Fusion proteins and uses thereof

Abstract

The application relates to a fusion protein and application thereof in the field of biotechnology. A polypeptide fragment for specifically recognizing a single-molecule phospholipid membrane and a polypeptide fragment for specifically recognizing a tissue or a cell are connected to form a fusion protein which can be specifically combined to a fat body surface in a non-covalent manner, so that precise targeting of the fat body is realized. The fat body targeting peptide segment can be widely used in the fields of precise drug delivery and vaccine preparation, so as to be used for treating diseases such as cancer, infectious diseases and metabolic diseases. Lung tissues and breast cancer cells are taken as target points, and good targeting of the fat body is realized.

Description

Fusion protein and its use

[0001] This application claims priority to the Chinese patent application filed with the China Patent Office on November 14, 2023, with application number 202311516762.2 and invention name “Fusion Protein and Its Uses”, the entire contents of which are incorporated by reference into this application. Technical Field

[0002] The present invention relates to the field of biotechnology, in particular to fusion proteins and uses thereof. Background Art

[0003] Adiposomes are nanospheres composed of a neutral lipid core encapsulated by a monomolecular phospholipid membrane. Their structure is similar to naturally occurring lipid droplets and lipoproteins. Adiposomes can recruit one or more intrinsic and / or functional proteins to create artificial lipid droplets, as well as one or more apolipoproteins to create artificial lipoproteins. Both of these play an important role in the preparation of drugs and / or drug carriers. As a novel nanocarrier, the neutral lipid core of adiposomes can load large quantities of lipophilic drugs and exhibits excellent biocompatibility.

[0004] Similar to the limitations of currently used nanocarriers for clinically administered nanodrugs, such as albumin and liposomes, adipocytes lack targeting. Consequently, drugs delivered via adipocytes are transported to healthy tissues, potentially leading to numerous side effects. Therefore, enabling targeted delivery of nanocarriers could effectively target diseased tissues or cells, minimizing damage to healthy tissues and potentially improving their potential for the treatment of diseases such as cancer, infectious diseases, and metabolic disorders.

[0005] Currently, the most commonly used method for achieving nanocarrier targeting is surface modification through covalent bonding, where targeting ligands or antibodies are covalently immobilized on the nanoparticle surface. For example, glycosylated PEGylated phospholipids and cell-penetrating peptide-modified PEGylated phospholipids are added to liposomes to target brain tissue. However, adipocytes, as a novel nanoparticle, are nanospheres with a neutral lipid core encapsulated by a monomolecular phospholipid membrane. Existing nanocarrier targeting techniques are not very effective for adipocytes. Therefore, methods and strategies specific to adipocyte targeting are still urgently needed.

[0006] Summary of the Invention

[0007] In light of this, the present invention aims to provide a fusion protein and its use. This invention achieves fat body targeting by constructing a fat body-targeted fusion protein and recruiting it to the fat body. The protein can be used in fields such as drug delivery and vaccine preparation, thereby treating diseases such as cancer, infectious diseases, and metabolic diseases.

[0008] The present invention provides a fusion protein comprising fragment A and fragment B; wherein:

[0009] Fragment A targets and recognizes the monolayer phospholipid membrane;

[0010] Fragment B targets and recognizes tissues or cells.

[0011] The fusion protein of the present invention comprises at least one fragment A and at least one fragment B, and the present invention does not limit the connection order thereof.

[0012] In the present invention, the structure of the fusion protein is:

[0013] [(fragment A)x-(fragment B)y]z or [(fragment B)x-(fragment A)y]z;

[0014] Wherein: x represents the number of repetitions of fragment A, y represents the number of repetitions of fragment B, and z represents the number of repetitions of the unit consisting of fragments A and B in the fusion protein. x, y, and z are independently selected from integers other than 0. For example, 1 ≤ x ≤ 10, 1 ≤ y ≤ 10, and 1 ≤ z ≤ 10. x, y, and z may or may not be equal, and this is not limited in the present invention. For example, if x = 1, y = 1, and z = 1, the structure of the fusion protein of the present invention is fragment A-fragment B or fragment B-fragment A.

[0015] When the number of fragment repeats is not 1, the sequences of the two repeated fragments can be the same or different, and the present invention does not limit this. For example, when y = 2, the two fragments B can target the same tissue or cell, or different tissues or cells, and the present invention does not limit this. When z = 2, the fragments in the two repeating units can be the same or different, and the present invention does not limit this. Taking x = 1, y = 2 and z = 2 as an example, the structure of the fusion protein of the present invention is:

[0016] [Fragment A-Fragment B2]2, that is: Fragment A-Fragment B-Fragment B-Fragment A-Fragment B-Fragment B;

[0017] Or [Fragment B-Fragment A2]2, that is: Fragment B-Fragment A-Fragment A-Fragment B-Fragment A-Fragment A.

[0018] In order to maintain the structural integrity of fragments A and B, the fusion protein may or may not include a linker, which is not limited in the present invention. In the present invention, the linker is located between two adjacent fragments, for example, between fragment A and fragment B, or between two fragments A, or between two fragments B, which is not limited in the present invention. Alternatively, the length of the linker is 2 to 20 amino acids, or 5 to 15 amino acids, or 10 to 15 amino acids, which is also not limited in the present invention. For example, the linker is (GGGGS)n, Gn, (EAAAK)n, (XP)n, A(EAAAK)4ALEA(EAAAK)4A, PAPAP, VSQTSKLTRAETVFPDV, PLGLWA, TRHRQPRGWE, AGNRVRRSVG, RVLAEA, EDVVCCSMSY, EDVVCCSMSY or GGIEGRGS, where n is a non-zero integer. Alternatively, 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.

[0019] In the present invention, fragment A targets the monomolecular phospholipid membrane. Since adipocytes have the remarkable characteristic of being monomolecular phospholipid membranes, fragment A can target the adipocytes.

