A Chinese herbal medicine extracellular vesicle composition, its preparation method and application

Abstract

The present invention relates to a traditional Chinese medicine extracellular vesicle composition, a preparation method thereof and an application. Among them, the traditional Chinese medicine extracellular vesicle composition includes at least four of turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles and dried ginger extracellular vesicles, and the traditional Chinese medicine extracellular vesicle composition contains turmeric extracellular vesicles. The traditional Chinese medicine extracellular vesicle composition provided by the present invention has the efficacy of promoting hair growth and preventing hair loss, is safe and non-toxic, and is easily taken up by cells.

Description

Technical Field

[0001] The present invention relates to the technical field of traditional Chinese medicine, and in particular to a traditional Chinese herbal medicine extracellular vesicle composition, a preparation method thereof, and an application thereof. Background Art

[0002] Currently, hair loss is commonly treated by surgery and drugs. Surgical treatment has disadvantages such as long time consumption, high cost, and limited acquisition of the number of hair follicle units. Drugs such as finasteride and minoxidil are restricted due to disadvantages such as long treatment cycles, limited efficacy, large adverse reactions, and easy recurrence. Therefore, finding safe and effective natural drugs has become a hot spot and a difficult point in the research of hair loss treatment. Traditional Chinese medicine in China has a relatively long history of being applied to hair loss, and has significant clinical efficacy. At present, many natural medicinal plants and active ingredients with high safety and remarkable efficacy for preventing hair loss and promoting hair growth have been discovered, such as Polygonum multiflorum, Dry Ginger, Black Sesame, and Polygonum cuspidatum.

[0003] Extracellular vesicles (EVs) are nanoscale (30 - 150 nm) membranous vesicles released from cells into the extracellular matrix, which participate in processes such as cell communication, migration, angiogenesis, and tumor cell growth. They are widely present in various body fluids and cell supernatants and stably carry important signaling molecules. Plant EVs, especially those from traditional Chinese herbal medicines (medicinal plants), are considered an important emerging field in recent years. Extracellular vesicles from traditional Chinese herbal medicines are derived from natural plants and have higher safety. There is little specific application of the vesicular membrane structure vesicles derived from plant cells. Although the hair growth promoting and hair loss preventing effects of extracts of Polygonum cuspidatum, Polygonum multiflorum, Platycladus orientalis, Dry Ginger, and Curcuma longa or their single components have been reported (such as patents CN117530901B, CN117180159A, CN110559418B, CN109674722B), there has been no report on the use of extracellular vesicles from traditional Chinese herbal medicines such as Curcuma longa, Polygonum multiflorum, and Dry Ginger for hair loss treatment.

[0004] Studies have shown that curcumin isolated from Curcuma longa can inhibit steroid 5-α reductase to achieve anti-androgenic activity, and the hair loss drugs finasteride and dutasteride mainly inhibit 5-α reductase to treat hair loss. In addition, curcumin has strong antioxidant and anti-inflammatory effects, which can reduce scalp inflammation and improve hair follicle health. Some animal studies have shown that curcumin can improve hair growth.

[0005] In view of this, there is an urgent need for a Chinese herbal medicine extracellular vesicle composition with good anti - hair - loss and hair - growth promoting effects, high safety, and few side effects. By adopting a combined formula and a multi - dimensional combination method of compounding turmeric with a variety of Chinese herbal medicine extracellular vesicles, it can achieve a multi - faceted and highly efficient hair - growth promotion and anti - hair - loss effect from aspects such as promoting the proliferation of hair follicle cells, promoting the secretion of hair follicle cell growth factors, regulating hair - growth related genes of hair follicle cells, clearing the reactive oxygen species (ROS) level in hair follicle cells, and anti - inflammation. Summary of the Invention

[0006] The object of the present invention is to overcome the defects of the above - mentioned prior art and provide a Chinese herbal medicine extracellular vesicle composition with hair - growth promotion and anti - hair - loss effects and safe without toxic side effects.

[0007] To achieve the above object, the technical solution adopted by the present invention is:

[0008] The present invention provides a Chinese herbal medicine extracellular vesicle composition, which includes at least four of turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles, and the Chinese herbal medicine extracellular vesicle composition contains turmeric extracellular vesicles.

[0009] Preferably, the Chinese herbal medicine extracellular vesicle composition includes turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles.

[0010] Preferably, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is (1 - 10):(0.1 - 2):(0.1 - 2):(0.5 - 5):(0.1 - 2).

[0011] Preferably, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is (1 - 6):(0.5 - 2):(0.5 - 2):(0.5 - 3):(0.5 - 2).

[0012] Preferably, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 4:1:1:2:1.

[0013] Preferably, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 2:1:1:2:1.

[0014] Preferably, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 1:1:1:2:1.

[0015] The present invention also provides a method for preparing the Chinese herbal medicine extracellular vesicle composition described in any one of the above. Respectively take turmeric, polygonum cuspidatum, fleece-flower root, biota orientalis leaf or dried ginger, soak, break the wall, centrifuge, and take the supernatant; perform ultracentrifugation on the obtained supernatant, take the precipitate, resuspend it with a buffer solution, and mix to prepare the extracellular vesicles, so as to obtain the Chinese herbal medicine extracellular vesicle composition.

[0016] The present invention also provides a composition with the efficacy of preventing hair loss and promoting hair growth. The composition includes the Chinese herbal medicine extracellular vesicle composition described in any one of the above or the Chinese herbal medicine extracellular vesicle composition prepared by the method described above.

[0017] Preferably, the concentration of the Chinese herbal medicine extracellular vesicle composition in the composition with the efficacy of preventing hair loss and promoting hair growth is 0.1 - 15 g / L.

