Pharmaceutical composition and preparation for treating alopecia as well as preparation method and application of pharmaceutical composition

Abstract

The invention provides a pharmaceutical composition for treating alopecia, a preparation as well as a preparation method and application thereof, and belongs to the technical field of medicines. The pharmaceutical composition comprises the following main components in parts by weight: 12-18 parts of cacumen biotae, 12-18 parts of szechwan chinaberry fruit and 8-12 parts of garden balsam stem, wherein the cacumen biotae, the szechwan chinaberry fruit and the garden balsam stem are used as main components. The invention further provides a preparation method and application of the compound platycladus orientalis tincture, cell and animal experiments prove that the compound platycladus orientalis tincture has the proliferation promoting capacity on human primary dermal papilla cells, meanwhile, hair growth can be promoted, the symptom of alopecia can be remarkably improved, and all the medicines in the prescription have a synergistic effect, so that the compound platycladus orientalis tincture has a good effect on treating alopecia. The effect is better than that of a positive drug minoxidil, and the application prospect is good.

Description

Technical Field

[0001] The present invention belongs to the field of medical technology, and in particular relates to a pharmaceutical composition for treating hair loss, a preparation method and an application thereof. Background Art

[0002] Hair loss has become a very common skin problem worldwide. In recent years, with the increasing pressure of work and study, many people are in a state of mental tension for a long time, which has led to an increase in the number of hair loss patients year by year. The trend of hair loss among young people is becoming more and more obvious, and young people are gradually becoming a high-risk group for hair loss.

[0003] Androgenetic alopecia (AGA) and alopecia areata (AA) are the two most common types of hair loss. Androgenetic alopecia, also known as seborrheic alopecia, usually begins in adolescence, and is mainly manifested by the gradual thinning of hair diameter and the continuous decrease in density, and eventually develops into varying degrees of baldness. Since this type of hair loss is often accompanied by increased oil secretion on the scalp, it has become the most common hair loss disease in clinical practice. Alopecia areata is the second most common hair loss disease. It is an autoimmune non-scarring hair loss related to genetics, emotional stress, endocrine disorders, autoimmunity and other factors. The pathogenesis is mainly lymphocyte infiltration around the hair follicles, which leads to impaired hair follicle stem cell function and impaired hair regeneration. Globally, its lifetime prevalence is about 1.7% to 2.1%. Although about 50% of patients may recover within 1 year, the recurrence rate is as high as 80%. The repeated nature of the disease causes patients to suffer long-term distress and bear both physical and psychological pressures.

[0004] Hair loss is not only related to aesthetics, but also brings a heavy mental burden to patients, affecting all aspects of life such as interpersonal communication, employment and job hunting, and may even cause psychological problems such as depression, anxiety and inferiority, seriously reducing the quality of life. At present, the treatment of hair loss faces many challenges. Although Western medicine has certain efficacy, it has prominent disadvantages. For example, minoxidil, glucocorticoids, cyclosporine, etc. are inconvenient to use and need to strictly follow complex specifications, which can easily affect medication compliance; there are many adverse reactions involving skin, metabolism, immunity, liver and kidney, etc.; the treatment effect varies from person to person, and most of them require long-term medication. Hair loss is prone to recurrence after stopping the medication. Traditional Chinese medicine has unique potential in the field of hair loss treatment. It can not only relieve hair loss, but also has advantages in preventing recurrence, stabilizing the condition, and improving the quality of life. However, the existing traditional Chinese medicine hair loss treatment scheme has shortcomings. For example, patent application CN104027395A discloses a drug for treating hair loss, which uses fresh medicinal materials, but it is difficult to preserve and obtain, cumbersome to use and the effect is poor; patent application CN109350589A discloses a traditional Chinese medicine hair growth liquid containing angelica extract, which has many Chinese medicine extracts, complex preparation and difficult to control, and the actual effective rate is only 67%; patent CN117122643A discloses a Tongqiao Shengfa tincture for treating seborrheic alopecia, which has many raw materials, complex formula, high cost, high preparation conditions, difficult quality control, large individual differences in anti-hair loss effects, and cannot solve the itching and greasy problems of seborrheic alopecia. So far, current hair loss treatment drugs are difficult to meet the diverse needs and individual differences of patients. In view of this, the development of new hair loss treatment drugs, especially the exploration of the potential of traditional Chinese medicine and the transformation into effective products, has become an urgent problem to be solved. Summary of the invention

[0005] The present invention provides a pharmaceutical composition for treating hair loss, a preparation thereof, and a preparation method and application thereof. The pharmaceutical composition aims to utilize the unique efficacy of natural plant ingredients, exert a synergistic effect through scientific proportioning, and effectively improve the condition of hair loss.

[0006] In order to achieve the above-mentioned object of the invention, the present invention provides the following technical solutions:

[0007] The invention provides a pharmaceutical composition for treating hair loss, which is composed of the following raw materials in parts by weight: 12 to 18 parts of Platycladus orientalis leaves, 12 to 18 parts of Toosendan seeds, and 8 to 12 parts of Herba Scutellariae.

[0008] Preferably, in the pharmaceutical composition, the weight parts of the raw materials are: 15 parts of Platycladus orientalis leaves, 15 parts of Toosendan fruits, and 10 parts of Herba Spermatophytae.

[0009] Preferably, in the pharmaceutical composition, the weight parts of the raw materials are: 12 parts of Platycladus orientalis leaves, 18 parts of Toosendan fruits, and 8 parts of Herba Spermatophytae.

[0010] Preferably, in the pharmaceutical composition, the weight parts of the raw materials are: 18 parts of Platycladus orientalis leaves, 12 parts of Toosendan fruits, and 12 parts of Herba Scutellariae spatholobi.

[0011] The present invention also provides the use of the above-mentioned pharmaceutical composition in preparing a preparation for treating hair loss.

[0012] Preferably, the hair loss includes androgenic alopecia and / or alopecia areata.

