Neogambogic acid adiposome, preparation method therefor, and use thereof

Abstract

A neogambogic acid adiposome formulation, a preparation method therefor, and use thereof. The neogambogic acid adiposome formulation comprises a monomolecular phospholipid membrane, and neogambogic acid and neutral lipid which are wrapped in the monomolecular phospholipid membrane. The adiposome formulation has good bioavailability and safety, thereby improving the anti-tumor effect.

Description

New gambogic acid fat body and its preparation method and application

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on March 5, 2024, with application number 202410251555.7 and invention name “New Garcinia Acid Fat Body and Its Preparation Method and Application”, the entire contents of which are incorporated herein by reference. Technical Field

[0002] The present invention relates to the field of pharmaceutical technology, in particular to a new gambogic acid fat body preparation and a preparation method and application thereof. Background Art

[0003] Neogambogic acid (NGA), a major component of the traditional Chinese medicine Garcinia cambogia, possesses antioxidant, anti-inflammatory, anticancer, and antibacterial properties. A growing body of research indicates that NGA can inhibit the development and progression of breast, lung, prostate, gastric, and head and neck cancers, primarily through inducing apoptosis, autophagy, and cell cycle arrest.

[0004] Neogamoic acid is a hydrophobic small molecule with a logP (oil-water partition coefficient) of 6.3 and an extremely low solubility in water, less than 0.0005 mg / mL. Its hydrophobicity significantly affects its absorption and bioavailability, while also leading to varying degrees of toxic side effects, thus severely limiting its clinical application.

[0005] In order to improve the solubility, efficacy and safety of neogamoic acid, many nanocarrier preparations have been used to encapsulate neogamoic acid, but these methods also have some disadvantages. For example, liposomes have a hydrophilic core, and only the hydrophobic environment between their phospholipid bilayers can be used to carry neogamoic acid, but their encapsulation capacity is low. In addition, the stability of liposomes carrying neogamoic acid is poor, and they are prone to fusion, rupture or drug leakage, which may affect its distribution in the body and drug release. Similarly, the amount of neogamoic acid encapsulated by polymer carriers is also low. In addition, polymers PEG and PLGA do not exist naturally in the body, which will cause the body to produce corresponding antibodies to resist their effects. Solid lipid nanoparticles require high temperature and high pressure steps during the preparation process, which easily destroys the structure of the neogamoic acid molecule.

[0006] The novel nanoparticle liposomes consist of a hydrophobic core composed of neutral lipids and are encapsulated by a monomolecular phospholipid membrane. Similar in structure to naturally occurring lipid droplets and lipoproteins, they can efficiently dissolve and encapsulate hydrophobic small molecules. Furthermore, the components of liposomes are naturally present in the body, resulting in excellent biocompatibility. Furthermore, their preparation is simple and efficient. If liposomes are used to carry neogamonic acid or other hydrophobic drugs, they are expected to enhance drug solubility, improve bioavailability, and enhance efficacy and safety, greatly facilitating treatment for patients.

[0007] Summary of the Invention

[0008] In view of this, the technical problem to be solved by the present invention is to provide a new gambogic acid fat body preparation and its preparation method and application, which has good bioavailability and safety and can carry targeting molecules.

[0009] The new gambogic acid fat body preparation provided by the present invention comprises: a single molecule phospholipid membrane and new gambogic acid and neutral lipid wrapped in the single molecule phospholipid membrane.

[0010] In the present invention, the neutral lipid is selected from one or more of triolein, fish oil, corn oil, tricaprylin, retinol esters, wax esters, sterol esters, sterol esters, castor oil, sunflower oil, soybean oil, peanut oil, clove oil, simethicone, cinnamon oil, tea oil, liquid paraffin, star anise oil, mixed fatty acid glycerides (stearin), hydrogenated vegetable oil, refined olive oil and fat-soluble vitamins.

[0011] In the present invention, the membrane material of the monomolecular phospholipid membrane comprises one or more of phospholipids or functional polar lipids.

[0012] In some embodiments, the phospholipid is selected from one or more of 2-bis-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine (DOPC), egg yolk lecithin, soybean lecithin, dioleoylphosphatidylethanolamine, distearoylphosphatidylcholine, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidic acid, sodium distearoylphosphatidylglycerol, dimyristoylphosphatidylcholine, 1-stearoyl-lysophosphatidylcholine, 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol 2000, phosphatidylethanolamine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, cardiolipin, and sphingomyelin.

[0013] In some embodiments, the functional polar lipid is selected from one or more of polyethylene glycol-modified sterols, biotin-modified sterols, amino acid-modified sterols, polypeptide-modified sterols, polysaccharide-modified sterols, nucleic acid-modified sterols, polyethylene glycol-modified phospholipids, biotin-modified phospholipids, amino acid-modified phospholipids, polypeptide-modified phospholipids, polysaccharide-modified phospholipids and nucleic acid-modified phospholipids.

[0014] In order to better encapsulate neogambogic acid, the present invention optimizes and screens the membrane materials of the single-molecule phospholipid membrane.

[0015] In some embodiments, the phospholipid is selected from egg yolk lecithin, soybean lecithin, or DOPC. The neutral lipid is selected from triolein, sunflower oil, or soybean oil.

[0016] In some specific embodiments, the membrane material of the monomolecular phospholipid membrane is egg yolk lecithin, and the neutral lipid is triolein.

[0017] Alternatively, the membrane material of the monomolecular phospholipid membrane is egg yolk lecithin, and the neutral lipid is sunflower oil.

[0018] Alternatively, the membrane material of the monomolecular phospholipid membrane is egg yolk lecithin, and the neutral lipid is soybean oil.

[0019] Alternatively, the membrane material of the monomolecular phospholipid membrane is soybean lecithin, and the neutral lipid is triolein.

[0020] Alternatively, the membrane material of the monomolecular phospholipid membrane is soybean lecithin, and the neutral lipid is sunflower oil.

[0021] Alternatively, the membrane material of the monomolecular phospholipid membrane is soybean lecithin, and the neutral lipid is soybean oil.

[0022] Alternatively, the membrane material of the monomolecular phospholipid membrane is DOPC, and the neutral lipid is triolein.

[0023] Alternatively, the membrane material of the monomolecular phospholipid membrane is DOPC, and the neutral lipid is sunflower oil.

[0024] Alternatively, the membrane material of the monomolecular phospholipid membrane is DOPC, and the neutral lipid is soybean oil.