[0018] Preferably, the composition with the efficacy of preventing hair loss and promoting hair growth further includes an emulsifier with a concentration of 1 - 15 g / L, a humectant with a concentration of 1 - 5 g / L, and a thickener with a concentration of 1 - 5 g / L;

[0019] The emulsifier is selected from one or more of glyceryl stearate, glyceryl stearate / PEG - stearate, cetearyl alcohol, cetearyl alcohol / PEG - 20 stearate, alkyl glucoside with 8 - 16 carbon atoms in the alkyl group, sodium isostearoyl lactyl lactate, etc.;

[0020] The humectant is selected from one or more of glycerol, propylene glycol, sodium PCA, 1,3 - propanediol, butanediol, hexanediol, 1,2 - pentanediol, octyldecanol, sorbitol, ethylhexylglycerin, trehalose, betaine, sodium hyaluronate, carbohydrate isomers, urea, allantoin, and panthenol, etc.;

[0021] The thickener is selected from one or more of xanthan gum, tara gum, acryloyldimethyltaurate / VP copolymer, sodium acryloyldimethyltaurate / hydroxyethyl acrylate copolymer, carbomer, and hydroxyethyl cellulose, etc.

[0022] The present invention also provides the application of the Chinese herbal medicine extracellular vesicle composition described in any one of the above, or the Chinese herbal medicine extracellular vesicle composition prepared by the method described above, or the composition with the efficacy of preventing hair loss and promoting hair growth described in any one of the above in the preparation of a product with the efficacy of preventing hair loss and promoting hair growth.

[0023] Preferably, the application is the application of the Chinese herbal medicine extracellular vesicles in the preparation of a product for promoting the secretion and expression level of VEGF in dermal papilla cells.

[0024] Preferably, the application is the application of the Chinese herbal medicine extracellular vesicles in the preparation of a product for inhibiting the expression levels of inflammatory factors IL - 6 and BMP4 in dermal papilla cells.

[0025] Effects of the Invention

[0026] The Chinese herbal extracellular vesicle composition provided by the present invention has a synergistic effect between the components, so that the Chinese herbal extracellular vesicle composition can effectively promote the proliferation of hair follicle cells, promote the secretion of hair follicle cell growth factors, regulate the expression of hair follicle cell growth-related genes, remove the level of hair follicle cell reactive oxygen ROS, and have antibacterial and anti-inflammatory effects, thereby achieving efficient hair growth and anti-hair loss effects. The Chinese herbal extracellular vesicle composition provided by the present invention is safe and effective, can be used for a long time, has good compatibility with most medicines or daily necessities raw materials, and has a simple process for adding it to medicines or daily necessities, and is suitable for industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 Electron microscopic images of extracellular vesicle particles of turmeric, Polygonum cuspidatum, Polygonum multiflorum, Platycladus orientalis leaves, and dried ginger (red arrows indicate extracellular vesicles);

[0028] Figure 2 The dermal papilla cells take up the Chinese herbal extracellular vesicle composition (red: PKH26-labeled Chinese herbal extracellular vesicle composition; blue: DAPI);

[0029] Figure 3 The ex vivo skin of 8-week-old Bama miniature pigs was infiltrated with a Chinese herbal extracellular vesicle composition (white arrows indicate hair follicles, red arrows represent PKH26-labeled Chinese herbal extracellular vesicles, and blue arrows represent DAPI-stained cell nuclei);

[0030] Figure 4 This is a bar graph showing that the extracellular vesicles prepared in Examples 1-2 promote the proliferation of hair papilla cells;

[0031] Figure 5 This is a bar graph showing that the extracellular vesicles prepared in Examples 1 and 2 promote the secretion of VEGF, a growth factor of hair papilla cells;

[0032] Figure 6 This is a bar graph showing that the extracellular vesicles prepared in Examples 1 and 2 inhibit the secretion of inflammatory factor IL-6 from hair papilla cells;

[0033] Figure 7 The histogram of the effect of the extracellular vesicles prepared in Examples 1-2 on the expression of the hair growth-related gene BMP4 in dermal papilla cells;

[0034] Figure 8 This is a bar graph showing the inhibition of ROS levels in dermal papilla cells by the extracellular vesicles prepared in Examples 1 and 2;

[0035] Figure 9 This is a test result diagram of the Chinese herbal extracellular vesicle anti-hair loss and hair growth essence on human body. DETAILED DESCRIPTION

[0036] In order to make the technical solutions and beneficial effects of the present invention more clearly understandable, the following is a detailed description by listing specific embodiments. The drawings are not necessarily drawn to scale, and local features may be enlarged or reduced to more clearly show the details of the local features; unless otherwise defined, the technical and scientific terms used herein have the same meanings as those in the technical field to which this application belongs.

[0037] A large number of studies have shown that extracellular vesicles produced by human cells (especially stem cells) can prevent and treat a variety of diseases. Studies have shown that stem cell extracellular vesicles can activate hair follicle dermal papilla cells and promote the transition of hair follicles from the resting phase to the growth phase, thereby starting the next growth phase and forming new hair shafts, allowing hair to regenerate. As the main mesenchymal component of the hair follicle unit, hair papilla cells play an important regulatory role in maintaining hair growth and inducing hair follicle regeneration, and extracellular vesicles can maintain the hair induction ability of hair papilla cells, thereby promoting hair regeneration. After extensive and in-depth research, the inventor unexpectedly discovered that a composition formed by extracellular vesicle particles derived from turmeric, Polygonum cuspidatum, Polygonum multiflorum, Platycladus orientalis, and dried ginger has the effect of preventing hair loss and promoting hair growth. More importantly, the inventor optimized the ratio of its components, and the obtained extracellular vesicle composition can promote hair follicle proliferation, promote the secretion of hair follicle cell growth factor, regulate the expression of hair follicle cell growth-related genes, remove the level of reactive oxygen ROS in hair follicle cells, and fight inflammation, so as to achieve a multi-faceted and efficient hair growth and hair loss prevention effect.