[0013] As an embodiment, the preparation includes tincture, solution, foam, powder, lotion, oil, emulsion, ointment, paste, film or gel.

[0014] The invention provides a compound Platycladus orientalis tincture, wherein the active component comprises the ethanol water extract of any one of the above-mentioned pharmaceutical compositions.

[0015] The present invention also provides a preparation method of the aforementioned compound Platycladus orientalis tincture, which comprises any one of a dissolution method, a dilution method, an immersion method, and a percolation method, and the specific steps are as follows: crushing each raw material of the pharmaceutical composition described in any one of the above items to obtain a medicinal material powder. Using any one of a dissolution method, a dilution method, an immersion method, and a percolation method, the medicinal material powder or the prepared fluid extract is mixed with an ethanol solution, filtered, and fixed to obtain a compound Platycladus orientalis tincture.

[0016] As an embodiment, the dissolution method comprises adding the powder or fluid extract into an ethanol solution for dissolution.

[0017] As an embodiment, the impregnation method comprises impregnating the powder in an ethanol solution, performing an immersion treatment, and filtering to obtain an ethanol extract.

[0018] As an embodiment, the percolation method includes wetting the powder and then loading it into a percolator, percolating and extracting with an ethanol solution to obtain a percolate, and concentrating the percolate at low temperature to obtain a fluid extract.

[0019] As an embodiment, the dilution method comprises diluting the fluid extract with an ethanol solution.

[0020] As an embodiment, the concentration of the ethanol solution is 50% to 90%.

[0021] As an embodiment, the immersion time of the immersion method is 5 to 9 days.

[0022] As an embodiment, the final crude drug content of the compound Platycladus orientalis tincture is 20% to 60%.

[0023] Compared with the prior art, the present invention has the following beneficial effects:

[0024] The present invention provides a pharmaceutical composition, preparation, preparation method and application for treating hair loss, and selects three natural plant raw materials, namely, Platycladus orientalis leaf, Toosendan fruit and Herba Spermatophytae, which have a wide range of sources. The formula specifies the weight of each raw material, and through scientific proportioning, each component acts synergistically. In the formula, Platycladus orientalis leaf is used as the main drug, which has the effects of nourishing the liver and kidney, clearing away heat and detoxifying, and invigorating qi and promoting hair growth; the auxiliary drug Toosendan fruit can soothe the liver and regulate qi, promote blood circulation and remove blood stasis, harmonize liver qi, improve qi and blood circulation, alleviate hair loss caused by liver qi stagnation, and can promote blood circulation of the scalp and enhance the nutritional supply of hair follicles; the adjuvant Herba Spermatophytae can dredge the meridians, promote blood circulation, and remove rheumatism, help improve the scalp obstruction caused by rheumatism or blood stasis, further promote the nutritional absorption and blood circulation of hair follicles, and enhance the growth ability of hair. The three complement each other, focusing on soothing the liver and regulating qi, tonifying the kidney and replenishing blood, and promoting blood circulation and removing blood stasis. They are particularly suitable for hair loss caused by qi stagnation and blood stasis, and liver and kidney deficiency. They can regulate the balance of qi and blood in the body, improve scalp microcirculation, and restore healthy hair growth, with good therapeutic effects. The present invention also provides a preparation method, which uses a mild extraction process to retain the activity of each component to the greatest extent, avoiding the loss or destruction of effective components that may be caused by traditional methods. The process is easy to operate and easy to achieve industrial production.

[0025] The present invention also provides a compound arborvitae tincture and a preparation method. Preliminary experiments have shown that the compound arborvitae tincture can effectively promote the proliferation of HDPCs, and its effect is better than minoxidil. It has a positive effect on the proliferation of key cells of hair follicles and provides a cellular basis for hair regeneration. In-depth animal experiments have found that the compound arborvitae tincture has a good therapeutic effect on androgenic alopecia and alopecia areata, and can significantly promote hair growth. The present invention brings a new and effective treatment plan to many hair loss patients and promotes the further development of hair loss treatment technology. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a bar graph showing the effect of compound Platycladus orientalis tincture on the proliferation ability of human primary dermal dermal papilla cells (HDPCs), from left to right are blank control group, minoxidil group, 0.05%, 0.1% and 0.25% dilution concentration of compound Platycladus orientalis tincture groups, where * indicates P < 0.05, ** indicates P < 0.01, **** indicates P < 0.0001.

[0027] Figure 2 The bar graph shows the effects of different tinctures on the proliferation of primary human dermal papilla cells (HDPCs), from left to right, blank control group, Platycladus orientalis tincture group, Toosendan tincture group, Herba scutellariae tincture group, Platycladus orientalis + Toosendan tincture group, Platycladus orientalis + Herba scutellariae tincture group, and Compound Platycladus orientalis tincture group. ** indicates P < 0.01, and *** indicates P < 0.001.

[0028] Figure 3The figure is a graph showing the efficacy of compound Platycladus orientalis tincture on the androgenic alopecia model of C57BL / J6 mice, from left to right are blank group, model group, positive group (minoxidil tincture treatment group), tincture group 1 (Example 1), tincture group 2 (Example 5), tincture group 3 (Example 6).

[0029] Figure 4 The figure is the result of HE staining of the back skin of androgenic alopecia model mice, from left to right are blank group, model group, positive group (minoxidil tincture treatment group), tincture group 1 (Example 1), tincture group 2 (Example 5), tincture group 3 (Example 6).

[0030] Figure 5 This is a graph showing the efficacy of compound Platycladus orientalis tincture on alopecia areata model in C57BL / J6 mice, from left to right are blank group, model group, positive group (minoxidil tincture treatment group), tincture group 1 (Example 1), tincture group 2 (Example 5), tincture group 3 (Example 6).

[0031] Figure 6 The figure is the result of HE staining of the back skin of alopecia areata model mice, from left to right are blank group, model group, positive group (minoxidil tincture treatment group), tincture group 1 (Example 1), tincture group 2 (Example 5), tincture group 3 (Example 6).

