeared milky white (Fig. 1A), with the average particle size of the EPA liposomes in the A1 / A5 / A6 samples being about 110 nm, and the particle size of the EPA liposomes in the A2 / A3 / A4 samples being larger, with an average particle size of about 200 nm (Fig. 1B). The PDI (polymer dispersity index) of all the EPA liposomes was no more than 0.3, indicating that the EPA liposomes prepared were uniform (Fig. 1C). The content of eicosapentaenoic acid ethyl ester was detected by high performance liquid chromatography (HPLC) (Fig. 1D), with the following HPLC detection conditions: the chromatographic column was Agilent Zorbax SB-C18, the mobile phase was acetonitrile: water = 95:5 (v / v), the column temperature was 25°C, the detection wavelength was 220 nm, and the flow rate was 1 ml / min. The concentration of EPA in the EPA liposomes of the A1 / A3 samples reached 15 mg / ml, the concentration of EPA in the EPA liposomes of the A2 / A4 samples reached 10 mg / ml, and the concentration of EPA in the EPA liposomes of the A5 samples was the lowest, at 5 mg / ml (Fig. 1E). The raw material utilization rate results showed that the raw material utilization rate of EPA in the EPA liposomes of the A3 / A4 / A5 samples was higher, at more than 50%, and the raw material utilization rate of EPA in the EPA liposomes of the A1 / A2 samples was about 30% (Fig. 1F). Further, TLC (thin layer chromatography) was used to detect whether the EPA liposomes contained EPA. The TLC detection method was as follows: the above prepared EPA liposomes were added with the same volume of methanol and 2 times the volume of chloroform to extract the lipids, the organic phase was collected and blown dry with nitrogen to obtain total lipids. The total lipids obtained were added with 100 μl of chloroform, 10 μl of which was loaded onto a silica gel plate, developed in a developing agent of n-hexane: diethyl ether: glacial acetic acid (volume ratio of 80:20:1), and then placed in an iodine chamber for color development. The TLC results showed that the EPA liposomes of the A1 / A2 / A3 / A4 / A5 samples all contained EPA (Fig. 1G). Optical microscopy was used to observe the morphological structure of the prepared EPA liposomes, which were all uniform spherical structures and were not contaminated with other membrane impurities (Fig. 1H). The above results show that under the first set of lipid conditions, the EPA liposomes containing eicosapentaenoic acid ethyl ester can be successfully prepared, with differences in particle size, concentration and EPA raw material utilization rate, which can be adjusted according to actual needs.

[0180] The EPA liposomes prepared under the second set of conditions all appeared milky white (Figure 2A), with the average particle size of the EPA liposomes in the B1 / B3 / B4 samples being about 110 nm, and the particle size of the EPA liposomes in the B2 sample being larger, with an average particle size of about 200 nm (Figure 2B). The PDI (polymer dispersity index) of all the EPA liposomes was no more than 0.3, indicating that the EPA liposomes were uniform (Figure 2C). The concentration of EPA in the EPA liposomes of the B1 / B2 / B3 / B4 samples could reach 30 mg / ml (Figure 2D). The raw material utilization rate of EPA in the EPA liposomes of the B1 / B2 / B3 / B4 samples was high, reaching 60% (Figure 2E). The TLC results showed that the EPA liposomes of the B1 / B2 / B3 / B4 samples all contained EPA (Figure 2F). The above results show that the EPA liposomes containing eicosapentaenoic acid ethyl ester can be successfully prepared under the second set of lipid conditions. The solubility of EPA in water is only 0.1 μg / mL, while the solubility of EPA in the EPA liposomes prepared under the lipid conditions is 30 mg / ml, which is 300,000 times that in the aqueous phase.

[0181] The EPA liposomes prepared under the third set of conditions all appeared milky white (Figure 3A), with the average particle size of the EPA liposomes in the C1 / C2 samples being about 110 nm, and the particle size of the EPA liposomes in the C3 sample being larger, with an average particle size of about 150 nm (Figure 3B). The PDI (polymer dispersity index) of all the EPA liposomes was no more than 0.3, indicating that the EPA liposomes were uniform (Figure 3C). The concentration of EPA in the EPA liposomes of the C1 sample reached 30 mg / ml, and the concentration of EPA in the EPA liposomes of the C2 / C3 samples reached 20 mg / ml (Figure 3D). The raw material utilization rate of EPA in the EPA liposomes of the C1 / C2 / C3 samples was high, all being about 50% (Figure 3E). The TLC results showed that the EPA liposomes of the C1 / C2 / C3 samples all contained EPA (Figure 3F). The above results show that the EPA liposomes containing eicosapentaenoic acid ethyl ester can be successfully prepared under the third set of lipid conditions, with differences in particle size, concentration and EPA raw material utilization rate, which can be adjusted according to actual needs.

