Flash release tablet containing plant exosome as well as preparation method and application of flash release tablet

Abstract

The invention belongs to the technical field of exosomes, and particularly relates to a flash release tablet containing a plant exosome as well as a preparation method and application of the flash release tablet. The preparation method of the flash release tablet containing the plant exosome comprises the following steps: selecting dried ginseng, angelica sinensis and astragalus root tablets in a mass ratio of 1: (1-3): (1-3); the preparation method comprises the following steps: cleaning and drying the raw materials, mixing, and crushing to 80-100 meshes to obtain plant powder; the method comprises the following steps: mixing plant powder with a precooled phosphate buffer solution, homogenizing, carrying out centrifugal ultrafiltration to obtain a plant exosome suspension, and preparing exosome freeze-dried powder; mixing the exosome freeze-dried powder with mannitol, gelatin, a disintegrating agent, an auxiliary material and an active protective agent according to a formula of the flash release tablet, adding magnesium stearate for lubrication, tabletting and freeze-drying to obtain the flash release tablet containing the plant exosome; wherein the active protective agent is sodium sulfate and lauryl sodium sulfate. The flash release tablet has the advantages of short disintegration time, stable exosome particle size after redissolution, high retention rate and high activity.

Description

Technical Field

[0001] This invention belongs to the field of exosome technology, specifically relating to a flash-release tablet containing plant exosomes, its preparation method, and its application. Background Technology

[0002] Plant-derived exosomes-like nanoparticles (PELNs) are natural nanoscale vesicles secreted by plant cells, rich in bioactive components such as miRNAs, proteins, and lipids. They possess various physiological functions, including immunomodulation, anti-inflammation, anti-oxidation, and anti-fatigue effects. Exosomes derived from traditional Chinese medicines such as ginseng, astragalus, and angelica are particularly noteworthy due to their significant bioactivity and good safety profile. The physiological functions of plant exosomes mainly manifest in the following aspects: signal transduction, immune regulation, and tissue repair. They have become a hot topic in natural drug research in recent years.

[0003] In existing technologies, plant exosomes are used to produce dosage forms including injectable solutions, lyophilized powders, oral liquids, and capsules. There are many types of dosage forms, each with its own applicable scenarios. For example, injectable solutions are often used for intravenous injection to achieve anti-inflammatory and anti-tumor effects; oral liquids are often used to regulate intestinal flora.

[0004] Because the application sites and requirements of medications differ, the quality requirements for different formulations also vary. However, plant exosomes, derived from plants, are natural active ingredients with relatively poor stability. Regardless of the formulation, exosomes are the main active ingredient; therefore, preserving the activity of plant exosomes is a problem that needs to be addressed. Summary of the Invention

[0005] To address the aforementioned technical problems, this invention provides a flash-release tablet containing plant exosomes, its preparation method, and its applications. The method of this invention is used to retain the activity of plant exosomes in a flash-release tablet form.

[0006] The purpose of this invention is to provide a method for preparing flash-release tablets containing plant exosomes, comprising the following steps: Dried ginseng, angelica, and astragalus root slices are selected as raw materials, with a mass ratio of ginseng, angelica, and astragalus root slices of 1:1~3:1~3; After the raw materials are cleaned and dried, they are mixed and pulverized to 80-100 mesh to obtain plant powder. Plant powder was mixed with pre-cooled phosphate buffer, homogenized, and then centrifuged and ultrafiltered to obtain a plant exosome suspension, which was then made into exosome lyophilized powder. According to the flash release tablet formulation, exosome lyophilized powder is mixed with mannitol, gelatin, disintegrant, excipients, and activity protectant, magnesium stearate is added for lubrication, tableting is performed, and lyophilization is carried out to obtain flash release tablets containing plant exosomes. The active protective agents are sodium sulfate and sodium dodecyl sulfate; By weight percentage, the formulation of flash-release tablets contains 10%~20% exosome lyophilized powder, 20%~30% gelatin, 12.5%~20% disintegrant, 2%~4% excipients, 0.2%~0.5% activity protectant, 2%~4% magnesium stearate, and mannitol to make up 100%.

[0007] Preferably, in the above-mentioned method for preparing flash-release tablets containing plant exosomes, when the active protective agents are sodium sulfate and sodium dodecyl sulfate, the mass ratio of the two is 1:1.