[0025] In the present invention, the monomolecular phospholipid membrane also includes a cationic lipid. Experiments in the present invention have shown that the addition of cationic lipids to the membrane material can further improve the bioavailability of the new gambogic acid fat body preparation, enhance anti-tumor activity, expand the scope of application of new gambogic acid, and ensure its safety. In the present invention, the type of cationic lipid is not limited; as long as it is a lipid with a positive charge, this improvement in effect can be achieved. In some embodiments, the cationic lipid is selected from one or more of DOTAP ((2,3-dioleoyl-propyl)-trimethylamine (chloride)), DODAP ((Z)-3-(dimethylamino)propane-1,2-diyl dioleate), DODMA (1,2-dioleyl-3-dimethylamino-propane), DOTMA (1,2-dioctadecenyloxy-3-methylammonium propane (chloride)), and D-Lin-MC3-DMA (4-(N,N-dimethylamino)butyric acid (dilinoleyl) methyl ester).

[0026] In some embodiments:

[0027] The membrane materials of the monomolecular phospholipid membrane are egg yolk phosphatidylcholine and DOTAP, and the neutral lipid is triolein.

[0028] Alternatively, the membrane material of the monomolecular phospholipid membrane is egg yolk lecithin and DOTAP, and the neutral lipid is sunflower oil.

[0029] Alternatively, the membrane material of the monomolecular phospholipid membrane is egg yolk lecithin and DOTAP, and the neutral lipid is soybean oil.

[0030] Alternatively, the membrane material of the monomolecular phospholipid membrane is soybean lecithin and DOTAP, and the neutral lipid is triolein.

[0031] Alternatively, the membrane materials of the monomolecular phospholipid membrane are soybean lecithin and DOTAP, and the neutral lipid is sunflower oil.

[0032] Alternatively, the membrane materials of the monomolecular phospholipid membrane are soybean lecithin and DOTAP, and the neutral lipid is soybean oil.

[0033] Alternatively, the membrane material of the monomolecular phospholipid membrane is DOPC and DOTAP, and the neutral lipid is triolein.

[0034] Alternatively, the membrane material of the monomolecular phospholipid membrane is DOPC and DOTAP, and the neutral lipid is sunflower oil.

[0035] Alternatively, the membrane material of the monomolecular phospholipid membrane is DOPC and DOTAP, and the neutral lipid is soybean oil.

[0036] Furthermore, the new gambogic acid fat body preparation of the present invention also includes a targeting molecule, which targets and recognizes organs, tissues or cells.

[0037] In the present invention, the organs, tissues or cells are from the human body or animal body;

[0038] The organs are endocrine organs, digestive organs, circulatory organs, urinary organs, reproductive organs, locomotor organs, the nervous system, and sensory organs. Endocrine organs include the thyroid gland and pancreas. Digestive organs include the stomach, liver, gallbladder, spleen, pancreas, small intestine, and large intestine. Respiratory organs include the lungs. Circulatory organs include the heart and blood vessels. Urinary organs include the kidneys, ureters, and bladder. Reproductive organs include the uterus and ovaries. Locomotor organs include muscles and bones. The nervous system includes the cerebrum and cerebellum. Sensory organs include the skin, eyes, and ears.

[0039] The tissue or cell is derived from a tumor of the 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, cervical cancer, oral or nasal mucosal cancer, bladder cancer, central nervous system cancer, cervical cancer, head and neck cancer, colon cancer, endometrial cancer, external genital cancer, esophageal cancer, gallbladder cancer, gastrointestinal cancer, genitourinary tract 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.

[0040] The targeting molecule can be embedded in the monomolecular phospholipid membrane, or it can be bound to the phospholipids on the monomolecular phospholipid membrane through the avidin-biotin system, or it can be connected to a substance that specifically targets phospholipids to bind to the monomolecular phospholipid membrane, or it can be a combination of any two or more of the above methods.

[0041] In the present invention, 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.

[0042] In some embodiments, the targeting molecule is linked to biotin, and a phospholipid molecule labeled with streptavidin is added during the preparation of the adipocytes. Alternatively, the targeting molecule is linked to streptavidin, and a phospholipid molecule labeled with biotin is added during the preparation of the adipocytes.

[0043] In some embodiments, the targeting molecule is connected to a peptide segment that targets and recognizes a single molecule of phospholipid membrane, which includes 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 of phospholipid membrane can be connected via a linker or directly connected without a linker, which is not limited by the present invention. The linker is a cleavable linker or a self-cleaving linker. The amino acid sequence of the cleavable linker is LEAGCKNFFPRSFTSCGSLE, and the self-cleaving linker is P2A, T2A, or E2A.

[0044] In the present invention, the membrane material of the preparation contains only phospholipids, or includes phospholipids and cationic lipids, or includes phospholipids and targeting molecules, or includes phospholipids, cationic lipids and targeting molecules.

[0045] in:

[0046] The mass ratio of phospholipid to cationic lipid is (0.1-10):1. In some embodiments, the mass ratio of phospholipid to cationic lipid is (1-10):1. In some embodiments, the mass ratio of phospholipid to cationic lipid is (0.1-1):1. In some embodiments, the mass ratio of phospholipid to cationic lipid is (0.5-5):1. In some specific embodiments, the mass ratio of phospholipid to cationic lipid is 0.5:1, 0.6:1, 0.7:1, 0.8:1, 0.9:1, 1:1, 2:1, 3:1, 4:1 or 5:1.

[0047] The mass ratio of phospholipids to targeting molecules is (10-100):(2-20). In some embodiments, the mass ratio of phospholipids to targeting molecules is (10-25):(2-10). More specifically, the mass ratio of phospholipids to targeting molecules is (15-20):(2-5). Preferably, the mass ratio of phospholipids to targeting molecules is 15:(2-5), or 16:(2-5), or 17:(2-5), or 18:(2-5), or 19:(2-5), or 20:(2-5). More preferably, the mass ratio of phospholipids to targeting molecules is (15-16):2, or (15-16):3, or (15-16):4, or (15-16):5; or (16-17):2, or (16-17):3, Or it is (16-17):4, or it is (16-17):5; or it is (17-18):2, or it is (17-18):3, or it is (17-18):4, or it is (17-18):5; or it is (18-19):2, or it is (18-19):3, or it is (18-19):4, or it is (18-19):5; or it is (19-20):2, or it is (19-20):3, or it is (19-20):4, or it is (19-20):5.