[0182] The EPA liposomes prepared under the fourth group of conditions were all milky white (Figure 4A). The average particle size of the EPA liposomes in samples A / B / 2 was around 150 nm, while the particle size of the EPA liposomes in samples 1 / 4 was larger, with an average particle size of around 220 nm. Sample 3 had the largest particle size, reaching 400 nm (Figure 4B). Except for sample B, whose PDI (Polymer Dispersibility Index) was around 0.4, the PDI of the EPA liposomes in the other samples did not exceed 0.3, indicating that the prepared EPA liposomes were homogeneous (Figure 4C). The EPA concentration in the EPA liposomes of samples 1 / 2 / 3 reached 30 mg / ml, the EPA concentration in sample 4 reached 20 mg / ml, while the EPA concentration in samples A / B was lower, less than 10 mg / ml (Figure 4D). These results indicate that liposomes containing ethyl eicosapentaenoic acid (EPA) can be successfully prepared under the fourth group of lipid conditions. The particle size, concentration, and EPA raw material utilization rate vary, and adjustments can be made according to actual needs.

[0183] The EPA liposomes prepared under the fifth group of conditions were all milky white and remained so after 5 weeks of storage at 4°C, indicating good solution stability (Figure 5A). The EPA concentration in samples 3-1 / 4-1 / 4-2 reached 30 mg / ml, while the concentration in samples 1-1 / 1-2 / 3-2 / 3-3 / 4-3 reached 20 mg / ml. Sample 1-3 showed a lower EPA concentration, around 15 mg / ml (Figure 5B). Furthermore, the average particle size of these EPA liposomes was below 250 nm, the PDI (polymer dispersibility index) did not exceed 0.3, and they were stable for 7 weeks (Figure 5C and D). These results indicate that liposomes containing ethyl eicosapentaenoic acid (EPA) can be successfully prepared under the fifth group of lipid conditions, and the EPA liposomes are stable for 7 weeks. Other groups also exhibited good stability.

[0184] The EPA liposomes prepared under the sixth group of conditions were all milky white and remained milky white after being stored at 4°C for 5 weeks, indicating good solution condition (Figure 6A). The EPA concentration in the EPA liposomes of samples P1-1 / P2-1 / P3-1 / P3-2 / P4-1 / P3-2 was between 30-40 mg / ml, while the concentration in the remaining EPA liposomes was between 20-30 mg / ml (Figure 6B). Furthermore, the average particle size of these EPA liposomes was below 300 nm, the PDI (polymer dispersibility index) did not exceed 0.3, and they were stable for 7 weeks (Figure 6C and D). These results indicate that liposomes containing ethyl eicosapentaenoic acid (EPA) can be successfully prepared under the sixth group of lipid conditions, and the EPA liposomes are stable for 7 weeks.

[0185] III. Bioavailability of EPA-containing fat bodies

[0186] To further compare the bioavailability of EPA after encapsulation in fat bodies and EPA without encapsulation, the following samples were selected for gavage in mice: EPA dissolved in corn oil (fat body not encapsulated, EPA group), 1-1 sample EPA fat body of the fifth group (1-1 group), 4-1 sample EPA fat body of the fifth group (4-1 group), P1-1 sample EPA fat body of the sixth group (P1-1 group), P3-1 sample EPA fat body of the sixth group (P3-1 group). The concentration and volume of EPA gavaged in each group of mice were the same, all 35.6 mg / ml, and the plasma of mice was taken 2 hours after gavage, and the concentration of EPA in the plasma of each mouse was detected by ELISA kit. The results showed that the concentration of EPA in the plasma of mice treated with EPA-containing fat bodies was increased, and the concentration of EPA in the plasma of mice treated with 4-1 group, P1-1 group, P3-1 group was significantly higher than that of EPA group without fat body encapsulation (EPA group) (Figure 7A). At the same time, compared with the group treated with EPA without fat body encapsulation, the concentration of EPA in the plasma of mice in 1-1 group, 4-1 group, P1-1 group, P3-1 group was increased by 6.3%, 58.7%, 43.5%, 38.9% respectively (Figure 7B). The above results show that EPA-containing fat bodies can significantly improve the bioavailability of EPA.