[0008] Preferably, in the above-mentioned method for preparing flash-release tablets containing plant exosomes, the disintegrant is hydroxypropyl methylcellulose and / or crospovidone.

[0009] Preferably, in the above-mentioned method for preparing flash-release tablets containing plant exosomes, when the disintegrants are hydroxypropyl methylcellulose and crospovidone, the mass ratio of the two is 1:1.

[0010] Preferably, in the above-mentioned method for preparing flash-release tablets containing plant exosomes, the excipients include flavoring agents and sweeteners.

[0011] Preferably, in the above-mentioned method for preparing flash-release tablets containing plant exosomes, the flavoring agent is menthol; and the sweetener is aspartame.

[0012] Preferably, in the above-mentioned method for preparing flash-release tablets containing plant exosomes, the conditions for centrifugation and ultrafiltration include: Centrifuge sequentially: 500×g, 10 min~15 min; 2,000×g, 20 min~30 min; 10,000×g, 30 min~50 min; Ultracentrifugation: 100,000×g, 2 h~2.5 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. Filtration using a 0.22 μm microporous membrane; Plant exosome suspension was obtained by centrifugation and concentration using a 100 kDa ultrafiltration tube.

[0013] This invention provides a flash-release tablet containing plant exosomes prepared by the preparation method described above.

[0014] Application of flash-release tablets containing plant exosomes in the preparation of immunomodulators, anti-fatigue agents and antioxidants.

[0015] The antioxidants mentioned refer to preparations that scavenge DPPH, hydroxyl radicals, and superoxide radicals.

[0016] Compared with the prior art, the present invention has the following beneficial effects: This invention first identifies the optimal plants: ginseng, angelica, and astragalus root slices in a mass ratio of 1:1 to 3:1 to 3. Subsequent processes are based on this ratio, primarily to preserve the exosomes from these medicinal materials. After washing and drying, the raw materials are mixed according to the mass ratio and pulverized to 80-100 mesh to obtain plant powder, thereby increasing subsequent extraction efficiency. The lyophilized exosome powder is mixed with mannitol, gelatin, disintegrant, excipients, and activity protectant according to the flash-release tablet formulation. Magnesium stearate is added for lubrication, and the mixture is compressed into tablets and lyophilized to obtain flash-release tablets containing plant exosomes. The activity protectant is sodium sulfate and / or sodium dodecyl sulfate. Sodium sulfate and sodium dodecyl sulfate serve as activity protectants for exosomes, maintaining their activity during the preparation of flash-release tablets. Finally, by combining improved differential centrifugation with ultrafiltration, lyophilization powder preparation process, and flash release tablet preparation process, the resulting flash release tablets require no water, disintegrate rapidly in the mouth, and are absorbed under the tongue / buccal mucosa, thus avoiding gastrointestinal destruction and the first-pass effect; improving bioavailability, making them suitable for travel and office use; and exerting immune regulation, anti-fatigue, and antioxidant effects.

[0017] The flash-release tablets of this invention have a disintegration time of as short as 8.5 seconds; the exosomes are stable after reconstitution, with high retention rate and high activity; after adding flavoring agents and sweeteners, the taste is refreshing and fruity, with high acceptability. Attached Figure Description

[0018] Figure 1 Results for plant exosome stability.

[0019] Figure 2 The results show the reconstitution time of the flash-release tablets.

[0020] Figure 3 Results of weight-bearing swimming time for mice.

[0021] Figure 4 DPPH removal rate.

[0022] Figure 5 This represents the hydroxyl radical scavenging rate. Detailed Implementation

[0023] To enable those skilled in the art to better understand and implement the technical solutions of the present invention, the present invention will be further described below in conjunction with specific embodiments and accompanying drawings.

[0024] Unless otherwise specified, all reagents used in this invention are commercially available, and all methods used are conventional techniques in the art.

[0025] The inventive concept of this invention is as follows: Traditional oral exosome formulations suffer from gastric acid degradation and first-pass effect, resulting in poor physiological activity stability and low absorption efficiency of plant exosomes.

