Traditional Chinese medicine compound composition for treating androgen-derived alopecia as well as preparation method and application of traditional Chinese medicine compound composition

Abstract

The invention discloses a traditional Chinese medicine compound composition for treating androgen-derived alopecia and a preparation method and application thereof. The traditional Chinese medicine compound composition is prepared from cacumen biotae, herba ecliptae, glossy privet fruit, radix polygoni multiflori and radix paeoniae alba according to a specific proportion. And after reflux extraction with 75% ethanol, the skin androgen level (Plt; 0.05) is significantly reduced through local smearing administration, the number of hair follicles is increased to 67 + / -5 / view, and hair regeneration in an alopecia area is promoted. The composition has the advantages of safety, effectiveness, low cost, avoidance of systemic side effects and the like, and is suitable for developing AGA external treatment medicines.

Description

Technical Field

[0001] The present invention relates to the technical field of biomedicine, and in particular to a traditional Chinese medicine compound composition for treating androgenic alopecia, and a preparation method and application thereof. Background Art

[0002] Androgenic alopecia (AGA), also known as seborrheic alopecia, is primarily caused by abnormalities in androgens and their receptors. Clinical manifestations include an oily scalp and hair loss, often accompanied by greasy, scalp-related scalp irritation, itching, and dandruff. In my country, the prevalence of AGA is approximately 1 in 5 men and 1 in 20 women. While hair loss does not affect physical health, it can cause psychological distress to patients. As living standards improve, the demand for treatment for seborrheic alopecia is also increasing significantly.

[0003] Western medicine believes that seborrheic alopecia is caused by genetics and hormonal abnormalities. Current treatments include medication, hair transplantation, low-intensity laser therapy, and mesotherapy. For medication, men typically take finasteride and 5% minoxidil topical application, while women take spironolactone or tanshinone capsules and 2% minoxidil topical application. If treatment is ineffective, external treatments such as surgery are available, but these are often expensive. Consequently, Traditional Chinese Medicine (TCM), known for its milder effects and nourishing properties, is gaining increasing popularity.

[0004] Traditional Chinese Medicine (TCM) believes that hair loss is closely related to the liver and kidneys, with emotions and diet being key contributing factors. TCM treatments for seborrheic alopecia include both internal and external therapies. Internal treatments utilize appropriate medications based on clinical diagnosis, while external treatments include acupuncture, topical applications, and washes.

[0005] Therefore, it is necessary to develop a Chinese medicine compound composition that can be used to treat androgenic alopecia. Summary of the Invention

[0006] The present invention aims to provide a Chinese medicinal compound composition for treating androgenic alopecia, which has an improving effect on alopecia.

[0007] In order to achieve the above object, the present invention adopts the following technical solutions: In a first aspect of the present invention, a Chinese herbal compound composition for treating androgenic alopecia is provided. The Chinese herbal compound composition for treating androgenic alopecia comprises the following raw materials in parts by weight: 10-20 parts of Platycladus orientalis leaves, 10-20 parts of Ecliptae herba, 10-20 parts of Ligustrum lucidum fruits, 10-20 parts of Polygonum multiflorum, and 10-20 parts of white peony root.

[0008] Furthermore, the Chinese herbal compound composition for treating androgenic alopecia comprises the following raw materials in parts by weight: 10 parts of Platycladus orientalis leaves, 10 parts of Ecliptae, 10 parts of Fructus Ligustri Lucidi, 10 parts of Polygonum multiflorum, 10 parts of the white paeonia lactiflora.

[0009] In a second aspect of the present invention, a method for preparing the Chinese herbal compound composition for treating androgenic alopecia is provided, the method comprising: Mixing the Platycladus orientalis leaves, Eclampsia chinensis, Ligustrum lucidum fruits, Polygonum multiflorum and Paeonia lactiflora to obtain a composition; The composition is crushed, ethanol is added for reflux extraction, and the extract is obtained after removing the medicinal residues by filtration.

[0010] Furthermore, the ethanol is 75% ethanol at a concentration of 6 times the weight of the composition.

[0011] Furthermore, the reflux extraction time is 3-5 h.

[0012] In a third aspect of the present invention, a pharmaceutical preparation is provided, comprising the composition or the extract obtained by the preparation method.

[0013] Furthermore, the medicine also includes pharmaceutically acceptable excipients and carriers.

[0014] Furthermore, the auxiliary materials include at least one of a filler, a disintegrant, a binder, an excipient, a diluent, a lubricant, a sweetener and a colorant.

