Composition containing hirudin and application of composition in preparation of product with effect of improving scalp absorption

Abstract

The invention relates to the technical field of biological medicines, and particularly discloses a hirudin-containing composition and application thereof in preparation of a product with a scalp absorption improving effect. The hirudin-containing composition is prepared from hirudin and a traditional Chinese medicine extract, the traditional Chinese medicine extract is prepared from the following raw materials: herba ecliptae, cacumen biotae and rhizoma drynariae. Researches show that the composition has the effect of promoting hair follicle cell proliferation; in addition, the absorption on the scalp can be improved; the important application value is realized.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, specifically to a composition containing hirudin and its application in the preparation of products that improve scalp absorption. Background Technology

[0002] Hair loss is a common dermatological condition with a complex pathogenesis involving multiple factors such as genetics, hormonal imbalances (e.g., elevated dihydrotestosterone (DHT) levels), hair follicle microcirculation disorders, chronic inflammation, oxidative stress, and functional decline of hair follicle stem cells. Currently, commonly used clinical treatments include minoxidil and finasteride. However, minoxidil has side effects such as local irritation and hirsutism, while finasteride may cause sexual dysfunction. Both require long-term use, and relapse is common after discontinuation, limiting patient adherence.

[0003] In recent years, traditional Chinese medicine (TCM) has received widespread attention in the field of hair loss prevention and treatment due to its multi-target, holistic regulatory, and relatively high safety profile. For example, herbs such as Platycladus orientalis leaves, Polygonum multiflorum, Angelica sinensis, and Salvia miltiorrhiza have been proven to have antioxidant, anti-inflammatory, 5α-reductase-inhibiting, or hair follicle cell proliferation-promoting effects. Besides its widespread use in antithrombotic therapy, recent studies have found that hirudin has a significant effect on improving microcirculation. Hirudin can reduce local blood viscosity and dilate microvessels by inhibiting thrombin-mediated platelet aggregation and fibrin deposition, thereby enhancing tissue perfusion. In topical skin applications, hirudin can effectively alleviate insufficient nutrient supply to hair follicles caused by microthrombus formation or blood stasis, creating a favorable microenvironment for hair follicle regeneration.

[0004] However, there are no research reports on the use of combining hirudin and traditional Chinese medicine extracts to promote hair follicle cell proliferation in the current technology. Summary of the Invention

[0005] In order to overcome at least one technical problem existing in the prior art, the present invention provides a composition containing hirudin and its application in the preparation of products that improve scalp absorption.

[0006] The technical solution of the present invention is as follows: The present invention first provides a composition containing hirudin, which includes hirudin and a traditional Chinese medicine extract; the raw materials for preparing the traditional Chinese medicine extract include Eclipta prostrata, Platycladus orientalis leaves and Drynaria fortunei.

[0007] The inventors discovered in their research that a composition obtained by combining hirudin with a traditional Chinese medicine extract obtained by the method described in this invention has the effect of promoting the proliferation of hair follicle cells.

[0008] Furthermore, in the composition described in this invention, hirudin can significantly optimize the microenvironment for the action of active ingredients by improving scalp microcirculation, inhibiting local inflammation and fibrosis, protecting hair follicle cell function, and synergistically regulating hair follicle cycle-related signaling pathways with traditional Chinese medicine extracts, thereby achieving functional improvement in scalp absorption.

[0009] Preferably, the weight ratio of hirudin to the traditional Chinese medicine extract is 1:3-6.

[0010] Most preferably, the weight ratio of hirudin to the traditional Chinese medicine extract is 1:5.

[0011] Preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei in the raw materials for preparing the traditional Chinese medicine extract is 4-6:2-4:2-4.

[0012] Most preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves and Drynaria fortunei in the raw materials for preparing the Chinese herbal extract is 5:3:3.

[0013] Preferably, the raw materials for preparing the traditional Chinese medicine extract also include Euonymus alatus.

[0014] Preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei and Euonymus alatus in the raw materials for preparing the Chinese herbal extract is 4-6:2-4:2-4:2-4.

[0015] Most preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei and Euonymus alatus in the raw materials for preparing the Chinese herbal extract is 5:3:3:3.

[0016] Further research by the inventors revealed that adding a Chinese herbal extract prepared from Euonymus alatus to the raw materials Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei could further enhance the hair follicle cell proliferation-promoting effect of the composition.

[0017] Preferably, the raw materials for preparing the traditional Chinese medicine extract also include Leonurus japonicus.

[0018] Preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei and Leonurus japonicus in the raw materials for preparing the Chinese herbal extract is 4-6:2-4:2-4:2-4.