[0020] In some embodiments, the monolayer phospholipid membrane targeted by fragment A comprises one or more of phospholipids, functional polar lipids, and cationic lipids;

[0021] The phospholipid is selected from one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, cardiolipin and sphingomyelin;

[0022] 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;

[0023] The cationic lipid is selected from one or more of (2,3-dioleoyl-propyl)-trimethylammonium-chloride, (2,3-dioleoyl-propyl)-trimethylamine, 2,3-dioleoyloxy-N-[2-(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propylamine hydrochloride, 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl] (nickel salt) and 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride.

[0024] Peptides targeting single-molecule phospholipid membranes are usually lipid droplet proteins, artificially designed peptides that can be localized on cellular lipid droplets, and a sequence in apolipoproteins. Apolipoproteins and peptides that can be localized on lipid droplets can be used as peptides (fragment A) that specifically recognize single-molecule phospholipid membranes. The following peptides can specifically bind to the surface of fat bodies in a non-covalent manner: including but not limited to 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.

[0025] Table 1 Name and amino acid sequence of fragment A

[0026] The amino acid sequence of fragment A of the present invention is as shown in any one of Table 1, or a sequence having at least 80% (preferably 85%, 90%, 95%, 98% or 99%) sequence identity with the amino acid sequence of the polypeptide described in Table 1.

[0027] In the present invention, fragment B targets and recognizes tissues or cells. The tissues or cells are derived from the human or animal body and may be tumors, pathogen-infected organs, lesions of metabolic diseases, and / or healthy tissues or cells. In the present invention, the choice of peptide segment for targeting specific tissues or cells depends on the specific protein on the surface of the tissue or cell being targeted. In some embodiments, fragment B includes, but is not limited to, at least one of Nrp-B, Trf-B, LDLR-B, ErbB2-B, CXCR4-B, GRP78-B, or Soma-B.

[0028] Table 2 Name and amino acid sequence of fragment B and its targeting protein

[0029] The amino acid sequence of fragment B of the present invention is shown in any one of Table 2, or a sequence having at least 80% (preferably 85%, 90%, 95%, 98% or 99%) sequence identity with the amino acid sequence of the polypeptide described in Table 2.

[0030] Alternatively, the fusion protein of the present invention is formed by connecting fragment A and fragment B. For example, the fusion protein is:

[0031] AAMB-Nrp-B、AAMB-Trf-B、AAMB-LDLR-B、AAMB-ErbB2-B、AAMB-CXCR4-B、AAMB-GRP78-B、AAMB-Soma-B、Nrp-B-AAMB、Trf-B-AAMB、LDLR-B-AAMB、ErbB2-B-AAMB、CXCR4-B-AAMB、GRP78-B-AAMB、Soma-B-AAMB、ALDI-Nrp-B、ALDI-Trf-B、ALDI-LDLR-B、ALDI-ErbB2-B、ALDI-CXCR4-B、ALDI-GRP78-B、ALDI-Soma-B、Nrp-B-ALDI、Trf-B-ALDI、LDLR-B-ALDI、ErbB2-B-ALDI、CXCR4-B-ALDI、GRP78-B-ALDI、Soma-B-ALDI、CYB5R3-N-Nrp-B、CYB5R3-N-Trf-B、CYB5R3-N-LDLR-B、CYB5R3-N-ErbB2-B、CYB5R3-N-CXCR4-B、CYB5R3-N-GRP78-B、CYB5R3-N-Soma-B、Nrp-B-CYB5R3-N、Trf-B-CYB5R3-N、LDLR-B-CYB5R3-N、ErbB2-B-CYB5R3-N、CXCR4-B-CYB5R3-N、GRP78-B-CYB5R3-N、Soma-B-CYB5R3-N、LDAMP1-Nrp-B、LDAMP1-Trf-B、LDAMP1-LDLR-B、LDAMP1-ErbB2-B、LDAMP1-CXCR4-B、LDAMP1-GRP78-B、LDAMP1-Soma-B、Nrp-B-LDAMP1、Trf-B-LDAMP1、LDLR-B-LDAMP1、ErbB2-B-LDAMP1、CXCR4-B-LDAMP1、GRP78-B-LDAMP1、Soma-B-LDAMP1、HSD17B13-N28-Nrp-B、HSD17B13-N28-Trf-B、HSD17B13-N28-LDLR-B、HSD17B13-N28-ErbB2-B、HSD17B13-N28-CXCR4-B、HSD17B13-N28-GRP78-B、HSD17B13-N28-Soma-B、Nrp-B-HSD17B13-N28、 Trf-B-HSD17B13-N28、LDLR-B-HSD17B13-N28、ErbB2-B-HSD17B13-N28、CXCR4-B-HSD17B13-N28、GRP78-B-HSD17B13-N28、Soma-B-HSD17B13-N28、MDT-28-P-Nrp-B、MDT-28-P-Trf-B、MDT-28-P-LDLR-B、MDT-28-P-ErbB2-B、MDT-28-P-CXCR4-B、MDT-28-P-GRP78-B、MDT-28-P-Soma-B、Nrp-B-MDT-28-P、Trf-B-MDT-28-P、LDLR-B-MDT-28-P、ErbB2-B-MDT-28-P、CXCR4-B-MDT-28-P、GRP78-B-MDT-28-P、Soma-B-MDT-28-P、MLDS-P-Nrp-B、MLDS-P-Trf-B、MLDS-P-LDLR-B、MLDS-P-ErbB2-B、MLDS-P-CXCR4-B、MLDS-P-GRP78-B、MLDS-P-Soma-B、Nrp-B-MLDS-P、Trf-B-MLDS-P、LDLR-B-MLDS-P、ErbB2-B-MLDS-P、CXCR4-B-MLDS-P、GRP78-B-MLDS-P、Soma-B-MLDS-P、DHS-3-P-Nrp-B、DHS-3-P-Trf-B、DHS-3-P-LDLR-B、DHS-3-P-ErbB2-B、DHS-3-P-CXCR4-B、DHS-3-P-GRP78-B、DHS-3-P-Soma-B、Nrp-B-DHS-3-P、Trf-B-DHS-3-P、LDLR-B-DHS-3-P、ErbB2-B-DHS-3-P、CXCR4-B-DHS-3-P、GRP78-B-DHS-3-P、Soma-B-DHS-3-P、HSD17B11-N28-Nrp-B、HSD17B11-N28-Trf-B、HSD17B11-N28-LDLR-B、HSD17B11-N28-ErbB2-B、HSD17B11-N28-CXCR4-B、HSD17B11-N28-GRP78-B、HSD17B11-N28-Soma-B、Nrp-B-HSD17B11-N28、Trf-B-HSD17B11-N28、LDLR-B-HSD17B11-N28、ErbB2-B-HSD17B11-N28、CXCR4-B-HSD17B11-N28、GRP78-B-HSD17B11-N28、Soma-B-HSD17B11-N28、PSPA-H1-Nrp-B、PSPA-H1-Trf-B、PSPA-H1-LDLR-B、PSPA-H1-ErbB2-B、PSPA-H1-CXCR4-B、PSPA-H1-GRP78-B、PSPA-H1-Soma-B、Nrp-B-PSPA-H1、Trf-B-PSPA-H1、LDLR-B-PSPA-H1、ErbB2-B-PSPA-H1、CXCR4-B-PSPA-H1、GRP78-B-PSPA-H1、Soma-B-PSPA-H1、VIPP1-H1-Nrp-B、VIPP1-H1-Trf-B、VIPP1-H1-LDLR-B、VIPP1-H1-ErbB2-B、VIPP1-H1-CXCR4-B、VIPP1-H1-GRP78-B、VIPP1-H1-Soma-B、Nrp-B-VIPP1-H1、Trf-B-VIPP1-H1、LDLR-B-VIPP1-H1、ErbB2-B-VIPP1-H1、CXCR4-B-VIPP1-H1、GRP78-B-VIPP1-H1、Soma-B-VIPP1-H1、SNF7-H1-Nrp-B、SNF7-H1-Trf-B、SNF7-H1-LDLR-B、SNF7-H1-ErbB2-B、SNF7-H1-CXCR4-B、SNF7-H1-GRP78-B、SNF7-H1-Soma-B、Nrp-B-SNF7-H1、Trf-B-SNF7-H1、LDLR-B-SNF7-H1、ErbB2-B-SNF7-H1、CXCR4-B-SNF7-H1、GRP78-B-SNF7-H1、Soma-B-SNF7-H1、CHMP1B-H1-Nrp-B、CHMP1B-H1-Trf-B、CHMP1B-H1-LDLR-B、CHMP1B-H1-ErbB2-B、CHMP1B-H1-CXCR4-B、CHMP1B-H1-GRP78-B、CHMP1B-H1-Soma-B、Nrp-B-CHMP1B-H1、Trf-B-CHMP1B-H1、LDLR-B-CHMP1B-H1、ErbB2-B-CHMP1B-H1、CXCR4-B-CHMP1B-H1、GRP78-B-CHMP1B-H1、Soma-B-CHMP1B-H1、PB-Nrp-B、PB-Trf-B、PB-LDLR-B、PB-ErbB2-B、PB-CXCR4-B、PB-GRP78-B、PB-Soma-B、Nrp-B-PB、Trf-B-PB、LDLR-B-PB、ErbB2-B-PB、CXCR4-B-PB、GRP78-B-PB, Soma-B-PB, PE-Nrp-B, PE-Trf-B, PE-LDLR-B, PE-ErbB2-B, PE-CXCR4-B, PE-GR P78-B, PE-Soma-B, Nrp-B-PE, Trf-B-PE, LDLR-B-PE, ErbB2-B-PE, CXCR4-B-PE, GRP78-B-PE , Soma-B-PE, PF-Nrp-B, PF-Trf-B, PF-LDLR-B, PF-ErbB2-B, PF-CXCR4-B, PF-GRP78-B, PF-S oma-B, Nrp-B-PF, Trf-B-PF, LDLR-B-PF, ErbB2-B-PF, CXCR4-B-PF, GRP78-B-PF, Soma-B-PF. ,

[0032] Furthermore, the present invention also provides a biomaterial related to the aforementioned fusion protein, including any of the following:

[0033] 1), a nucleic acid encoding the fusion protein as described above;

[0034] II), an expression unit comprising the nucleic acid described in I);

[0035] III), a vector containing the nucleic acid described in I) or the expression unit described in II);

[0036] IV), transforming or transfecting a host with the vector described in III);

[0037] V) and IV) the expression product of the host.

[0038] The present invention also provides a method for preparing the aforementioned fusion protein. The fusion protein can be prepared by genetic engineering or chemical synthesis.

[0039] In some embodiments, the preparation method comprises:

[0040] The method comprises: culturing the host as described above to obtain the fusion protein;

[0041] Or comprising: coupling fragment A and fragment B by biological and / or chemical methods;

[0042] Or comprising: synthesizing the fusion protein as described above by chemical synthesis.

[0043] The coupling may optionally include a linker.

[0044] The chemical synthesis method described in the present invention includes solid phase synthesis, liquid phase synthesis, and solid phase-liquid phase combined synthesis, which are not limited in the present invention.

[0045] The fusion protein described herein can specifically bind to the surface of adipocytes through a non-covalent mechanism, thereby achieving specific targeting of adipocytes. Targeted adipocytes constructed using this targeting peptide segment can be widely used in fields such as precision drug delivery and vaccine preparation, thereby treating diseases such as cancer, infectious diseases, and metabolic disorders. Based on this, the present invention also provides the use of the aforementioned fusion protein in the preparation of targeted adipocytes.