[0187] Example 2: Construction of fat body containing retinol (vitamin A) palmitate (VA fat body)

[0188] Retinol palmitate is a derivative of vitamin A, which is a storage form of vitamin A, mainly exists in animal food and supplements. Retinol palmitate plays an important role in maintaining normal vision, promoting cell growth and immune regulation, and is widely used in beauty and skin care and treatment of skin diseases. In order to improve its stability and solubility, we prepared a fat body containing retinol (vitamin A) palmitate (VA fat body). The solubility of retinol palmitate in aqueous phase is only 7.58 μg / mL, while the solubility of retinol palmitate in VA fat body can reach 400 μg / mL, which is 52 times that in aqueous phase. The following is the specific experimental process and results:

[0189] 1) Preparation of neutral fat: triolein (TAG) and retinol (vitamin A) palmitate are prepared according to the volume ratio of 4:1 (the ratio can be adjusted).

[0190] 2) Take 80 μl 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (containing 2 mg DOPC) and add it to a microcentrifuge tube, and dry the solvent with high-purity nitrogen.

[0191] 3) Add 100 μΐ PBS and 5 μΐ neutral lipid prepared in step 1) into a microcentrifuge tube, vortex for 4 min (vortex for 10 s, stop for 10 s, vortex condition is 4000 rpm), get a milky white lipid mixture 1, centrifuge the lipid mixture 1 at 1000 g for 5 min, after centrifugation, the liquid phase system presents two layers of stratification, the lower layer milky white solution is collected by suction, vortex, get a milky white lipid mixture 2.

[0192] 4) Centrifuge the lipid mixture 2 obtained in step 3) at 20000 g for 5 min. After centrifugation, remove the precipitate component at the bottom of the microcentrifuge tube, vortex, get a milky white lipid mixture 3, which is the final fat body carrying retinol (vitamin A) palmitate (VA fat body).

[0193] The morphological structure of the constructed VA fat body was observed by optical microscope and electron microscope, which was a uniform spherical structure and had no other membrane impurity pollution (Figure 8 A). In addition, the average particle size of the VA fat body was detected by dynamic light scattering instrument, which was 146.5 nm, and the PDI (polymer dispersity index) was 0.099 (Figure 8 B). Further, the concentration of retinol (vitamin A) palmitate in the VA fat body was detected by HPLC (high performance liquid chromatography) (Figure 8 C). The detection conditions of HPLC were as follows: the chromatographic column was Agilent Zorbax SB-C18, the mobile phase was methanol: water = 92.5:2.5 (volume ratio), the column temperature was 25℃, the detection wavelength was 313 nm, and the flow rate was 1.5 ml / min. The concentration of vitamin A palmitate in the VA fat body was 0.4 mg / ml. The above results showed that the fat body containing vitamin A palmitate was successfully prepared, and it had high purity and good uniformity. In addition, the detection results showed that the bioavailability and stability of the fat body prepared in this example were not lower than the effect of example 1.

[0194] Example 3: Construction of fat body containing lutein (Lu fat body)

[0195] Lutein is a natural pigment belonging to carotenoids, commonly found in many plants and microorganisms. Lutein is an important natural antioxidant with various health benefits, including protection of eye, cardiovascular and skin health, anti-cancer and anti-inflammatory effects. In order to improve its stability and solubility, we prepared a fat body containing lutein (Lu fat body). The solubility of lutein in aqueous phase is only 0.73 μg / mL, while the solubility of lutein in Lu fat body can reach 15 μg / mL, which is 20 times that of aqueous phase. The following is the specific experimental procedure and results:

[0196] 1) Neutral lipid condition: soybean oil, castor oil, triolein, mixed oil (soybean oil: castor oil: triolein = 1:1:1 by volume).

[0197] 2) Formulation of neutral lipids containing lutein: 1 mg lutein was added to 1 ml of the above neutral lipids, respectively, and heated at 55°C for 10 min, then sonicated twice (power: 216 W, water temperature: 25°C, 3 min), and then centrifuged at 20,000 g for 5 min. After centrifugation, the supernatant was taken, which was the neutral lipid containing lutein, namely lutein-soybean oil, lutein-castor oil, lutein-triolein, and lutein-mixed oil, respectively.

