Perilla leaf-derived nanovesicle and use thereof in preparation of product with Anti-inflammatory efficacy

Abstract

A perilla leaf-derived nanovesicle and the use thereof in the preparation of a product with anti-inflammatory efficacy. The nanovesicle can be phagocytosed and internalized by HaCaT cells, reduce the levels of ROS and inflammatory factors in HaCaT cells, and exert an anti-inflammatory effect. Meanwhile, the nanovesicle has excellent transdermal properties, and thus can effectively alleviate and treat psoriatic dermatitis symptoms in mice. The nanovesicle can also be added as an active ingredient to different matrices to form compositions.

Description

Perilla leaf-derived nanovesicles and their application in the preparation of anti-inflammatory products Technical Field

[0001] This invention belongs to the field of plant extracellular vesicle technology, and relates to nanovesicles derived from perilla leaves and their application in the preparation of anti-inflammatory products. Background Technology

[0002] The skin is the largest organ in the human body, playing a vital role in protecting the body, resisting external pathogens, and maintaining homeostasis. Its importance lies in its role as the first line of defense against external threats.

[0003] Dermatitis is a general term for inflammatory skin diseases caused by various internal and external infectious or non-infectious factors, characterized by itching, skin redness, and rashes. Its etiology and clinical manifestations are complex and diverse, and it is prone to recurrence, making clinical treatment difficult. Topical application of corticosteroids, moisturizers, and ointments is commonly used. Severe cases require systemic treatment such as oral antihistamines or immunosuppressants, and biologics. Other treatment methods include phototherapy and traditional Chinese medicine. While existing treatments offer some relief for dermatitis, each therapy has potential drawbacks and limitations. For example, topical treatments may cause allergies or irritation at the application site and are prone to drug resistance; systemic treatments may produce systemic side effects, and their long-term safety is unclear; long-term phototherapy may increase the risk of skin cancer. Therefore, there is an urgent need to develop treatments with proven efficacy and high safety.

[0004] Psoriasis is one of the most common skin diseases in clinical practice, characterized by well-defined erythematous and scaly lesions on the skin. Its etiology is complex, involving multiple factors such as immunity, metabolism, genetics, and environment. It is clinically difficult to cure and prone to relapse, placing immense psychological and physiological stress on patients. Currently, psoriasis treatments mainly include topical therapy, phototherapy, and systemic therapy. Topical therapy is the first-line treatment, but commonly used drugs have serious adverse reactions, limiting their long-term use. Therefore, developing new drugs for the treatment of psoriasis with significant efficacy and high safety is of great importance.

[0005] Mild skin inflammation leading to skin barrier damage is a major concern in daily life. While it may not cause serious health problems, it can easily lead to discomfort, itching, pain, and negatively impact appearance, thus reducing quality of life. Furthermore, in modern life, skin problems caused by environmental pollution, ultraviolet radiation, stress, and other factors are becoming increasingly common, leading to a growing demand for skincare products that can alleviate skin inflammation and repair the skin barrier.

[0006] With consumers increasingly favoring natural, green, and non-irritating cosmetics, products using natural plant extracts are becoming more popular.

[0007] Summary of the Invention

[0008] To address the shortcomings of existing technologies, the present invention aims to provide nanovesicles derived from perilla leaves and their application in the preparation of anti-inflammatory products.

[0009] This invention studies nanovesicles derived from perilla leaves and finds that they can be phagocytosed and internalized by HaCaT cells, reducing ROS and inflammatory factor levels in HaCaT cells and exerting a strong anti-inflammatory effect. Simultaneously, using a mouse model of psoriasis, its superior transdermal properties were demonstrated, making it more effective in alleviating and treating psoriatic dermatitis symptoms in mice, reducing spleen index and PASI score, and exhibiting good biocompatibility. Furthermore, these nanovesicles can be added as active ingredients to different matrices to form compositions to meet diverse needs. This indicates that nanovesicles derived from perilla leaves have broad application prospects in the preparation of products (such as cosmetics or pharmaceuticals) for alleviating skin inflammation and preventing and treating inflammatory skin diseases.

