Electuary containing natural plant lipid nano vesicles and large-scale preparation method

Abstract

The invention relates to an electuary containing natural plant lipid nano-vesicles and a large-scale preparation method. The electuary comprises the natural plant lipid nano-vesicles used for carrying bioactive small molecules. The electuary containing the natural plant lipid nano-vesicles is prepared from plants, the natural plant lipid nano-vesicles in the electuary are loaded with bioactive small molecules, the problems that the bioactive small molecules are poor in stability, low in absorption efficiency, easy to decompose and the like in gastrointestinal tracts can be solved, and the nano-vesicles have the effects of resisting inflammation, resisting cancer and the like; the invention provides a method for preparing natural plant lipid nano-vesicles on a large scale, which can solve the problems of long preparation time and low yield of the natural plant nano-vesicles and reduce the production cost.

Description

Technical Field

[0001] The invention relates to the technical field of health care products and functional foods, and more specifically to an granule containing natural plant lipid nanovesicles and a large-scale preparation method thereof. Background Art

[0002] Bioactive small molecules refer to a variety of small molecules with high reactivity that are synthesized by living organisms themselves. Edible plants and medicinal plants contain a variety of bioactive small molecules, such as catechins, epicatechin, epigallocatechin gallate, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanins, proanthocyanidins, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, naringin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, etc. These ingredients have been proven to have multiple nutritional values. However, most bioactive small molecules have the disadvantages of being easily oxidized, easily decomposed, having poor stability or being almost insoluble in water.

[0003] Tea contains a variety of active ingredients, such as epigallocatechin gallate (EGCG), bioactive proteins and polysaccharides. EGCG is a polyphenolic compound and a catechin monomer isolated from tea. EGCG has a variety of biological activities, including anti-inflammatory, antioxidant, anti-apoptotic, hypoglycemic, antibacterial, anti-cancer, neuroprotective, etc., and has been widely used in nutritional health care products. However, EGCG has the defects of unstable chemical properties, easy oxidation and decomposition, and low bioavailability. It cannot be directly taken orally, which limits the application of EGCG. In order to achieve the controlled release of EGCG and improve its stability and bioavailability, various biomaterials are used to load EGCG, such as proteins, polysaccharides, lipids, and inorganic materials;

[0004] Natural plant lipid nanovesicles can be isolated from various freshly squeezed fruit juices, vegetable juices, and Chinese herbal juices. Their morphology and composition are similar to mammalian exosomes. Natural plant lipid nanovesicles are loaded with a variety of bioactive ingredients such as proteins, polysaccharides, various secondary metabolites, and small RNAs, and have multiple biological functions. The common method for isolating natural plant lipid nanovesicles is density gradient separation by differential ultracentrifugation. In addition, there are also PEG precipitation method, size exclusion chromatography, kit, ultrafiltration membrane separation, etc. These methods have problems such as low extraction yield, long time consumption, and complex preparation process, and are not suitable for large-scale production.

[0005] Therefore, there is a need for a granule containing natural plant lipid nanovesicles and a large-scale preparation method that can solve the problems of long preparation time and low yield of natural plant lipid nanovesicles, poor stability of bioactive small molecules in the gastrointestinal tract, low absorption efficiency, and easy decomposition. Summary of the invention

[0006] The technical problem to be solved by the present invention is to provide an granule containing natural plant lipid nanovesicles and a large-scale preparation method of the natural plant lipid nanovesicles in view of the above-mentioned defects of the prior art.

[0007] The technical solution adopted by the present invention to solve its technical problem is:

[0008] A granule containing natural plant lipid nanovesicles is constructed, wherein the granule contains natural plant lipid nanovesicles for carrying bioactive small molecules.

[0009] The granules containing natural plant lipid nanovesicles of the present invention, wherein the natural plant lipid nanovesicles in the granules are loaded with biologically active small molecules.

[0010] The granules containing natural plant lipid nanovesicles of the present invention, wherein the natural plant lipid nanovesicles in the granules are prepared by the following method:

[0011] Collect plants and store them at set temperatures;

[0012] After washing the plants, place them in a grinder, add ddH2O at a set ratio, and grind them;

[0013] The grinding liquid is centrifuged and the supernatant is collected;

[0014] The supernatant is filtered through a filter membrane with a set pore size to obtain natural plant lipid nanovesicles;

[0015] The obtained natural plant lipid nanovesicles are freeze-dried and stored at a set temperature.