[0026] This invention aims to produce flash-release tablets that rapidly disintegrate and release nutrients in the oral cavity without the need for drinking water. These tablets are suitable for sublingual or buccal absorption, avoiding gastric acid degradation and the first-pass effect, thereby improving absorption efficiency. Furthermore, the flash-release tablet formulation can maintain the long-term physiological activity of plant exosomes.

[0027] In this invention, the preparation method of flash-release tablets containing plant exosomes is as follows: The first step is to select a suitable plant-based formula for extracting plant exosomes. Specifically, Select dried ginseng ( Panax ginseng ), Angelica sinensis ( Angelica sinensis Astragalus ( ) Astragalus membranaceus ) root slices, with a mass ratio of 1:1~3:1~3.

[0028] Because different plants have different genome sequences and medicinal effects, the active ingredients contained in exosomes are different, and their structures are different, the requirements for preparation processes are also different. For example, ginseng exosomes contain ginsenosides (Rg1, Rb1, Re, etc.), polysaccharides, flavonoids, amino acids and other substances; angelica exosomes contain ferulic acid, ligustilide, volatile oil, coumarin and other substances; astragalus exosomes contain astragaloside A, verbascoside, polysaccharides, flavonoids and other substances.

[0029] Therefore, this invention first determines the optimal plant formulation, and all subsequent processes are based on this formulation, primarily focusing on preserving the exosomes of these medicinal materials. Note that, according to the control experiments based on this invention, if other plant species are used, the exosome stability preservation effect of the process may decrease. Therefore, this invention has not yet investigated other plant formulations.

[0030] The second step is to crush it; After the raw materials are cleaned and dried, they are mixed according to the mass ratio and pulverized to 80-100 mesh to obtain plant powder, which increases the efficiency of subsequent extraction.

[0031] The third step involves preparing a plant exosome suspension using centrifugation and ultrafiltration. Step 4: Determining the formulation of flash-release tablets (orally dissolving, no water required) By weight percentage, the flash-release tablet formulation contains 10%–20% exosome lyophilized powder, 20%–30% gelatin, 12.5%–20% disintegrant, 2%–4% excipients, 0.2%–0.5% activity protectant, 2%–4% magnesium stearate, and mannitol to make up 100%. The activity protectant is sodium sulfate and / or sodium dodecyl sulfate.

[0032] In the above-mentioned flash-release tablet formulation, sodium sulfate and sodium lauryl sulfate act as exosome activity protectants to maintain exosome activity during tablet preparation. Attention should be paid to the dosage of both. Studies in this invention show that when the mass percentage of sodium lauryl sulfate or sodium sulfate exceeds 0.15%, exosome activity is lost. This is because excessively high concentrations of sodium lauryl sulfate or sodium sulfate affect the exosome membrane potential, leading to membrane damage and disrupting the physiological structure of exosomes, resulting in loss of exosome activity. However, within the recommended mass percentage range of the flash-release tablet formulation of this invention, the exosome membrane potential is appropriate, thereby protecting exosome activity.

[0033] The method for preparing exosome lyophilized powder is as follows: Mannitol and gelatin were added to the plant exosome suspension as freeze-drying protectants. The suspension was first pre-frozen and then freeze-dried under vacuum to obtain a loose exosome freeze-dried powder.

[0034] Step 5: Preparation of flash-release tablets (direct compression + freeze-drying method) According to the flash release tablet formulation, exosome lyophilized powder is mixed with mannitol, gelatin, disintegrant, excipients, and activity protectant, magnesium stearate is added for lubrication, tableting is performed, and lyophilization is carried out to obtain flash release tablets containing plant exosomes; aluminum-plastic blister packaging is used for light protection and airtight storage.

[0035] Preferably, the present invention employs a modified differential centrifugation combined with ultrafiltration: ① Mix the plant powder with phosphate buffer solution pre-cooled at 4℃ and homogenize. The formula for each liter of phosphate buffer solution is as follows: 8g sodium chloride, 0.2g potassium chloride, 2.9g disodium hydrogen phosphate, 0.24g potassium dihydrogen phosphate, and deionized water to make up to 1L, pH 7.4.