[0019] Most preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei and Leonurus japonicus in the raw materials for preparing the Chinese herbal extract is 5:3:3:3.

[0020] Further research by the inventors revealed that adding Leonurus japonicus extract to the traditional Chinese medicine raw materials Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei can further enhance the hair follicle cell proliferation promoting effect of the composition.

[0021] Preferably, the raw materials for preparing the traditional Chinese medicine extract also include Euonymus alatus and Leonurus japonicus.

[0022] Preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, Euonymus alatus, and Leonurus japonicus in the raw materials for preparing the herbal extract is 4-6:2-4:2-4:2-4:2-4.

[0023] Most preferably, the weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, Euonymus alatus, and Leonurus japonicus in the raw materials for preparing the Chinese herbal extract is 5:3:3:3:3.

[0024] Further research by the inventors revealed that the herbal extracts prepared by simultaneously adding Euonymus alatus and Leonurus japonicus to the raw materials Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei can significantly enhance the hair follicle cell proliferation-promoting effect of the composition. The degree of enhancement in promoting hair follicle cell proliferation is significantly higher than that of herbal extracts prepared by adding Euonymus alatus or Leonurus japonicus alone. Furthermore, the herbal extracts prepared by simultaneously adding Euonymus alatus and Leonurus japonicus to the raw materials Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei can synergistically enhance the hair follicle cell proliferation-promoting effect of the composition. Preferably, the herbal extract is prepared by the following method: the herbal raw materials are mixed and extracted with a solvent, the extract is separated after extraction, and the extract is concentrated and dried to obtain a solvent extract; the solvent extract is then used to obtain the herbal extract.

[0025] Preferably, the ratio of the solvent to the total weight of the Chinese herbal raw materials is 5-10L:1Kg.

[0026] Most preferably, the ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg.

[0027] Preferably, the extraction is performed by heating and reflux extraction; the extraction time is 1-3 hours.

[0028] Preferably, the solvent is selected from water or an organic solvent.

[0029] Preferably, the organic solvent is selected from ethanol or ethyl acetate.

[0030] More preferably, the preparation method of the traditional Chinese medicine extract further includes the following steps: loading the solvent extract onto a silica gel column, first eluting with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:5-8 to remove impurities; then eluting with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:25-28, collecting the eluent eluted by the mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:25-28, concentrating and drying to obtain the traditional Chinese medicine extract.

[0031] Most preferably, the preparation method of the traditional Chinese medicine extract further includes the following steps: loading the solvent extract onto a silica gel column, first eluting with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:7 to remove impurities; then eluting with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:27, collecting the eluent eluted by the mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:27, concentrating and drying to obtain the traditional Chinese medicine extract.

[0032] In further research, the inventors discovered that the preparation method of the present invention is crucial; when the extract prepared under the silica column elution conditions described above is added to the composition, its effect on promoting hair follicle cell proliferation is significantly enhanced compared to the extract prepared without the silica column elution conditions described above.

[0033] Furthermore, the inventors have discovered that the silica column elution conditions of the present invention are crucial; the extract prepared under the silica column elution conditions described above must be added to the composition for its effect of promoting hair follicle cell proliferation to be further significantly enhanced; however, the addition of extracts prepared under other silica column elution conditions does not result in a further significant enhancement of its effect of promoting hair follicle cell proliferation. The present invention also provides the use of the above-mentioned composition containing hirudin in the preparation of products that improve scalp absorption.

[0034] The present invention also provides the use of the above-mentioned composition containing hirudin in the preparation of products that promote the proliferation of hair follicle cells.

[0035] Beneficial effects: This invention provides a novel composition containing hirudin and traditional Chinese medicine extracts; studies have shown that the composition promotes hair follicle cell proliferation; in addition, it can improve absorption by the scalp; it has important application value. Detailed Implementation

[0036] The present invention will be further explained below with reference to specific embodiments, but the embodiments do not limit the scope of protection of the present invention.

[0037] Example 1: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: After mixing the Chinese herbal raw materials, the mixture is heated to boiling with a solvent and extracted for 1 hour. After extraction, the extract is separated, concentrated and dried to obtain the solvent extract. The solvent extract is the Chinese herbal extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg; the solvent is ethyl acetate. The Chinese medicinal materials consist of Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei in a weight ratio of 5:3:3.

[0038] Example 2: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: After mixing the Chinese herbal raw materials, the mixture is heated to boiling with a solvent and extracted for 1 hour. After extraction, the extract is separated, concentrated and dried to obtain the solvent extract. The solvent extract is the Chinese herbal extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg; the solvent is ethyl acetate. The Chinese medicinal materials consist of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, and Euonymus alatus in a weight ratio of 5:3:3:3.