[0048] The mass ratio of phospholipid, cationic lipid and targeting molecule is (1-10):(1-10):(0.2-2)

[0049] In some embodiments, the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.01-0.2):(0.5-4):(2-10); specifically, the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.01-0.1):(0.5-4):(2-10), or the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.01-0.2):(1-4):(2-10), or the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.01-0.2): (0.5-4): (2-7), more specifically, the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.01-0.1): (1-4): (2-7), preferably, the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.05-0.1): (1-3): (4-6), more preferably, the mass ratio of the neogamoic acid, phospholipids and neutral lipids is (0.05-0.1): 2: 5, or 0.09: (1-3): 5, or 0.09: 2: (4-6). In some specific embodiments, the phospholipid is egg yolk lecithin and the neutral lipid is triolein, then the mass ratio of the neogamoic acid, egg yolk lecithin and triolein is 0.09: 2: 5.

[0050] In some embodiments, the mass ratio of the neogarcinia acid, phospholipids, neutral lipids and cationic lipids is (0.01-0.2): (0.5-4): (2-10): (0.1-4). Specifically, the mass ratio of the neogarcinia acid, phospholipids, neutral lipids and cationic lipids is (0.01-0.1): (0.5-4): (2-10): (0.1-4), or the mass ratio of the neogarcinia acid, phospholipids, neutral lipids and cationic lipids is (0.01-0.2): (1-4): (2-10): (0.1-4), or the mass ratio of the neogarcinia acid, phospholipids, neutral lipids and cationic lipids is (0.01-0.2): (0.5-4): (2-7): (0.1-4), or the mass ratio of the neogarcinia acid, phospholipids, neutral lipids and cationic lipids is (0.01-0.2): (0.5-4): (2-10): (0. 1~1); more specifically, the mass ratio of the new gambogic acid, phospholipids, neutral lipids and cationic lipids is (0.01~0.1):(1~4):(2~7):(0.1~1), preferably, the mass ratio of the new gambogic acid, phospholipids, neutral lipids and cationic lipids is (0.05~0.1):(1~3):(4~6):(0.1~1), more preferably, the mass ratio of the new gambogic acid, phospholipids, neutral lipids and cationic lipids is (0.05~0.1):1.5:5:0.5, or 0.09:(1~3):5:0.5, or 0.09:2:(4~6):0.5, or 0.09:2:1.5:(0.1~1). In some specific embodiments, the phospholipid is egg yolk lecithin, the neutral lipid is triolein, and the cationic lipid is DOTAP, and the mass ratio of the neogambogic acid, egg yolk lecithin, triolein and DOTAP is 0.09:1.5:5:0.5.

[0051] Compared with other cases, the phospholipid is egg yolk lecithin, the neutral lipid is triolein, the cationic lipid is DOTAP, and the mass ratio of the new gamboge acid, egg yolk lecithin, triolein and DOTAP is 0.09:1.5:5:0.5, which can obtain better encapsulation effect, better bioavailability, and can further improve the anti-tumor effect.

[0052] Furthermore, the present invention also provides a method for preparing the above-mentioned preparation, comprising:

[0053] The neogambogic acid solution and neutral lipid are mixed, and then mixed with a buffer solution and a monolayer phospholipid membrane material, and the neogambogic acid-encapsulated fat body is obtained after repeated vortexing and centrifugation;

[0054] The membrane material comprises one or more of phospholipids or functional polar lipids. In some embodiments, the membrane material further comprises cationic lipids and / or targeting molecules.

[0055] That is, the membrane material of the present invention includes phospholipids, or includes phospholipids and cationic lipids, or includes phospholipids and targeting molecules, or includes phospholipids, cationic lipids and targeting molecules.

[0056] The buffer solution is PBS buffer solution, HEPES buffer solution, sucrose solution, NaCl solution, KCl solution or MgCl2 solution.

[0057] The membrane material contains more than just phospholipids, and the various components of the membrane material are dissolved in an organic solvent, mixed, and then the organic solvent is evaporated. The organic solvent is a mixture of methanol and at least one of the following solvents: anhydrous ethanol, chloroform, benzene, toluene, xylene, butanol, isopropanol, ethyl ether, acetone, cyclohexanone, methyl isobutyl ketone, ethyl acetate, butyl acetate, cyclohexanone, or petroleum ether, preferably a mixture of methanol and chloroform.

[0058] As described above, the repeated vortexing and centrifugation include: after mixing, vortexing to obtain mixture 1, centrifuging the mixture 1 to collect the lower layer solution, and vortexing again to obtain mixture 2; centrifuging the mixture 2 to remove the precipitate, and vortexing again to obtain mixture 3; centrifuging the mixture 3 to collect the lower layer solution, and vortexing again to obtain mixture 4 containing the fat body.

[0059] Specifically:

[0060] After mixing, the mixture was vortexed to obtain Mixture 1.

[0061] In this step, the parameters of the vortex include: vortexing at 3000-4000 rpm for 3-7 minutes, operating for 1-10 seconds, and resting for 1-10 seconds. Preferably, the vortex speed is 3000, 3200, 3400, 3500, 3600, 3700, 3800, 3900, or 4000 rpm. Preferably, the vortex duration is 3 minutes, 4 minutes, 5 minutes, 6 minutes, or 7 minutes. Preferably, the vortex stops for 1 second every working 1 second, or stops for 2 seconds every working 2 seconds, or stops for 2 seconds every working 2 seconds, or stops for 3 seconds every working 3 seconds, or stops for 4 seconds every working 4 seconds, or stops for 5 seconds every working 5 seconds, or stops for 6 seconds every working 6 seconds, or stops for 7 seconds every working 7 seconds, or stops for 8 seconds every working 8 seconds, or stops for 9 seconds every working 9 seconds, or stops for 10 seconds every working 10 seconds, or stops for 6 seconds every working 5 seconds, or stops for 7 seconds every working 5 seconds, or stops for 8 seconds every working 5 seconds, or stops for 9 seconds every working 5 seconds, or stops for 10 seconds every working 6 seconds, or stops for 7 seconds every working 6 seconds, or stops for 8 seconds every working 6 seconds, or stops for 9 seconds every working 6 seconds, or stops for 10 seconds every working 6 seconds, or stops for 8 seconds every working 7 seconds, or stops for 9 seconds every working 7 seconds, or stops for 10 seconds every working 8 seconds, or stops for 10 seconds every working 8 seconds, or stops for 10 seconds every working 9 seconds. In some specific embodiments, the vortex parameters in this step include 4000 rpm, vortexing for 10 seconds, stopping for 10 seconds, and vortexing for 4 minutes. Alternatively, the vortex parameters in this step include 4000 rpm, vortexing for 10 seconds, stopping for 5 seconds, and vortexing for 3 minutes.