[0046] Furthermore, the present invention also provides a targeted fat body, the raw materials for preparing the fat body include: the fusion protein as described above, polar lipids and neutral lipids.

[0047] Optionally, the polar lipids include one or more of phospholipids, functional polar lipids and cationic lipids;

[0048] The phospholipid is selected from one or more of phosphatidylcholine, phosphatidylethanolamine, phosphatidylinositol, phosphatidylserine, phosphatidylglycerol, phosphatidic acid, cardiolipin and sphingomyelin;

[0049] 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;

[0050] The cationic lipid is selected from one or more of (2,3-dioleoyl-propyl)-trimethylammonium-chloride, (2,3-dioleoyl-propyl)-trimethylamine, 2,3-dioleoyloxy-N-[2-(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propylamine hydrochloride, 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl](nickel salt) and 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride;

[0051] The neutral lipid is selected from one or more of triglycerides, wax esters, sterol esters, sterol esters, retinol esters and fat-soluble vitamins.

[0052] Alternatively, the raw material preparation further comprises a solvent, and the solvent comprises an organic solvent and a buffer. The organic solvent includes but is not limited to methanol, ethanol, isopropanol, ether, chloroform, ethyl acetate, dichloromethane, chloroform, benzene, toluene, xylene, methane, acetonitrile, pyridine or phenol. The buffer includes but is not limited to citrate buffer, phosphate buffer, acetate buffer.

[0053] The targeted fat bodies described herein can deliver therapeutic drugs to target locations. Optionally, the raw materials used in the preparation also include drugs. In this invention, the drugs are substances used to prevent, treat, and diagnose diseases, including but not limited to drugs for preventing and treating cancer, infectious diseases, cardiovascular diseases, and metabolic diseases such as diabetes.

[0054] In addition, a marker with an indicator function, such as a fluorescent protein, may also be added.

[0055] Furthermore, the present invention also provides a method for preparing the targeted fat body, which comprises:

[0056] Dissolve the fusion protein as described above to prepare solution A;

[0057] Dissolve polar lipids to prepare solution B;

[0058] Solution A and solution B are mixed, dried, redissolved, mixed with neutral lipids, and then centrifuged to obtain a product containing targeted fat bodies.

[0059] In an embodiment of the present invention, the molar ratio of the polar lipid to the fusion protein is (100-100000): (1-10). In some embodiments, the molar ratio of the polar lipid to the fusion protein is (100-10000): (1-10), or (100-1000): (1-10), or (1000-100000): (1-10), or (10000-100000): (1-10), or 100: (1-10), or 1000: (1-10), or 10000:(1~10), or 100000:(1~10), or 100:1, or 1000:1, or 10000:1, or 100000:1, or 100:2, or 1000:2, or 10000:2, or 100:3, or 1000:3, or 10000:3, or 100 000:3, or 100:4, or 1000:4, or 10000:4, or 100000:4, or 100:5, or 1000:5, or 10000:5, or 100000:5, or 100:6, or 1000:6, or 10000:6, or 100000:6, or 100:7, or 1000:7, or It is 10000:7, or 100000:7, or 100:8, or 1000:8, or 10000:8, or 100000:8, or 100:9, or 1000:9, or 10000:9, or 100000:9, or 100:10, or 1000:10, or 10000:10, or 100000:10.

[0060] The solvent of solution A is a mixed solution of methanol and chloroform, wherein the volume ratio of methanol to chloroform is (1-10):(1-10). For example, the volume ratio is 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9 or 1:10. The concentration of the fusion protein in solution A is 0.1-10 mg / mL, for example, 0.1 mg / ml, 0.2 mg / ml, 0.3 mg / ml, 0.4 mg / ml, 0.5 mg / ml, 1 mg / ml, 2 mg / ml, 3 mg / ml, 4 mg / ml, 5 mg / ml, 6 mg / ml, 7 mg / ml, 8 mg / ml, 9 mg / ml, or 10 mg / ml.

[0061] The solvent of solution B is 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution. Each 80 μL of solution B contains 2 mg of phospholipid. The volume ratio of solution A to solution B is 1:4.

[0062] The re-dissolution step and / or the centrifugal separation step may be performed with reference to Chinese patent application number 201510974956.6.

[0063] The method for preparing the targeted fat body of the present invention has simple and concise steps, does not require complicated instruments, and has strong operability.

[0064] Furthermore, the present invention also provides the use of the aforementioned targeted fat bodies or the targeted fat bodies prepared by the aforementioned preparation method in the preparation of drugs or vaccines for preventing and treating diseases.

[0065] In some embodiments, the disease is cancer, a pathogenic infectious disease, or a metabolic disease. The disease is preferably cancer, and the cancer is selected from the group consisting of breast cancer, lung cancer, kidney cancer, laryngeal cancer, liver cancer, muscle tissue cancer, blood cancer, bone cancer, brain cancer, neck cancer, oral or nasal mucosal cancer, bladder cancer, central nervous system cancer, cervical 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, testicular cancer, and / or thyroid cancer.

[0066] Furthermore, the present invention provides a drug or vaccine for preventing or treating a disease, comprising a pharmaceutically acceptable excipient and the targeted adipocytes described above or the targeted adipocytes prepared by the aforementioned preparation method. The drug or vaccine of the present invention is in the form of an oral preparation, an inhalant, or an injection.

[0067] 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.

[0068] In some embodiments provided herein, the capsule is a hard capsule or a soft capsule.

[0069] In some embodiments provided herein, the tablet is an oral tablet or buccal tablet.

[0070] 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.

[0071] 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.

[0072] The medicine or vaccine is an injection, for example, an injection solution or an injection powder.