[0032] Figure 7 The figures are before and after comparison of two patients treated with the compound Platycladus orientalis tincture prepared in Example 1 for 3 months. DETAILED DESCRIPTION

[0033] The invention provides a pharmaceutical composition for treating hair loss. The pharmaceutical composition is composed of the following raw materials in parts by weight: 12 to 18 parts of Platycladus orientalis leaves, 12 to 18 parts of Toosendan seeds, and 8 to 12 parts of Herba Scutellariae.

[0034] In a specific embodiment, the present invention provides a pharmaceutical composition for treating hair loss, wherein the weight parts of the raw materials are: 15 parts of Platycladus orientalis leaves, 15 parts of Toosendan fruits, and 10 parts of Herba Scutellariae spatholobi.

[0035] In a specific embodiment, the present invention provides a pharmaceutical composition for treating hair loss, wherein the weight parts of the raw materials are: 12 parts of Platycladus orientalis leaves, 18 parts of Toosendan fruits, and 8 parts of Herba Spermatophytae.

[0036] In a specific embodiment, the present invention provides a pharmaceutical composition for treating hair loss, wherein the weight parts of the raw materials are: 18 parts of Platycladus orientalis leaves, 12 parts of Toosendan seeds, and 12 parts of Herba Scutellariae spatholobi.

[0037] In the present invention, the Platycladus orientalis leaves are the dried branches and leaves of Platycladus orientalis (L.) Franco, a plant of the Cupressaceae family. They are mostly harvested in summer and autumn and dried in the shade. The medicinal material is preferably tender, green and free of broken pieces. It is bitter, astringent and cold. It enters the lung, liver and spleen meridians. It has the effects of cooling blood and stopping bleeding, resolving phlegm and relieving cough, and promoting hair growth and blackening hair. In the present invention, the Platycladus orientalis leaves are used raw, and the original medicinal material is taken, and impurities, thick stems and fruits are removed, and the ash is sieved out before use. The Toosendan fruit is the dried mature fruit of the Meliaceae plant Toosendan Meliatoosendan Sieb.et Zucc. It is harvested in winter when the fruit is ripe, and impurities are removed and dried. It is bitter, cold and slightly toxic. It enters the liver, small intestine and bladder meridians. It has the effects of soothing the liver and relieving heat, promoting qi and relieving pain, and killing insects. In the present invention, the Toosendan fruit is a raw product. When used, the original medicinal material is taken, impurities are removed, and the raw material is crushed when used. The herb is also called "impatiens stalk". "Compendium of Materia Medica" records that "Impatiens balsamina, also known as herb, is sharp and can soften hard objects, so it has this name". It is the dried stem of Impatiens balsamina L., a plant of the Impatiens family. In the present invention, the herb is represented by the name of herb. When the plant grows vigorously in summer and autumn, the above-ground part is cut, the leaves and flowers and fruits are removed, washed, and dried. The reddish brown one is preferred. It tastes spicy and is warm in nature. It has the effects of dispelling wind and dampness, promoting blood circulation and relieving pain. In the present invention, the herb is used in a raw product, that is, the original medicinal material is taken, impurities are removed, washed, slightly moistened, cut, dried, and sieved to remove the ash. There is no special restriction on the source of the raw materials used in the present invention. It can be obtained by conventional technical means in the art, or purchased from commercial products in the art.

[0038] In the present invention, the use of the above-mentioned pharmaceutical composition for treating hair loss in the preparation of a hair loss treatment preparation is provided. In the present invention, the hair loss includes androgenic alopecia and / or alopecia areata. The androgenic alopecia includes male pattern alopecia and female pattern alopecia. In traditional Chinese medicine, the internal causes of androgenic alopecia are mainly damp-heat in the body and liver and kidney disorders, and the external causes are exogenous wind, dampness, and heat evil as inducements. The internal and external evils are combined, resulting in hair malnutrition. The kidney stores essence, and its beauty is in the hair. The liver stores blood, and the hair is the rest of the blood, and its glory is in the hair. If the disease is not cured for a long time, the essence of the liver and kidney will be damaged, and the hair will have no source of growth, resulting in hair loss. Alopecia areata is caused by liver and kidney deficiency, qi stagnation and blood stasis, insufficient endowment, or overwork and long-term illness, resulting in liver and kidney deficiency, essence and blood deficiency, hair loss, worry and depression, falls or long-term illness, which block the head and cause hair to fall off in pieces. In the present invention, Platycladus orientalis leaves, as the main drug, have the effects of nourishing the liver and kidney, clearing away heat and detoxifying, and invigorating qi and promoting hair growth; Toosendan fruit, as the ministerial drug, can soothe the liver and regulate qi, promote blood circulation and remove blood stasis, regulate liver qi, improve the circulation of qi and blood, alleviate the hair loss caused by liver qi stagnation, and can also promote scalp blood circulation and enhance the nutritional supply of hair follicles; Herba Polygoni Multiflori, as the adjuvant, plays the role of promoting blood circulation and removing rheumatism, helps to improve the scalp meridian obstruction caused by rheumatism or blood stasis, further promotes the absorption of nutrition and blood circulation by hair follicles, and enhances the growth ability of hair. The three complement each other and play the role of soothing the liver and regulating qi, invigorating the kidney and replenishing blood, and promoting blood circulation and removing blood stasis. The composition of the present invention can regulate the balance of qi and blood in the body, improve the microcirculation of the scalp, restore the healthy growth of hair, and has a good therapeutic effect. After cell and animal experiments, it was verified that the pharmaceutical composition of the present invention can effectively promote the proliferation of primary human dermal hair papilla cells. In the formula of the pharmaceutical composition, there is a significant synergistic effect between the various drugs. This synergistic effect makes its effect of promoting cell proliferation significantly better than the effect of using Platycladus orientalis leaves alone and the effect of simply compounding Platycladus orientalis leaves and Herba Scutellariae. Compared with minoxidil, a commonly used clinical drug for treating hair loss, the pharmaceutical composition of the present invention also shows more significant advantages in promoting the proliferation of primary human dermal hair papilla cells. And by establishing a mouse androgenic alopecia model induced by testosterone propionate and a mouse alopecia model induced by cyclophosphamide, it was proved that the pharmaceutical composition of the present invention can promote hair growth in a shorter time, regulate the growth of hair papilla cells, and significantly improve hair loss. Moreover, by comparing with traditional chemical drugs, it was confirmed that the composition has no irritation, high efficiency, and its effect is far superior to chemical drugs.