[0016] The granules containing natural plant lipid nanovesicles of the present invention have a set ratio of (2-4):1 and a grinding time of 4-5 minutes.

[0017] The granules containing natural plant lipid nanovesicles of the present invention, wherein the pore size of the filter membrane is 0.22 or 0.45 μm.

[0018] The granules containing natural plant lipid nanovesicles of the present invention are characterized in that the storage temperature of the plants is -20 degrees; and the freeze-dried storage temperature of the natural plant lipid nanovesicles is 4 degrees.

[0019] The granules containing natural plant lipid nanovesicles of the present invention, wherein the plants are one or more of tea, mulberry, tea, camellia, ginger, tomato, grape, grapefruit, lemon, apple, broccoli, carrot, onion, turmeric, strawberry, shiitake mushroom, salvia miltiorrhiza, ginseng, bitter melon, cabbage, aloe, corn, watermelon, astragalus, blueberry, garlic, soybean, cantaloupe, pear, pea, coconut, kiwi, citrus, sunflower, wheat, cherry, rehmannia, knotweed, and gastrodia.

[0020] The granules containing natural plant lipid nanovesicles of the present invention, wherein the bioactive small molecules include one or more of catechins, gallocatechin, catechin gallate, gallocatechin gallate, epicatechin, epigallocatechin gallate, epigallocatechin, epicatechin gallate, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanidins, proanthocyanidins, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, naringin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, isorhamnetin, and hyperoside.

[0021] A large-scale preparation method of natural plant lipid nanovesicles is applied to the above-mentioned granules containing natural plant lipid nanovesicles, which comprises the following steps:

[0022] Collect plants and store them at set temperatures;

[0023] After washing the plants, place them in a grinder, add ddH2O at a set ratio, and grind them;

[0024] The grinding liquid is centrifuged and the supernatant is collected;

[0025] The supernatant is filtered through a filter membrane with a set pore size to obtain natural plant lipid nanovesicles;

[0026] The obtained natural plant lipid nanovesicles are freeze-dried and stored at a set temperature.

[0027] The large-scale preparation method of natural plant lipid nanovesicles of the present invention, wherein the set ratio is (2-4): 1, the grinding time is 4-5 minutes; the pore size of the filter membrane is 0.22 or 0.45 μm; the storage temperature of the plant is -20 degrees; the freeze-dried storage temperature of the natural plant lipid nanovesicles is 4 degrees;

[0028] The plant is one or more of tea, mulberry, tea, camellia, ginger, tomato, grape, grapefruit, lemon, apple, broccoli, carrot, onion, turmeric, strawberry, mushroom, salvia miltiorrhiza, ginseng, bitter melon, cabbage, aloe, corn, watermelon, astragalus, blueberry, garlic, soybean, cantaloupe, pear, pea, coconut, kiwi, citrus, sunflower, wheat, cherry, rehmannia root, knotweed, and gastrodia elata;

[0029] The bioactive small molecules include one or more of catechins, gallocatechins, catechin gallates, gallocatechin gallates, epicatechins, epigallocatechin gallates, epigallocatechins, epicatechin gallates, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanins, proanthocyanidins, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, nobiletin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, isorhamnetin, and hyperoside.

[0030] The beneficial effects of the present invention are as follows: the present invention utilizes plants to prepare an granule containing natural plant lipid nanovesicles, and the natural plant lipid nanovesicles in the granule are loaded with bioactive small molecules, which can solve the problems of poor stability, low absorption efficiency, and easy decomposition of the bioactive small molecules in the gastrointestinal tract; the nanovesicles have anti-inflammatory, anti-cancer and other effects. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the present invention will be further described below in conjunction with the accompanying drawings and embodiments. The drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative work:

[0032] Figure 1a This is a schematic diagram of the determination of tea natural lipid nanovesicles loaded with bioactive small molecules in gastric simulated fluid;