[0010] The technical solution of the present invention is as follows:

[0011] This invention discloses nanovesicles derived from perilla leaves, which are prepared by perilla leaves and have a tea tray-like shape with a diameter of 30-300 nm.

[0012] Plant nanovesicles, similar in morphology, composition, and function to mammalian vesicles, exhibit a classic saucer-like shape with a diameter of 30–300 nm and possess multiple functions, playing a crucial role in regulating plant immunity. Plant-derived nanovesicles possess potential biological activities, most notably anti-inflammatory, anticancer, and antioxidant effects. Perilla (Perilla frutescens (L.) Britt.) is a traditional Chinese medicinal and edible plant. Perilla leaves, the commonly used medicinal part, are warm in nature and pungent in taste, entering the spleen and lung meridians. As a commonly used pungent and warm exterior-releasing herb, perilla leaves have been used clinically for over a thousand years, and in modern clinical practice, they are frequently used to treat inflammatory diseases such as exogenous wind-cold syndrome, cough and asthma, vomiting, sores, and arthralgia. Plant NEs have a nanoscale size, which determines their excellent transdermal ability. Therefore, perilla leaf-derived nanovesicles have great potential in the preparation of products (such as cosmetics or pharmaceuticals) for relieving skin inflammation and preventing and treating inflammatory skin diseases.

[0013] The nanovesicles derived from perilla leaves involved in this invention can be prepared by various methods, with the following ultracentrifugation method being more preferred. This method has a large yield, is simple to operate, and has low cost.

[0014] The specific steps are as follows: Wash fresh perilla leaves with deionized water and air dry them. Extract the juice using a slow juicer, then centrifuge and collect the supernatant at least three times: 2000–5000×g for 10–30 min, 8000–15000×g for 35–65 min, and 10000–15000×g for 35–65 min. Then, ultracentrifuge the supernatant at 100000–200000×g for 60–120 min, remove the supernatant, and resuspend the precipitate in deionized water to obtain perilla leaf-derived nanovesicles. Store at -20℃ to -80℃.

[0015] The nanovesicles derived from perilla leaves involved in this invention can also be extracted and separated using other methods, including density gradient ultracentrifugation, ultrafiltration, immunoaffinity capture, microfluidics, affinity chromatography, polymer precipitation, immunomagnetic beads, polyethylene glycol precipitation, anion exchange, ultrabuoyancy centrifugation, size exclusion chromatography, and combinations of the above methods.

[0016] This invention also discloses the application of perilla leaf-derived nanovesicles in the preparation of anti-inflammatory products.

[0017] Furthermore, the anti-inflammatory product is a transdermal anti-inflammatory product, including pharmaceuticals or cosmetics.

[0018] Furthermore, the medicine is a medicine used for the prevention or treatment of inflammatory skin diseases.

[0019] Furthermore, the inflammatory skin diseases include psoriasis and atopic dermatitis.

[0020] Furthermore, the dosage form of the drug is a skin delivery dosage form, including topical solutions, emulsions, transdermal patches, liniments, ointments, plasters, adhesive plasters, lotions, pastes, gels, coatings, films, microneedles, suspensions, foams, powders, sprays, or films.

[0021] Furthermore, the preferred dosage of the drug is 0.2–0.5 mg / mL.

[0022] Furthermore, the cosmetic is used to relieve mild skin inflammation, such as acne and pimples.

[0023] Furthermore, the types of cosmetics include toners, lotions, moisturizing creams, nourishing creams, massage creams, lotions, face creams, serums, ampoules, gels, eye creams, makeup removers, facial cleansers, makeup removers, face masks, sprays, body lotions, hand creams, gels, lip balms, or sunscreens.

[0024] Furthermore, the preferred dosage of the cosmetic is 0.2–0.5 mg / mL.