[0073] The present invention also provides a method for preventing and treating a disease, comprising administering the aforementioned drug or vaccine, wherein the administration comprises oral administration, inhalation, and / or injection.

[0074] The present invention connects a polypeptide fragment that targets and recognizes single-molecule phospholipid membranes with a polypeptide fragment that targets and recognizes tissues or cells. The resulting fusion protein can be specifically and non-covalently bound to the surface of fat bodies, thereby achieving precise targeting of fat bodies. Targeted fat bodies constructed using this fat body-targeting peptide segment can be widely used in fields such as precision drug delivery and vaccine preparation, thereby being used for the prevention, treatment, and diagnosis of diseases such as cancer, infectious diseases, and metabolic diseases. In the examples of the present invention, lung tissue and breast cancer cells were used as targets, achieving good targeting of fat bodies. BRIEF DESCRIPTION OF THE DRAWINGS

[0075] Figure 1 shows a schematic diagram of the adipocyte-targeted peptide technology;

[0076] Figure 2 shows the preparation of the adipocyte-targeting peptide LTA-P33;

[0077] FIG3 shows the construction of a fat body containing the fat body targeting peptide LTA-P33, wherein:

[0078] A shows the analysis of fat bodies containing the fat body targeting peptide LTA-P33 by SDS-PAGE protein gel electrophoresis. The arrow in the LTA fat body lane indicates the fat body targeting peptide LTA-P33 adsorbed on the fat body.

[0079] B shows the confocal laser microscopy results of adipocytes containing the adipocyte-targeting peptide LTA-P33. The bright ring-shaped structure shown in a is the adipocyte-targeting peptide LTA-P33. b shows a adipocyte stained red with the neutral lipid-specific dye LipidTox Red (shown in light gray). The bright ring-shaped structure on the surface is the adipocyte-targeting peptide LTA-P33. The scale bar is 5 microns. Silver staining and confocal laser microscopy results indicate the successful construction of adipocytes containing the adipocyte-targeting peptide LTA-P33, namely, lung-targeted adipocytes.

[0080] FIG4 shows a fat body targeting peptide LTA-P33 targeting the lungs, wherein:

[0081] A shows the expression level of protein Neuropilin-1 in heart, lung, spleen, kidney and liver tissues;

[0082] B shows that fluorescently labeled LTA-free fat bodies (fat bodies without the fat body targeting peptide LTA-P33) and LTA fat bodies (fat bodies with the fat body targeting peptide LTA-P33) were injected into mice through the tail vein. Three hours after injection, various tissues were dissected and identified; a is the distribution of LTA-free fat bodies in various tissues; b is the distribution of LTA fat bodies in various tissues;

[0083] C shows the fluorescence statistics of LTA-free fat bodies and LTA fat bodies in lung tissue relative to the blank group;

[0084] D shows the distribution of LTA-free fat bodies and LTA fat bodies in various tissues 24 hours (a) and 54 hours (b) after injection;

[0085] FIG5 shows the preparation of the adipocyte-targeting peptide TRTA-P33;

[0086] FIG6 shows the construction of a fat body containing the fat body targeting peptide TRTA-P33; wherein:

[0087] A shows that fat bodies containing the fat body targeting peptide TRTA-P33 were analyzed by SDS-PAGE protein gel electrophoresis. The arrow in the TRTA fat body lane indicates the fat body targeting peptide TRTA-P33 adsorbed on the fat body;

[0088] B shows confocal laser scanning microscopy of a fat body containing the adipocyte-targeting peptide TRTA-P33. The bright ring-shaped structure shown in a is the adipocyte-targeting peptide TRTA-P33. b shows a fat body stained red with the neutral lipid-specific dye LipidToxRed (shown in light gray). The bright ring-shaped structure on the surface is the adipocyte-targeting peptide TRTA-P33. Scale bar is 5 μm. Silver staining and confocal laser scanning microscopy results demonstrate the successful construction of a fat body containing the adipocyte-targeting peptide TRTA-P33, i.e., a transferrin receptor-targeted fat body.

[0089] FIG7 shows a fat body targeting peptide TRTA-P33 targeting MCF7 breast cancer cells with high transferrin receptor expression; wherein:

[0090] A shows: the expression level of Transferrin receptor 1 protein in different cancer cells;

[0091] Figure B shows: laser confocal microscopy results of fat bodies without the fat body targeting peptide TRTA-P33 (fat bodies without TRTA, a) and fat bodies with the fat body targeting peptide TRTA-P33 (fat bodies with TRTA, b) after incubation with MCF7 cells; bright dots are fat bodies containing fluorescent labels; scale bar is 5 μm; a shows the signal intensity of fluorescently labeled fat bodies in MCF7 cells; b shows the statistical results of the signal intensity of fluorescently labeled fat bodies in MCF7 cells;

[0092] C shows the flow cytometry results of TRTA-free fat bodies (a) and TRTA fat bodies (b) after incubation with MCF7 cells. DETAILED DESCRIPTION

[0093] The present invention provides fusion proteins and uses thereof. Those skilled in the art can refer to the contents of this article and appropriately improve the process parameters to achieve the desired results. 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, and relevant personnel can obviously 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.

[0094] 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.

[0095] 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.

[0096] 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.

[0097] 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.

[0098] The term "nucleic acid" herein refers to a nucleic acid encoding a fusion protein as described above, which may be single-stranded or double-stranded, and may be DNA, RNA, cDNA or PNA. The nucleic acid may include nucleotide sequences having different functions, such as coding regions and non-coding regions such as regulatory sequences (e.g., promoters or transcription terminators). The nucleic acid may be linear or circular in topology. The nucleic acid may be, for example, a portion of a vector (e.g., an expression or cloning vector), or a fragment or a combination of multiple fragments. The nucleic acid may be obtained directly from a natural source, or may be prepared by recombinant, enzymatic or chemical techniques. The RNA form may be mRNA obtained by gene transcription, etc.