[0198] 3) Polar lipid condition: chloroform was used to formulate egg yolk phosphatidylcholine (Egg PC) at a concentration of 25 mg / ml.

[0199] 4) The phospholipids in step 2) were added to 1.5 ml microcentrifuge tubes and blown dry with high-purity nitrogen (see Table 3 for ingredients).

[0200] 5) 100 μl ddH2O and 5 μl of the lutein-containing neutral lipid in step 2) were added to the dried microcentrifuge tube in step 3) (see Table 3 for ingredients), and sonicated 5 times (power: 216 W, water temperature: 25°C, 3 min) to obtain a milky white lipid mixture 1. The lipid mixture 1 was centrifuged at 1,000 g for 5 min, and after centrifugation, the liquid phase system showed two layers of stratification. The lower layer of milky white solution was collected by suction, vortexed, and a milky white lipid mixture 2 was obtained, which was the final lutein-containing fat body (Lu fat body).

[0201] Table 5 Lutein fat bodies of different lipid formulations

[0202] Lutein has very low solubility in water, so it appears turbid precipitate, while lutein liposomes prepared under different neutral lipid conditions all present a milky yellow state (Fig. 9A), in which the average particle size of Lu liposomes in E1 / E2 / E3 samples is about 150 nm, while the particle size of Lu liposomes in E4 / E5 samples is larger, with an average particle size of about 250 nm (Fig. 9B), and the PDI (polymer dispersity index) of all Lu liposomes does not exceed 0.3, indicating that the prepared Lu liposomes are uniform and stable after being placed at room temperature for 5 days (Fig. 9C). The lutein content was detected by high performance liquid chromatography (HPLC) (Fig. 9D), and the HPLC detection conditions were as follows: the chromatographic column was Agilent Zorbax SB-C18, the mobile phase was methanol: acetonitrile = 10:90 (v / v), the column temperature was 25°C, the detection wavelength was 446 nm, and the flow rate was 1 ml / min. Among them, the concentration of lutein in Lu liposomes of E4 / E3 samples reached 15 μg / mL, and the concentration of lutein in Lu liposomes of E1 / E2 samples was 10 μg / mL, and the concentration was basically unchanged after being placed at room temperature for 5 days (Fig. 9E). The morphology of the constructed Lu liposomes was observed by optical microscope, which was uniform spherical structure and had no contamination of other membrane impurities (Fig. 9F). The above results show that lutein-containing liposomes are successfully prepared. The detection results show that the stability of the liposomes prepared in this example is not lower than that of Example 1.

[0203] Example 4: Construction of Ginsenoside-containing Liposomes (Gi Liposomes)

[0204] Ginsenoside is an active ingredient in ginseng, mainly present in the roots of ginseng. Ginsenoside has a variety of important functions and effects, including improving immune function, anti-fatigue, improving cognitive function, regulating blood sugar levels, protecting cardiovascular health, and anti-tumor activity. In order to improve its stability and solubility, we prepared ginsenoside-containing liposomes (Gi liposomes). The solubility of ginsenoside in aqueous phase is only 0.18 μg / mL, while the solubility of ginsenoside in Gi liposomes can reach 800 μg / mL, which is 4,444 times that of aqueous phase. The following is the specific experimental procedure and results:

[0205] 1) Neutral lipid conditions: soybean oil, fish oil, trioctanoin, mixed oil (soybean oil: fish oil: trioctanoin = 1:1:1 by volume).

[0206] 2) Formulation of neutral lipids containing ginsenosides: 5 mg of ginsenosides was added into 1 ml of absolute ethanol, 500 μl of ginsenosides prepared above was added into 500 μl of neutral lipids, respectively, and heated at 55 °C for 10 min, then sonicated twice (power: 216 W, water temperature: 25 °C, 3 min), and then removed ethanol by nitrogen blowing, and centrifuged at 20,000 g for 5 min. After centrifugation, the supernatant was collected, which was neutral lipids containing ginsenosides, and was ginsenosides-soybean oil, ginsenosides-fish oil, ginsenosides-tricaprylin, and ginsenosides-mixed oil, respectively.

[0207] 3) Polar lipids condition: chloroform was used to prepare egg yolk phosphatidylcholine (Egg PC) at a concentration of 25 mg / ml.