[0039] In the present invention, the pharmaceutical composition for treating hair loss is used in the preparation of a preparation for treating hair loss, and the preparation includes but is not limited to tincture, solution, foam, powder, lotion, oil, emulsion, ointment, paste, film coating, gel. The preparation also includes pharmaceutically acceptable excipients, including but not limited to solvents, solubilizers, cosolvents, preservatives, antioxidants, pH regulators, moisturizers, thickeners, emulsifiers, penetration enhancers, colorants, and flavoring agents.

[0040] In the present invention, the pharmaceutical composition for treating hair loss is used in the preparation of a preparation for treating hair loss, and each raw material in the pharmaceutical composition needs to extract its effective ingredients to exert the subsequent therapeutic effect. The present invention has no special restrictions on the extraction method, and the technical means well known in the art can be used, and the method includes but is not limited to decoction, percolation, reflux extraction, continuous reflux extraction, steam distillation, supercritical fluid extraction, ultrasonic extraction, and microwave extraction. The following lists the extraction methods selected when extracting the effective ingredients of each raw material, among which the extraction methods applicable to Platycladus orientalis leaves include ethanol immersion, supercritical CO2 extraction, and decoction. Preferably, the extraction method is ethanol immersion. Applicable to Radix Polygoni Multiflori include ethanol immersion, supercritical CO2 extraction, ultrasonic extraction, and decoction. Preferably, the extraction method is ethanol immersion. The extraction methods applicable to Toosendan fruit include ethanol immersion, ethanol percolation, microwave-assisted extraction, and decoction. Preferably, the extraction method is ethanol immersion. To summarize the extraction methods of the above Chinese medicine raw materials, some methods are suitable for the extraction of several raw materials at the same time. Therefore, the methods can be combined to extract the effective ingredients of multiple raw materials at the same time to improve the extraction efficiency.

[0041] In the present invention, a compound oriental arborvitae tincture is also provided, and the active ingredient includes the ethanol water extract of any of the aforementioned pharmaceutical compositions. The present invention also provides a preparation method of the aforementioned compound oriental arborvitae tincture, the preparation method includes any one of a dissolution method, a dilution method, an immersion method, and a percolation method, and the specific steps are as follows: crushing each raw material of any of the aforementioned pharmaceutical compositions to obtain a medicinal material powder, using any one of a dissolution method, a dilution method, an immersion method, and a percolation method, mixing the medicinal material powder or the prepared fluid extract with an ethanol solution, filtering, and constant volume to obtain a compound oriental arborvitae tincture. In the present invention, the dissolution method is to add a certain concentration of ethanol solution to the powder or fluid extract of oriental arborvitae leaves, herba scutellariae and fructus toosendan, dissolve or dilute, stand, and filter to obtain a compound oriental arborvitae tincture. The dilution method includes diluting the fluid extract with a certain concentration of ethanol solution, standing, and filtering to obtain a compound oriental arborvitae tincture. Wherein, the concentration of the ethanol solution is 70% to 90%. Preferably, the concentration of the ethanol solution is 75% to 85%, and further preferably, the concentration of the ethanol solution is 80%. The final crude drug content of the compound Platycladus orientalis tincture is 20% to 60%. Preferably, the final crude drug content is 30% to 50%, and further preferably, the final crude drug content is 40%. In the present invention, the source of the fluid extract is not particularly limited, and it can be purchased from commercial products in the field, or it can be prepared by conventional technical means in the field, that is, using decoction, reflux or percolation to prepare the extract, and all the extracts are concentrated to a thick paste at low temperature to obtain a fluid extract. In the present invention, the percolation method is to crush Platycladus orientalis leaves, Herba Polygoni Multiflori and Fructus Toosendan, respectively, add a certain concentration of ethanol to uniformly wet them, place them in a closed manner for a period of time, then load them into a percolator, flatten them evenly, add ethanol to immerse the medicinal materials, and maintain a certain liquid level height for impregnation, then open the lower outlet of the percolator, percolate at a flow rate of 1 to 3 mL / kg of medicinal materials per minute, and collect the percolate. During the percolation process, ethanol needs to be continuously replenished to ensure that the solvent always immerses the medicinal materials. The percolation liquids of Platycladus orientalis leaves, Herba Scutellariae and Fructus Toosendan are mixed, filtered and fixed to volume to obtain the compound Platycladus orientalis tincture. Platycladus orientalis leaves, Herba Scutellariae and Fructus Toosendan medicinal materials can also be crushed and mixed, and percolated at the same time, and the obtained percolation liquid is added with ethanol to fix the volume to obtain the compound Platycladus orientalis tincture. Wherein, the ethanol is an ethanol solution with a concentration of 70% to 90%. Preferably, the concentration of the ethanol solution is 75% to 85%, and further preferably, the concentration of the ethanol solution is 80%. The sealing time is 15min to 8h. Preferably, the sealing time is 30min to 6h, more preferably 1 to 5h, and further preferably 2 to 4h. The immersion time is 24 to 96h. Preferably, the immersion time is 36 to 72h, and further preferably 48 to 60h. The flow rate is 1 to 3mL / kg of medicinal materials per minute, preferably 2mL / kg of medicinal materials per minute. The final crude drug content of the compound Platycladus orientalis tincture is 20% to 60%.Preferably, the final crude drug content is 30% to 50%, and more preferably, the final crude drug content is 40%. In the present invention, the immersion method is to crush Platycladus orientalis leaves, Herba Scutellariae and Toosendan fruits, immerse them in an ethanol solution, filter and fix the volume to obtain the compound Platycladus orientalis tincture. Platycladus orientalis leaves, Herba Scutellariae and Toosendan fruits can be crushed, immersed separately, and the obtained extracts can be mixed and prepared, or the three can be mixed and immersed at the same time. In the present invention, it is preferred that the three are mixed and immersed at the same time. Wherein, the concentration of the ethanol solution is 50% to 90%. Preferably, the concentration of the ethanol solution is 60% to 80%, and more preferably, the concentration of the ethanol solution is 65% to 75%. The immersion time is 5 to 9 days, preferably, the immersion time is 6 to 8 days, and more preferably 7 days. The final crude drug content of the compound Platycladus orientalis tincture is 20% to 60%. Preferably, the final crude drug content is 30% to 50%, and more preferably, the final crude drug content is 40%. In the present invention, the preparation method of the compound Platycladus orientalis tincture is preferably an immersion method.