[0099] The term "expression unit" herein includes the nucleic acid, promoter, and terminator of the present invention. In order to regulate the expression level or protein folding, the expression unit may also include other elements.

[0100] The term "vector" herein refers to a nucleic acid molecule capable of amplifying another nucleic acid to which it is linked. The term includes vectors that are self-replicating nucleic acid structures as well as vectors that integrate into the genome of a host cell into which the vector has been introduced. Certain vectors are capable of directing the expression of nucleic acids to which they are operably linked. Such vectors are referred to herein as "expression vectors." In the present invention, the vectors are used for amplification and preservation of nucleic acids, or for expression of mutants in a host. The vectors described in the present invention include a backbone and a nucleic acid. Specifically, the backbone may be derived from plants, animals, bacteria, fungi, bacteriophages, or viruses, although this is not a limitation of the present invention.

[0101] The term "host cell" herein refers to a cell into which an exogenous nucleic acid has been introduced, including the progeny of such a cell. Host cells include "transformants" and "transformed cells," which include the original transformed cell and its progeny, regardless of the number of passages. Progeny may not be completely identical to the parent cell in nucleic acid content, but may contain mutations. Mutant progeny having the same function or biological activity as that screened or selected for in the initially transformed cell are included herein.

[0102] 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.

[0103] 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.

[0104] 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.

[0105] The test materials used in the present invention are all common commercial products and can be purchased in the market.

[0106] In the embodiment of the present invention, the following two combinations were used to verify the fat body targeting peptide segment: 1) Fragment A selected the peptide segment AAMB in Table 1 that specifically recognizes the single-molecule phospholipid membrane, and fragment B selected the peptide segment Nrp-B in Table 2 that targets the highly expressed protein Neuropilin-1 in lung tissue. The fat body targeting peptide segment LTA-P33 (lung-targeted adiposome peptide 33) was constructed by chemical synthesis, and the sequence was: MELTIFILRLAIYILTFPLYLLNFLGLWCRGDK. The fat body constructed by this peptide segment LTA-P33 can specifically target lung tissue. 2) Fragment A selected the peptide PspA-H1 in Table 1 that specifically recognizes single-molecule phospholipid membranes, and fragment B selected the peptide Trf-B in Table 2 that targets the Transferrin receptor 1 protein highly expressed in MCF7 breast cancer cells. The adiposome-targeting peptide TRTA-P33 (Transferrin receptor 1-targeted adiposome peptide 33) was constructed by chemical synthesis. The sequence is MGIFSRFADIVNANINALLEKTHRPPMWSPVWP. The adiposome constructed by this peptide TRTA-P33 can specifically target MCF7 breast cancer cells with highly expressed transferrin receptors.

[0107] The present invention will be further described below in conjunction with the embodiments:

[0108] Example 1: Adipocyte-specific targeting of lung tissue by adipocyte-targeting peptide LTA-P33

[0109] 1. Preparation of the adipocyte-targeting peptide LTA-P33

[0110] First, the adiposome-targeted peptide LTA-P33 (lung-targeted adiposome peptide 33) was constructed through chemical synthesis by Shanghai Jier Peptide Co., Ltd. The sequence is: MELTIFILRLAIYILTFPLYLLNFLGL WCRGDK, where the amino acid sequence MELTIFILRLAIYILTFPLYLLNFLGLW represents AAMB and the amino acid sequence CRGDK represents Nrp-B, which specifically recognizes the protein Neuropilin-1 (UniProt ID: P97333). The HPLC (high-performance liquid chromatography) results of the chemically synthesized adiposome-targeted peptide LTA-P33 are shown by the arrow, with a peak elution time of 12.5 minutes and a purity of 95.7%.

[0111] 2. Construction of Fat Bodies Containing the Fat Bodies-Targeting Peptide LTA-P33

[0112] First, adipocytes containing the adipocyte targeting peptide LTA-P33 were constructed according to the following method, and then the constructed adipocytes containing the adipocyte targeting peptide LTA-P33 were further verified by silver staining and laser confocal microscopy.

[0113] (1) Dissolve LTA-P33 in a mixture of methanol and chloroform (1:1, v / v) to a final concentration of 0.5 mg / ml.

[0114] (2) Take 20 μl of the solution in (1) and 80 μl of 2-di-(9Z-octadecenoyl)-sn-glycero-3-phosphocholine solution (containing 2 mg DOPC), mix them evenly, and add the total volume of 100 μl into a microcentrifuge tube. Blow dry the solvent with high-purity nitrogen gas.

[0115] (3) After completing step (2), add 100 μl of phosphate buffered saline (PBS) and 5 mg of triglyceride (TAG) or other neutral lipids to a microcentrifuge tube and vortex for 5 min (vortex for 10 s, stop for 5 s) to obtain a milky white lipid mixture 1 (i.e., the initial preparation component). Centrifuge the lipid mixture 1 at 1000 g for 5 min to remove the upper white band and the precipitate at the bottom of the tube.

[0116] (4) After completing step (3), the lipid mixture 1 is centrifuged at 20,000 g for 5 min (in practice, 18,000-22,000 g for 3-7 min is acceptable). After centrifugation, the sediment at the bottom of the tube is removed and the remaining emulsion in the tube is resuspended to obtain a milky white lipid mixture 2.

[0117] (5) After completing step (4), repeat step 3 again to obtain the final fat body containing the fat body targeting peptide LTA-P33 (abbreviated as LTA fat body, i.e. lung-targeted fat body, Figure 3).