[0015] Furthermore, the dosage form of the drug includes at least one of granules, tablets, pills, capsules, injections or dispersions.

[0016] In a third aspect of the present invention, there is provided use of the Chinese herbal compound composition or the pharmaceutical preparation according to claims 6 to 9 in the preparation of a medicament for treating androgenic alopecia.

[0017] One or more technical solutions in the embodiments of the present invention have at least the following technical effects or advantages: 1. The present invention provides a traditional Chinese medicine compound composition for treating androgenic alopecia, which uses Platycladus orientalis leaves, Ecliptae macrophylla, Ligustrum lucidum fruit, Polygonum multiflorum and Paeonia lactiflora as raw materials. It can effectively promote hair growth in the androgenic alopecia area of ​​mice, is safe and effective, and has no adverse reactions. Moreover, the various traditional Chinese medicines of the present invention have a wide range of sources, low cost, economical and affordable, and the preparation method is simple.

[0018] 2. The Chinese herbal compound solution prepared by the present invention can be directly applied to the skin for transdermal administration. Compared with the oral tablets of anti-androgenic alopecia drugs on the market, it can reduce systemic adverse reactions and enhance patient compliance, providing a new idea for the design of topical preparations for the treatment of androgenic alopecia. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following is a brief introduction to the drawings required for use in the description of the embodiments. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0020] Figure 1 It is the construction of AGA mouse model; Figure 2 To study the effects of compound Chinese medicine on the skin moisture and oil content of AGA mice; Figure 3 To study the effect of compound Chinese medicine on hair growth in AGA mice; Figure 4 The effect of compound Chinese medicine on androgen levels in the skin of AGA mice; Figure 5 Figure 3 shows the effect of compound Chinese medicine on the skin tissue structure of AGA mice. A is the blank control group; B is the AGA model group; C is the Chinese medicine treatment group; and D is the positive control group. DETAILED DESCRIPTION

[0021] The present invention will be described in detail below in conjunction with specific embodiments and examples, and the advantages and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific embodiments and examples are for illustrating the present invention, rather than for limiting the present invention.

[0022] Throughout this specification, unless otherwise specified, the terms used herein should be understood as having the same meaning as commonly used in the art. Therefore, unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In the event of any conflict, the present specification shall take precedence.

[0023] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or obtained through existing methods.

[0024] The following is a detailed description of a Chinese herbal compound composition for treating androgenic alopecia of the present application in combination with examples, comparative examples and experimental data.

[0025] Example 1. Preparation of Chinese herbal compound solution 1. The composition of the Chinese herbal compound: 10 parts each of Platycladus orientalis leaves, Ecliptae macrophylla, Ligustrum lucidum fruit, Polygonum multiflorum and Paeonia lactiflora.

[0026] 2. Preparation of Chinese medicine compound solution: Take the above-mentioned Chinese medicine composition in proportion, crush it and wrap it in gauze, add 6 times the amount of 75% ethanol and reflux and extract for 4 hours. After filtering with gauze to remove the residue, place it in a rotary evaporator and concentrate it to 0.5 g / mL (total amount of crude drug), that is, the final volume is 150 mL.

[0027] Example 2: Construction and grouping of AGA mouse model 1. Construction of AGA mouse model Several C57BL / 6 male mice, 4 weeks old, weighing 10-15 g were anesthetized with an intraperitoneal injection of 4% chloral hydrate. The hair on the back was removed using a depilatory cream, covering as uniform an area as possible. Testosterone propionate was injected subcutaneously at multiple sites for 28 days at a dose of 0.1 ml / 10 g body weight. The AGA mouse model was considered established successfully when no hair grew back in the depilated area on the back of the mouse (e.g., Figure 1 ).

[0028] 2. Grouping and Dosing Mice with successful modeling were randomly divided into three groups: an AGA model group, a TCM compound treatment group, and a positive control group (minoxidil treatment). The AGA model group received daily application of normal saline to the back. The TCM treatment group received daily application of 200 μL of the TCM compound extract (the extract was diluted 10-fold with normal saline before use) directly after depilation. The positive control group received daily application of 200 μL of minoxidil to the back for 21 consecutive days.

[0029] Example 3: Effects of a Chinese herbal compound on skin moisture, oil content, and hair growth in AGA mice 1. Determination of skin moisture and oil content Before and after administration, the moisture and oil content of the back skin of mice in each group was measured using a Zhixian skin tester. Changes in moisture and oil content in the skin tissue of mice in each group were analyzed on the 1st, 7th, 14th, and 21st days of administration.