[0208] 4) The phospholipids in step 2) were added into 1.5 ml microcentrifuge tubes, and dried by high-purity nitrogen blowing (see Table 4 for ingredients).

[0209] 5) 100 μl of ddH2O and 5 μl of neutral lipids containing ginsenosides in step 2) were added into the dried microcentrifuge tubes in step 3) (see Table 3 for ingredients), and sonicated 5 times (power: 216 W, water temperature: 25 °C, 3 min) to obtain a milky white lipid mixture 1, and the sonication instrument was SBL-10DT. The lipid mixture 1 was centrifuged at 1,000 g for 5 min, and after centrifugation, the liquid phase system showed two layers of stratification, and the lower layer of milky white solution was collected by suction, and vortexed to obtain a milky white lipid mixture 2, which was the final ginsenoside-loaded fat body (Gi fat body).

[0210] Table 6 Ginsenoside fat bodies of different lipid formulations

[0211] The solubility of ginsenosides in water is very low, so there is a turbid precipitate, and the ginsenoside fat bodies prepared under different neutral lipid conditions all show a milky white state (Fig. 10A). The average particle size of the Gi fat body in the G1 / G2 / G3 / G4 sample is about 150 nm (Fig. 10B), and the PDI (polymer dispersity index) of all Gi fat bodies is not more than 0.3, indicating that the prepared Gi fat bodies are uniform (Fig. 10C). The ginsenoside content was detected by high performance liquid chromatography (HPLC) (Fig. 10D). The HPLC detection conditions are as follows: the chromatographic column is Agilent Zorbax SB-C18, the mobile phase is methanol: acetonitrile = 70:30 (v / v), the column temperature is 25°C, the detection wavelength is 203 nm, and the flow rate is 0.5 ml / min. Among them, the concentration of ginsenosides in the Gi fat body of the G3 sample is the highest, reaching 0.8 mg / ml, the concentration of ginsenosides in the Gi fat body of the G1 sample is the second, reaching 0.7 mg / ml, and the concentration of ginsenosides in the Gi fat body of the G2 / G4 sample is lower, less than 0.2 mg / ml (Fig. 10E). The morphological structure of the constructed Gi fat body was observed under an optical microscope, which was a uniform spherical structure and was not contaminated by other membrane impurities (Fig. 10F). The above results show that the ginsenoside-containing fat body is successfully prepared. The detection results show that the bioavailability and stability of the fat body prepared in this example are not lower than the effect of Example 1.

[0212] Example 5: Construction of a fat body containing berberine (Be fat body)

[0213] Berberine, also known as yellow lotus, is the main component of Chinese herbal medicine yellow lotus and also exists in many other plants, and has been used to inhibit the growth of parasites and inhibit fungal infection. In order to improve its stability and solubility, we prepared a fat body containing berberine (Be fat body). The solubility of berberine in water is only 0.35 μg / mL, while the solubility of berberine in Be fat body can reach 90 μg / mL, which is 250 times that of water. The following is the specific experimental procedure and results:

[0214] 1) Neutral lipid conditions: castor oil, medium-chain triglyceride.

[0215] 2) Preparation of ethanol containing berberine: 5 mg of berberine was added to 1 ml of ethanol, heated at 65°C for 10 min, then ultrasonicated twice (power: 216 W, water temperature: 25°C, 3 min), and then centrifuged at 20,000g for 5 min. After centrifugation, the supernatant was taken.

[0216] 3) Polar lipid conditions: egg yolk lecithin (Egg PC) and polyoxyethylene ether castor oil. Neutral lipid conditions: castor oil, fish oil, medium-chain triglyceride.

[0217] 4) The raw materials in step 2) and step 3) were added into 1.5 ml microcentrifuge tubes according to the following ingredient table 7.

[0218] 5) The prepared solution system was ultrasonicated 5 times (power: 216 W, water temperature: 25℃, 3 min) to obtain a milky white lipid mixture 1, and the ultrasonic instrument model was SBL-10DT. The lipid mixture 1 was centrifuged at 500 g for 5 min, and after centrifugation, the liquid phase system showed two layers of stratification, and the lower layer of milky white solution was collected by suction, vortexed, to obtain a milky white lipid mixture 2, which was the final berberine carrying fat body (Be fat body).

