CN115779028B - Traditional Chinese medicine composition and preparation method and application thereof - Google Patents
Traditional Chinese medicine composition and preparation method and application thereof Download PDFInfo
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Abstract
The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition and a preparation method and application thereof. The invention provides a traditional Chinese medicine composition which comprises the following components in parts by weight: 0.01 to 15 parts of black sesame, 2.5 to 100 parts of vine of multiflower knotweed, 2.5 to 100 parts of eclipta, 2.5 to 100 parts of cacumen biotae, 0.4 to 10 parts of herba lycopi, 2.5 to 100 parts of rhizoma dioscoreae, 1.5 to 60 parts of radix scutellariae, 1.5 to 60 parts of cortex phellodendri, 1.5 to 50 parts of cassia twig, 0.75 to 30 parts of pseudo-ginseng, 0.75 to 30 parts of rosewood and 0.075 to 3 parts of storax. The traditional Chinese medicine composition extract provided by the invention can promote hair growth and is applied to the aspects of hair growth and hair loss adjuvant therapy. The main functions of the traditional Chinese medicine composition extract provided by the invention are as follows: nourishing liver and kidney, clearing heat and drying dampness.
Description
Technical Field
The invention relates to the field of traditional Chinese medicines, in particular to a traditional Chinese medicine composition and a preparation method and application thereof.
Background
Androgenetic alopecia (Androgenetic alopecia, AGA), also known as seborrheic alopecia (Seborrheic alopecia, SA), is one of the most common alopecia disorders, and is a progressive pattern type of alopecia characterized by progressive loss of hair density. According to hamilton's study, the average prevalence of AGA was 30% at age 30 and 50% at age 50. AGA is commonly found in the adult male population and increases in prevalence with age, with typical clinical manifestations of forehead and hairline withdrawal, with progression of the disease, the patient's head hair gradually falls off until the hair follicle is collapsed, no new hair or vellus hair is seen, and finally only occipital and temporal hair remains, forming typical horseshoe hair loss. The female suffers from alopecia with hidden disease, mainly including hair loss at the top of the head, which is not easily perceived in the early stage and generally does not involve hairline. Since AGA is one of the common lossy diseases, severely affecting the physical and mental health of the patient, more effective treatments are sought.
It is currently widely recognized that AGA may be genetically related, involving multiple genes, most notably the androgen receptor gene on the X chromosome, and that androgens and androgen receptors (Androgen receptor, AR) may play an important role in AGA. Studies have shown that AR is specifically expressed in the scalp of AGA patients, elevated in the forehead and top of the head, and normal in the occipital region. In addition, AGA is associated with a differential expression of type II 5α reductase, an enzyme present in the inner root sheath of hair follicles that converts testosterone (Testoste rone, T) to dihydrotestosterone (Dihydrotestosterone, DHT), which binds 5 times more affinity to AR than T. Type II 5α reductase is expressed more in the hair follicles of the forehead than in the back occipital, and therefore the hair follicles of the forehead are more sensitive to androgens and DHT is higher, a phenomenon which is more pronounced in the scalp of alopecia patients. When DHT is combined with DHT, an AR signal channel is activated, so that AR enters a promoter region of a downstream target gene combined in a cell nucleus to start transcription of the gene, wherein DKK-1 (D ickkopf, DKK-1) is a typical downstream target gene, and after the DKK-1 is transcribed and synthesized, a Wnt channel is further negatively regulated, so that proliferation of cells is inhibited, and hair shedding is promoted. Therefore, the reduction of DKK-1 secretion can exert a certain alopecia preventing effect.
Key pathophysiological features of AGA are inflammation, changes in hair growth cycle, and hair follicle micromation. The three interactions, the follicular micro-inflammation may cause apoptosis, microminiaturization and perifollicular fibrosis of the hair follicle stem cells, which may affect the hair cycle change, and the hair follicle of the AGA patient is shortened with the cycle of the hair cycle, and the hair follicle is gradually microminiaturized, so that the microminiaturization of the AGA hair follicle is characterized by gradually decreasing the number of hair follicles in the growth phase, and increasing the number of hair follicles in the catagen phase and the telogen phase. Therefore, by shortening the telogen phase of hair follicles, extending the anagen phase, and promoting the hair follicles from telogen phase to anagen phase, it is possible to improve AGA.