[0038] The present invention provides a Chinese herbal medicine extracellular vesicle composition, which includes at least four of turmeric extracellular vesicles, Polygonum cuspidatum extracellular vesicles, Polygonum multiflorum extracellular vesicles, Platycladus orientalis leaf extracellular vesicles and dried ginger extracellular vesicles, wherein the Chinese herbal medicine extracellular vesicle composition includes turmeric extracellular vesicles. The Chinese herbal medicine extracellular vesicle composition of the present invention uses pure Chinese medicinal plants as raw materials, and animal toxicology proves that it is safe, non-toxic and has no side effects. The diameter of each extracellular vesicle of the present invention is 20-500nm, which is a nano-scale raw material with good permeability, and is more easily absorbed and ingested by cells and skin to exert its efficacy.

[0039] In certain embodiments, the Chinese herbal extracellular vesicle composition includes turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, polygonum multiflorum extracellular vesicles, and platycladus orientalis leaf extracellular vesicles.

[0040] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, and oriental arborvitae leaf extracellular vesicles is (1-10):(0.1-2):(0.1-2):(0.5-5), such as 1:1:1:1, 2:1:1:1, 3:1:1:1, 4:1:1:1, 5:1:1:1, 6:1:1:1, 7:1:1:1, 8:1:1:1, 9:1:1:1, 10:1:1:1, 1:2:1:1, 2:2:1:1, 3:2:1:1, 4:2:1:1, 5:2:1:1, 6:2:1:1, 7:2:1:1, 8:2:1:1, 9:2:1:1, 10:2:1:1, 1:1:2:1, 2:1:2:1, 3:1:2:1, 4:1:2:1, 5:1:2:1, 6:1:2:1, 7:1:2:1, 8:1:2:1, 9:1:2:1, 10:1:2:1, 1:1:1:2, 2:1:1:2, 3:1:1:2, 4:1:1:2, 5:1:1:2, 6:1:1:2, 7:1:1:2, 8:1:1:2, 9:1:1:2, 10:1:1:2, etc.

[0041] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, and oriental arborvitae leaf extracellular vesicles is (1-6):(0.5-2):(0.5-3):(0.5-2).

[0042] In some embodiments, the traditional Chinese medicine extracellular vesicle composition includes turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, and dried ginger extracellular vesicles.

[0043] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, and dried ginger extracellular vesicles is (1-10):(0.1-2):(0.1-2):(0.1-2), such as 1:1:1:1, 2:1:1:1, 3:1:1:1, 4:1:1:1, 5:1:1:1, 6:1:1:1, 7:1:1:1, 8:1:1:1, 9:1:1:1, 10:1:1:1, 1:2:1:1, 2:2:1:1, 3:2:1:1, 4:2:1:1, 5:2:1:1, 6:2:1:1, 7:2:1:1, 8:2:1:1, 9:2:1:1, 10:2:1:1, 1:1:2:1, 2:1:2:1, 3:1:2:1, 4:1:2:1, 5:1:2:1, 6:1:2:1, 7:1:2:1, 8:1:2:1, 9:1:2:1, 10:1:2:1, 1:1:1:2, 2:1:1:2, 3:1:1:2, 4:1:1:2, 5:1:1:2, 6:1:1:2, 7:1:1:2, 8:1:1:2, 9:1:1:2, 10:1:1:2, etc.

[0044] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, and dried ginger extracellular vesicles is (1-6):(0.5-2):(0.5-2):(0.5-2).

[0045] In some embodiments, the traditional Chinese medicine extracellular vesicle composition includes turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles.

[0046] In some embodiments, the mass ratio of curcuma extracellular vesicles, polygonum cuspidatum extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles is (1 to 10):(0.1 to 2):(0.5 to 5):(0.1 to 2), such as 1:1:1:1, 2:1:1:1, 3:1:1:1, 4:1:1:1, 5:1:1:1, 6:1:1:1, 7:1:1:1, 8:1:1:1, 9:1:1:1, 10:1:1:1, 1:2:1:1, 2:2:1:1, 3:2:1:1, 4:2:1:1, 5:2:1:1, 6:2:1:1, 7:2:1:1, 8:2:1:1, 9:2:1:1, 10:2:1:1, 1:1:2:1, 2:1:2:1, 3:1:2:1, 4:1:2:1, 5:1:2:1, 6:1:2:1, 7:1:2:1, 8:1:2:1, 9:1:2:1, 10:1:2:1, 1:1:1:2, 2:1:1:2, 3:1:1:2, 4:1:1:2, 5:1:1:2, 6:1:1:2, 7:1:1:2, 8:1:1:2, 9:1:1:2, 10:1:1:2, etc.

[0047] In some embodiments, the mass ratio of curcuma extracellular vesicles, polygonum cuspidatum extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles is (1 to 6):(0.5 to 2):(0.5 to 3):(0.5 to 2).

[0048] In some embodiments, the Chinese herbal medicine extracellular vesicle composition includes curcuma extracellular vesicles, fleece-flower root extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles.

[0049] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum multiflorum extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles is (1 to 10):(0.1 to 2):(0.5 to 5):(0.1 to 2), such as 1:1:1:1, 2:1:1:1, 3:1:1:1, 4:1:1:1, 5:1:1:1, 6:1:1:1, 7:1:1:1, 8:1:1:1, 9:1:1:1, 10:1:1:1, 1:2:1:1, 2:2:1:1, 3:2:1:1, 4:2:1:1, 5:2:1:1, 6:2:1:1, 7:2:1:1, 8:2:1:1, 9:2:1:1, 10:2:1:1, 1:1:2:1, 2:1:2:1, 3:1:2:1, 4:1:2:1, 5:1:2:1, 6:1:2:1, 7:1:2:1, 8:1:2:1, 9:1:2:1, 10:1:2:1, 1:1:1:2, 2:1:1:2, 3:1:1:2, 4:1:1:2, 5:1:1:2, 6:1:1:2, 7:1:1:2, 8:1:1:2, 9:1:1:2, 10:1:1:2, etc.

[0050] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum multiflorum extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles is (1 to 6):(0.5 to 2):(0.5 to 3):(0.5 to 2).

[0051] In some embodiments, the Chinese herbal medicine extracellular vesicle composition includes turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, polygonum multiflorum extracellular vesicles, platycladus orientalis extracellular vesicles, and dried ginger extracellular vesicles.