[0036] ② Centrifuge sequentially: 500×g, 10 min~15 min; 2,000×g, 20 min~30 min; 10,000×g, 30 min~50 min, to remove large particles and collect the supernatant; ③ Ultracentrifugation: 100,000×g, 2 h~2.5 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. ④ Filtration using a 0.22 μm microporous membrane; The plant exosome suspension was concentrated by centrifugation using a 100 kDa ultrafiltration tube to remove small molecule impurities, yielding a high-purity suspension. The obtained plant exosome suspension can be used directly in the next experimental step or stored at -80℃ for later use.

[0037] The centrifugation speed is increased sequentially to remove large particles of different sizes in stages, preventing different particles from adhering to each other and encapsulating exosomes, thus allowing the exosomes to remain better in the suspension.

[0038] The following are specific embodiments of the present invention.

[0039] Example 1 A method for preparing a flash-release tablet containing plant exosomes includes the following steps: The first step is to select a suitable plant-based formula for extracting plant exosomes. Specifically, Dried ginseng, angelica, and astragalus root slices were selected in a mass ratio of 1:1:1.

[0040] The second step is to crush it; After the raw materials are cleaned and dried, they are mixed according to the mass ratio and pulverized to 80 mesh to obtain plant powder.

[0041] The third step involves using a modified differential centrifugation method combined with ultrafiltration: ① Mix the plant powder with phosphate buffer solution pre-cooled to 4℃ and homogenize. The phosphate buffer solution formula per liter is as follows: 8g sodium chloride, 0.2g potassium chloride, 2.9g disodium hydrogen phosphate, 0.24g potassium dihydrogen phosphate, and deionized water to make up to 1L, pH 7.4. This phosphate buffer solution formula was used in the following examples and control group, and will not be described again.

[0042] ② Centrifuge sequentially: 500×g, 10 min; 2000×g, 20 min; 10000×g, 30 min, to remove large particles and collect the supernatant; ③ Ultracentrifugation: 100,000×g, 2 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. ④ Filtration using a 0.22 μm microporous membrane; The plant exosome suspension was concentrated by centrifugation using a 100 kDa ultrafiltration tube to remove small molecule impurities.

[0043] Step 4: Determine the formulation of the flash-release tablets (orally dissolves instantly, no water required). The composition of the flash release tablets is shown in Table 1.

[0044] Table 1. Composition of the flash-release tablets in Example 1

[0045] Note: The raw material sources for other embodiments and control groups are the same as those in Table 1, and will not be repeated here.

[0046] The method for preparing exosome lyophilized powder is as follows: Mannitol (5% by weight of plant exosome suspension) and gelatin (2% by weight of plant exosome suspension) were added as freeze-drying protectants. The mixture was pre-frozen at -80℃ for 12 h and then freeze-dried under vacuum for 24 h to obtain loose freeze-dried exosome powder.

[0047] Step 5: Preparation of flash-release tablets (direct compression + freeze-drying method) ① Premix: Mix the exosome freeze-dried powder with mannitol, gelatin, HPMC, PVPP, sweetener, and flavoring through an 80-mesh sieve, and mix with the ingredients except magnesium stearate according to the formula of the flash release tablets in Table 1. ② Lubrication: Add magnesium stearate according to the formulation of the flash-release tablets in Table 1, and mix for 5 min; ③ Tableting: Using a 6 mm shallow concave die, press into plain tablets; ④ Freeze-drying: Place the tablets in a freeze dryer, pre-freeze at -40℃ for 4 h, and vacuum dry for 12 h to obtain flash release tablets; ⑤ Packaging: Aluminum-plastic blister packaging, stored in a light-proof and airtight container.

[0048] Example 2 A method for preparing a flash-release tablet containing plant exosomes includes the following steps: The first step is to select a suitable plant-based formula for extracting plant exosomes. Specifically, Dried ginseng, angelica, and astragalus root slices were selected in a mass ratio of 1:1:1.

[0049] The second step is to crush it; After the raw materials are cleaned and dried, they are mixed according to the mass ratio and pulverized to 80 mesh to obtain plant powder, which increases the efficiency of subsequent extraction.