[0039] Example 3: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: After mixing the Chinese herbal raw materials, the mixture is heated to boiling with a solvent and extracted for 1 hour. After extraction, the extract is separated, concentrated and dried to obtain the solvent extract. The solvent extract is the Chinese herbal extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg; the solvent is ethyl acetate. The Chinese medicinal materials consist of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, and Leonurus japonicus in a weight ratio of 5:3:3:3.

[0040] Example 4: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: After mixing the Chinese herbal raw materials, the mixture is heated to boiling with a solvent and extracted for 1 hour. After extraction, the extract is separated, concentrated and dried to obtain the solvent extract. The solvent extract is the Chinese herbal extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg; the solvent is ethyl acetate. The Chinese medicinal materials consist of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, Euonymus alatus, and Leonurus japonicus in a weight ratio of 5:3:3:3:3.

[0041] Example 5: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: (1) After mixing the Chinese herbal raw materials, heat them to boiling with a solvent and extract for 1 hour. After extraction, separate the extract and concentrate and dry the extract to obtain the solvent extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg. The solvent is ethyl acetate. The Chinese herbal raw materials are composed of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, Euonymus alatus and Leonurus japonicus in a weight ratio of 5:3:3:3:3. (2) The solvent extract is loaded onto a silica gel column (the silica gel column is filled with silica gel of 30 times the weight of the solvent extract; the silica gel is 200-300 mesh silica gel). First, it is eluted with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:7 with a volume ratio of 5 column volumes to remove impurities. Then, it is eluted again with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:27 with a volume ratio of 5 column volumes. The eluent eluted with the mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:27 is collected, concentrated and dried to obtain the Chinese herbal extract.

[0042] Comparative Example 1: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: (1) After mixing the Chinese herbal raw materials, heat them to boiling with a solvent and extract for 1 hour. After extraction, separate the extract and concentrate and dry the extract to obtain the solvent extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg. The solvent is ethyl acetate. The Chinese herbal raw materials are composed of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, Euonymus alatus and Leonurus japonicus in a weight ratio of 5:3:3:3:3. (2) The solvent extract is loaded onto a silica gel column (the silica gel column is filled with silica gel of 30 times the weight of the solvent extract; the silica gel is 200-300 mesh silica gel). First, it is eluted with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:5, which is 5 column volumes, to remove impurities. Then, it is eluted again with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:15, which is 5 column volumes. The eluent eluted by the mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:15 is collected, concentrated and dried to obtain the Chinese herbal extract.

[0043] Comparative Example 2: Preparation of a composition containing hirudin The composition containing hirudin is obtained by mixing hirudin and traditional Chinese medicine extract at a weight ratio of 1:5. The herbal extract mentioned above is prepared by the following method: (1) After mixing the Chinese herbal raw materials, heat them to boiling with a solvent and extract for 1 hour. After extraction, separate the extract and concentrate and dry the extract to obtain the solvent extract. The ratio of the solvent to the total weight of the Chinese herbal raw materials is 8L:1Kg. The solvent is ethyl acetate. The Chinese herbal raw materials are composed of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei, Euonymus alatus and Leonurus japonicus in a weight ratio of 5:3:3:3:3. (2) The solvent extract is loaded onto a silica gel column (the silica gel column is filled with silica gel of 30 times the weight of the solvent extract; the silica gel is 200-300 mesh silica gel). First, it is eluted with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:20 (5 column volumes) to remove impurities. Then, it is eluted again with a mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:40 (5 column volumes). The eluent eluted with the mixed solvent of cyclohexane and ethyl acetate in a volume ratio of 100:40 is collected, concentrated and dried to obtain the Chinese herbal extract.

[0044] Experimental Example: Experiment to Promote Hair Follicle Cell Growth Ninety-six KM mice (half male and half female) were selected. Hair was removed from the backs of the mice, covering an area of ​​2×2 cm. The mice were randomly divided into seven experimental groups and one control group, with 12 mice in each group. The experimental group mice were coated with an equal amount of glycerol containing 1% of the test sample, while the control group mice were coated with an equal amount of glycerol without the test sample. The treatment was carried out twice a day for seven consecutive days, after which the mice were sacrificed. Skin samples from each group were collected and stained. The number of hair follicle cells in the skin of each group was counted under a microscope. The results are shown in Table 1.

[0045] Among them, the test samples applied to experimental groups 1-7 were compositions containing hirudin prepared in Examples 1-5 and Comparative Examples 1-2, respectively.