[0062] The mixture 1 was centrifuged to collect the lower layer solution, and then vortexed again to obtain a mixture 2.

[0063] In this step, the vortexing parameters include 1000-4000 rpm, and the centrifugation parameters include: centrifugation at 800-1200×g for 3-7 minutes at room temperature. Preferably, the centrifugation speed is 800×g, 900×g, 1000×g, 1100×g, or 1200×g, and the centrifugation time is 3 minutes, 4 minutes, 5 minutes, 6 minutes, or 7 minutes. In some embodiments, the centrifugation conditions include centrifugation at 1000×g for 5 minutes at room temperature.

[0064] After the mixture 2 is centrifuged to remove the precipitate, it is vortexed again to obtain the mixture 3;

[0065] In this step, the vortexing parameters include 1000-4000 rpm, and the centrifugation parameters include: in actual applications, centrifugation at 18,000-22,000 × g for 3-7 minutes. Preferably, the centrifugation speed is 18,000 × g, 19,000 × g, 20,000 × g, 21,000 × g, or 22,000 × g, and the centrifugation time is 3 minutes, 4 minutes, 5 minutes, 6 minutes, or 7 minutes. In some embodiments, the centrifugation conditions include centrifugation at 20,000 × g for 5 minutes at room temperature.

[0066] The mixture 3 is centrifuged to collect the lower layer solution, and vortexed again to obtain the mixture 4 containing the fat body. In this step, the parameters of the vortex include 1000-4000rpm, and the parameters of the centrifugation include: in actual application, centrifugation at 800-1200×g for 3-7min. Preferably, the centrifugation speed is 800×g, 900×g, 1000×g, 1100×g or 1200×g, and the centrifugation time is 3min, 4min, 5min, 6min or 7min. In some embodiments, the centrifugation conditions include centrifugation at 1000×g for 5min at room temperature.

[0067] The preparation method provided by the present invention is simple and easy to operate, and the obtained preparation has good drug loading and bioavailability. It has been verified that the content of neogambogic acid in the neogambogic acid fat body preparation prepared by this method reaches 1 mg / ml, which is 2000 times the concentration in water. At the same time, the neogambogic acid fat body has a good anti-tumor effect, which can inhibit the growth of various cancer cells such as colorectal cancer, breast cancer, and liver cancer, and the effect is significantly better than that of free neogambogic acid.

[0068] Furthermore, the present invention provides the use of the above-mentioned preparation or the preparation obtained by the above-mentioned method in the preparation of medicines or vaccines.

[0069] The efficacy of the drug and / or vaccine includes at least one of anti-oxidation, anti-inflammatory, anti-cancer and antibacterial.

[0070] In some embodiments, the cancer comprises breast cancer, lung cancer, prostate cancer, stomach cancer, head and neck cancer, kidney cancer, laryngeal cancer, liver cancer, muscle tissue cancer, blood tumor, bone cancer, brain cancer, oral or nasal mucosal cancer, bladder cancer, central nervous system cancer, cervical cancer, head and neck cancer, colon cancer, endometrial cancer, external genital cancer, esophageal cancer, gallbladder cancer, gastrointestinal cancer, genitourinary tract cancer, ovarian cancer, pancreatic cancer, skin cancer, spleen cancer, small intestine cancer, large intestine cancer, melanoma, testicular cancer and / or thyroid cancer.

[0071] Furthermore, the present invention provides a medicine or vaccine, which includes the preparation as described above or the preparation prepared by the method.

[0072] The medicine or vaccine of the present invention further comprises an effective amount of a tumor inhibitor;

[0073] The tumor suppressors include: cisplatin, carboplatin, oxaliplatin, 5-fluorouracil (5-FU), methotrexate, daunorubicin, dactinomycin-D, irinotecan (CPT-11), mitoxantrone, estramustine, vincristine, dexamethasone, prednisone, lomustine, methotrexate, pirarubicin, doxorubicin, gemcitabine, quizartinib or bevacizumab.

[0074] The medicine or vaccine of the present invention further includes pharmaceutically acceptable excipients.

[0075] In the present invention, the pharmaceutically acceptable excipients are selected according to the requirements of the dosage form.

[0076] The dosage form of the medicine or vaccine of the present invention is oral preparation, inhalant or injection.

[0077] Optionally, the drug or vaccine is in the form of an oral preparation, for example, a tablet, a pill, an oral solution, a capsule, a syrup, a dropper or a granule.

[0078] In some embodiments provided herein, the capsule is a hard capsule or a soft capsule.

[0079] In some embodiments provided herein, the tablet is an oral tablet or buccal tablet.

[0080] Oral tablets are tablets for oral administration. Most of the drugs in these tablets are absorbed through the gastrointestinal tract to exert their effects, while some drugs in other tablets exert their effects locally in the gastrointestinal tract. In some embodiments provided herein, the oral tablets are conventional compressed tablets, dispersible tablets, effervescent tablets, chewable tablets, coated tablets, or sustained-release tablets.

[0081] The medicine or vaccine is inhaled, and optionally, it is an inhalation aerosol, an inhalation powder, or a liquid preparation for use with a nebulizer.

[0082] The medicine or vaccine is an injection, for example, an injection solution or an injection powder.

[0083] The present invention also provides a method for preventing and treating tumors, comprising administering the aforementioned drug or vaccine, wherein the administration comprises oral administration, inhalation, and / or injection.

[0084] The subject of the method is a human or, the subject of the method is a primate or a non-primate mammal.