[0033] Figure 1b This is a schematic diagram of the determination of tea natural lipid nanovesicles loaded with bioactive small molecules in small intestine simulated fluid;

[0034] Figure 1c This is a schematic diagram of the determination of tea natural lipid nanovesicles loaded with bioactive small molecules in colon simulated fluid;

[0035] Figure 2 It is the chromatogram of natural lipid nanovesicles from tea leaves;

[0036] Figure 3 The pictures are freeze-dried powders of tea natural lipid nanovesicles prepared by two preparation methods;

[0037] Figure 4 Schematic diagram of the anti-inflammatory activity assay of tea natural lipid nanovesicles prepared by two preparation methods;

[0038] Figure 5 Schematic diagram of the anticancer activity assay of tea natural lipid nanovesicles prepared by two preparation methods;

[0039] Figure 6 It is a flow chart of the large-scale preparation method of natural plant lipid nanovesicles. DETAILED DESCRIPTION

[0040] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the following will be described clearly and completely in combination with the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are partial embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work belong to the protection scope of the present invention.

[0041] Embodiment 1

[0042] The granules containing natural plant lipid nanovesicles in a preferred embodiment of the present invention contain natural plant lipid nanovesicles for carrying bioactive small molecules;

[0043] Preferably, the natural plant lipid nanovesicles in the granules are loaded with biologically active small molecules.

[0044] The present invention utilizes plants to prepare an granule containing natural plant lipid nanovesicles. The natural plant lipid nanovesicles in the granule are loaded with bioactive small molecules. Using the natural plant lipid nanovesicles as carriers to load the bioactive small molecules can solve the problems of poor stability, low absorption efficiency, and easy decomposition of the bioactive small molecules in the gastrointestinal tract; the nanovesicles have anti-inflammatory, anti-cancer and other effects.

[0045] Preferably, the natural plant lipid nanovesicles are prepared by:

[0046] Collect fresh plants and store them at -20°C;

[0047] Wash the fresh plants with water and place them in a grinder. Add ddH2O in a ratio of 3:1 and grind for 4-5 minutes.

[0048] The grinding solution was centrifuged at 12000g for 1.5h, and the supernatant was collected;

[0049] The supernatant was filtered through a 0.45 μm filter membrane;

[0050] The obtained natural plant lipid nanovesicles are freeze-dried;

[0051] The obtained natural plant lipid nanovesicle freeze-dried powder was stored at 4°C.

[0052] The natural plant lipid nanovesicles prepared by the above method are used as carriers to load bioactive small molecules, which can solve the problems of poor stability, low absorption efficiency, and easy decomposition of bioactive small molecules in the gastrointestinal tract; experimental verification shows that the natural plant lipid nanovesicles prepared by the above method also have anti-inflammatory and anti-cancer effects.

[0053] It should be noted that the above-mentioned components are only used for illustrative purposes and are not intended to be limiting. For those skilled in the art, they can be easily modified into other proportions. Simple modification of the proportions based on this principle also falls within the scope of protection of this application.

[0054] Preferably, the plant of the present invention is one or more of tea, mulberry, tea, camellia, ginger, tomato, grape, grapefruit, lemon, apple, broccoli, carrot, onion, turmeric, strawberry, shiitake mushroom, salvia miltiorrhiza, ginseng, bitter melon, cabbage, aloe, corn, watermelon, astragalus, blueberry, garlic, soybean, cantaloupe, pear, pea, coconut, kiwi, citrus, sunflower, wheat, cherry, rehmannia, knotweed, and gastrodia.

[0055] Preferably, the bioactive small molecules of the present invention comprise one or more of catechin, gallocatechin, catechin gallate, gallocatechin gallate, epicatechin, epigallocatechin gallate, epigallocatechin, epicatechin gallate, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanidin, proanthocyanidin, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, nobiletin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, isorhamnetin, and hyperoside.