[0025] A composition containing nanovesicles derived from perilla leaves, the composition further comprising excipients, which are any one or more of the following: humectants, emulsifiers, surfactants, diluents, dispersants, binders, fillers, thickeners, lubricants, disintegrants, pH adjusters, flavor masking agents, colorants, plasticizers, antioxidants, antibacterial agents, preservatives, fragrances, pigments, and sunscreens.

[0026] Application of a composition containing nanovesicles derived from perilla leaves in the preparation of anti-inflammatory products.

[0027] Based on the anti-inflammatory research results of this invention, those skilled in the art can prepare nanovesicle compositions containing perilla leaf source according to different needs, in order to form anti-inflammatory products, especially transdermal anti-inflammatory products, such as cosmetics or pharmaceuticals.

[0028] Compared with the prior art, the present invention has the following beneficial effects:

[0029] This invention studies nanovesicles derived from perilla leaves and finds that they can be phagocytosed and internalized by HaCaT cells, reducing ROS and inflammatory factor levels in HaCaT cells and exerting a strong anti-inflammatory effect. Simultaneously, using a mouse model of psoriasis, its superior transdermal properties were demonstrated, making it more effective in alleviating and treating psoriatic dermatitis symptoms in mice, reducing spleen index and PASI score, and exhibiting good biocompatibility. Based on the in vitro and in vivo anti-inflammatory research results of this invention, those skilled in the art can prepare perilla leaf-derived nanovesicles into various forms of anti-inflammatory products, especially topical (transdermal) anti-inflammatory products, such as pharmaceuticals or cosmetics, according to specific needs. Attached Figure Description

[0030] Figure 1 shows the morphology of the PLNEs prepared in Preparation Example 1;

[0031] Figure 2 shows the average particle size of the PLNEs prepared in Preparation Example 1;

[0032] Figure 3 shows the average potential of the PLNEs prepared in Preparation Example 1;

[0033] Figure 4 shows a comparison of proteins in perilla juice and PLNEs prepared in Example 1.

[0034] Figure 5 shows the toxic effects of PLNEs prepared in Example 1 on HaCaT cells;

[0035] Figure 6 shows the effect of PLNEs prepared in Example 1 on the levels of cellular inflammatory factors IL-6 and IL-1β: (a) IL-6; (b) IL-1β;

[0036] Figure 7 shows the effect of PLNEs prepared in Example 1 on cell ROS levels.

[0037] Figure 8 shows the results of two-photon microscopy detection of the transdermal permeability of PLNEs prepared in Example 1;

[0038] Figure 9 shows the curves and line graphs obtained from the topical application of PLNEs in mice prepared in Example 1: (A) Body weight curve after PLNEs; (B) Changes in skin thickness on the back of the mouse; (C) PASI score line graph; (D) Spleen index line graph.

[0039] Figure 10 shows the skin on the back of a mouse and the size of its spleen after applying PLNEs to the mouse in Example 1.

[0040] Figure 11 shows the HE staining results of skin tissue sections obtained from Example 1 after topical application of PLNEs to psoriasis mice.

[0041] Figure 12 shows the skin appearance of healthy mice after local application of the PLNEs solution obtained in Example 1.

[0042] Figure 13 shows the changes in body weight and skin thickness of healthy mice after topical application of the PLNEs solution prepared in Example 1: (a) Changes in body weight; (b) Changes in skin thickness.

[0043] Figure 14 shows the morphology of PLNEs in the PLNEs-containing gel obtained in Preparation Example 2;

[0044] Figure 15 shows the particle size distribution of PLNEs in the PLNEs-containing gel obtained in Preparation Example 2;

[0045] Figure 16 shows the restorative effect of topical application of the PLNEs solution obtained in Example 1 on SDS-induced barrier function damage (elevated TEWL) in mice. Detailed Implementation

[0046] The technical solution of the present invention will be further illustrated below through specific embodiments. Those skilled in the art should understand that the examples described are merely illustrative of the invention and should not be construed as limiting the invention.