[0085] The novel nanoparticle fat body (having a hydrophobic core) provided by the present invention encapsulates neogamonic acid to construct a neogamonic acid fat body, and the neogamonic acid fat body is modified by different phospholipids to increase its effectiveness or safety. At the same time, the present invention also provides a preparation method thereof. The content of neogamonic acid in the prepared neogamonic acid fat body preparation reaches 1 mg / ml, which is 2000 times its concentration in water. Its bioavailability and scope of application are significantly improved. At the same time, the neogamonic acid fat body has a good anti-tumor effect and can inhibit the growth of various cancer cells such as colorectal cancer, breast cancer, and liver cancer, and the effect is significantly better than that of free neogamonic acid. BRIEF DESCRIPTION OF THE DRAWINGS

[0086] Figure 1 shows the encapsulation of neo-gambogic acid to construct neo-gambogic acid fat bodies;

[0087] Figure 2 shows the preparation of new gambogic acid fat bodies, wherein: A, optical microscope photos (ac) and electron microscope photos (d) of fat bodies without NGA (empty fat bodies, i.e., Blank-adiposome, Blank-AD); B, optical microscope photos (ac) and electron microscope photos (d) of fat bodies with NGA but without cations (new gambogic acid fat bodies, i.e., NGA-adiposome without cationic, NGA-AD); C, optical microscope photos (ac) and electron microscope photos (d) of fat bodies with NGA with cations (new gambogic acid cationic fat bodies, i.e., NGA-adiposome with cationic, NGA-C-AD); a is the result of DIC, b is the result of neutral lipid dye Nile Figure 3 is the staining result of Red, Figure c is the result of the fusion of Figure a and Figure b, and Figure d is the electron microscopy result; the scale bars in Figures ac and d are 5 microns, and the scale bar in d is 10 nm; D, dynamic light scattering instrument detected the average particle size and PDI (polymer dispersibility index) of these fat bodies; E, TLC detected the lipid composition and NGA signal in these fat bodies; lanes 1-3 are standards, namely Egg PC, DOTAP, and NGA; lanes 4-6 are Blank-AD, NGA-AD, and NGA-C-AD, respectively; F, HPLC detection of free NGA standards in ethanol and NGA in new gambogic acid fat bodies; G, Ztea potential on the surface of prepared new gambogic acid fat bodies;

[0088] Figure 3 shows that the new gambogic acid fat body has anti-tumor activity, and the effect is significantly better than that of free new gambogic acid. The different new gambogic acid fat bodies prepared in Example 1 were treated with the corresponding cells according to the concentrations shown, and the cell survival rate was measured by CCK8; A, rectal cancer cell CT26; B, breast cancer cell 4T1; C, human hematologic malignant tumor cell H929; D, human hematologic malignant tumor cell Raji; where ns indicates not significant, *p<0.05, **p<0.01, ***p<0.001;

[0089] Figure 4 shows that neogambogic acid fat bodies inhibit the growth of colorectal cancer in mice, and the effect is better than that of free neogambogic acid, where: A, growth curve of tumors in mice in different treatment groups; B, weight changes of mice in different treatment groups; C, photos of tumors in mice in different treatment groups; D, weight and tumor inhibition rate of tumors in mice in different treatment groups; E, safety evaluation of tumors in mice in different treatment groups; RBC, red blood cell count; HGB, hemoglobin, a blood test indicator; Cre, creatinine, an indicator of kidney inflammation; AST, aspartate aminotransferase, an indicator of liver inflammation;

[0090] Figure 5 shows that the neogamoic acid fat body inhibits the growth of liver cancer in mice, and its effect is better than that of free neogamoic acid, among which: A, photos of liver tumors in mice in different treatment groups; B, weights and tumor inhibition rates of liver tumors in mice in different treatment groups; C, weight changes of mice in different treatment groups; D, safety evaluation of tumors in mice in different treatment groups; RBC, red blood cell count; HGB, hemoglobin, a blood routine indicator; Cre, creatinine, an indicator of kidney inflammation; AST, aspartate aminotransferase, an indicator of liver inflammation. DETAILED DESCRIPTION

[0091] The present invention provides a new gambogic acid fat body preparation and its preparation method and application. Those skilled in the art can refer to the content of this article and appropriately improve the process parameters to achieve it. It should be noted that all similar substitutions and modifications are obvious to those skilled in the art and are considered to be included in the present invention. The methods and applications of the present invention have been described through preferred embodiments. It is obvious that relevant personnel can modify or appropriately change and combine the methods and applications herein without departing from the content, spirit and scope of the present invention to implement and apply the technology of the present invention.

[0092] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as understood by those of ordinary skill in the art. For definitions and terminology in this field, professionals are specifically referred 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 used L-amino acids.

[0093] In this application, the term "and / or" describes the association relationship between associated objects, indicating that three relationships may exist. For example, A and / or B can mean: A exists alone, A and B exist at the same time, and B exists alone. A and B can be singular or plural.

[0094] The terms "include," "comprising," and "having" are used interchangeably herein and are intended to indicate the inclusiveness of a solution, meaning that the solution may contain other elements in addition to the listed elements. It should also be understood that the use of "include," "comprising," and "having" in this document also provides a "consisting of" solution.

[0095] In this application, "at least one" means one or more, and "more than one" means two or more. "At least one of the following" or similar expressions refers to any combination of these items, including any combination of single or plural items.

[0096] The term "drug" herein refers to a preparation that is in a form that permits the biological activity of the active ingredient contained therein to be effective and that contains no additional ingredients that are unacceptably toxic to a subject to which the pharmaceutical composition is administered.

[0097] The term "prevent and treat" herein includes prevention and / or treatment. The "treatment" refers to surgical or pharmaceutical treatment, the purpose of which is to prevent, slow down (reduce) undesirable physiological changes or lesions in the treated subject, such as cancer and tumors. Beneficial or desired clinical results include, but are not limited to, alleviation of symptoms, weakening of the disease extent, stabilization of the disease state (i.e., no worsening), delay or slowing of disease progression, improvement or alleviation of the disease state, and relief (whether partial relief or complete relief), whether detectable or undetectable. Objects in need of treatment include objects already suffering from a condition or disease, as well as objects susceptible to a condition or disease, or objects intended to prevent a condition or disease. When referring to terms such as slowing down, alleviating, weakening, alleviating, and alleviating, their meanings also include situations such as elimination, disappearance, and non-occurrence.

[0098] The subject of the term "administered" herein refers to an organism that receives treatment for a specific disease or condition as described herein. Exemplarily, the subject receiving treatment for a disease or condition is a mammal, such as a human, a primate (e.g., a monkey), or a non-primate mammal.

[0099] The term "subject" herein refers to an organism that is being treated for a particular disease or condition as described herein. Exemplarily, a "subject" includes a mammal, such as a human, primate (e.g., monkey), or non-primate mammal, being treated for a disease or condition.