[0118] Figure 3, A, shows SDS-PAGE protein gel electrophoresis analysis of adipocytes containing the adipocyte-targeting peptide LTA-P33. The arrow in the LTA adipocyte lane indicates adipocyte-targeting peptide LTA-P33 adsorbed onto adipocytes. Figure 3, B, shows confocal laser scanning microscopy of adipocytes containing the adipocyte-targeting peptide LTA-P33. The bright ring-like structure shown in Figure A is the adipocyte-targeting peptide LTA-P33. Figure 3, B, shows adipocytes stained red with the neutral lipid-specific dye LipidTox Red (shown in light gray). The bright ring-like structure on the surface is the adipocyte-targeting peptide LTA-P33. Scale bar is 5 μm. Silver staining and confocal laser scanning microscopy results demonstrate the successful construction of adipocytes containing the adipocyte-targeting peptide LTA-P33, i.e., lung-targeted adipocytes.

[0119] 3. Validation of a lung-targeted adipocyte targeting peptide containing LTA-P33 (abbreviated as LTA adipocyte, i.e. lung-targeted adipocyte)

[0120] The adipocyte-targeting peptide LTA-P33 specifically adsorbs to the adipocyte surface via fragment A and recognizes the specific protein Neuropilin-1 via fragment B, thereby achieving adipocyte-specific targeting. The amino acid sequence of fragment B is CRGDK, which specifically recognizes the protein Neuropilin-1 (UniProt ID: P97333). Neuropilin-1 is highly expressed in the lungs, but is expressed at low levels in other tissues such as the heart and liver (Figure 4A). We injected mice with adipocytes without the adipocyte-targeting peptide LTA-P33 (LTA-free adipocytes) and with the LTA-targeting peptide LTA-P33 (LTA-free adipocytes). Three hours after injection, tissues were dissected and analyzed to determine the tissue distribution of LTA-free and LTA-free adipocytes. Almost all LTA-free fat bodies accumulated in the liver (Figure 4, Ba), while almost all LTA fat bodies accumulated in the lung tissue (Figure 4, Bb). Lung fluorescence statistics showed that LTA fat bodies had specific targeting properties and could be targeted to the lung tissue (Figure 4, C). Furthermore, LTA fat bodies were still retained in the lung tissue 24 hours after injection (Figure 4, D).

[0121] Example 2: Adipocyte-specific targeting of MCF7 breast cancer cells with high transferrin receptor expression using the adipocyte-targeted peptide TRTA-P33

[0122] 1. Preparation of the adipocyte-targeting peptide LTA-P33

[0123] First, the adiposome-targeting peptide TRTA-P33 (Transferrin receptor 1-targeted adiposome peptide 33) was constructed through chemical synthesis by Shanghai Jier Peptide Co., Ltd. The peptide sequence is MGIFSRFADIVNANINALLEKTHRPPMWSPVWP. Adiposomes constructed with this peptide, TRTA-P33, can specifically target MCF7 breast cancer cells, which overexpress transferrin receptors. The amino acid sequence MGIFSRFADIVNANINALLEK represents PspA-H1, and the amino acid sequence THRPPMWSPVWP represents Trf-B, a peptide that specifically recognizes the Transferrin receptor 1 protein (UniProtID: P02786). HPLC (high-performance liquid chromatography) analysis of the chemically synthesized adiposome-targeting peptide TRTA-P33 shows a peak peak (arrowed) at 12.8 minutes and a purity of 97.6%.

[0124] 2. Construction of adipocytes containing the adipocyte-targeting peptide TRTA-P33 (abbreviated as TRTA adipocytes, i.e. transferrin receptor-targeted adipocytes).

[0125] The preparation method is the same as that in Example 1, and the final fat body containing the fat body targeting peptide TRTA-P33 is obtained (Figure 6). In Figure 6, A is the SDS-PAGE protein gel electrophoresis analysis of the fat body containing the fat body targeting peptide TRTA-P33. The arrow in the TRTA fat body lane indicates the fat body targeting peptide TRTA-P33 adsorbed on the fat body. B is the laser confocal microscopy result of the fat body containing the fat body targeting peptide TRTA-P33. The bright ring-shaped structure shown in a is the fat body targeting peptide TRTA-P33. b is a fat body stained red by the neutral lipid-specific dye LipidTox Red (light gray in the figure), and the bright ring-shaped structure on the surface is the fat body targeting peptide TRTA-P33. The scale bar is 5 microns. The results of silver staining and laser confocal microscopy show that the fat body containing the fat body targeting peptide TRTA-P33, that is, the transferrin receptor targeted fat body, was successfully constructed.

[0126] 3. Validation of the adipocyte targeting peptide TRTA-P33 targeting MCF7 breast cancer cells with high transferrin receptor expression (abbreviated as TRTA adipocytes, i.e. transferrin receptor targeted adipocytes)

[0127] The adipocyte-targeting peptide TRTA-P33 specifically adsorbs to the adipocyte surface through its fragment A sequence and recognizes the specific protein Transferrin receptor 1 through its fragment B, thus achieving adipocyte-specific targeting. The amino acid sequence of fragment B is THRPPMWSPVWP, which specifically recognizes the protein Transferrin receptor 1 (UniProtID: P02786). Transferrin receptor 1 is highly expressed in MCF7 breast cancer cells, but is expressed at low levels in 231 and 4T1 cancer cells (Figure 7A). Adipocytes without the TRTA-targeting peptide TRTA-P33 (TRTA-free adipocytes) and with the TRTA-targeting peptide (TRTA adipocytes) were incubated with MCF7 cells. After two hours, the culture medium was removed and the cells were washed three times with PBS. The cells were then observed under laser confocal microscopy and fluorescence signal was analyzed by flow cytometry. TRTA-free adipocytes were barely visible in MCF7 cells (Figure 7B), while TRTA adipocytes clearly accumulated in MCF7 cells (Figure 7Bb). At the same time, flow cytometry results showed that the fluorescence peak of MCF7 cells treated with TRTA fat bodies shifted significantly to the right (Figure 7, Ca). Statistically, the signal of TRTA fat bodies in MCF7 cells was significantly higher than that of fat bodies without TRTA (Figure 7, Cb). These results indicate that fat bodies containing the fat body targeting peptide TRTA-P33 specifically target MCF7 breast cancer cells with high transferrin receptor expression.