[0042] In the embodiments of the present invention, all data were statistically analyzed using SPSS 19.0 software, and the quantitative data were expressed in the form of mean ± standard deviation, * indicates P < 0.05, ** indicates P < 0.01, *** indicates P < 0.001, and **** indicates P < 0.0001.

[0043] In the following embodiments, unless otherwise specified, all of them are conventional methods.

[0044] Unless otherwise specified, the materials and reagents used in the following examples can be obtained from commercial sources.

[0045] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0046] Example 1

[0047] Take 15g of Platycladus orientalis leaves, 15g of Toosendan fruit, and 10g of Radix Scutellariae Radix, clean them, crush them, mix them, put them into a container together, add 70% ethanol solution to immerse the medicinal materials, seal them and immerse them at room temperature for 7 days, and shake them every day. After immersion, filter them, transfer the filtrate to a volumetric flask, rinse the filter residue and make it constant to obtain a compound Platycladus orientalis tincture containing about 40% of the crude drug.

[0048] Example 2

[0049] Take 12g, 18g and 8g of Platycladus orientalis leaves, Toosendan fruit and Herba Scutellariae Radix scutellariae respectively, and moisten them with 80% ethanol solution. Seal for 3 hours and then put them into a percolator. Add 80% ethanol to immerse the medicinal materials for 48 hours, and percolate at a flow rate of 2mL / kg of medicinal materials per minute. Continuously add ethanol during the process. Collect the percolated liquid, mix it, filter it and make it to volume, and prepare a compound Platycladus orientalis tincture containing about 60% of the crude drug.

[0050] Example 3

[0051] The fluid extract prepared by percolation of 18g Platycladus orientalis leaf, 12g Toosendan fruit, and 12g Herba Spatholobi is that Platycladus orientalis leaf, Herba Spatholobi and Herba Spatholobi are crushed, and a small amount of 75% ethanol solution is added respectively to uniformly wet, and the mixture is placed in a closed manner for 3h, and then loaded into a percolator, uniformly flattened, ethanol is added to immerse the medicinal materials, and a certain liquid level is maintained for impregnation, and the mixture is impregnated for 54h. Subsequently, the lower outlet of the percolator is opened, and percolation is performed at a flow rate of 2mL / kg of medicinal materials per minute, and the percolation liquid is collected. During the percolation process, ethanol needs to be continuously supplemented to ensure that the solvent always immerses the medicinal materials. The obtained Platycladus orientalis leaf, Herba Spatholobi and Herba Spatholobi percolation liquids are mixed, and condensed at low temperature to a thick paste to obtain a fluid extract. Take an appropriate amount of the fluid extract and put it into a container, add 75% ethanol solution to dissolve, filter after standing, and transfer the filtrate to a volumetric flask. After rinsing the container and filter paper, the volume is constant to obtain a compound Platycladus orientalis tincture containing about 20% of the crude drug amount.

[0052] Example 4

[0053] Take 15g of Platycladus orientalis leaves, 15g of Toosendan seeds, and 10g of Radix Scutellariae Radix, clean them, crush them, mix them, put them into a container, add 75% ethanol solution to immerse the medicinal materials, seal them and immerse them at room temperature for 5 days, and shake them every day. After immersion, filter them, transfer the filtrate to a volumetric flask, rinse the filter residue and make it constant to obtain a compound Platycladus orientalis tincture containing about 45% of the crude drug.

[0054] Example 5

[0055] Take 12g of Platycladus orientalis leaves, 18g of Toosendan seeds, and 8g of Radix Scutellariae Radix, clean them, crush them, mix them, put them into a container together, add 70% ethanol solution to immerse the medicinal materials, seal them and immerse them at room temperature for 6 days, and shake them every day. After immersion, filter them, transfer the filtrate to a volumetric flask, rinse the filter residue and make it constant to obtain a compound Platycladus orientalis tincture containing about 50% of the crude drug.

[0056] Example 6

[0057] Take 18g of Platycladus orientalis leaves, 12g of Toosendan seeds, and 12g of Radix Scutellariae Radix, clean them, crush them, mix them, put them into a container, add 65% ethanol solution to immerse the medicinal materials, seal them and immerse them at room temperature for 8 days, and shake them every day. After immersion, filter them, transfer the filtrate to a volumetric flask, rinse the filter residue and make it constant to obtain a compound Platycladus orientalis tincture containing about 30% of the crude drug.

[0058] Comparative Example 1

[0059] Take 15g of Platycladus orientalis leaves to prepare Platycladus orientalis leaves tincture containing about 15% of crude drug. The remaining operation method is the same as that of Example 1.

[0060] Comparative Example 2

[0061] Take 15g of Toosendan Fructus to prepare Toosendan Fructus tincture containing about 15% of crude drug. The remaining operation method is the same as that of Example 1.