[0052] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is (1 to 10):(0.1 to 2):(0.1 to 2):(0.5 to 5):(0.1 to 2), such as 1:1:1:1:1, 2:1:1:1:1, 3:1:1:1:1, 4:1:1:1:1, 5:1:1:1:1, 6:1:1:1:1, 7:1:1:1:1, 8:1:1:1:1, 9:1:1:1:1, 10:1:1:1:1, 1:2:1:1:1, 2:2:1:1:1, 3:2:1:1:1, 4:2:1:1:1, 5:2:1:1:1, 6:2:1:1:1, 7:2:1:1:1, 8:2:1:1:1, 9:2:1:1:1, 10:2:1:1:1, 1:1:2:1:1, 2:1:2:1:1, 3:1:2:1:1, 4:1:2:1:1, 5:1:2:1:1, 6:1:2:1:1, 7:1:2:1:1, 8:1:2:1:1, 9:1:2:1:1, 10:1:2:1:1, 1:1:1:2:1, 2:1:1:2:1, 3:1:1:2:1, 4:1:1:2:1, 5:1:1:2:1, 6:1:1:2:1, 7:1:1:2:1, 8:1:1:2:1, 9:1:1:2:1, 10:1:1:2:1, etc.

[0053] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is (1 to 6):(0.5 to 2):(0.5 to 2):(0.5 to 3):(0.5 to 2).

[0054] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 4:1:1:2:1.

[0055] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 2:1:1:2:1.

[0056] In some embodiments, the mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleeceflower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 1:1:1:2:1.

[0057] The present invention also provides a method for preparing the Chinese herbal medicine extracellular vesicle composition described in any one of the above. Turmeric, polygonum cuspidatum, fleece-flower root, biota orientalis leaf or dried ginger are respectively taken, soaked, broken, centrifuged, and the supernatant is taken; the obtained supernatant is ultracentrifuged, the precipitate is resuspended with a buffer solution, and the extracellular vesicles prepared by mixing are obtained to obtain the Chinese herbal medicine extracellular vesicle composition.

[0058] The present invention also provides a composition having the efficacy of preventing hair loss and promoting hair growth. The composition includes the Chinese herbal medicine extracellular vesicle composition described in any one of the above or the Chinese herbal medicine extracellular vesicle composition prepared by the method described above.

[0059] In some embodiments, the concentration of the Chinese herbal medicine extracellular vesicle composition in the composition having the efficacy of preventing hair loss and promoting hair growth is 0.1-15 g / L, such as 1 g / L, 1.5 g / L, 2 g / L, 2.5 g / L, 3 g / L, 3.5 g / L, 4 g / L, 4.5 g / L, 5 g / L, 5.5 g / L, 6 g / L, 6.5 g / L, 7 g / L, 7.5 g / L, 8 g / L, 8.5 g / L, 9 g / L, 9.5 g / L, 10 g / L, etc. Compared with surgical treatment, which has disadvantages such as long single treatment time, high price, and limited acquisition of the number of hair follicle units, the Chinese herbal medicine extracellular vesicle composition of the present invention has low cost, can be used as a daily necessity, and has good compatibility with other drug or daily necessity raw materials.

[0060] In some embodiments, the content of the Chinese herbal medicine extracellular vesicle composition in the composition having the efficacy of preventing hair loss and promoting hair growth is 5-15 g / L.

[0061] In some embodiments, the content of the Chinese herbal medicine extracellular vesicle composition in the composition having the efficacy of preventing hair loss and promoting hair growth is 8-12 g / L.

[0062] In some embodiments, the composition having the efficacy of preventing hair loss and promoting hair growth further includes an emulsifier with a concentration of 1-15 g / L, a humectant with a concentration of 1-5 g / L, and a thickener with a concentration of 1-5 g / L;

[0063] The emulsifier is selected from one or more of glyceryl stearate, glyceryl stearate / PEG, stearate, cetearyl alcohol, cetearyl alcohol / PEG-20 stearate, alkyl glycoside with 8-16 carbon atoms in the alkyl group, sodium isostearoyl lactyl lactate, etc.;

[0064] The humectant is selected from one or more of glycerol, propylene glycol, sodium PCA, 1,3-propanediol, butanediol, hexanediol, 1,2-pentanediol, octyldecanol, sorbitol, ethylhexylglycerin, trehalose, betaine, sodium hyaluronate, carbohydrate isomers, urea, allantoin, and panthenol, etc.;

[0065] The thickener is selected from one or more of xanthan gum, tara gum, acryloyldimethyltauramide / VP copolymer, hydroxyethyl acrylate / sodium acryloyldimethyltaurate copolymer, carbomer, hydroxyethyl cellulose, etc.

[0066] In some embodiments, the emulsifier is selected from alkyl glycosides having 8 to 16 carbon atoms in the alkyl group.

[0067] In a preferred embodiment, the emulsifier is selected from alkyl glycosides having 10 to 16 carbon atoms in the alkyl group.

[0068] In some embodiments, the humectant is selected from glycerol.

[0069] In some embodiments, the thickener is selected from carbomer.

[0070] In some embodiments, the concentration of the emulsifier is 5 to 15 g / L, such as 5 g / L, 5.5 g / L, 6 g / L, 6.5 g / L, 7 g / L, 7.5 g / L, 8 g / L, 8.5 g / L, 9 g / L, 9.5 g / L, 10 g / L, etc.

[0071] In some embodiments, the concentration of the humectant is 1 to 4 g / L, such as 1 g / L, 1.5 g / L, 2 g / L, 2.5 g / L, 3 g / L, 3.5 g / L, 4 g / L, etc.

[0072] In some embodiments, the concentration of the thickener is 1 to 4 g / L, such as 1 g / L, 1.5 g / L, 2 g / L, 2.5 g / L, 3 g / L, 3.5 g / L, 4 g / L, etc.