[0050] The third step involves using a modified differential centrifugation method combined with ultrafiltration: ① Mix plant powder with phosphate buffer solution pre-cooled at 4℃ and homogenize; ② Centrifuge sequentially: 500×g, 15min; 2000×g, 30min; 10000×g, 50min, to remove large particles and collect the supernatant; ③ Ultracentrifugation: 100,000×g, 2 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. ④ Filtration using a 0.22 μm microporous membrane; The plant exosome suspension was concentrated by centrifugation using a 100 kDa ultrafiltration tube to remove small molecule impurities.

[0051] Step 4: Determine the formulation of the flash-release tablets (orally dissolves instantly, no water required). The composition of the flash release tablets is shown in Table 2.

[0052] Table 2. Composition of the flash-release tablets in Example 2

[0053] The method for preparing exosome lyophilized powder is as follows: Mannitol (5% by weight of plant exosome suspension) and gelatin (2% by weight of plant exosome suspension) were added as freeze-drying protectants. The mixture was pre-frozen at -80℃ for 12 h and then freeze-dried under vacuum for 24 h to obtain loose freeze-dried exosome powder.

[0054] Step 5: Preparation of flash-release tablets (direct compression + freeze-drying method) ① Premix: Mix the exosome freeze-dried powder with mannitol, gelatin, HPMC, PVPP, sweetener, and flavoring through an 80-mesh sieve, and mix with the ingredients except magnesium stearate according to the formula of the flash release tablets in Table 2. ② Lubrication: Add the amount of magnesium stearate specified in Table 2 for the flash release tablet formulation, and mix for 5 min; ③ Tableting: Using a 6 mm shallow concave die, press into plain tablets; ④ Freeze-drying: Place the tablets in a freeze dryer, pre-freeze at -40℃ for 4 h, and vacuum dry for 12 h to obtain flash release tablets; ⑤ Packaging: Aluminum-plastic blister packaging, stored in a light-proof and airtight container.

[0055] Example 3 A method for preparing a flash-release tablet containing plant exosomes includes the following steps: The first step is to select a suitable plant-based formula for extracting plant exosomes. Specifically, Dried ginseng, angelica, and astragalus root slices were selected in a mass ratio of 1:3:3.

[0056] The second step is to crush it; After the raw materials are cleaned and dried, they are mixed according to the mass ratio and pulverized to 80 mesh to obtain plant powder, which increases the efficiency of subsequent extraction.

[0057] The third step involves using a modified differential centrifugation method combined with ultrafiltration: ① Mix plant powder with phosphate buffer solution pre-cooled at 4℃ and homogenize; ② Centrifuge sequentially: 500×g, 10 min; 2000×g, 20 min; 10000×g, 30 min, to remove large particles and collect the supernatant; ③ Ultracentrifugation: 100,000×g, 2 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. ④ Filtration using a 0.22 μm microporous membrane; The plant exosome suspension was concentrated by centrifugation using a 100 kDa ultrafiltration tube to remove small molecule impurities.

[0058] Step 4: Determine the formulation of the flash-release tablets (orally dissolves instantly, no water required). The composition of the flash release tablets is shown in Table 1.

[0059] The method for preparing exosome lyophilized powder is as follows: Mannitol (5% by weight of plant exosome suspension) and gelatin (2% by weight of plant exosome suspension) were added as freeze-drying protectants. The mixture was pre-frozen at -80℃ for 12 h and then freeze-dried under vacuum for 24 h to obtain loose freeze-dried exosome powder.

[0060] Step 5: Preparation of flash-release tablets (direct compression + freeze-drying method) ① Premix: Mix the exosome freeze-dried powder with mannitol, gelatin, HPMC, PVPP, sweetener, and flavoring through an 80-mesh sieve, and mix with the ingredients except magnesium stearate according to the formula of the flash release tablets in Table 1. ② Lubrication: Add magnesium stearate according to the formulation of the flash-release tablets in Table 1, and mix for 5 min; ③ Tableting: Using a 6 mm shallow concave die, press into plain tablets; ④ Freeze-drying: Place the tablets in a freeze dryer, pre-freeze at -40℃ for 4 h, and vacuum dry for 12 h to obtain flash release tablets; ⑤ Packaging: Aluminum-plastic blister packaging, stored in a light-proof and airtight container.

[0061] Example 4 A method for preparing a flash-release tablet containing plant exosomes includes the following steps: The first step is to select a suitable plant-based formula for extracting plant exosomes. Specifically, Dried ginseng, angelica, and astragalus root slices were selected in a mass ratio of 1:1:1.