[0056] Embodiment 2

[0057] A large-scale preparation method of natural plant lipid nanovesicles, such as Figure 6 As shown, the instant granules containing natural plant lipid nanovesicles as described above include the following steps:

[0058] S01: Collect plants and store them at set temperature;

[0059] S02: After washing the plants, place them in a grinder, add ddH2O in a set ratio, and grind them;

[0060] S03: centrifuging the grinding liquid and collecting the supernatant after centrifugation;

[0061] S04: filtering the supernatant with a filter membrane of set pore size to obtain natural plant lipid nanovesicles;

[0062] S05: freeze-drying the obtained natural plant lipid nanovesicles and storing them at a set temperature;

[0063] The natural plant lipid nanovesicles prepared by the above method have high yield and low production cost, and can solve the problem of long preparation time and low yield of natural plant nanovesicles; experimental verification shows that the natural plant lipid nanovesicles prepared by the above method have anti-inflammatory and anti-cancer effects.

[0064] A preferred implementation example is as follows:

[0065] Collect fresh plants and store them at -20°C;

[0066] Wash the fresh plants with water and place them in a grinder. Add ddH2O in a ratio of 3:1 and grind for 4-5 minutes.

[0067] The grinding solution was centrifuged at 12000g for 1.5h, and the supernatant was collected;

[0068] The supernatant was filtered through a 0.45 μm filter membrane;

[0069] The obtained natural plant lipid nanovesicles are freeze-dried;

[0070] The obtained natural plant lipid nanovesicle freeze-dried powder was stored at 4°C;

[0071] It should be noted that the above components are only used for illustration and are not intended to be limiting. For those skilled in the art, they can be easily modified to other proportions. The modification of simple proportions based on this principle also falls within the scope of protection of this application.

[0072] The plants in the large-scale preparation method of natural plant lipid nanovesicles are one or more of tea, mulberry, tea, camellia, ginger, tomato, grape, grapefruit, lemon, apple, broccoli, carrot, onion, turmeric, strawberry, shiitake mushroom, salvia miltiorrhiza, ginseng, bitter melon, cabbage, aloe, corn, watermelon, astragalus, blueberry, garlic, soybean, cantaloupe, pear, pea, coconut, kiwi, citrus, sunflower, wheat, cherry, rehmannia root, knotweed, and gastrodia elata;

[0073] The bioactive small molecules in the large-scale preparation method of natural plant lipid nanovesicles include one or more of catechins, gallocatechin, catechin gallate, gallocatechin gallate, epicatechin, epigallocatechin gallate, epigallocatechin, epicatechin gallate, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanins, proanthocyanidins, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, nobiletin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, isorhamnetin, and hyperoside.

[0074] Experimental description:

[0075] Experiment 1: Stability test of tea natural lipid nanovesicles loaded with bioactive small molecules in gastrointestinal simulated fluid

[0076] (1) Collect fresh tea leaves and store them at -20°C;

[0077] (2) Wash the fresh tea leaves prepared in step (1) with water and place them in a grinder, add ddH2O in a ratio of 3:1, and grind for 4-5 minutes;

[0078] (3) centrifuging the grinding solution from step (2) at 12000 g for 1.5 h and collecting the supernatant;

[0079] (4) Filter the supernatant from step (3) using a 0.45 μm filter membrane;

[0080] (5) freeze-drying the natural tea lipid nanovesicles obtained in step (4);

[0081] (6) storing the freeze-dried powder of natural tea lipid nanovesicles obtained in step (5) at 4° C.;

[0082] (7) Take 10 mg of the lyophilized powder of the nanovesicles prepared in step (6), add 1 mL of ddH2O to disperse it, take 100 μL and add it to 2 mL of simulated fluids of the stomach, small intestine, and colon, respectively, and mix well. Then use a DLS particle size analyzer to measure the particle size and potential.

[0083] Experiment 2: Analysis of active ingredients in tea natural lipid nanovesicles

[0084] (1) Take 300 μL of nanovesicles and 700 μL of methanol to make the final concentration of methanol 70%;

[0085] (2) extracting the liquid in step (1) at 4° C. for 8 h;

[0086] (3) Centrifuge the liquid in step (2) at 11000 g for 10 min and collect the supernatant;

[0087] (4) filtering the supernatant in step (3) through a 0.22 μm filter membrane to remove large particles;

[0088] (5) The filtrate in step (4) is subjected to qualitative analysis of the active ingredients by UHPLC-MS / MS.