[0073] The present invention also provides a method for preparing the composition having the efficacy of preventing hair loss and promoting hair growth, the method comprising the following steps: mixing the thickener and water, adding the emulsifier after stirring evenly, heating to 75 - 85°C, stirring, adding the humectant to control the temperature at 75 - 85°C, stirring evenly, adding the Chinese herbal medicine extracellular vesicle composition after homogenization is completed, mixing evenly and then discharging to obtain the composition having the efficacy of preventing hair loss and promoting hair growth.

[0074] In some embodiments, the homogenization temperature is controlled at 75 - 85°C and the homogenization time is 20 - 40 min.

[0075] The present invention also provides the application of the Chinese herbal medicine extracellular vesicle composition described in any one of the above, or the Chinese herbal medicine extracellular vesicle composition prepared by the method as described above, or the composition having the efficacy of preventing hair loss and promoting hair growth described in any one of the above in the preparation of a product having the efficacy of preventing hair loss and promoting hair growth.

[0076] In some embodiments, the application is the application of the Chinese herbal medicine extracellular vesicles in the preparation of a product for promoting the secretion and expression level of VEGF in dermal papilla cells.

[0077] In some embodiments, the application is the application of the Chinese herbal medicine extracellular vesicles in the preparation of a product for inhibiting the expression levels of inflammatory factors IL-6 and BMP4 in dermal papilla cells.

[0078] The present invention will be further described below through specific examples. Unless otherwise specified herein, "%" represents mass percentage. The materials and reagents in the following examples, unless otherwise specified, are common materials or reagents in the art and can be obtained from commercial sources or synthesized by known methods. The experimental methods without specified conditions in the following implementation cases are usually carried out according to conventional experimental conditions or the conditions recommended by the manufacturers of relevant reagents (kits). Example 1

[0079] Weigh 100 g of turmeric, polygonum cuspidatum, fleece-flower root, biota orientalis leaf, and dried ginger respectively, wash them 3 times with ultrapure water to remove impurities, and place them in a clean beaker; add 500 mL of PBS to each respectively, soak overnight at 4°C; then wash 3 times with sterile water, transfer to a blender, add 500 mL of PBS, tighten the lid, and blend for 8 - 20 min to ensure no obvious lumpy fragments; transfer the juice to 50 mL sterile centrifuge tubes respectively, centrifuge at 500×g for 10 min with a low-speed centrifuge at 4°C, discard the precipitate, and take the supernatant; centrifuge at 2000×g for 20 min with a low-speed centrifuge at 4°C, discard the precipitate, and take the supernatant; centrifuge at 4000×g for 30 min with a low-speed centrifuge at 4°C, discard the precipitate, and take the supernatant; centrifuge at 10000×g for 1 h with a high-speed centrifuge at 4°C, discard the precipitate, and take the supernatant; centrifuge at 110000×g for 70 min with an ultra-high-speed centrifuge at 4°C, discard the supernatant, and resuspend the precipitate with an appropriate amount of PBS to obtain extracellular vesicle particles of turmeric, polygonum cuspidatum, fleece-flower root, biota orientalis leaf, and dried ginger respectively (labeled as CL-Exo, PCM-Exo, PM-Exo, PLA-Exo, ZO-Exo). Example 2

[0080] The extracellular vesicle particles of turmeric, polygonum cuspidatum, fleece-flower root, biota orientalis leaf, and dried ginger from Example 1 are compounded according to different mass ratios to obtain different Chinese herbal medicine extracellular vesicle compositions with anti-hair loss and hair growth-promoting effects (as shown in Table 1). If used in the short term, they can be directly stored in a -80°C refrigerator. For convenient long-term use and transportation, a lyoprotectant can be added for lyophilization.

[0081] Table 1

[0082] Example 3

[0083] The Chinese herbal extracellular vesicle composition derived from Example 2 was added to the essence matrix liquid to prepare the anti-hair loss and hair growth essence liquid. The specific operation was as follows: 100 mL of deionized water and 2 g of carbomer were added to a stirring pot and stirred for 3-10 min. After the thickener was completely dissolved, 10 g of an alkyl glycoside emulsifier with a carbon number of 14 was added and heated to 75-85° C.; after the emulsifier was basically dissolved, 3 g of glycerol was added and continued to stir for 5-10 min. The temperature was maintained at 75-85° C. After the material was completely dissolved, homogenized for 8-15 min, and continued to keep warm and stir for 20-40 min; 10 g of the Chinese herbal extracellular vesicle composition derived from Example 2 was added and stirred for 20-30 min, then PB was added to make the volume 1 L, the material was discharged, and various indicators were tested. HF-Mix1-8 corresponded to formulas 1-8 respectively.

[0084] PB formula: weigh 8 g NaCl, 0.2 g KCl, 1.44 g Na2HPO4 and 0.24 g KH2PO4, dissolve in 800 mL distilled water, adjust the pH value of the solution to 7.4 with HCl, and finally add distilled water to make up to 1 L.

[0085] Test Example 1 Electron microscopy results of extracellular vesicle particles

[0086] The shapes of the extracellular vesicle particles of each Chinese herbal medicine prepared in Example 1 were analyzed by transmission electron microscopy (TEM).

[0087] According to the sample conditions, the extracellular vesicle particles from Example 1 were adjusted to a suitable concentration or viscosity. Use a pipette to draw about 15 μL of extracellular vesicles onto the copper mesh and let it stand for 1 min. Use filter paper to blot the extracellular vesicles on the copper mesh dry, and draw about 15 μL of 2% uranyl acetate staining solution to stain at room temperature for 1 min. If obvious adsorbents are visible on the copper mesh, pure water can be dripped onto the surface, quickly blotted off, and repeatedly washed several times. Use filter paper to blot the extracellular vesicles on the copper mesh dry. Observe and take pictures, and save the pictures. The results show ( Figure 1 ): Saucer-cup-shaped vesicles can be observed in turmeric, Polygonum cuspidatum, Polygonum multiflorum, Platycladus orientalis leaves, and dried ginger.