[0062] The second step is to crush it; After the raw materials are cleaned and dried, they are mixed according to the mass ratio and pulverized to 100 mesh to obtain plant powder, which increases the efficiency of subsequent extraction.

[0063] The third step involves using a modified differential centrifugation method combined with ultrafiltration: ① Mix plant powder with phosphate buffer solution pre-cooled at 4℃ and homogenize; ② Centrifuge sequentially: 500×g, 15 min; 2000×g, 30 min; 10000×g, 50 min, to remove large particles and collect the supernatant; ③ Ultracentrifugation: 100,000×g, 2.5 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. ④ Filtration using a 0.22 μm microporous membrane; The plant exosome suspension was concentrated by centrifugation using a 100 kDa ultrafiltration tube to remove small molecule impurities.

[0064] Step 4: Determine the formulation of the flash-release tablets (orally dissolves instantly, no water required). The composition of the flash release tablets is shown in Table 1.

[0065] The method for preparing exosome lyophilized powder is as follows: Mannitol (5% by weight of plant exosome suspension) and gelatin (2% by weight of plant exosome suspension) were added as freeze-drying protectants. The mixture was pre-frozen at -80℃ for 12 h and then freeze-dried under vacuum for 24 h to obtain loose freeze-dried exosome powder.

[0066] Step 5: Preparation of flash-release tablets (direct compression + freeze-drying method) ① Premix: Mix the exosome freeze-dried powder with mannitol, gelatin, HPMC, PVPP, sweetener, and flavoring through an 80-mesh sieve, and mix with the ingredients except magnesium stearate according to the formula of the flash release tablets in Table 1. ② Lubrication: Add magnesium stearate according to the formulation of the flash-release tablets in Table 1, and mix for 5 min; ③ Tableting: Using a 6 mm shallow concave die, press into plain tablets; ④ Freeze-drying: Place the tablets in a freeze dryer, pre-freeze at -40℃ for 4 h, and vacuum dry for 12 h to obtain flash release tablets; ⑤ Packaging: Aluminum-plastic blister packaging, stored in a light-proof and airtight container.

[0067] Control group 1 Compared to Example 1, the difference is that the traditional Chinese medicine was replaced with Codonopsis pilosula. This was used to explore the effects of different plant species on exosome activity.

[0068] Control group 2 (blank control) Compared to Example 1, the active protectant has been removed. The composition of the flash release tablets is shown in Table 3.

[0069] Table 3. Composition of flash-release tablets in control group 2

[0070] Control group 3 Compared to Example 1, only one active protective agent, sodium dodecyl sulfate, was added. The composition of the flash release tablets is shown in Table 4.

[0071] The composition of the flash release tablets is shown in Table 4.

[0072] Table 4. Composition of the flash-release tablets in the control group.

[0073] Control group 4 Compared to Example 1, only one active protective agent, sodium sulfate, was added. The composition of the flash release tablets is shown in Table 5.

[0074] The formulation composition of the flash release tablets is shown in Table 5.

[0075] Table 5. Composition of flash-release tablets in control group 4

[0076] It should be noted that gelatin, mannitol, disintegrants (such as hydroxypropyl methylcellulose and crospovidone), and excipients (such as menthol, aspartame, and magnesium stearate) are essential components of flash-release tablets. This invention aims to produce flash-release tablets for oral mucosal absorption. The main innovation lies in how to create a flash-release tablet that maintains high exosome physiological activity; therefore, experiments involving the direct combination of active preservatives and lyophilized plant exosome powder are not conducted. This invention primarily compares control groups and examples to determine the impact of the addition or absence of active preservatives and their dosage on the physiological activity of plant exosomes.

[0077] Control group 5 Compared to Example 1, the active preservative was removed, and the Chinese herbal raw material was replaced with Codonopsis pilosula. This was used to compare the correlation between different Chinese herbal materials and the exosome preparation process, as well as the effect of adding or removing the active preservative on different plant raw materials.