[0089] Experiment 3: Comparison of the yields of tea-derived natural lipid nanovesicles prepared by two preparation methods

[0090] Traditional preparation method:

[0091] (1) Collect fresh tea leaves and store them at -20°C;

[0092] (2) 100 g of fresh tea leaves prepared in step (1) were washed with water and placed in a grinder, ddH2O was added in a ratio of 3:1, and ground for 4-5 min;

[0093] (3) centrifuging the grinding liquid in step (2) at 1000 g for 20 min, 5000 g for 40 min, and 10000 g for 60 min to remove residues and collect the tea supernatant;

[0094] (4) centrifuging the tea leaves from step (3) at 100,000 g for 2 h at 4°C, removing the supernatant, and resuspending the precipitate in ddH2O;

[0095] (5) Add the resuspension in step (4) to a sucrose density gradient (8% / 15% / 30% / 45% / 60%), centrifuge at 100,000 g for 2 h at 4°C, and resuspend the precipitate in ddH2O;

[0096] (6) Centrifuge the resuspension in step (5) at 100,000 g for 2 h at 4° C., and repeat this step once to remove the residual sucrose solution;

[0097] (7) freeze-drying the natural tea lipid nanovesicles obtained in step (6);

[0098] (6) The tea natural lipid nanovesicle freeze-dried powder obtained in step (7) is stored at 4°C.

[0099] Large-scale preparation method:

[0100] (1) Collect fresh tea leaves and store them at -20°C;

[0101] (2) 100 g of fresh tea leaves prepared in step (1) were washed with water and placed in a grinder, ddH2O was added in a ratio of 3:1, and ground for 4-5 min;

[0102] (3) centrifuging the grinding solution from step (2) at 12000 g for 1.5 h and collecting the supernatant;

[0103] (4) Filter the supernatant from step (3) using a 0.45 μm filter membrane;

[0104] (5) freeze-drying the natural tea lipid nanovesicles obtained in step (4);

[0105] (6) The tea natural lipid nanovesicle freeze-dried powder obtained in step (5) is stored at 4°C.

[0106] Experiment 4: Determination of the anti-inflammatory activity of tea natural lipid nanovesicles prepared by two preparation methods

[0107] Inflammatory factor determination:

[0108] (1) Plating: Raw 264.7 cells were plated in 12-well plates. 1 mL of cell suspension was added to each well. The cell density was 1.0 × 10 5 cells / mL, overnight culture;

[0109] (2) After culturing the cells in step (1) overnight, remove the culture medium and wash once with PBS containing calcium and magnesium;

[0110] (3) Adding drugs to the cells in step (2): the nanoparticles were prepared in a serum-free basal medium, positive control (500 μL of basal medium), negative control (500 μL of basal medium), experimental group (500 μL of nanoparticle suspension)

[0111] (4) Incubate the cells in step (3) at 37°C for 4 h, then add 500 μL of complete culture medium and continue incubation for 20 h;

[0112] (5) After the cells are incubated in step (4), the culture medium is removed and the cells are washed once with PBS containing calcium and magnesium. 500 μL (0.5 μg / mL) of LPS is added to all wells except the negative control wells and the cells are incubated for another 6 h.

[0113] (6) After the cell incubation in step (5) is completed, the culture medium is collected and centrifuged at 3000 g for 10 min at 4°C. The supernatant is taken and the inflammatory factor content in the supernatant is detected using the corresponding ELISA kit.

[0114] Experiment 5: Determination of the anticancer activity of tea-derived natural lipid nanovesicles prepared by two preparation methods

[0115] Cell apoptosis experiment:

[0116] (1) Plating: CT-26 cells were plated in a 12-well plate, 1 mL of cell suspension was added to each well, and the cell density was 1.0×10 5 cells / mL, overnight culture;

[0117] (2) After culturing the cells in step (1) overnight, remove the culture medium, wash once with PBS containing calcium and magnesium, and add 500 μL of the prepared drug solution according to the experimental group;

[0118] (3) After incubating the cells and drugs in step (2) for 24 h, the cell culture medium was aspirated into a 15 mL centrifuge tube;