With the development of modern society, the mass dependence degree is reduced, the demand of people on beauty is higher and higher, AGA is a progressive alopecia disease, the personal image and the self-confidence are seriously influenced, no radical treatment method exists at present, the affected group of AGA is huge, the illness of the AGA is in a trend of younger in recent years, and the treatment means of the AGA is far insufficient to meet the strong treatment demand of patients. The current FDA approval for finasteride and minoxidil treatment of AGA is limited in its therapeutic effect and has certain side effects, far from meeting the urgent therapeutic needs of the vast diseased population. Therefore, it is highly necessary to find an alternative therapy for AGA. The traditional Chinese medicine treatment method is various, has fewer side effects, is simple and convenient, thereby arousing great interest of people, has great market potential and exploration space for AGA treatment, and deserves further research and popularization, however, the existing research formulas for growing and developing the traditional Chinese medicine have more prescription curative effect, longer action time and no related in-vivo efficacy verification experiment, and have certain limitation.
Therefore, the development of the traditional Chinese medicine formula for treating AGA has important significance for the treatment and clinical application of AGA patients.
Disclosure of Invention
In view of this, the invention provides a traditional Chinese medicine composition, a preparation method and application thereof. The invention provides a traditional Chinese medicine composition extract which can promote hair growth and is applied to the aspects of hair growth and hair loss adjuvant therapy. The main functions of the traditional Chinese medicine composition extract provided by the invention are as follows: nourishing liver and kidney, clearing heat and drying dampness.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides a traditional Chinese medicine composition which comprises the following components in parts by weight:
In some embodiments of the present invention, the above-mentioned Chinese medicinal composition comprises the following components in parts by weight:
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Or (b)
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Or (b)
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Or (b)
The invention also provides a preparation method of the traditional Chinese medicine composition, wherein the components with the formula amount are crushed, extracted by alcohol, and the extracting solution is collected.
In some embodiments of the invention, the collecting the extract in the above preparation method further comprises concentrating and/or freeze-drying;
The concentration temperature is 70 ℃, and the concentration ratio is 1:15; the pressure of freeze drying is 1Pa, the temperature is-65 ℃, and the freezing time is 24 hours.
In some embodiments of the present invention, the alcohol extraction in the above preparation method comprises soaking in 90% ethanol for 30min.
In some embodiments of the invention, the alcohol extraction in the above preparation method comprises soaking in 50% ethanol, wherein the soaking time is not less than 72 hours.
In some embodiments of the invention, the alcohol extraction in the above preparation method employs ultrasonic extraction; the ultrasonic extraction time is 30min, and the times are 3 times.
The invention also provides a preparation method of the traditional Chinese medicine composition extract, which comprises the steps of mixing the traditional Chinese medicine composition with ethanol, filtering, and mixing with butanediol and glycerol aqueous solution to obtain the traditional Chinese medicine composition extract.
In the preparation method, the volume fraction of the glycerol aqueous solution is 50%.
In the preparation method, the mixing time of the traditional Chinese medicine composition and ethanol is 72 hours.
The invention also provides application of the traditional Chinese medicine composition and/or the traditional Chinese medicine composition obtained by the preparation method in preparation of medicines and/or preparations for growing and/or nourishing hair.
The invention also provides application of the traditional Chinese medicine composition and/or the traditional Chinese medicine composition obtained by the preparation method in preparation of medicines and/or preparations for treating and/or preventing androgenetic alopecia.
The invention also provides application of the traditional Chinese medicine composition and/or the traditional Chinese medicine composition obtained by the preparation method in preparation of medicines and/or preparations for regulating and controlling DKK-1 gene expression.
The invention also provides a medicine or a preparation, which comprises the traditional Chinese medicine composition and/or the traditional Chinese medicine composition obtained by the preparation method and pharmaceutically acceptable auxiliary materials.
In some embodiments of the invention, the dosage form of the above-described medicament or formulation comprises: one or more of oral decoction, pill, and topical spray.