[0078] As can be seen from control groups 5 and 1, as well as Example 1, the activity of Codonopsis pilosula exosomes was worse than that of the exosomes in Example 1 of this invention, regardless of whether an activity protectant was added. This demonstrates that the overall approach to different species and preparation processes is crucial to the activity of exosomes. The results prove that the formulation of the traditional Chinese medicine flash-release tablets of this invention is highly correlated with the exosome preparation process. The mechanistic explanation for this phenomenon is that different plants have significant species differences, resulting in different genome sequences, different efficacy, different active ingredients in the exosomes, and different structures, thus requiring different preparation processes.

[0079] Control group 6 Compared to Example 1, a higher dose of sodium sulfate was used.

[0080] Compared to Example 1, the proportion of sodium sulfate was changed to 0.15%.

[0081] Control group 7 Compared to Example 1, a higher dose of sodium dodecyl sulfate was used.

[0082] Compared to Example 1, the proportion of sodium dodecyl sulfate was changed to 0.15%.

[0083] Experiment 1: Stability Test of Plant Exosomes The flash-release tablets were reconstituted in phosphate buffer at a concentration of 100 μg / mL. The number of vesicles (A) with a size between 30 nm and 150 nm was measured. An accelerated assay was performed at 40°C in the dark for 3 months, and the number of vesicles (B) with a size between 30 nm and 150 nm was measured. The ratio of B to A was calculated to evaluate the stability of plant exosomes; a higher ratio indicates less exosome loss and higher activity retention.

[0084] Specifically, for the stability of plant exosomes in different embodiments and control groups, please refer to [link to relevant documentation]. Figure 1 The exosome stability of the present invention can reach over 88.3%.

[0085] Experiment 2, Disintegration Experiment Refer to the 2025 edition of the Chinese Pharmacopoeia. For reconstitution time of flash-release tablets, please refer to [reference needed]. Figure 2 The results showed that the disintegration time of the flash-release tablets in this embodiment was less than or equal to 8.5 seconds, which met the flash-release standard.

[0086] Experiment 3, Fatigue Resistance Experiment (1) Grouping Male 5-week-old ICR mice were randomly assigned to groups, n=5: Mouse blank control: An equal volume of blank flash-release tablet powder. The blank flash-release tablet powder did not contain plant exosome powder, and was otherwise the same as in Example 1; Treatment for each experimental group: 2 mg / kg body weight, flash-release tablets were mixed in 0.2 mL of water beforehand and administered by gavage.

[0087] (2) Administration Once daily for 14 consecutive days; testing begins 30 minutes after the last dose.

[0088] (3) Weighted swimming A lead plate equal to 5% of the rat's body weight was tied to the base of the rat's tail; the water depth was 30 cm and the water temperature was 25 ± 1 ℃; the "exhaustion time" was recorded from the moment the rat entered the water until it could not resurface after 10 seconds of sinking.

[0089] (4) Results In this example, a time to exhaustion ≥40% longer than that in the mouse control group was considered an effective initial screening result. (See results below.) Figure 3 .

[0090] Experiment 4. Antioxidant effect (evaluated by free radical scavenging efficiency) The flash-release tablets were reconstituted in phosphate buffer at a concentration of 100 μg / mL to obtain the sample solution.

[0091] (1) Free radical scavenging experiment - DPPH method: Weigh DPPH in the dark and prepare a 0.1 mmol / L DPPH working solution with anhydrous ethanol. Prepare and use immediately, and store in the dark.

[0092] Add the relevant reagents to each well of the 96-well plate as follows: A0: Absorbance of 100 μL anhydrous ethanol + 100 μL DPPH working solution, i.e. blank control; A1: Absorbance of 100 μL sample solution + 100 μL DPPH working solution; Set up 3 duplicate wells for each group, and gently tap to mix. Let stand at room temperature in the dark for 30 min. Measure the absorbance at 517 nm and record A0 and A1. Calculate the clearance rate.

[0093] Clearance rate (%) = [(A0 - A1) / A0] × 100.

[0094] The free radical scavenging rate of the blank control was 0.

[0095] (2) Free radical scavenging experiment - hydroxyl radical: Blank tube: Add 1 mL buffer solution + 0.5 mL salicylic acid + 0.5 mL FeSO4 + 0.5 mL H2O2 in sequence, and mix well.