[0219] Table 6 Berberine fat bodies of different lipid formulations

[0220] The fat body without adding berberine showed a milky white color (Be1 / Be3 / Be 5), while the fat body prepared under the condition of containing berberine showed a milky yellow state (Be2 / Be4 / Be 6) (Figure 11A). The average particle size of the fat body without containing berberine and the fat body containing berberine was about 100 nm (Figure 11B), and the PDI (polymer dispersity index) was not more than 0.3, indicating that the prepared berberine fat body was uniform (Figure 11C). The berberine content was detected by high performance liquid chromatography (HPLC) (Figure 11D), and the detection conditions of HPLC were as follows: the chromatographic column was Agilent Zorbax SB-C18, the mobile phase was methanol: 0.1% phosphoric acid solution (triethylamine was used to adjust the pH to 6.5) = 50:50 (v / v), the column temperature was 35℃, the detection wavelength was 346 nm, and the flow rate was 1 ml / min. Among them, the concentration of berberine in the berberine fat body of Be2 / Be4 / Be6 samples was 89.6, 96.3, and 99.0 μg / mL, respectively (Figure 11E). The morphology of the constructed Be fat body was observed by optical microscope, which was a uniform spherical structure and was not contaminated by other membrane impurities (Figure 11F). The above results showed that the berberine-containing fat body was successfully prepared.

[0221] The above is only a preferred embodiment of the present application, and it should be noted that for ordinary skilled persons in the art, several improvements and refinements can be made without departing from the principles of the present application, and these improvements and refinements should also be considered as the protection scope of the present application.

Claims

1. A fat body formulation for encapsulating a fat-soluble nutrient comprising: The single-molecule phospholipid film and the fat-soluble nutritional ingredient and the neutral lipid wrapped inside the single-molecule phospholipid film.

2. The fatty body preparation of claim 1, wherein, The fat-soluble nutritional ingredient includes at least one of unsaturated fatty acid, fat-soluble vitamin, steroid hormone and / or fat-soluble extract.

3. The fat body preparation according to claim 2, wherein, The unsaturated fatty acid includes at least one of Omega-3, Omega-6, Omega-9 unsaturated fatty acid, and the unsaturated fatty acid includes at least one of oleic acid, linoleic acid, linolenic acid, eicosapentaenoic acid, docosapentaenoic acid, docosahexaenoic acid, fish oil, snakegourd seed oil, soybean oil, castor oil, corn oil. The fat-soluble vitamin includes at least one of vitamin A, vitamin D, vitamin E or vitamin K. The steroid hormone includes at least one of androgen, estrogen, progestogen, anti-estrogen, anti-progestogen or non-steroidal estrogen. The fat-soluble extract includes at least one of berberine, coenzyme Q10, retinol, ginsenoside, lutein, quercetin, anthocyanin, lycopene, notoginseng saponin, puerarin, chlorophyll, carotenoid, polyphenol or phytosterol.

4. The preparation according to claim 1, characterized in that, The neutral lipid is selected from one or more of glyceryl trioleate, fish oil, castor oil, corn oil, glyceryl trioctanoate, retinol ester, wax ester, sterol ester, stanols ester, sunflower oil, soybean oil, peanut oil, clove oil, simethicone, cinnamon oil, tea oil, liquid paraffin, star anise oil, mixed fatty acid glyceride (stearin), hydrogenated vegetable oil, refined olive oil, refined olive oil, retinol ester and fat-soluble vitamin.

5. The preparation according to claim 4, characterized in that, The neutral lipid further includes an antioxidant selected from at least one of vitamin E, carotenoid, ubiquinone, guaiac resin, gallic acid, butylated hydroxyanisole.

6. The formulation of claim 1, wherein, The mass fraction of the antioxidant in the neutral lipid is 1% to 10%.

7. The formulation of claim 1, wherein, The membrane material of the single-molecule phospholipid film includes one or more of phospholipid or functional polar lipid. The phospholipid is selected from one or more of egg yolk lecithin, glycocholic acid, soybean phospholipid, trimethyl-2,3-dioleoylpropyl ammonium bromide, dioleoyl phosphatidyl ethanolamine, distearoyl phosphatidyl choline, egg yolk lecithin, dipalmitoyl phosphatidyl choline, dipalmitoyl phosphatidic acid, distearoyl phosphatidyl glycerol sodium, dimyristoyl phosphatidyl choline, 1-stearoyl-hydrolyzed phosphatidyl choline, 1,2-dimyristoyl-rac-glycerol-3-methoxypolyethylene glycol 2000, phosphatidyl ethanolamine, phosphatidyl choline, phosphatidyl ethanolamine, phosphatidylinositol, phosphatidyl serine, phosphatidyl glycerol, phosphatidic acid, cardiolipin and sphingomyelin. The functional polar lipid is selected from one or more of cholesterol, polyethylene glycol modified sterol, biotin modified sterol, amino acid modified sterol, polypeptide modified sterol, polysaccharide modified sterol, nucleic acid modified sterol, polyethylene glycol modified phospholipid, biotin modified phospholipid, amino acid modified phospholipid, polypeptide modified phospholipid, polysaccharide modified phospholipid and nucleic acid modified phospholipid.