[0128] 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 fusion protein comprising fragment A and fragment B; in: Fragment A targets and recognizes the monomolecular phospholipid membrane; Fragment B targets tissues or cells for recognition.

2. The fusion protein according to claim 1, characterized in that Its structure is: [(fragment A)x-(fragment B)y]z or [(fragment B)x-(fragment A)y]z; Wherein: x, y, z are independently selected from integers other than 0.

3. The fusion protein according to claim 1 or 2, characterized in that This also includes the linker fragment.

4. The fusion protein according to claim 3, characterized in that The linker is selected from (GGGGS)n, Gn, (EAAAK)n, (XP)n, A(EAAAK)4ALEA(EAAAK)4A, PAPAP, VSQTSKLTRAETVFPDV, PLGLWA, TRHRQPRGWE, AGNRVRRSVG, RVLAEA, EDVVCCSMSY, EDVVCCSMSY or GGIEGRGS, where n is a non-zero integer.

5. The fusion protein according to any one of claims 1 to 4, characterized in that The monomolecular phospholipid membrane contains one or more of phospholipids, functional polar lipids and cationic lipids; The phospholipid is selected from one or more of 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; The cationic lipid is selected from one or more of (2,3-dioleoyl-propyl)-trimethylammonium-chloride, (2,3-dioleoyl-propyl)-trimethylamine, 2,3-dioleoyloxy-N-[2-(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propylamine hydrochloride, 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl](nickel salt) and 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride.

6. The fusion protein according to any one of claims 1 to 4, characterized in that The fragment A is 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.

7. The fusion protein according to any one of claims 1 to 4, characterized in that The tissues or cells are from the human or animal body: tumors, organs infected by pathogens, lesions of metabolic diseases and / or healthy tissues or cells.

8. The fusion protein according to claim 7, characterized in that The fragment B is at least one of Nrp-B, Trf-B, LDLR-B, ErbB2-B, CXCR4-B, GRP78-B or Soma-B.

9. The fusion protein according to any one of claims 1 to 4, characterized in that: It is AAMB-Nrp-B or PspA-H1-Trf-B.

10. Biological materials: including any of the following: I), a nucleic acid encoding the fusion protein according to any one of claims 1 to 9; II), an expression unit containing the nucleic acid described in I); III), a vector containing the nucleic acid described in I) or the expression unit described in II); IV), transforming or transfecting a host with the vector described in III); V) and IV) the expression product of the host.

11. A method for preparing the fusion protein according to any one of claims 1 to 9, It is characterized in that The method comprises: culturing the host according to claim 10 to obtain the fusion protein; Or comprising: coupling fragment A and fragment B by biological and / or chemical methods; Or comprising: synthesizing the fusion protein according to any one of claims 1 to 9 by chemical synthesis.

12. Use of the fusion protein according to any one of claims 1 to 9 in preparing targeted fat bodies.

13. Targeted fat body, the raw materials for its preparation include: The fusion protein, polar lipid and neutral lipid according to any one of claims 1 to 9.

14. The targeted fat body according to claim 13, characterized in that: The polar lipids include one or more of phospholipids, functional polar lipids and cationic lipids; The phospholipid is selected from one or more of 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; The cationic lipid is selected from one or more of (2,3-dioleoyl-propyl)-trimethylammonium-chloride, (2,3-dioleoyl-propyl)-trimethylamine, 2,3-dioleoyloxy-N-[2-(sperminecarboxamido)ethyl]-N,N-dimethyl-1-propylamine hydrochloride, 1,2-dioleoyl-sn-glycero-3-[(N-(5-amino-1-carboxypentyl)iminodiacetic acid)succinyl](nickel salt) and 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol hydrochloride; The neutral lipid is selected from one or more of triglycerides, wax esters, sterol esters, sterol esters, retinol esters and fat-soluble vitamins.

15. The targeted fat body according to claim 13 or 14, characterized in that: The preparation raw materials also include a solvent, and the solvent includes an organic solvent and a buffer solution.

16. The targeted fat body according to claim 13 or 14, characterized in that: The preparation raw materials also include drugs.

17. The method for preparing targeted fat bodies according to any one of claims 13 to 16, characterized in that: It includes: Dissolving the fusion protein according to any one of claims 1 to 9 to prepare solution A; Dissolving polar lipids to prepare solution B; Solution A and solution B are mixed, dried, redissolved, mixed with neutral lipids, and then centrifuged to obtain a product containing targeted fat bodies.

18. The preparation method according to claim 17, characterized in that: The molar ratio of the polar lipid to the fusion protein is (100-100000): (1-10).

19. Use of the targeted fat body according to any one of claims 13 to 16 or the targeted fat body prepared by the preparation method according to claim 17 or 18 in the preparation of drugs or vaccines for preventing and treating diseases.

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

21. A medicine or vaccine comprising a pharmaceutically acceptable excipient and the targeted fat body according to any one of claims 13 to 16 or the targeted fat body prepared by the preparation method according to claim 17 or 18.

22. The drug or vaccine according to claim 21, characterized in that Its dosage form is oral preparation, inhalation or injection; The oral preparation is a tablet, a pill, an oral liquid, a capsule, a syrup, a drop pill or a granule; The inhalation agent is an inhalation aerosol, an inhalation powder, and a liquid preparation for use in a nebulizer; The injection is a liquid injection or a powder injection.

23. A method for preventing and treating a disease, comprising administering the drug or vaccine according to claim 22.

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