[0047] The perilla leaf raw material involved in the following preparation examples has been identified as the leaf of the perilla plant of the Lamiaceae family.

[0048] Preparation Example 1: Preparation of Perilla Leaf-Derived Nanovesicles (PLNEs) by Ultracentrifugation

[0049] (1) Take fresh perilla leaves from the local area, wash them with deionized water, and squeeze out the juice using a juicer.

[0050] (2) Gradient centrifugation (2500g×30min, 10000g×60min, 10000g×60min) to remove large particle precipitates.

[0051] (3) Take the supernatant and centrifuge at 100,000 × g for 60 min; discard the supernatant and resuspend the precipitate in deionized water to obtain PLNEs. Store at -20℃ to -80℃ for later use.

[0052] Test Example 1: Characterization of PLNEs obtained in Preparation Example 1

[0053] (1) Morphological observation by transmission electron microscopy (TEM): The PLNEs solution obtained in Preparation Example 1 was diluted with an appropriate amount of pure water and dropped onto a copper grid. After drying naturally, it was negatively stained with phosphotungstic acid and its morphology was observed by transmission electron microscopy. As shown in Figure 1, the PLNEs are in a typical teacup shape, which is consistent with the morphological structure of general plant nanovesicles.

[0054] (2) Particle size distribution was detected by Malvern Nanoparticle Size Analyzer (NTA): The PLNEs solution was diluted 1000 times and measured. As shown in Figure 2, the average particle size was 161.1 ± 3.1 nm. The particle size is consistent with the particle size range of general plant extracellular vesicles.

[0055] (3) Dynamic light dispersive spectroscopy (DLS) detection of zeta potential: The PLNEs solution was appropriately diluted and placed in the sample measurement cell for detection. As shown in Figure 3, the average potential of PLNEs was -13.56mV. The negative charge is consistent with the charged properties of plant extracellular vesicles.

[0056] (3) Comparative analysis of protein spectra: SDS-PAGE electrophoresis experiments were performed on perilla leaf juice and PLNEs solution. As shown in Figure 4, perilla leaf juice contains more types of proteins and is more complex, while the molecular weight of proteins in PLNEs is mainly distributed below 72kDa.

[0057] Test Example 2: The toxic and anti-inflammatory effects of PLNEs obtained in Preparation Example 1 on HaCaT cells

[0058] (1) CCK-8 assay for the cytotoxic effect of PLNEs on HaCaT cells: PLNEs obtained in Preparation Example 1 were diluted with complete culture medium to different concentrations (2, 5, 10, 25, 50 μg / mL) and analyzed by 1×10⁻⁶ ppm. 4 pcs / cm 2 HaCaT cells were seeded in 96-well plates at various densities. After 24 hours, different concentrations of PLNEs were introduced, and after another 24 hours of co-culture, the cell viability was measured using the CCK-8 assay at 450 nm. As shown in Figure 5, PLNEs at concentrations of 2–10 μg / mL showed no significant cytotoxicity to HaCaT cells, indicating good biocompatibility. However, a concentration of 25 μg / mL of PLNEs led to a decrease in cell viability.

[0059] (2) Evaluation of in vitro anti-inflammatory effects:

[0060] ①Inflammatory factor detection

[0061] HaCaT cells were treated with 25 ng / mL IL-6 for 24 h, and then co-cultured with different concentrations of PLNEs prepared in Example 1 (2, 5, 10, 25, 50 μg / mL). After 24 h, cell supernatants were collected, and the levels of inflammatory factors IL-6 and IL-1β were detected using an ELISA kit (Hangzhou Linke Biotechnology Co., Ltd.). The results are shown in Figure 6. PLNEs effectively inhibited IL-6-induced cellular inflammatory responses, and the 10 μg / mL PLNEs treatment group showed the best effect.