[0096] Sample tube: Replace the buffer solution with 0.5 mL of sample solution, otherwise the same as the blank tube; total volume 2.5 mL.

[0097] Water bath at 37℃ for 30 minutes.

[0098] Measure absorbance at 510 nm: blank A0, sample A1.

[0099] Calculate the clearance rate: Clearance rate (%) = [(A0 - A1) / A0] × 100.

[0100] The average value was taken from three parallel trials. The free radical scavenging rate of the blank tube was 0.

[0101] See results Figure 4 and Figure 5 The free radical scavenging rate of each embodiment of the present invention can reach over 90%.

[0102] It should be noted that when numerical ranges are involved in this invention, it should be understood that the two endpoints of each numerical range and any value between the two endpoints can be selected. Since the steps and methods used are the same as in the embodiments, preferred embodiments are described in this invention to avoid redundancy. Although preferred embodiments of this invention have been described, those skilled in the art, once they understand the inventive concept of this invention, can make other changes and modifications to these embodiments, and all such changes and modifications fall within the scope of this invention.

[0103] Obviously, those skilled in the art can make various modifications and variations to this invention without departing from its spirit and scope. If such modifications and variations fall within the scope of equivalents of this invention, then this invention also intends to include these modifications and variations.

Claims

1. A method for preparing a flash-release tablet containing plant exosomes, characterized in that, Includes the following steps: Dried ginseng, angelica, and astragalus root slices were selected as raw materials, with a mass ratio of 1:1~3:1~3. After the raw materials are cleaned and dried, they are mixed and pulverized to 80-100 mesh to obtain plant powder. Plant powder was mixed with pre-cooled phosphate buffer, homogenized, and then centrifuged and ultrafiltered to obtain a plant exosome suspension, which was then made into exosome lyophilized powder. According to the flash release tablet formulation, exosome lyophilized powder is mixed with mannitol, gelatin, disintegrant, excipients, and activity protectant, magnesium stearate is added for lubrication, tableting is performed, and lyophilization is carried out to obtain flash release tablets containing plant exosomes. The active protective agents are sodium sulfate and sodium dodecyl sulfate; By weight percentage, the formulation of flash-release tablets contains 10%~20% exosome lyophilized powder, 20%~30% gelatin, 12.5%~20% disintegrant, 2%~4% excipients, 0.2%~0.5% activity protectant, 2%~4% magnesium stearate, and mannitol to make up 100%.

2. The method for preparing flash-release tablets containing plant exosomes according to claim 1, characterized in that, When the active protective agents are sodium sulfate and sodium dodecyl sulfate, their mass ratio is 1:

1.

3. The method for preparing flash-release tablets containing plant exosomes according to claim 1, characterized in that, The disintegrant is hydroxypropyl methylcellulose and / or crospovidone.

4. The method for preparing flash-release tablets containing plant exosomes according to claim 3, characterized in that, When the disintegrants are hydroxypropyl methylcellulose and crospovidone, their mass ratio is 1:

1.

5. The method for preparing flash-release tablets containing plant exosomes according to claim 1, characterized in that, The excipients include flavoring agents and sweeteners.

6. The method for preparing flash-release tablets containing plant exosomes according to claim 5, characterized in that, The flavoring agent is menthol; the sweetener is aspartame.

7. The method for preparing flash-release tablets containing plant exosomes according to claim 1, characterized in that, The conditions for centrifugal ultrafiltration include: Centrifuge sequentially: 500×g, 10 min~15 min; 2,000×g, 20 min~30 min; 10,000×g, 30 min~50 min; Ultracentrifugation: 100,000×g, 2 h~2.5 h, 4℃; discard the supernatant and resuspend the precipitate in phosphate buffer. Filtration using a 0.22 μm microporous membrane; Plant exosome suspension was obtained by centrifugation using a 100 kDa ultrafiltration tube.

8. Flash-release tablets containing plant exosomes prepared by the preparation method according to any one of claims 1 to 7.

9. The use of the flash-release tablets containing plant exosomes as described in claim 8 in the preparation of immunomodulators, anti-fatigue agents, and antioxidants.

10. The application according to claim 9, characterized in that, The antioxidants mentioned refer to preparations that scavenge DPPH, hydroxyl radicals, and superoxide radicals.

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