[0119] (4) Wash the cells in step (3) once with PBS, and add an appropriate amount of trypsin cell digestion solution to digest the cells (the digestion process can be performed while incubating and tapping);

[0120] (5) Add complete medium to the cells in step (4) to terminate digestion, transfer the liquid to a 15 mL centrifuge tube, centrifuge at 1000 g for 5 minutes, discard the supernatant, collect the cells, and gently resuspend the cells with PBS;

[0121] (6) Resuspend the cells from step (5), centrifuge at 1000 g for 5 min, discard the supernatant, add 1× binding buffer and gently resuspend the cells to adjust the concentration to 5×10 6 / mL; Take 100μL of cell suspension, add 5μL of Annexin V / PE, and mix gently; incubate at room temperature in the dark for 5min;

[0122] (7) Add 10 μL of 20 μg / mL 7AAD to the cell suspension in step (6) and add 400 μL of PBS;

[0123] (8) Immediately perform flow cytometry on the cells stained in step (7).

[0124] Experimental results analysis:

[0125] Figure 1a , Figure 1b , Figure 1c This is the particle size potential distribution diagram of tea natural lipid nanovesicles loaded with bioactive small molecules in Experiment 1. Figure 1a It is the determination of tea natural lipid nanovesicles loaded with bioactive small molecules in gastric simulated fluid. Figure 1b It is the determination of tea natural lipid nanovesicles loaded with bioactive small molecules in small intestine simulated fluid. Figure 1c This is the determination of tea leaf natural lipid nanovesicles loaded with bioactive small molecules in colon simulated fluid. As can be seen from the figure, tea leaf natural lipid nanovesicles are stable in the gastrointestinal tract.

[0126] Figure 2 This is the flavonoid and polyphenol composition diagram of tea natural lipid nanovesicles in Experiment 2. As can be seen from the figure, tea natural lipid nanovesicles contain a variety of flavonoids and polyphenols.

[0127] Figure 3 This is a comparison of freeze-dried powders of tea natural lipid nanovesicles prepared by two preparation methods in Experiment 3. In the figure, the left side is freeze-dried powder of tea natural lipid nanovesicles obtained by the traditional preparation method, and the right side is freeze-dried powder of tea natural lipid nanovesicles obtained by the large-scale preparation method. It can be seen from the figure that the yield of tea natural lipid nanovesicles prepared by the large-scale preparation method is much higher than that of the traditional preparation method.

[0128] Figure 4 This is a graph showing the inflammatory factor content in Raw264.7 cells treated with tea natural lipid nanovesicles prepared by two preparation methods in Experiment 4 for 24 hours. TNF-α and IL-6 are typical pro-inflammatory cytokines. As can be seen from the figure, the tea natural nanovesicles prepared by the large-scale method showed a similar ability to inhibit the secretion of pro-inflammatory cytokines as the tea natural nanovesicles prepared by the traditional method.

[0129] Figure 5 This is a cell apoptosis diagram of CT-26 cells treated with natural tea lipid nanovesicles prepared by two preparation methods in Experiment 5 for 24 hours. It can be seen from the figure that the natural tea nanovesicles prepared by the large-scale method show a similar ability to promote tumor cell apoptosis as the natural tea nanovesicles prepared by the traditional method.

[0130] In summary, the present invention utilizes plants to prepare a granule containing biologically active small molecules. The granule contains natural plant lipid nanovesicles. The natural plant lipid nanovesicles are used as carriers to load the biologically active small molecules, which can solve the problems of poor stability, low absorption efficiency, and easy decomposition of the biologically active small molecules in the gastrointestinal tract; the granule has anti-inflammatory and anti-cancer effects; the present invention provides a method for large-scale preparation of natural plant lipid nanovesicles, which can solve the problems of long time and low yield in the preparation of natural plant nanovesicles and reduce production costs.

[0131] It should be understood that those skilled in the art can make improvements or changes based on the above description, and all these improvements and changes should fall within the scope of protection of the appended claims of the present invention.

Claims

1. A granule containing natural plant lipid nanovesicles, characterized in that: The granules contain natural plant lipid nanovesicles for carrying bioactive small molecules.