[0062] ② Detection of reactive oxygen species (ROS)

[0063] HaCaT cells were treated with 25 ng / mL IL-6 for 24 h, and then co-cultured with different concentrations of PLNEs prepared in Example 1 (2, 5, 10, 25, 50 μg / mL). After 24 h, the culture medium was removed, and the DCFH-DA fluorescent probe was added. The cells were incubated in the dark for 30 min, and then washed 2-3 times with PBS to remove free dye. After trypsin digestion, the cells were resuspended in PBS. Intracellular fluorescence intensity was detected by flow cytometry. The results are shown in Figure 7. PLNEs can effectively inhibit the increase in ROS caused by IL-6 stimulation, and 10 μg / mL PLNEs showed the best effect.

[0064] The above experiments demonstrate that PLNEs have good in vitro anti-inflammatory effects.

[0065] Test Example 3: The therapeutic effect of PLNEs obtained in Preparation Example 1 on psoriatic skin inflammation in mice.

[0066] (1) Establishment of a psoriatic dermatitis model:

[0067] One day before modeling, the backs of mice were shaved, covering an area of ​​approximately 2.5cm × 2.5cm. For the next 7 days, 62.5mg of imiquimod (IMQ) ointment was evenly applied to the shaved area daily to establish a mouse psoriasis model.

[0068] (2) Grouping and administration methods:

[0069] Thirty-five healthy 4-6 week old BALB / c mice were used, divided into four groups of seven: a control group, a model group, a low-dose group (0.2 mg / mL PLNEs), a medium-dose group (0.5 mg / mL PLNEs), and a high-dose group (1.0 mg / mL PLNEs). Starting on day 3, mice were given IMQ cream daily for 4 hours, followed by application of 200 μl of different doses of PLNEs to the treatment groups. The control and model groups received an equal amount of PBS applied to their backs. Treatment continued for 5 days, and the mice were euthanized on day 8.

[0070] (3) Back lesion scoring: The severity of psoriatic inflammation in mice was assessed using the Psoriasis Area and Severity Index (PASI) scoring system. The principle was to assess the severity of skin inflammation daily, using a 5-point scale (0-4). Erythema, scaling, and thickening were scored from 0 to 4 points: 0 points, none; 1 point, mild; 2 points, moderate; 3 points, severe; 4 points, very severe. The total score of the three indicators represented the severity of psoriatic dermatitis (0-12 points).

[0071] (4) Evaluation of in vivo transdermal effect: Healthy mice were shaved and, after one day of recovery, were uniformly coated with FITC (fluorescein isothiocyanate)-labeled PLNEs or free FITC solution. The skin was scanned along the z-axis using a two-photon microscope at a depth of 50 μm, and images were taken at 5, 10, 20, 30, and 40 μm. The results are shown in Figure 8. Compared with free FITC, FITC-labeled PLNEs were able to penetrate deeper into the skin layer at the same time point, indicating that PLNEs have superior transdermal performance.

[0072] (5) Evaluation results of each indicator:

[0073] ① Mouse weight change: Mouse weight was recorded daily, and weight change curves were plotted, as shown in Figure 9A. After continuous IMQ stimulation, the weight of mice in the model group decreased significantly, showing a significant difference compared to the control group. However, after administration of 0.2 mg / mL and 0.5 mg / mL PLNEs, the weight of mice showed a clear recovery trend compared to the model group.

[0074] ② Skin thickness changes: The skin thickness on the backs of mice was measured daily, and a skin thickness change curve was plotted. The results are shown in Figure 9(B). The skin thickness of the model group mice showed a significant increasing trend, while this process gradually slowed down after administration of PLNEs, and the 0.2 mg / mL and 0.5 mg / mL PLNEs groups showed a significant decreasing trend. This indicates that PLNEs can effectively inhibit the abnormal proliferation of keratinocytes and alleviate skin thickening, and 0.5 mg / mL PLNEs showed the best therapeutic effect.