[0030] 2. Evaluation of hair growth After hair removal, hair growth was recorded in detail on days 1, 7, 14, and 21 of treatment, with the exception of the blank group. The exact time point at which the skin color of the depilated area changed from pink to black, as well as the time point at which new hair began to grow in the depilated area, was recorded for each mouse. Hair growth was photographed and recorded daily in the same environment after treatment, and scored.

[0031] 3. Results The results are as follows Figure 2As shown in the results, after 21 days of treatment, applying the Chinese herbal compound extract can increase the moisture content and reduce the oil content of the skin tissue of the model mice. The hair growth in the treatment group was obvious, and the hair growth scores of the Chinese herbal compound group and the positive control group were the same, indicating that the improvement effect of the Chinese herbal compound extract on androgenic alopecia is close to that of minoxidil.

[0032] The results are as follows Figure 3 As shown, 14 days after dosing, sparse hair began to grow on the backs of the model group mice. After 21 days, the hair in the model group covered two-thirds of the hair loss area and was light gray. Seven days after dosing, mice treated with the Chinese herbal compound grew noticeable but sparse hair on their backs. After 14 days, abundant new hair was observed, similar to that in the positive control group. After 21 days, the hair on the backs of the mice almost covered the entire hair loss area and was black in color.

[0033] Example 4: Effect of a Chinese herbal compound on androgen levels in the skin tissue of AGA mice 1. ELISA detection of androgen levels in mouse skin tissue: After the administration and observation, the skin tissues on the back of the mice were cut and homogenized, and the androgen levels in the skin of the mice in each group were detected using an ELISA kit.

[0034] The specific operation method is: (1) Move all reagents to room temperature (18-25°C) and equilibrate for at least 30 minutes. Prepare the reagents according to the above method and set aside.

[0035] (2) Take out the ELISA plate and set up a blank control well without adding any liquid; set up three wells for each standard point in sequence, add 50μl of the sample to each well, and directly add 50μl of the sample to be tested to each of the remaining detection wells.

[0036] (3) Add 50 μl of incubation solution to each well and shake gently for 10 minutes.

[0037] (4) Add 50 μl of enzyme conjugate to each well (except the blank control well), mix thoroughly, attach a self-adhesive sealant, and incubate at 37°C for 1 hour.

[0038] (5) Use a hand brush to clean the liquid in the pores. Inject the cleaning solution into each hole, let it stand for 10 seconds, pour it out and shake it dry, pat it dry with a paddle, and repeat this step 5 times.

[0039] (6) Add 100 μl of substrate solution to each well, shake and mix thoroughly, then color develop for 20 min at ambient temperature (18-25°C) in the dark, and add 50 μl of stop solution to each well.

[0040] (7) Within 10 minutes after stopping the reaction, the optical density (OD value) of each sample well was measured in sequence using a microplate reader at a wavelength of 450 nm.

[0041] 2. Results The results are as follows Figure 4 As shown in the results, the androgen level in the skin tissue of mice in the AGA model group was significantly higher than that in the control group (P<0.05). After 21 days of treatment, the androgen level in the skin tissue of AGA mice was significantly downregulated (P<0.05), indicating that the Chinese herbal compound extract can improve the excessive androgen level in the scalp tissue of AGA mice. Example 5: Effect of Chinese herbal compound on skin tissue structure of AGA mice 1. HE staining to observe the changes in mouse skin tissue structure: (1) Sampling and fixation: Cut 2x2cm of mouse back skin tissue and fix it with 2% paraformaldehyde for more than 24 hours.

[0042] (2) Dehydration and transparency: Dehydrate the tissue blocks with 70% to 90% alcohol. Immerse the mouse tissue blocks in xylene to make them transparent and use it to displace the alcohol in the mouse tissue blocks.

[0043] (3) Wax immersion and embedding: Place the mouse tissue block in the melted paraffin and keep it in an incubator for 24 hours. Place the wax-immersed mouse tissue block in the melted paraffin and wait for it to cool down and form a block.

[0044] (4) Sectioning and dewaxing: Use a paraffin slicer to slice the mouse skin tissue into thin slices. Then place the slices in heated water for spreading. After processing, place them in a 45°C constant temperature oven to dry. Use xylene to wash away the paraffin in the mouse tissue slices. Then, wash away the xylene with alcohol from high concentration (90%) to low concentration (70%), and finally wash away the alcohol in distilled water.

[0045] (5) HE staining: Place the processed skin tissue sections in hematoxylin solution for 5 minutes. Rinse with running water for 2 minutes, then soak in distilled water for 5 minutes. Dehydrate in low concentration (70%) and high concentration (90%) alcohol for 5 minutes each, then stain with eosin staining solution for 2 minutes. After staining, the sections are first dehydrated and then transparentized, and finally sealed with neutral gum.