2. The granule containing natural plant lipid nanovesicles according to claim 1, characterized in that: The natural plant lipid nanovesicles in the granules are loaded with biologically active small molecules.

3. The granule containing natural plant lipid nanovesicles according to claim 1 or 2, characterized in that: The preparation method of the natural plant lipid nanovesicles in the granules is as follows: Collect plants and store them at set temperatures; After washing the plants, place them in a grinder, add ddH2O at a set ratio, and grind them; The grinding liquid is centrifuged and the supernatant is collected; The supernatant is filtered through a filter membrane with a set pore size to obtain natural plant lipid nanovesicles; The obtained natural plant lipid nanovesicles are freeze-dried and stored at a set temperature.

4. The granule containing natural plant lipid nanovesicles according to claim 3, characterized in that: The setting ratio is (2-4):1, and the grinding time is 4-5 minutes.

5. The granule containing natural plant lipid nanovesicles according to claim 3, characterized in that: The pore size of the filter membrane is 0.22 or 0.45 μm.

6. The granule containing natural plant lipid nanovesicles according to claim 3, characterized in that: The storage temperature of the plant is -20 degrees; the freeze-dried storage temperature of the natural plant lipid nanovesicles is 4 degrees.

7. The granule containing natural plant lipid nanovesicles according to claim 3, characterized in that: The plants are one or more of tea, mulberry, tea, camellia, ginger, tomato, grape, grapefruit, lemon, apple, broccoli, carrot, onion, turmeric, strawberry, shiitake mushroom, salvia miltiorrhiza, ginseng, bitter melon, cabbage, aloe, corn, watermelon, astragalus, blueberry, garlic, soybean, cantaloupe, pear, pea, coconut, kiwi, citrus, sunflower, wheat, cherry, rehmannia, knotweed, and gastrodia.

8. The granule containing natural plant lipid nanovesicles according to claim 1 or 2, characterized in that: The bioactive small molecules include one or more of catechins, gallocatechins, catechin gallates, gallocatechin gallates, epicatechins, epigallocatechin gallates, epigallocatechins, epicatechin gallates, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanins, proanthocyanidins, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, nobiletin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, isorhamnetin, and hyperoside.

9. A large-scale preparation method of natural plant lipid nanovesicles, applied to the granules containing natural plant lipid nanovesicles as claimed in any one of claims 1 to 8, characterized in that: The following steps are involved: Collect plants and store them at set temperatures; After washing the plants, place them in a grinder, add ddH2O at a set ratio, and grind them; The grinding liquid is centrifuged and the supernatant is collected; The supernatant is filtered through a filter membrane with a set pore size to obtain natural plant lipid nanovesicles; The obtained natural plant lipid nanovesicles are freeze-dried and stored at a set temperature.

10. The large-scale preparation method of natural plant lipid nanovesicles according to claim 9, characterized in that: The set ratio is (2-4): 1, the grinding time is 4-5 minutes; the pore size of the filter membrane is 0.22 or 0.45 μm; the storage temperature of the plant is -20 degrees; the freeze-dried storage temperature of the natural plant lipid nanovesicles is 4 degrees; The plant is one or more of tea, mulberry, tea, camellia, ginger, tomato, grape, grapefruit, lemon, apple, broccoli, carrot, onion, turmeric, strawberry, mushroom, salvia miltiorrhiza, ginseng, bitter melon, cabbage, aloe, corn, watermelon, astragalus, blueberry, garlic, soybean, cantaloupe, pear, pea, coconut, kiwi, citrus, sunflower, wheat, cherry, rehmannia root, knotweed, and gastrodia elata; The bioactive small molecules include one or more of catechins, gallocatechins, catechin gallates, gallocatechin gallates, epicatechins, epigallocatechin gallates, epigallocatechins, epicatechin gallates, curcumin, resveratrol, quercetin, rutin, vitamins, cucurbitacin, anthocyanins, proanthocyanidins, lycopene, sulforaphane, kaempferol, hesperidin, hesperidin, luteolin, asiatica glycoside, nobiletin, glycyrrhizic acid, glycyrrhizin, bisabolol, chlorogenic acid, isorhamnetin, and hyperoside.

Images