[0062] Comparative Example 3

[0063] Take 10g of Radix Angelicae Pubescentis to prepare Radix Angelicae Pubescentis tincture containing about 10% of the crude drug. The remaining operation method is the same as that of Example 1.

[0064] Comparative Example 4

[0065] Take 15g of Platycladus orientalis leaves and 15g of Toosendan fruit to prepare Platycladus orientalis leaves + Toosendan fruit tincture containing about 30% of the crude drug amount. The remaining operation method is the same as that of Example 1.

[0066] Comparative Example 5

[0067] Take 15g of Platycladus orientalis leaves and 10g of Herba Scutellariae Radix and prepare Platycladus orientalis leaves + Herba Scutellariae Radix tincture containing about 25% of the crude drug. The remaining operation method is the same as that of Example 1.

[0068] Test Example 1

[0069] Effects of Compound Platycladus orientalis Tincture on Primary Human Dermal Papilla Cells (HDPCs)

[0070] The compound arborvitae tincture prepared in Example 1 is taken for use. The compound arborvitae tincture is diluted with a culture medium into different concentrations of medicinal solutions, that is, diluted to prepare medicinal solutions with concentrations of 0.05%, 0.1%, and 0.25%, respectively, for use. The culture medium is a mixture of 89% high-glucose DMEM culture medium, 10% fetal bovine serum, and 1% double antibody.

[0071] Using CCK-8 method, human dermal papilla cells (HDPCs) in logarithmic growth phase were cultured at 4.5×10 3 / well cells were inoculated on a 96-well plate, and the edge wells were filled with sterile PBS and placed in a constant temperature culture at 37°C and 5% CO2. After the cells adhered to the wall, the above-mentioned drug solutions of different concentrations were replaced respectively, and a blank control group (Control) and a minoxidil group were set up as controls and cultured for 24 hours. After the drug effect ended, the supernatant was discarded, 100 μL / well of the diluted CCK8 working solution was added, and the culture plate was gently shaken several times, and the culture was continued at 37°C and 5% CO2 for 2 hours. The absorbance value of each well was measured at 450nm to calculate the cell proliferation rate.

[0072] The results are as follows Figure 1As shown in the figure, the effect of compound arborvitae tincture on the proliferation ability of HDPCs shows that, compared with the blank control group, minoxidil and compound arborvitae tincture at different dilution concentrations can effectively promote the proliferation of HDPCs. When the dilution concentration is 0.25%, the ability of compound arborvitae tincture to promote cell proliferation is significantly stronger than that of minoxidil, indicating that the compound arborvitae tincture prepared by the present invention has a good and outstanding ability to promote HDPCs cell proliferation.

[0073] Test Example 2

[0074] Effects of the separated prescriptions of Platycladus orientalis tincture on the proliferation of human primary dermal dermal papilla cells (HDPCs)

[0075] Take the tinctures prepared in Example 1 (compound Platycladus orientalis tincture), Comparative Example 1 (Platycladus orientalis leaf tincture), Comparative Example 2 (Tougulianzi tincture), Comparative Example 3 (Piercinga oleracea tincture), Comparative Example 4 (Platycladus orientalis leaf + Tougulianzi tincture) and Comparative Example 5 (Platycladus orientalis leaf + Tougulianzi tincture), dilute each group of tinctures to 0.25% with a culture medium for standby use. The culture medium is a mixture of 89% high-glucose DMEM culture medium, 10% fetal bovine serum and 1% double antibody.

[0076] Using CCK-8 method, human dermal papilla cell lines in logarithmic growth phase were taken and cultured at 4.5×10 3 Cells were inoculated in 96-well plates, and the edge wells were filled with sterile PBS and cultured at 37°C and 5% CO2. After the cells adhered to the wall, the above-mentioned different drug solutions were replaced, and a blank control group (Control) was cultured for 24 hours. After the drug effect ended, the supernatant was discarded, 100 μL / well of diluted CCK8 working solution was added, and the culture plate was gently shaken several times. The culture was continued at 37°C and 5% CO2 for 2 hours. The absorbance value of each well was measured at 450nm to calculate the cell proliferation rate.

[0077] The results are as follows Figure 2 As shown in the results, the compound Platycladus orientalis tincture group showed significant advantages in promoting the proliferation of human primary dermal dermal papilla cells (HDPCs). Compared with the blank control group, the compound Platycladus orientalis tincture group, Platycladus orientalis leaf tincture group, Toosendan tincture group, Herba Tougu tincture group, Platycladus orientalis leaf + Toosendan tincture group, and Platycladus orientalis leaf + Herba Tougu tincture group can promote the proliferation of HDPCs to a certain extent. In addition, the cell proliferation rate of the compound Platycladus orientalis tincture group was significantly higher than that of the other split groups. Compared with a single medicinal material and a split formula composed of only two medicinal materials, the complete formula of the compound Platycladus orientalis tincture was more effective in stimulating the growth of dermal papilla cells. This result shows that there is a synergistic effect between the medicinal materials in the complete compound Platycladus orientalis tincture formula, and after cooperating with each other, it can more effectively promote the proliferation of human primary dermal dermal papilla cells. When matched in a certain proportion, Toosendan tincture and Herba Tougu can better assist the main drug Platycladus orientalis leaf to play its role.

[0078] Test Example 3

[0079] Experimental study on the efficacy of compound Platycladus orientalis tincture on androgenic alopecia model in C57BL / J6 mice

[0080] The experimental animals were male C57BL / J6, 6 weeks old, weighing 18-20g. C57BL / J6 mice were raised under normal conditions, with a temperature of 25±2℃, a humidity of 50%-70%, and alternating light and dark for 12h each. They were free to eat and drink, and the corncob bedding was changed every two days. The experiment began 7 days after they adapted to the breeding environment.

[0081] Thirty-six C57BL / J6 mice were randomly divided into a blank control group (Control), a model group (Model), a minoxidil tincture group (Minoxidil, i.e., a positive group), a tincture group 1 (Example 1), a tincture group 2 (Example 5), and a tincture group 3 (Example 6), with 6 mice in each group.