[0100] As used herein, the term "effective amount" refers to an amount of a therapeutic agent that, when administered alone or in combination with another therapeutic agent to a cell, tissue, or subject, is effective in preventing or ameliorating a disease symptom or the progression of that disease. "Effective amount" also refers to an amount of a compound sufficient to alleviate symptoms, e.g., to treat, cure, prevent, or alleviate a related medical condition, or to increase the rate of treatment, cure, prevention, or alleviation of such a condition. When an active ingredient is administered alone to a subject, a therapeutically effective dose refers to that ingredient alone. When a combination is used, a therapeutically effective dose refers to the combined amounts of the active ingredients that produce a therapeutic effect, whether administered in combination, sequentially, or simultaneously.

[0101] As used herein, the term "cancer" refers to or describes the physiological condition in mammals that is typically characterized by unregulated cell growth. Both benign and malignant cancers are included in this definition. As used herein, the terms "tumor" or "neoplasm" refer to all neoplastic cell growth and proliferation, whether malignant or benign, and all precancerous and cancerous cells and tissues. The terms "cancer" and "tumor" are not mutually exclusive when used herein.

[0102] The term "IC50" in this article refers to the half-inhibitory concentration of the antagonist being measured. It can be understood that a certain concentration of a drug induces 50% tumor cell death. This concentration is called the 50% inhibitory concentration, that is, the concentration corresponding to the ratio of dead cells to total cells is equal to 50%. The IC50 value can be used to measure the ability of a drug to induce death. That is, the stronger the induction ability, the lower the value.

[0103] It should be understood that in the various embodiments of the present application, the size of the serial numbers of the above-mentioned processes does not mean the order of execution. Some or all of the steps can be executed in parallel or sequentially. The execution order of each process should be determined by its function and internal logic, and should not constitute any limitation on the implementation process of the embodiments of the present application.

[0104] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0105] The fragment names and sequences involved in this article are as follows:

[0106] Table 1 Names and amino acid sequences of peptides targeting single-molecule phospholipid membranes

[0107] Table 2 Targeting molecule names and amino acid sequences and targeting proteins

[0108] The present invention uses nanoparticle fat bodies to encapsulate neogamoic acid to construct neogamoic acid alcohol fat bodies. The content of neogamoic acid in the prepared neogamoic acid fat body preparation reaches 1 mg / ml, and the concentration is 2000 times that in water. At the same time, the neogamoic acid fat body has a good anti-tumor effect and can inhibit the growth of various cancer cells such as colorectal cancer, breast cancer, and liver cancer, and the effect is significantly better than that of free neogamoic acid. Therefore, the present invention provides a new gambogic acid fat body and a preparation method thereof, thereby significantly improving the solubility of neogamoic acid and effectively inhibiting tumor growth, expanding the application range of neogamoic acid, laying the foundation for the clinical transformation of neogamoic acid, and having a broad and promising application market in cancer treatment. The present invention is further described below in conjunction with the examples.

[0109] Example 1 Preparation of New Gambogic Acid Adiposome (NGA-AD)

[0110] 1) Neutral fat: triolein.

[0111] 2) NGA was directly dissolved in triolein to saturation. High-performance liquid chromatography (HPLC) determined the NGA content in the triolein to be 18.3 mg / ml. HPLC conditions: Agilent Zorbax SB-C18 column, mobile phase: methanol:acetonitrile:water = 6:3:1 (v / v / v, 0.1% trichloroacetic acid, m%), column temperature 35°C, detection wavelength 360 nm, flow rate 1 ml / min.

[0112] 3) Phospholipids: Egg yolk phosphatidylcholine (EggPC) and cationic lipid (2,3-dioleoyl-propyl)-trimethylamine (chloride) (DOTAP) were prepared in chloroform at a concentration of 25 mg / ml.

[0113] 4) 60 μl of the above-prepared Egg PC and 20 μl of the above-prepared DOTAP were added to a microcentrifuge tube, and the solvent was blown dry with high-purity nitrogen gas.

[0114] 5) Add 100 μl of PBS and 5 μl of the neutral lipid containing NGA prepared in step 2) to a microcentrifuge tube and vortex for 4 minutes (vortex for 10 seconds, rest for 10 seconds) (in actual application, vortex for 3-7 minutes, vortexing conditions are 3000-4000 rpm, and 4000 rpm is used in this example) to obtain a milky white lipid mixture 1. The lipid mixture 1 is centrifuged at 1000 × g for 5 minutes (in actual application, centrifugation at 800-1200 × g for 3-7 minutes is acceptable). After centrifugation, the liquid phase system exhibits two stratifications. The lower milky white solution is collected by extraction and vortexed to obtain a milky white lipid mixture 2.

[0115] 6) The lipid mixture 2 obtained in step 5) was centrifuged at 20,000×g for 5 minutes (in actual application, 18,000-22,000×g for 3-7 minutes is acceptable). After centrifugation, the precipitate at the bottom of the microcentrifuge tube was removed and vortexed to obtain a milky white lipid mixture 3.

[0116] 7) The lipid mixture 3 obtained in step 6) was centrifuged at 1000×g for 5 minutes (in actual application, 800-1200×g for 3-7 minutes is acceptable). 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 4, which was the final cationic adipome carrying the hydrophobic small molecule compound neogamoic acid (neogamoic acid cationic adipome, i.e., NGA-adiposome with cationic, NGA-C-AD). Using the same production method, we prepared adipomes with NGA but without cationic (neogamoic acid adipome, i.e., NGA-adiposome without cationic, NGA-AD). Using the same production method, we prepared adipomes without NGA (blank adipome, i.e., blank adipome, blank-AD).

[0117] The morphological structures of the three constructed fat bodies were observed using optical and electron microscopy, revealing uniform spherical structures free of contamination from other membrane impurities (Figure 2, A, B, and C). Furthermore, dynamic light scattering revealed an average particle size of 120 nm and a PDI (dispersity index) of 0.15 (Figure 2, D). The presence of neogambogic acid in the neogambogic acid fat bodies was further determined by TLC (thin-layer chromatography) and HPLC (high-performance liquid chromatography). The TLC assay involved adding an equal volume of methanol and two volumes of chloroform to the constructed neogambogic acid fat bodies to extract lipids. The organic phase was collected and dried with nitrogen to obtain total lipids. The total lipids were then added to 100 μl of chloroform, and 10 μl was loaded onto a silica gel plate. The plate was then developed in a solvent consisting of n-hexane: ether: glacial acetic acid (80:20:1 by volume) to separate triolein. The silica gel plate was then developed in a solvent consisting of chloroform:methanol:glacial acetic acid:water (volume ratio: 75:13:9:3) to separate EggPC, DOTAP, and NGA. TLC results indicated the presence of NGA signals in the fat bodies based on the position of the standard. In lane 4 (Blank-AD), no NGA signal was detected. In lane 5 (NGA-AD), an NGA signal was detected, but no DOTAP signal was detected. In lane 6 (NGA-C-AD), both NGA and DOTAP signals were detected. However, all of these fat bodies contained triolein signals (Figure 2, E). HPLC results showed that the elution time of NGA in the neogambogic acid fat bodies coincided with that of the free NGA standard in ethanol (Figure 2, F). At the same time, the Ztea potential results showed that NGA-C-AD was positively charged at about 60 mV, while Blank-AD and NGA-AD were not positively charged, at 0 mV and -20 mV, respectively (G in Figure 2). The above results indicate that the fat body containing neogamonic acid was successfully prepared with high purity and good uniformity.