[0088] Test Example 2: The situation of hair papilla cells taking up the Chinese herbal extracellular vesicle composition with the effect of preventing hair loss and promoting hair growth

[0089] The Chinese herbal extracellular vesicle composition obtained from Example 2 was labeled with PKH26 to track the uptake of Chinese herbal extracellular vesicles by dermal papilla cells.

[0090] Dermal papilla cells were added to a 3.5-cm confocal dish at a density of 200,000 cells / well and cultured for 24 h for later use. Take 1 mg of the Chinese herbal medicine extracellular vesicle composition, and use diluent C in the kit (Sigma, PKH26 Red Fluorescent Cell Labeling Kit, MINI26-1KT) to make up the volume of the sample to 1 mL. Take 6 μL of the PKH26 dye in the kit (MINI26-1KT from Sigma) and add it to a test tube containing 1 mL of diluent C. Gently pipette and mix continuously for 30 seconds. Let it stand at room temperature for 5 minutes. Add 2 mL of PBS containing 10% BSA for quenching. Use serum-free medium to make up the volume to 30 mL. Centrifuge at 110,000 g at 2 - 8 °C for 2 hours. Gently pipette and resuspend the extracellular vesicle particles precipitate in 1 mL of serum-free medium for later use.

[0091] The PKH26-labeled Chinese herbal medicine extracellular vesicle composition was added to dermal papilla cells at a concentration of 50 μg / mL and incubated for 12 h. After incubation, the cells were fixed with 4% paraformaldehyde at room temperature for 10 min. After washing with PBS, the cell skeleton was stained with phalloidin for 30 min, and then DAPI was added to stain the cell nucleus. Then, photographs were taken using a confocal microscope. The results showed ( Figure 2 ): The Chinese herbal medicine extracellular vesicle composition could be taken up by dermal papilla cells into the cells.

[0092] Detection Example 3 Skin Penetration of the Chinese Herbal Medicine Extracellular Vesicle Composition

[0093] The skin on the back or abdomen of 8-week-old Bama mini-pigs was cut into 1 cm × 1 cm pieces for later use. Take 1 mg of the Chinese herbal medicine extracellular vesicle composition HF-Mix2 prepared in Example 2, and use diluent C in the kit (Sigma, PKH26 Red Fluorescent Cell Labeling Kit MINI26-1KT) to make up the volume of the sample to 1 mL. Take 6 μL of the PKH26 dye in the kit and add it to a test tube containing 1 mL of diluent C. Gently pipette and mix continuously for 30 seconds. Let it stand at room temperature for 5 minutes. Add 2 mL of PBS containing 10% BSA for quenching. Use serum-free medium to make up the volume to 30 mL. Centrifuge at 110,000 g at 2 - 8 °C for 2 hours. Gently pipette and resuspend the extracellular vesicle precipitate in 1 mL of serum-free medium for later use.

[0094] The 1 cm × 1 cm skin of 8-week-old Bama mini-pigs was placed in an environment at 32 °C. Take 50 μL of the PKH26-labeled Chinese herbal medicine extracellular vesicle composition at 1 mg / mL and drop it onto the 1 cm × 1 cm skin of 8-week-old Bama mini-pigs, evenly covering the entire epidermis, and incubate for 2 h and 6 h. After incubation, cryosectioning was performed, and the section thickness was 10 μm. After staining the cell nucleus with DAPI, photographs were taken using a fluorescence microscope. The results showed (Figure 3 ): The extracellular vesicle composition of Chinese herbal medicines of the present invention can rapidly penetrate into the skin of 8-week-old Bama minipigs within 2 h and accumulate in large amounts in hair follicles (white arrows).

[0095] Detection Example 4: The extracellular vesicle composition of Chinese herbal medicines can promote the proliferation of dermal papilla cells

[0096] Dermal papilla cells were seeded into 96-well plates at a density of 5000 cells / 100 μL / well. The cells were cultured for 24 h. The cell supernatant was discarded, and 100 μL of negative control reagent (NC: T4 medium, Beso Biological, catalog number 04304P05), positive control group reagent (PC: T4 medium + 10% fetal bovine serum), and experimental group reagent (extracellular vesicle particles from different sources of HF-Mix1 and HF-Mix2 in Examples 1 and 2 (diluted to a protein concentration of 0.5 mg / mL with T4 medium)) were added respectively and cultured for 24 h. After discarding the supernatant, 100 μL of basal medium containing 10% CCK8 was added and incubated at 37 °C for 2-4 h, and then detected by an enzyme-linked immunosorbent assay (ELISA) reader.

[0097] The results showed ( Figure 4 ): Extracellular vesicle particles from turmeric (CL-Exo), Platycladus orientalis (PLA-Exo), and Polygonum cuspidatum (PCM-Exo) could significantly promote the proliferation of dermal papilla cells. The extracellular vesicle composition HF-Mix2 of Chinese herbal medicines of the present invention could significantly promote the proliferation of dermal papilla cells, and its effect was better than that of HF-Mix1 and each single extracellular vesicle particle of turmeric, Platycladus orientalis, and Polygonum cuspidatum.

[0098] Detection Example 5: The extracellular vesicle composition of Chinese herbal medicines promotes the secretion of growth factors in androgenetic alopecia models

[0099] Dermal papilla cells were seeded into 96-well plates at a density of 5000 cells / 100 μL / well. The cells were cultured for 24 h. The cell supernatant was discarded, and 100 μL of medium containing 1 μM dihydrotestosterone (DHT) was added to each well and cultured for 24 h to establish an androgenetic alopecia cell model. After discarding the PBS, 100 μL of negative control reagent (NC: T4 medium), positive control group reagent (PC: 1 μM finasteride, diluted with T4 medium), and experimental group reagent (extracellular vesicle particles from different sources of HF-Mix1 and HF-Mix2 in Examples 1 and 2 (diluted to a protein concentration of 0.5 mg / mL with T4 medium)) were added respectively. The basal medium (BC) was added to the group without DHT treatment, and the cells were cultured for 24 h. The supernatant was taken, and the secretion of growth factor VEGF was detected by Jianglai Bio vascular endothelial growth factor (VEGF) ELISA detection kit.