[0046] (6) After the gum has dried slightly, observe the changes in skin thickness, hair follicle size, and number under a microscope, and take photos for preservation.

[0047] 2. Results The results are as follows Figure 5As shown, the average number of hair follicles per field of view in the blank group was 35, with a relatively even distribution of follicles. The majority of visible follicles were medium-hair follicles with darker hair shaft pigmentation, indicating that some follicles were in the growth phase. Compared with the blank group, skin sections from the model group showed reduced hair follicle volume and number, and thinner skin thickness. The average number of hair follicles per field of view was 24, with a sparse distribution of follicles and reduced hair shaft pigmentation. The lighter hair shaft pigmentation suggests that the follicles are in the resting phase. After 21 days of topical treatment, the number and diameter of hair follicles in the skin tissue of mice in the compound Chinese medicine group increased significantly. The average number of hair follicles per field of view was 67, essentially returning to normal levels. Follicle density increased and became evenly distributed, and hair shaft pigmentation also increased significantly, indicating that the hair follicles have entered an active growth phase.

[0048] In summary, the traditional Chinese medicine compound solution prepared by the present invention can effectively promote hair growth in the androgenic alopecia area of ​​mice, reduce the androgen level in skin tissue, is safe and effective, and has no adverse reactions; it is low in cost, economical and affordable, and the preparation method is simple; the traditional Chinese medicine compound solution prepared by the present invention can be directly applied to the skin for transdermal administration, which can reduce systemic adverse reactions and enhance patient compliance, providing a new idea for the design of topical preparations for the treatment of androgenic alopecia.

[0049] Finally, it should be noted that the terms "comprises," "includes," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that includes a list of elements includes not only those elements, but also other elements not expressly listed, or elements inherent to such process, method, article, or apparatus.

[0050] Although the preferred embodiments of the present invention have been described, those skilled in the art may make additional changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0051] Obviously, those skilled in the art may make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if such changes and modifications fall within the scope of the claims and their equivalents, the present invention is intended to include such changes and modifications.

Claims

1. A Chinese medicinal compound composition for treating androgenic alopecia, characterized in that: The Chinese medicinal compound composition for treating androgenic alopecia comprises the following raw materials in parts by weight: 10-20 parts of Platycladus orientalis leaves, 10-20 parts of Ecliptae herba, 10-20 parts of Ligustrum lucidum fruits, 10-20 parts of Polygonum multiflorum, and 10-20 parts of white peony root.

2. The Chinese herbal compound composition for treating androgenic alopecia according to claim 1, characterized in that: The Chinese medicinal compound composition for treating androgenic alopecia comprises the following raw materials in parts by weight: 10 parts of Platycladus orientalis leaves, 10 parts of Ecliptae, 10 parts of Fructus Ligustri Lucidi, 10 parts of Polygonum multiflorum, 10 parts of the white paeonia lactiflora.

3. A method for preparing the Chinese medicinal compound composition for treating androgenic alopecia according to any one of claims 1 to 2, characterized in that: The method comprises: Mixing the Platycladus orientalis leaves, Eclampsia chinensis, Ligustrum lucidum fruit, Polygonum multiflorum and Paeonia lactiflora to obtain a composition; The composition is crushed, ethanol is added for reflux extraction, and the extract is obtained after removing the medicinal residue by filtration.

4. The preparation method according to claim 3, characterized in that The ethanol is 75% ethanol with a concentration of 6 times the weight of the composition.

5. The preparation method according to claim 3, characterized in that The reflux extraction time is 3-5 h.

6. A pharmaceutical preparation, characterized in that: The invention comprises the composition according to claim 1 or the extract obtained by the preparation method according to claims 3-5.

7. The pharmaceutical preparation according to claim 6, characterized in that The medicine also includes pharmaceutically acceptable excipients and carriers.

8. The pharmaceutical preparation according to claim 7, characterized in that The auxiliary materials include at least one of a filler, a disintegrant, a binder, an excipient, a diluent, a lubricant, a sweetener and a colorant.

9. The pharmaceutical preparation according to claim 6, characterized in that The dosage form of the drug includes at least one of granules, tablets, pills, capsules, injections or dispersions.

10. Use of the Chinese herbal compound composition according to any one of claims 1 to 2 or the pharmaceutical preparation according to claims 6 to 9 in the preparation of a medicament for treating androgenic alopecia.

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