[0118] Example 2 Neogambogic acid fat bodies have anti-tumor activity, and the effect is significantly better than free neogambogic acid

[0119] We further tested whether the new garcinia fat bodies have biological activity and whether they can kill tumor cells. Different new garcinia acid fat bodies prepared in Example 1 were selected to treat rectal cancer cells CT26 (Figure 3 A), breast cancer cells 4T1 (Figure 3 B) and liver cancer cells Hepa1-6 (Figure 3 C). The results showed that compared with the equal volume of normal saline group and Blank-AD group, as the concentration of NGA increased, the effect of NGA fat bodies in killing cancer cells became better and better, significantly better than free NGA. In addition, compared with fat bodies with NGA but no cations (NGA-AD), fat bodies with NGA and cations (NGA-C-AD) had the best effect in killing various cancer cells.

[0120] Cell viability assay: 3000 cells / well of the cells to be treated were plated 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 1 hour of incubation, the absorbance at 450 nm was read using a microplate reader. Cell viability was calculated using the following formula:

[0121] Cell Viability=(Ae-Ab) / (Ac-Ab)*100%

[0122] Wherein, Ae represents the absorbance value of the well containing cells after drug treatment.

[0123] Ac represents the absorbance value of the wells containing cells without drug treatment.

[0124] Ab represents the absorbance value of blank wells containing only culture medium and CCK8 reagent, which is used to correct background noise.

[0125] Example 3 Neogambogic acid fat bodies inhibit the growth of colorectal cancer in mice, and the effect is better than that of free neogambogic acid

[0126] Previous in vitro cell experiments have shown that neogambogic acid liposomes can inhibit the growth of various cancer cells, including colorectal cancer cells, breast cancer cells, and liver cancer cells, with an inhibitory effect superior to that of free neogambogic acid. We further compared the efficacy of neogambogic acid liposomes with that of free neogambogic acid in the treatment of colorectal cancer in mice in vivo.

[0127] First, mouse colorectal cancer cells, CT26, were injected subcutaneously into the backs of BALB / c female mice for tumor growth. Eleven days later, the mice were randomly divided into five groups: saline, blank adipocytes (Blank-AD), free NGA, adipocytes containing NGA but no cations (NGA-AD), and adipocytes containing both NGA and cations (NGA-C-AD), with seven mice per group. NGA was administered once weekly at a dose of 10 mg / kg per mouse. Tumor size and body weight changes were measured during the dosing period. As shown in Figure 4, NGA-AD and NGA-C-AD demonstrated even greater tumor inhibition, with rates of 59.8% and 80.5%, respectively, compared to free NGA, which inhibited tumors by 47.7%. There was no difference in body weight between mice treated with neogambogic acid adipocytes, nor in blood counts such as hemoglobin (HGB), red blood cell count (RBC), or creatinine (Cre), a marker of renal inflammation. However, NGA-C-AD-treated mice showed a significant decrease in aspartate aminotransferase (AST), a marker of liver inflammation, indicating that NGA-C-AD has lower hepatotoxicity than NGA and NGA-AD. These results indicate that neogambogic acid fat bodies inhibit the growth of colorectal cancer in mice, significantly outperforming free neogambogic acid and demonstrating a good safety profile, laying the foundation for its clinical translation.

[0128] Example 4 Neogambogic acid fat bodies inhibit the growth of mouse liver cancer, and the effect is better than free neogambogic acid

[0129] Previous in vitro cell experiments have shown that neogambogic acid liposomes can inhibit the growth of various cancer cells, including colorectal cancer cells, breast cancer cells, and liver cancer cells, with an inhibitory effect superior to that of free neogambogic acid. We further compared the efficacy of neogambogic acid liposomes with that of free neogambogic acid in treating liver cancer in mice in vivo.

[0130] We constructed a mouse liver cancer tumor model to explore the therapeutic effect of the new gambogic acid fat body. First, a C57BL / 6 mouse liver cancer model was constructed by combining diethylnitrosamine (DEN) with carbon tetrachloride. The construction method is as follows: Prepare DEN solution: take a small amount of DEN, dilute it to 4 mg / ml with normal saline, and inject it intraperitoneally at 10 μl / g body weight. Prepare CCl4 (20%) solution: measure 10 ml CCl4, add 40 ml corn oil to make a 20% CCl4 solution, and inject it intraperitoneally at 5 μl / g body weight. Two-week-old mice were intraperitoneally injected with DEN (40 mg / kg) once. After the mice were raised for 6 weeks (8-week-old mice), they were injected with 20% CCl4 solution twice a week. The administration was continued for 16 weeks, and the liver was dissected and observed to check for primary liver cancer. If yes, it indicates that the liver cancer model was successfully constructed, and then the mice were randomly divided into 4 groups, namely normal saline group, empty fat body group (Blank-AD), free NGA, and fat body with NGA and cations (NGA-C-AD), with 5 mice in each group. The drug was administered twice a week, with a dose of 18 mg / kg per mouse each time. Compared with the tumor inhibition rate of 22.3% of free NGA, NGA-C-AD had a better effect in inhibiting mouse tumors, with a tumor inhibition rate of 76.2%. This result is consistent with the results of colorectal cancer mice, and the new gambogic acid fat body has good biocompatibility in mice. These results show that the new gambogic acid fat body inhibits the growth of mouse liver cancer, and the effect is significantly better than that of free new gambogic acid, and it has good safety, which lays the foundation for its clinical transformation.

[0131] The above are only preferred embodiments of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principles of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A new gambogic acid fat body preparation comprising: Monomolecular phospholipid membrane and neogamoic acid and neutral lipids wrapped inside the monomolecular phospholipid membrane.