[0082] After weighing the mice, they were anesthetized with isoflurane. After satisfactory anesthesia, the back skin of the mice was shaved short with an electric razor. The depilatory cream was evenly applied to the back and wiped off after about 1 minute. The hair removal area on the back of the mouse was about 2cm×3cm. Testosterone propionate was diluted with olive oil, mixed with ultrasound for 10 minutes to make a 5mg / mL solution, and used as a modeling agent to model mice. The blank control group was subcutaneously injected with 20μL of normal saline behind the neck for 28 consecutive days. The model group, minoxidil tincture group and compound arborvitae tincture group were subcutaneously injected with testosterone propionate behind the neck for 28 consecutive days, and multiple subcutaneous injections were performed every day at a dose of 5mg / kg, i.e. 20μL / mouse, to establish a mouse model of sex hormone alopecia.

[0083] From the first day to the 28th day, the mice were given the drug once a day in the depilatory area on the back of the mice for 28 consecutive days. Tincture group 1, tincture group 2 and tincture group 3 were given the tincture solutions prepared in Example 1, Example 5 and Example 6, respectively, with a dosage of 200 μL per mouse. The positive group was given commercially available minoxidil tincture with a concentration of 5% and a dosage of 200 μL per mouse. The blank control group and the model group were not treated.

[0084] The results are as follows Figure 3 As shown, HE staining results of the back skin of androgenic alopecia mice are as follows Figure 4 As shown in the figure, the data and images show that the compound arborvitae tincture has a significant therapeutic effect on the C57BL / J6 mouse androgenic alopecia model. Compared with the blank control group, the compound arborvitae tincture group can significantly promote hair growth, and there are significant improvements in the number of hair follicles and hair density. Compared with the positive drug minoxidil, the compound arborvitae tincture group also showed a significant effect on promoting hair growth, and played an outstanding role in promoting the proliferation of hair follicle cells and inducing hair to enter the growth phase.

[0085] Test Example 4

[0086] The experimental animals were female C57BL / J6, 6 weeks old, weighing 18-20g. C57BL / J6 mice were raised under normal conditions, with a temperature of 25±2℃, a humidity of 50%-70%, and alternating light and dark for 12h each. They were free to eat and drink, and the corncob bedding was changed every two days. The experiment began 7 days after they adapted to the breeding environment.

[0087] Thirty-six C57BL / J6 mice were randomly divided into a blank control group (Control), a model group (Model), a minoxidil tincture group (Minoxidil, i.e., a positive group), a tincture group 1 (Example 1), a tincture group 2 (Example 5), and a tincture group 3 (Example 6), with 6 mice in each group.

[0088] After the mice were weighed, they were anesthetized with isoflurane. After satisfactory anesthesia, the skin on the back of the mice was shaved short with an electric razor. The depilatory cream was evenly applied to the back and wiped off after about 1 minute. The growth phase hair was induced in the area of ​​about 2cm×3cm on the back of the mouse. On the 9th day after depilation, the growth phase hair follicles were induced in the back skin of the mice. At this time, the blank control group was intraperitoneally injected with a single dose of 100μL of normal saline, and the model group, minoxidil tincture group and each tincture group were intraperitoneally injected with a single dose of cyclophosphamide at an amount of 150mg / kg. The cyclophosphamide was prepared into a 30mg / ml cyclophosphamide normal saline dilution, as a modeling agent, and each mouse was intraperitoneally injected with 100μL to establish a mouse alopecia areata model.

[0089] From the 9th day to the 27th day, the mice were given the drug once a day in the depilatory area on the back of the mice for 19 consecutive days. Tincture group 1, tincture group 2 and tincture group 3 were given the tincture solutions prepared in Example 1, Example 5 and Example 6, respectively, with a dosage of 200 μL per mouse. The positive group was given commercially available minoxidil tincture with a concentration of 5% and a dosage of 200 μL per mouse. The blank control group and the model group were not treated.

[0090] The results are as follows Figure 5 As shown in the figure, the compound Platycladus orientalis tincture has a significant therapeutic effect on the C57BL / J6 mouse alopecia areata model. The hair growth of the mouse back skin began to be obvious on the 9th day, and within one month, it was no different from the blank group. The HE staining results of the alopecia areata mouse back skin are shown in the figure. Figure 6 As shown in the results, compared with the blank control group, the compound arborvitae tincture group significantly promoted hair growth. Even compared with the positive drug minoxidil, the compound arborvitae tincture group also performed well in promoting hair growth and had a significant effect.

[0091] Test Example 1

[0092] Two female patients with hair loss symptoms were selected. The patients had not received other hair loss-related treatments in the past 3 months and had no serious liver and kidney dysfunction, immune system diseases, skin allergies, and other diseases that could affect the test results.

[0093] The compound arborvitae tincture prepared in Example 1 was used for treatment. The patient took an appropriate amount of the compound arborvitae tincture and applied it directly to the scalp in the hair loss area once in the morning and evening, and gently massaged with the fingertips for 3-5 minutes. The massage force was moderate, and the massage was performed in circles from the edge of the scalp to the center of the top of the head to allow the drug to fully penetrate. After application, it was dried naturally and contact with water was avoided as much as possible within 4 hours. During the treatment period, the patient was required to maintain a regular daily routine, a balanced diet, and avoid excessive mental stress.

[0094] The test period lasted for 3 months, and detailed records of hair loss were taken before treatment and 3 months after treatment, including the measurement of the area of ​​hair loss.

[0095] The results are as follows Figure 7 As shown in the figure, compared with before treatment, after 3 months of treatment with compound arborvitae tincture, the hair loss symptoms of the two patients were significantly improved. The area of ​​hair loss was significantly reduced, and it can be clearly observed that the new hair gradually increased, and the texture of the new hair was thicker and tougher than before treatment, indicating that the compound arborvitae tincture of the present invention has shown a good therapeutic effect in the treatment of hair loss, can effectively promote hair growth, significantly increase the number and length of hair, and improve the hair coverage rate of patients with hair loss.