8. The formulation according to any one of claims 1 to 7, characterized in that, The monomolecular phospholipid film further comprises a cationic lipid selected from one or more of DOTAP, DODAP, DODMA, DOTMA, D-Lin-MC3-DMA.

9. The formulation according to any one of claims 1 to 8, characterized in that, The targeting molecule targets an organ, tissue or cell.

10. The preparation according to claim 9, characterized in that The organ, tissue or cell is from a human or animal body.

11. Formulation according to claim 9 or 10, characterized in that The targeting molecule is: The targeting molecule is embedded in the monomolecular phospholipid film, or is connected to the phospholipid through avidin-biotin interaction, or is connected to a substance that specifically targets the phospholipid, or a combination thereof.

12. The preparation according to claim 9 or 10, characterized in that The targeting molecule is: At least one of LTA-P33, ApoE, BCMA antibody, Nrp-B, Trf-B, LDLR-B, ErbB2-B, CXCR4-B, GRP78-B or Soma-B.

13. Formulation according to any one of claims 9 to 12, characterized in that One or more of the phospholipid, functional polar lipid and cationic lipid in the monomolecular phospholipid film is labeled with biotin or streptavidin.

14. The formulation of claim 1, wherein, The lipid-soluble nutritional ingredient is eicosapentaenoic acid ethyl ester, the monomolecular phospholipid film comprises at least one of egg yolk lecithin, glycocholic acid, trimethyl-2,3-dioleoyloxypropyl ammonium bromide, cholesterol, trimethyl-2,3-dioleoyloxypropyl ammonium bromide and / or phosphatidyl ethanolamine-polyethylene glycol 2000; and the neutral fat is triolein and / or soybean oil.

15. The preparation according to claim 14, wherein, The monomolecular phospholipid film comprises egg yolk lecithin, and eicosapentaenoic acid ethyl ester and triolein are wrapped inside the monomolecular phospholipid film; The monomolecular phospholipid film comprises egg yolk lecithin and glycocholic acid, and eicosapentaenoic acid ethyl ester is wrapped inside the monomolecular phospholipid film; The monomolecular phospholipid film comprises egg yolk lecithin and glycocholic acid, and eicosapentaenoic acid ethyl ester and triolein are wrapped inside the monomolecular phospholipid film; The monomolecular phospholipid film comprises egg yolk lecithin, and eicosapentaenoic acid ethyl ester, vitamin E and soybean oil are wrapped inside the monomolecular phospholipid film; The monomolecular phospholipid film comprises egg yolk lecithin and glycocholic acid, and eicosapentaenoic acid ethyl ester, vitamin E and soybean oil are wrapped inside the monomolecular phospholipid film; The monomolecular phospholipid film comprises egg yolk lecithin, DOTAP, cholesterol and DSPE-PEG2000, and eicosapentaenoic acid ethyl ester, vitamin E and soybean oil are wrapped inside the monomolecular phospholipid film; The monomolecular phospholipid film comprises egg yolk lecithin, DOTAP, cholesterol, DSPE-PEG2000 and glycocholic acid, and eicosapentaenoic acid ethyl ester, vitamin E and soybean oil are wrapped inside the monomolecular phospholipid film.

16. The preparation according to claim 14 or 15, wherein, The volume ratio of eicosapentaenoic acid ethyl ester to neutral fat is (1-5):(1-5); The mass ratio of egg yolk lecithin to glycocholic acid is (10-40):(2-40); The volume ratio of egg yolk lecithin, DOTAP, cholesterol and DSPE-PEG2000 is 300:(30-300):(30-150):(50-300); The volume ratio of the egg yolk lecithin, DOTAP, cholesterol, DSPE-PEG2000 and glycocholic acid is 300:(30-300):(30-150):(50-300):(1-10).