[0100] The results showed ( Figure 5): In the cellular androgenetic alopecia model, extracellular vesicle particles derived from Zingiber officinale (ZO-Exo), Curcuma longa (CL-Exo), Platycladus orientalis (PLA-Exo), and Polygonum cuspidatum (PCM-Exo) can significantly promote the secretion of vascular endothelial growth factor (VEGF), a growth factor of dermal papilla cells. The Chinese herbal medicine extracellular vesicle composition HF-Mix2 of the present invention can also significantly promote the secretion of VEGF, a growth factor of dermal papilla cells, and its effect is better than that of HF-Mix1 and each single extracellular vesicle particle of Zingiber officinale, Curcuma longa, Platycladus orientalis, and Polygonum cuspidatum.

[0101] Detection Example 6: Inhibition of the secretion of inflammatory factors by the Chinese herbal medicine extracellular vesicle composition in the androgenetic alopecia model

[0102] Dermal papilla cells were seeded into 96-well plates at a density of 5000 cells / 100 μL / well. The cells were cultured for 24 h. The cell supernatant was discarded, and 100 μL of medium containing 1 μM dihydrotestosterone (DHT) was added to each well and cultured for 24 h to establish a cellular androgenetic alopecia model. After discarding the PBS, 100 μL of negative control reagent (NC: T4 medium), positive control group reagent (PC: 1 μM finasteride, diluted with T4 medium), experimental group reagent (extracellular vesicle particles from different sources of HF-Mix1 and HF-Mix2 in Examples 1 and 2 (diluted with T4 medium to a protein concentration of 0.5 mg / mL)) were added respectively. For the group not treated with DHT, basal medium (BC) was added and cultured for 24 h. The supernatant was taken, and the secretion of the pro-inflammatory factor interleukin-6 (IL-6) was detected by the ELISA kit for interleukin-6 (IL-6) of Lianke Biotech.

[0103] The results showed ( Figure 6 ): In the cellular androgenetic alopecia model, extracellular vesicle particles derived from Polygonum multiflorum (PM-Exo) can significantly inhibit the secretion of the inflammatory factor IL-6 in dermal papilla cells. Although the Chinese herbal medicine extracellular vesicle compositions HF-Mix1 and HF-Mix2 of the present invention can significantly inhibit the secretion of the inflammatory factor IL-6 in dermal papilla cells, the effect of HF-Mix2 is better than that of HF-Mix1, Curcuma longa, and extracellular vesicle particles of Polygonum multiflorum.

[0104] Detection Example 7: Influence of the Chinese herbal medicine extracellular vesicle composition on the expression of hair growth-related genes in the androgenetic alopecia model

[0105] Dermal papilla cells were seeded into 24-well plates at a density of 20,000 cells / 500 μL / well. The cells were cultured for 24 h. The cell supernatant was discarded, and 500 μL of medium containing 1 μM dihydrotestosterone (DHT) was added to each well and cultured for 24 h to establish a cellular androgenetic alopecia model. The supernatant was discarded, and 500 μL of negative control reagent (NC: T4 medium), positive control group reagent (PC: 1 μM finasteride, diluted with T4 medium), experimental group reagent (extracellular vesicle particles from different sources of HF-Mix1 and HF-Mix2 in Example 1 and Example 2 (diluted with T4 medium to a protein concentration of 0.5 mg / mL)) were added respectively. For the group without DHT treatment, basal medium (BC) was added, and the cells were cultured for 24 h. The supernatant was discarded, the cells were collected, and the RNA of the cells was extracted to detect the expression of the hair growth-related gene BMP4.

[0106] The results showed that in the cellular androgenetic alopecia model, Polygonum multiflorum (PM-Exo) and Curcuma longa (CL-Exo) could significantly inhibit the expression of the hair growth-negative related gene BMP4 in dermal papilla cells ( Figure 7 ). The Chinese herbal medicine extracellular vesicle composition HF-Mix2 of the present invention could significantly inhibit the expression of the hair growth-negative related gene BMP4 in dermal papilla cells ( Figure 7 ), and the effect of HF-Mix2 was better than that of single extracellular vesicles and also better than that of HF-Mix1.

[0107] Detection Example 8 Effect of Chinese herbal medicine extracellular vesicle composition on the level of reactive oxygen species in oxidative stress alopecia model

[0108] Dermal papilla cells were seeded into 24-well plates at a density of 20,000 cells / 500 μL / well. The cells were cultured for 24 h. The supernatant was discarded, and 100 μL of negative control reagent (NC: T4 medium), positive control group reagent (PC: 1 μM finasteride, diluted with T4 medium), experimental group reagent (extracellular vesicle particles from different sources of HF-Mix1 and HF-Mix2 in Example 1 and Example 2 (diluted with T4 medium to a protein concentration of 0.5 mg / mL)) were added respectively, and the cells were cultured for another 24 h. 250 μM hydrogen peroxide (H2O2) was added to each well and cultured for 30 min to establish a cellular oxidative stress alopecia model. The DCFH-DA fluorescent probe was used to determine the effect of different samples on the level of reactive oxygen species in dermal papilla cells.

[0109] The results showed ( Figure 8In the cell oxidative stress-induced alopecia model, extracellular vesicle particles derived from dried ginger (ZO-Exo) and polygonum cuspidatum (PCM-Exo) can significantly inhibit the level of reactive oxygen species (ROS) in dermal papilla cells. The herbal extracellular vesicle composition HF-Mix2 of the present invention can significantly inhibit the ROS level in dermal papilla cells, and the effect of HF-Mix2 is better than that of HF-Mix1, curcuma longa, dried ginger, and polygonum cuspidatum extracellular vesicle particles.