[0075] ③PASI score: The PASI scores of mice in each group were counted daily, and the change curves were plotted. The results are shown in Figure 9(C). The effect of IMQ caused the PASI score to gradually increase, while after administration of PLNEs, the upward trend of the PASI score slowed down significantly, and the scores of mice in the 0.5 mg / mL PLNEs group began to show a downward trend.

[0076] ④ Spleen Index: The spleen weight and body weight of mice in each group were measured. The spleen index of each group was calculated according to the formula: Spleen Index = Spleen weight (g) / Mouse body weight (g) × 100. The results are shown in Figure 9(D). Compared with the control group, the spleen index of mice in the model group was significantly increased. However, there was no significant difference in spleen index between the 0.2 mg / mL and 1 mg / mL PELNs groups and the model group. Only after treatment with 0.5 mg / mL PELNs did the spleen index of mice decrease significantly, indicating that this was the optimal therapeutic dose.

[0077] (6) Comparison of back skin and spleen size in mice: Mice were euthanized on day 8, and their back skin was photographed. The spleens were also photographed for comparison. The results are shown in Figure 10. The model group mice exhibited significant dryness, erythema, increased scaling, and thickened skin, indicating that IMQ successfully induced psoriatic dermatitis symptoms in mice. After administration of PLNEs, the skin lesions in the mice showed varying degrees of improvement compared to the model group, indicating that PLNEs can effectively alleviate psoriatic dermatitis symptoms. Furthermore, the spleens of the mice in the PLNEs group were significantly smaller than those in the model group, indicating that PLNEs successfully reduced the level of inflammatory response in the mice. Among the observations, the 0.5 mg / mL PLNEs treatment showed the best therapeutic effect.

[0078] (7) Histopathological examination of back skin tissue: Skin or lesion tissue from the back of mice in each group was cut, fixed with 4% paraformaldehyde, dehydrated with alcohol and xylene, embedded in paraffin, sectioned, placed on glass slides, dewaxed, and stained with hematoxylin and eosin using H&E staining. The results are shown in Figure 11. The skin tissue structure of the control group mice was intact, with no epidermal thickening or inflammatory infiltration. In contrast, the skin tissue of the model group showed significant epidermal thickening and significant inflammatory cell infiltration in the dermis. Compared with the model group, the PELNs group mice showed significantly reduced responses in all aspects, with only mild inflammatory cell infiltration.

[0079] Test Example 4: In vivo safety of PLNEs obtained from Preparation Example 1

[0080] Healthy mice underwent hair removal on their backs, followed by daily application of 200 μL of PLNEs solution for five consecutive days. Administration was then stopped for five days until the skin recovered, after which 200 μL of PLNEs solution was applied daily again for five consecutive days. The control group received an equal volume of PBS. Skin appearance (see Figure 12), mouse weight, and skin thickness changes were observed daily (see Figure 13). These results indicate that PLNEs have good in vivo safety profiles for both short-term and relatively long-term use.

[0081] Preparation Example 2: Preparation of PLNE-containing gel compositions

[0082] The PLNEs obtained in Preparation Example 1 were dispersed in a carbomer gel matrix: 0.2% carbomer 940 was taken, and after swelling for 4 hours, it was completely dissolved under magnetic stirring. PLNEs were added under low temperature conditions of 4°C, and magnetic stirring was used to ensure uniform dispersion. The pH was adjusted to increase the viscosity for easy drug administration.

[0083] Test Example 5: Characterization of the PLNE-containing gel composition obtained in Preparation Example 2

[0084] (1) TEM observation of morphology and structure: Take an appropriate amount of the PLNEs-containing gel composition obtained in Example 2 and dilute it. TEM observation shows that, as shown in Figure 14, the PLNEs in the hydrogel are similar in morphology to free PLNEs and have a typical vesicle structure.

[0085] (2) NTA determination of particle size distribution: An appropriate amount of the PLNEs-containing gel composition obtained in Preparation Example 2 was diluted 1000 times and then subjected to NTA detection. The results are shown in Figure 15. The average particle size of PLNEs in the gel was 162.8 ± 2.5 nm, which is consistent with the particle size of nanovesicles.