[0096] Test Example 2

[0097] 96 patients diagnosed with androgenic alopecia and 86 patients diagnosed with alopecia areata were selected, including 120 males and 62 females. All patients had not received other hair loss-related treatments in the past 3 months, and had no severe liver and kidney dysfunction, immune system diseases, skin allergies, and other diseases that could affect the test results.

[0098] The random number table method was used for grouping. The androgenic alopecia group (AGA) was divided into the androgenic alopecia minoxidil tincture group (48 cases, 35 males and 13 females) and the androgenic alopecia compound arborvitae tincture group (48 cases, 36 males and 12 females). The alopecia areata group (AA) was divided into the alopecia areata minoxidil tincture group (43 cases, 30 males and 13 females) and the alopecia areata compound arborvitae tincture group (43 cases, 29 males and 14 females).

[0099] Minoxidil tincture group: All patients in the minoxidil group took an appropriate amount of 5% minoxidil liniment and applied it evenly to the scalp in the hair loss area once a day in the morning and evening. After application, gently massage with the fingertips for 3 to 5 minutes to promote drug absorption, but be careful to use moderate massage force to avoid damaging the scalp. During use, keep the scalp clean and dry, and avoid washing your hair immediately after application.

[0100] Compound Platycladus orientalis tincture group: Compound Platycladus orientalis tincture prepared in Example 1 was used. Patients took an appropriate amount of compound Platycladus orientalis tincture and applied it directly to the scalp in the hair loss area once in the morning and evening, and gently massaged with the fingertips for 3 to 5 minutes to allow the drug to fully penetrate. Make sure the scalp is clean before application, dry naturally after application, and avoid contact with water as much as possible within 4 hours.

[0101] Trial period: The entire trial period is set at 6 months. During the trial period, patients maintain good living habits, regular work and rest, avoid staying up late, eat a balanced diet, and reduce mental stress.

[0102] Judgment criteria: Significantly effective: For patients with androgenic alopecia, after 6 months of treatment, the hair loss area is significantly reduced, a large amount of new hair grows, and the new hair is coarser in texture and darker in color, and the hair density increases significantly; for patients with alopecia areata, the hair loss area is completely covered by new hair, and the hair returns to normal growth.

[0103] Ineffective: For patients with androgenic alopecia, after 6 months of treatment, there was no significant change in the hair loss area, the new hair was sparse, and the hair density did not increase significantly; for patients with alopecia areata, there was no significant reduction in the hair loss area, no new hair or very little new hair.

[0104] Adverse reactions: scalp itching, skin allergic reactions, such as redness, swelling, rash, etc.; or local skin irritation causing pain or burning sensation; or systemic reactions, such as difficulty breathing, dizziness, nausea, etc. caused by allergies.

[0105] The experimental results are shown in Table 1 below:

[0106]

[0107] The results show that the compound Platycladus orientalis tincture of the present invention has a good therapeutic effect in treating androgenic alopecia and alopecia areata. In an expanded population experiment, it can effectively promote hair growth, significantly increase the number and length of hair, and improve the hair coverage of patients with alopecia areata. At the same time, the tincture has fewer adverse reactions, high safety, and little difference in therapeutic effect between different individuals, and has high clinical application value and promotion prospects.

[0108] The above is only a preferred embodiment of the present invention. It should be pointed out that for ordinary technicians in this technical field, several improvements and modifications can be made without departing from the principle of the present invention. These improvements and modifications should also be regarded as the scope of protection of the present invention.

Claims

1. A pharmaceutical composition for treating hair loss, characterized in that: The invention is composed of the following raw materials in parts by weight: 12-18 parts of Platycladus orientalis leaves, 12-18 parts of Toosendan seeds and 8-12 parts of Herba speranskiae radiatae.

2. The pharmaceutical composition according to claim 1, characterized in that The weight parts of the raw materials are respectively: 15 parts of Platycladus orientalis leaves, 15 parts of Toosendan fruits, and 10 parts of Herba Spermatophytae.

3. The pharmaceutical composition according to claim 1, characterized in that The weight parts of the raw materials are respectively: 12 parts of Platycladus orientalis leaves, 18 parts of Toosendan fruits, and 8 parts of Herba Spermatophytae.

4. The pharmaceutical composition according to claim 1, characterized in that The weight parts of the raw materials are respectively: 18 parts of Platycladus orientalis leaves, 12 parts of Toosendan fruits, and 12 parts of Herba Spermatophytae.

5. Use of the pharmaceutical composition according to any one of claims 1 to 4 in the preparation of a preparation for treating hair loss.

6. The use according to claim 5, characterized in that: The hair loss includes androgenic alopecia and / or alopecia areata.

7. The use according to claim 5, characterized in that: The preparations include tinctures, solutions, foams, powders, lotions, oils, emulsions, ointments, pastes, paints or gels.

8. A compound Platycladus orientalis tincture, characterized in that: The active ingredient comprises the ethanol water extract of the pharmaceutical composition according to any one of claims 1 to 4. The preparation method of the compound Platycladus orientalis tincture according to claim 8 is characterized in that the preparation method comprises any one of a dissolution method, a dilution method, an immersion method, and a percolation method, and the specific steps are as follows: crushing the raw materials of the pharmaceutical composition according to any one of claims 1 to 4 to obtain medicinal material powder; using any one of a dissolution method, a dilution method, an immersion method, and a percolation method, mixing the medicinal material powder or the prepared fluid extract with an ethanol solution, filtering, and fixing the volume to obtain the compound Platycladus orientalis tincture.

9. The method for preparing the compound Platycladus orientalis tincture according to claim 8, characterized in that: The concentration of the ethanol solution is 50% to 90%; the immersion time of the immersion method is 5 to 9 days; and the final crude drug content of the compound Platycladus orientalis tincture is 20% to 60%.

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