17. The formulation of claim 1, wherein, The fat-soluble nutritional ingredient is vitamin A palmitate, the single-molecule phospholipid membrane comprises DOPC, and the triolein and vitamin A palmitate are wrapped inside the single-molecule phospholipid membrane.

18. The formulation of claim 17, wherein, The volume ratio of the triolein and vitamin A palmitate is (1-10):

1.

19. The formulation of claim 1, wherein, The fat-soluble nutritional ingredient is lutein, the single-molecule phospholipid membrane comprises egg yolk lecithin, and the neutral fat is at least one of soybean oil, castor oil or tricaprylin.

20. The formulation of claim 19, wherein, The neutral fat is soybean oil, castor oil, tricaprylin, or a mixture of soybean oil, castor oil and tricaprylin.

21. The preparation according to claim 20, wherein, The mass ratio of lutein to egg yolk lecithin is (1-10):(1-10); The mass-volume ratio of the lutein to the neutral fat is (0.1-10) mg:(0.1-10) mL; The volume ratio of the soybean oil, castor oil and tricaprylin in the mixture is (1-10):(1-10):(1-10).

22. The formulation of claim 1, wherein, The fat-soluble nutritional ingredient is ginsenoside, the single-molecule phospholipid membrane comprises egg yolk lecithin, and the neutral fat is at least one of soybean oil, fish oil or tricaprylin.

23. The formulation of claim 22, wherein, The neutral fat is soybean oil, castor oil, tricaprylin, or a mixture of soybean oil, castor oil and tricaprylin.

24. The preparation according to claim 23, wherein, The mass ratio of ginsenoside to egg yolk lecithin is (1-50):(1-10); The mass-volume ratio of the ginsenoside to the neutral fat is (0.1-10) mg:(0.1-5) mL; The volume ratio of the soybean oil, castor oil and tricaprylin in the mixture is (1-10):(1-10):(1-10).

25. The formulation of claim 1, wherein, The fat-soluble nutritional ingredient is berberine, the single-molecule phospholipid membrane comprises egg yolk lecithin and / or polyoxyethylene ether castor oil, and the neutral fat is at least one of castor oil, fish oil or medium-chain triglyceride.

26. The formulation of claim 25, wherein, The castor oil, fish oil or medium-chain triglyceride, and the single-molecule phospholipid membrane are a mixture of egg yolk lecithin and polyoxyethylene ether castor oil.

27. The preparation according to claim 25 or 26, wherein, The mass ratio of berberine to castor oil is (0.001-0.01):(1-10); The mass ratio of egg yolk lecithin to polyoxyethylene ether castor oil in the mixture is (1-10):(1-10).

28. A method for preparing the preparation according to any one of claims 1-27, comprising: mixing the fat-soluble nutritional ingredient and the neutral fat, mixing with the medium and the membrane material of the single-molecule phospholipid membrane, performing ultrasonic treatment and centrifugation, taking the lower-layer milky-white solution, performing vortex treatment to obtain the fat body preparation for loading the fat-soluble nutritional ingredient. or the fat-soluble nutritional ingredient is mixed with the medium and the membrane material of the monomolecular phospholipid film, and after repeated vortexing, a fat body preparation containing the fat-soluble nutritional ingredient is obtained; the medium is water or a buffer solution.

29. The method of claim 28, wherein, the buffer solution is a PBS buffer solution, a HEPES buffer solution, a sucrose solution, a NaCl solution, a KCl solution or a MgCl2 solution.

30. The method of claim 28, wherein, the parameters of the ultrasonic treatment include: power: 216 W, water temperature: 25℃, 3 min, ultrasonic treatment for three times.

31. The method of claim 28, wherein, the repeated vortexing and centrifugation include: after mixing, vortexing is performed to obtain a mixture 1, the mixture 1 is centrifuged to collect the lower solution, and vortexing is performed again to obtain a mixture 2; the mixture 2 is centrifuged to remove the precipitate, and vortexing is performed again to obtain a mixture 3; the mixture 3 is centrifuged to collect the lower solution, and vortexing is performed again to obtain a mixture 4 containing the fat body.

32. Use of the preparation of any one of claims 1-28 or the preparation obtained by the method of any one of claims 29-32 in the preparation of health food, cosmetics and / or drugs.

33. Health food, cosmetics and / or drugs comprising the preparation of any one of claims 1-8 or the preparation obtained by the method of any one of claims 29-32.

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