[0046] Table 1. Experimental results of the hair follicle cell growth promotion effect of the hirudin-containing composition of the present invention. As can be seen from the experimental results in Table 1, the number of hair follicle cells in experimental group 1 was significantly higher than that in the blank group; this indicates that the composition obtained by combining hirudin with the traditional Chinese medicine extract obtained by the method described in this invention has the effect of promoting the proliferation of hair follicle cells.

[0047] As can be seen from the experimental results in Table 1, the number of hair follicle cells in experimental group 2 was higher than that in experimental group 1. This indicates that the addition of Euonymus alatus extract to the raw materials Eclipta prostrata, Platycladus orientalis leaves and Drynaria fortunei can further enhance the hair follicle cell proliferation promoting effect of the composition.

[0048] As can be seen from the experimental results in Table 1, the number of hair follicle cells in experimental group 3 was higher than that in experimental group 1. This indicates that the Chinese herbal extract prepared by adding Leonurus japonicus seeds to the raw materials Eclipta prostrata, Platycladus orientalis leaves and Drynaria fortunei can further enhance the hair follicle cell proliferation promoting effect of the composition.

[0049] The experimental results in Table 1 show that the number of hair follicle cells in experimental group 4 was significantly higher than that in experimental group 1, and also significantly higher than that in experimental groups 2 and 3. This indicates that the herbal extract prepared by simultaneously adding Euonymus alatus and Leonurus japonicus to the raw materials Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei can significantly enhance the hair follicle cell proliferation-promoting effect of the composition. The degree of enhancement of the hair follicle cell proliferation-promoting effect of the herbal extract is significantly higher than that prepared by adding Euonymus alatus or Leonurus japonicus alone. The herbal extract prepared by simultaneously adding Euonymus alatus and Leonurus japonicus to the raw materials Eclipta prostrata, Platycladus orientalis leaves, and Drynaria fortunei can synergistically enhance the hair follicle cell proliferation-promoting effect of the composition. As can be seen from the experimental results in Table 1, the number of hair follicle cells in experimental group 5 was significantly higher than that in experimental group 4. This indicates that the preparation method of the present invention is crucial. When the extract prepared by the above-mentioned silica column elution conditions of the present invention is added to the composition, its effect on promoting hair follicle cell proliferation is further significantly improved compared with the extract prepared without the above-mentioned silica column elution conditions.

[0050] As can be seen from the experimental results in Table 1, although the number of hair follicle cells in experimental groups 6 and 7 was increased compared with that in experimental group 4, the increase was not significant and much smaller than that in experimental group 5. This indicates that the silica column elution conditions of the present invention are crucial. The extract prepared under the silica column elution conditions of the present invention must be added to the composition to further significantly enhance its effect on promoting hair follicle cell proliferation. However, adding extracts prepared under other silica column elution conditions does not further significantly enhance its effect on promoting hair follicle cell proliferation.

Claims

1. A composition containing hirudin, characterized in that, It contains hirudin and traditional Chinese medicine extracts; the raw materials for preparing the traditional Chinese medicine extracts include Eclipta prostrata, Platycladus orientalis leaves and Drynaria fortunei.

2. The composition containing hirudin according to claim 1, characterized in that, The weight ratio of hirudin to traditional Chinese medicine extract is 1:3-6.

3. The composition containing hirudin according to claim 1, characterized in that, The weight ratio of Eclipta prostrata, Platycladus orientalis leaves and Drynaria fortunei in the raw materials for preparing the Chinese herbal extract is 4-6:2-4:2-4.

4. The composition containing hirudin according to claim 1, characterized in that, The raw materials used in the preparation of the herbal extract also include Euonymus alatus.

5. The composition containing hirudin according to claim 1, characterized in that, The weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei and Euonymus alatus in the raw materials for preparing the Chinese herbal extract is 4-6:2-4:2-4:2-4.

6. The composition containing hirudin according to claim 1, characterized in that, The raw materials used in the preparation of the herbal extract also include Leonurus japonicus seeds.

7. The composition containing hirudin according to claim 6, characterized in that, The weight ratio of Eclipta prostrata, Platycladus orientalis leaves, Drynaria fortunei and Leonurus japonicus in the raw materials for preparing the Chinese herbal extract is 4-6:2-4:2-4:2-4.

8. The composition containing hirudin according to claim 1, characterized in that, The herbal extract is prepared by the following method: the herbal raw materials are mixed and extracted with a solvent. After extraction, the extract is separated, and the extract is concentrated and dried to obtain the solvent extract. The solvent extract is then used to obtain the herbal extract.

9. The use of the hirudin-containing composition according to any one of claims 1-8 in the preparation of a product having the effect of improving scalp absorption.

10. The use of the hirudin-containing composition according to any one of claims 1-8 in the preparation of a product having the effect of promoting hair follicle cell proliferation.

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