2. The preparation according to claim 1, characterized in that The neutral lipid is selected from one or more of triolein, fish oil, corn oil, tricaprylin, retinol esters, wax esters, sterol esters, sterol esters, castor oil, sunflower oil, soybean oil, peanut oil, clove oil, simethicone, cinnamon oil, tea oil, liquid paraffin, star anise oil, mixed fatty acid glycerides (stearin), hydrogenated vegetable oil, refined olive oil and fat-soluble vitamins.

3. The preparation according to claim 1, characterized in that The membrane material of the monomolecular phospholipid membrane comprises one or more of phospholipids or functional polar lipids; The phospholipid is selected from one or more of egg yolk lecithin, soybean lecithin, dioleoylphosphatidylethanolamine, distearoylphosphatidylcholine, egg yolk lecithin, dipalmitoylphosphatidylcholine, dipalmitoylphosphatidic acid, sodium distearoylphosphatidylglycerol, dimyristoylphosphatidylcholine, 1-stearoyl-lysophosphatidylcholine, 1,2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol 2000, phosphatidylethanolamine, phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, cardiolipin and sphingomyelin; The functional polar lipid is selected from one or more of polyethylene glycol-modified sterols, biotin-modified sterols, amino acid-modified sterols, polypeptide-modified sterols, polysaccharide-modified sterols, nucleic acid-modified sterols, polyethylene glycol-modified phospholipids, biotin-modified phospholipids, amino acid-modified phospholipids, polypeptide-modified phospholipids, polysaccharide-modified phospholipids and nucleic acid-modified phospholipids.

4. The preparation according to any one of claims 1 to 3, characterized in that The monomolecular phospholipid membrane further comprises a cationic lipid, and the cationic lipid is selected from one or more of DOTAP, DODAP, DODMA, DOTMA, and D-Lin-MC3-DMA.

5. The preparation according to any one of claims 1 to 4, characterized in that Also included are targeting molecules that target and recognize organs, tissues, or cells.

6. The preparation according to claim 5, characterized in that The organs, tissues or cells are from the human body or animal body.

7. The preparation according to claim 5 or 6, characterized in that The targeting molecule is: The targeting molecule is embedded in a single molecule phospholipid membrane, or is connected to phospholipids through avidin-biotin interaction, or is connected to a substance that specifically targets phospholipids, or a combination of the above methods.

8. The preparation according to claim 5 or 6, 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.

9. The preparation according to any one of claims 5 to 8, characterized in that One or more of the phospholipids, functional polar lipids and cationic lipids in the monomolecular phospholipid membrane is labeled with biotin or streptavidin.

10. The preparation according to any one of claims 1 to 9, characterized in that The mass ratio of the neogambogic acid, phospholipid and neutral lipid is (0.01-0.2):(0.5-4):(2-10); The mass ratio of the neogambogic acid, phospholipid, neutral lipid and cationic lipid is (0.01-0.2):(0.5-4):(2-10):(0.1-4).

11. The preparation according to any one of claims 1 to 10, characterized in that The phospholipid is egg yolk lecithin, the neutral lipid is triolein, and the positive lipid is DOTAP.

12. The preparation according to claim 11, characterized in that The mass ratio of the neogamoic acid, egg yolk lecithin and triolein is (0.01-0.1):(1-4):(2-7); The mass ratio of the neogamoic acid, egg yolk lecithin, triolein and DOTAP is (0.01-0.1):(1-4):(2-7):(0.1-1).

13. A method for preparing the preparation according to any one of claims 1 to 12, comprising: The neogambogic acid solution and neutral lipid are mixed, and then mixed with a buffer solution and a monolayer phospholipid membrane material, and the neogambogic acid-encapsulated fat body is obtained after repeated vortexing and centrifugation; The membrane material includes one or more of phospholipids or functional polar lipids.

14. The preparation method according to claim 13, characterized in that The membrane material also includes cationic lipids and / or targeting molecules.

15. The preparation method according to claim 13 or 14, characterized in that: The buffer solution is PBS buffer solution, HEPES buffer solution, sucrose solution, NaCl solution, KCl solution or MgCl2 solution.

16. The preparation method according to claim 13, characterized in that The repeated vortexing and centrifugation include: After mixing, the mixture was vortexed to obtain a mixture 1. The mixture 1 is centrifuged to collect the lower layer solution, and then vortexed again to obtain a mixture 2; After the mixture 2 is centrifuged to remove the precipitate, it is vortexed again to obtain the mixture 3; The mixture 3 is centrifuged to collect the lower layer solution, which is vortexed again to obtain the mixture 4 containing the fat body.

17. Use of the preparation according to any one of claims 1 to 12 or the preparation obtained by the method according to any one of claims 13 to 16 in the preparation of drugs and / or vaccines.

18. The use according to claim 17, characterized in that The efficacy of the drug and / or vaccine includes at least one of anti-oxidation, anti-inflammatory, anti-cancer and antibacterial.

19. The use according to claim 17, characterized in that The cancers include: breast cancer, lung cancer, prostate cancer, stomach cancer, head and neck cancer, kidney cancer, laryngeal cancer, liver cancer, muscle tissue cancer, blood tumors, bone cancer, brain cancer, oral or nasal mucosal cancer, bladder cancer, central nervous system cancer, cervical cancer, head and neck cancer, colon cancer, endometrial cancer, external genital cancer, esophageal cancer, gallbladder cancer, gastrointestinal cancer, genitourinary tract cancer, ovarian cancer, pancreatic cancer, skin cancer, spleen cancer, small intestine cancer, large intestine cancer, melanoma, testicular cancer and / or thyroid cancer.

20. A medicine or vaccine comprising the preparation according to any one of claims 1 to 12 or the preparation prepared by the method according to any one of claims 13 to 16.

21. The drug or vaccine according to claim 20, characterized in that It also includes an effective amount of a tumor suppressor; The tumor suppressors include: cisplatin, carboplatin, oxaliplatin, 5-fluorouracil (5-FU), methotrexate, daunorubicin, dactinomycin-D, irinotecan (CPT-11), mitoxantrone, estramustine, vincristine, dexamethasone, prednisone, lomustine, methotrexate, pirarubicin, doxorubicin, gemcitabine, quizartinib or bevacizumab.

22. A method for preventing and treating tumors, comprising administering the drug or vaccine according to claim 20 or 21.

23. The method according to claim 22, characterized in that The subject of the method is a human.

24. The method according to claim 22, characterized in that The subject of the method is a primate or a non-primate mammal.

Images