[0086] All the above results indicate that the PLNE-containing gel composition has no significant effect on the stability of PLNEs.

[0087] Example 6: PLNEs-containing gel composition for restoring skin barrier function in mice. Sodium dodecyl sulfate (SDS)-induced skin irritation leads to increased transepidermal water loss (TEWL), which can be used to indicate damage to skin barrier function. Hair was removed from the backs of mice in each group. After 3 days of recovery, TEWL was measured using a transepidermal water loss meter (ASCH VAPO SCAN AS-VT100RS, imported from Japan). Subsequently, 200 μL of 1% SDS was uniformly applied to the bare skin. Four hours later, mice were administered high (1 mg / mL), medium (0.5 mg / mL), and low (0.2 mg / mL) concentrations of PLNEs-containing gel composition, while the control group received an equal volume of PBS. TEWL was measured at the same time point the following day, and the difference between the two concentrations was used to calculate the change in TEWL. The results are shown in Figure 16. The change in TEWL in the model group was significantly higher than that in the control group, while the change in TEWL was significantly reduced after administration of the PLNEs-containing gel composition. The low- and medium-concentration PLNEs-containing gel compositions showed more significant performance, indicating that 0.2–0.5 mg / mL PLNEs can effectively restore the barrier function damage caused by SDS in mice and can be used to develop cosmetics with the function of repairing skin barrier damage caused by inflammation.

Claims

1. A perilla leaf-derived nanovesicle, characterized by, The perilla leaf-derived nanovesicles are prepared from perilla leaves, have a tea tray-like shape, and have a diameter of 30-300 nm.

2. Use of the perilla leaf-derived nanovesicles in the preparation of an anti-inflammatory efficacy product.

3. Use according to claim 2, wherein the compound is ###0002### The anti-inflammatory efficacy product is a transdermal anti-inflammatory efficacy product, including a pharmaceutical product or a cosmetic product.

4. Use according to claim 3, wherein the compound is ###0002### The pharmaceutical product is a pharmaceutical product for preventing or treating an inflammatory skin disease.

5. The use according to claim 4, wherein the compound is ###0002### The inflammatory skin disease includes psoriasis, atopic dermatitis.

6. Use according to any one of claims 3 to 5, wherein the compound is ###0002### The pharmaceutical product has a skin administration dosage form, including an external solution, an emulsion, a transdermal patch, a liniment, an ointment, a plaster, a paste, a tape, a lotion, a paste, a gel, a film, a membrane, a micro-needle, a suspension, a foam, a powder, a spray, or a film.

7. The use according to claim 3, wherein the compound is ###0002### The cosmetic product is used to relieve mild skin inflammatory reactions.

8. Use according to claim 3 or 7, wherein the compound is ###0002### The cosmetic product includes cosmetic water, toner, moisturizing cream, nutrient cream, massage cream, emulsion, face cream, essence, ampoule, gel, eye cream, makeup remover, facial cleanser, makeup remover, facial mask, spray, body milk, hand cream, jelly, lip balm, or sunscreen.

9. A composition comprising nanovesicles derived from perilla leaf, characterized in that, The composition has perilla leaf-derived nanovesicles as an active ingredient, and further includes an auxiliary material, which includes any one or more of a humectant, an emulsifier, a surfactant, a diluent, a dispersant, a binder, a filler, a thickening agent, a lubricant, a disintegrant, a pH adjuster, a taste masking agent, a coloring agent, a plasticizer, an antioxidant, a bacteriostatic agent, a preservative, a fragrance, a pigment, and a sunscreen.

10. Use of a composition comprising nanovesicles of perilla leaf origin for the preparation of a product with anti-inflammatory efficacy, characterized in that, The anti-inflammatory efficacy product is a transdermal anti-inflammatory efficacy product, including a pharmaceutical product for preventing or treating an inflammatory skin disease, and a cosmetic product for relieving mild skin inflammatory reactions.

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