[0110] Detection Example 9: Promotion of hair growth of isolated hair follicles of mice by herbal extracellular vesicle composition

[0111] The back hair of 10-week-old C57BL / 6 mice was shaved off. After removing the skin, it was washed 3-5 times with PBS, and then dissected into tissue blocks containing 3-4 hair follicles under a microscope and placed into 24-well plates respectively. 500 μL of negative control reagent (NC: T4 medium) and experimental group reagents (extracellular vesicle particles from different sources in Example 1 and Example 2 (diluted to a protein concentration of 0.5 mg / mL with T4 medium)) were added respectively, and cultured for 5 days. The culture medium was changed every two days, and the hair growth conditions on the 2nd and 5th days were recorded respectively (N = 10).

[0112] The results are shown in Table 2: Extracellular vesicle particles derived from fleece-flower root (PM-Exo), curcuma longa (CL-Exo), biota orientalis (PLA-Exo), polygonum cuspidatum (PCM-Exo), and the herbal extracellular vesicle composition can all promote the hair growth of isolated hair follicles. Among them, HF-Mix2, HF-Mix7, and HF-Mix8 can significantly promote the hair growth of isolated hair follicles, and especially HF-Mix2 has the best effect.

[0113] Table 2

[0114]

[0115] Detection Example 10: Human anti-hair loss and hair growth promotion test of herbal extracellular vesicle composition

[0116] 270 experiencers aged 30-50 with hair loss problems and who had not used hair loss treatment drugs recently were enrolled and randomly divided into 9 groups. The experimental group applied the anti-hair loss and hair growth promotion essence containing herbal extracellular vesicles from Example 3 once in the morning and once in the evening every day, and the control group applied the essence matrix (i.e., the essence without the extracellular vesicle composition) once in the morning and once in the evening every day. The hair growth was tracked by taking pictures every month. The effective rate was calculated as follows: (((number of significantly improved people * 100%) + (number of slightly improved people * 50%)) / 30) * 100%. The results of the human anti-hair loss and hair growth promotion test of the herbal extracellular vesicle composition are shown in Table 3.

[0117] Table 3

[0118]

[0119] When using the Chinese herbal medicine extracellular vesicle composition for 3 months, hair growth can be observed. More hair grows in the hair loss area on the top of the head, and the hair loss area shrinks; those with severe hair loss grow a lot of hair, most of which are villous hairs; in addition, the female hairline becomes significantly narrower (such as Figure 9 ).

[0120] It should be understood that the above embodiments are all exemplary and do not cover all possible implementation manners included in the claims. Without departing from the scope of the present disclosure, various modifications and changes can also be made based on the above embodiments. Similarly, any combination of the technical features of the above embodiments can also be made to form additional embodiments of the present invention that may not be clearly described. Therefore, the above embodiments only represent several implementation manners of the present invention and do not limit the protection scope of the present invention patent.

Claims

1. A Chinese herbal medicine extracellular vesicle composition with the efficacy of preventing hair loss and promoting hair growth, characterized in that, The Chinese herbal medicine extracellular vesicle composition includes turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles. The mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is (1-6):(0.5-2):(0.5-2):(0.5-3):(0.5-2).

2. The extracellular vesicle composition of Chinese herbal medicine according to claim 1, wherein The mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 4:1:1:2:1; or, The mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 2:1:1:2:1; or, The mass ratio of the turmeric extracellular vesicles, polygonum cuspidatum extracellular vesicles, fleece-flower root extracellular vesicles, oriental arborvitae leaf extracellular vesicles, and dried ginger extracellular vesicles is 1:1:1:2:

1.

3. A method for preparing the Chinese herbal medicine extracellular vesicle composition according to any one of claims 1 to 2, characterized in that, Take turmeric, polygonum cuspidatum, fleece-flower root, oriental arborvitae leaf, or dried ginger respectively, soak, break the cell wall, centrifuge, and take the supernatant; subject the obtained supernatant to ultracentrifugation, take the precipitate, resuspend it with a buffer solution, and mix to prepare extracellular vesicles, thereby obtaining the Chinese herbal medicine extracellular vesicle composition.

4. A composition with the efficacy of preventing hair loss and promoting hair growth, characterized in that, The composition includes the Chinese herbal medicine extracellular vesicle composition according to any one of claims 1-2 or the Chinese herbal medicine extracellular vesicle composition prepared by the method according to claim 3.

5. The composition with anti - hair loss and hair growth promoting efficacy according to claim 4, characterized in that, The concentration of the Chinese herbal medicine extracellular vesicle composition in the composition having the efficacy of preventing hair loss and promoting hair growth is 0.1-15 g / L.

6. The composition with anti - hair - loss and hair - growth promoting efficacy according to claim 4 or 5, characterized in that, The composition having the efficacy of preventing hair loss and promoting hair growth further includes an emulsifier with a concentration of 1-15 g / L, a humectant with a concentration of 1-5 g / L, and a thickener with a concentration of 1-5 g / L; The emulsifier is selected from one or more of glyceryl stearate, glyceryl stearate / PEG-stearate, cetearyl alcohol, cetearyl alcohol / PEG-20 stearate, alkyl glucoside with 8-16 carbon atoms in the alkyl group, sodium isostearoyl lactyl lactate, etc.; The humectant is selected from one or more of glycerol, propylene glycol, sodium PCA, 1,3-propanediol, butanediol, hexanediol, 1,2-pentanediol, octyldecanol, sorbitol, ethylhexylglycerin, trehalose, betaine, sodium hyaluronate, carbohydrate isomers, urea, allantoin, and panthenol, etc.; The thickener is selected from one or more of xanthan gum, tara gum, acryloyldimethyltaurate / VP copolymer, sodium acryloyldimethyltaurate / acrylate copolymer, carbomer, and hydroxyethyl cellulose, etc.

7. Use of the Chinese herbal medicine extracellular vesicle composition according to any one of claims 1-2 or the Chinese herbal medicine extracellular vesicle composition prepared by the method according to claim 3 or the composition having the efficacy of preventing hair loss and promoting hair growth according to any one of claims 4-6 in the preparation of a product having the efficacy of preventing hair loss and promoting hair growth.

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