The invention also provides a pharmaceutical composition comprising the traditional Chinese medicine composition and/or the traditional Chinese medicine composition obtained by the preparation method and other optional effective components.
The invention also provides a using method of the medicine or the preparation, and the medicine or the preparation is administrated by sampling.
In some embodiments of the invention, the dosage administered in the methods of use described above is from 0.01 to 4g/kg.
In some embodiments of the invention, the dosage administered in the methods of use described above is from 0.01 to 0.44g/kg (human).
In some embodiments of the invention, the dosage administered in the methods of use described above is from 0.064 to 2.8g/kg (rat).
In some embodiments of the invention, the dosage administered in the methods of use described above is from 0.091 to 4g/kg (mouse).
In some embodiments of the invention, the route of administration described in the methods of use described above comprises: rinsing or spraying.
The invention provides a traditional Chinese medicine composition which comprises the following components in parts by weight:
The beneficial effects of the invention include:
(1) The extracts of the traditional Chinese medicine composition provided by the invention are prepared from the single medicines of black sesame, vine of multiflower knotweed, eclipta, cacumen biotae, herba lycopi, rhizoma dioscoreae, radix scutellariae, cortex phellodendri, cassia twig, pseudo-ginseng, rosewood and storax by selecting a certain part, so that the hair growth of a C57BL/6 mouse can be promoted, the expression of a key cell factor DKK-1 gene in DPC cells involved in the androgen alopecia occurrence process can be regulated, and the effect of hair growth and development can be exerted.
(2) The traditional Chinese medicine composition extract provided by the invention can promote hair growth and is applied to the aspects of hair growth and hair loss adjuvant therapy.
(3) The main functions of the traditional Chinese medicine composition extract provided by the invention are as follows: nourishing liver and kidney, clearing heat and drying dampness.
(4) The traditional Chinese medicine composition extract provided by the invention is suitable for patients suffering from alopecia or people suffering from alopecia, and clinically presents such as few and sparse hair, increased hair loss, greasy and moist scalp, thin, soft and yellow hair, and the like.
(5) The traditional Chinese medicine composition group Fang Fangjie and the action mechanism provided by the invention are as follows: the group Fang Zi has the functions of nourishing liver and kidney, clearing heat and drying dampness. The hair is the remainder of blood, the herba lycopi and the biota orientalis have the effects of promoting blood circulation by removing blood stasis, cooling blood and stopping bleeding, and have the effects of growing and blackening hair, and are all monarch drugs. Black sesame and eclipta have the effects of nourishing liver and kidney, cooling blood and stopping bleeding, and firstly have the effects of benefiting essence and blood, growing hair and blackening hair, and secondly have the effects of cooling blood and stopping bleeding of cacumen biotae; 37. the rosewood heart-fire herb has the effects of activating blood circulation to dissipate blood stasis, relieving swelling and pain, supplementing with the herba lycopi and jointly playing the roles of activating blood circulation to dissipate blood stasis; the baikal skullcap root, the amur corktree bark and the rhizoma dioscoreae septemlobae clear heat and remove dampness; the upper drugs are ministerial drugs. Ramulus Cinnamomi, radix Polygoni Multiflori, has effects of warming and dredging channels and collaterals, nourishing blood and tranquillizing mind, and above are adjuvant drugs. A small amount of storax is used as a guiding drug, and can be used for inducing resuscitation and relieving pain. The medicines are combined to achieve the effects of promoting blood circulation to remove blood stasis, cooling blood to stop bleeding, nourishing liver and kidney, tonifying essence and blood to strengthen body resistance and tonify deficiency, removing blood stasis to stop bleeding, clearing heat and drying dampness to relieve local damp-heat symptoms, nourishing blood and tranquillizing, inducing resuscitation with fragrance, and treating both principal and secondary aspect of disease.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below.
FIG. 1 shows the hair growth of each group of mice;
FIG. 2 shows neonatal hair growth status scores for each group of mice;
FIG. 3 shows the effect of the Chinese medicinal composition (example 9, comparative example 3, comparative example 4) on DKK-1 gene expression in DPC cells.
Detailed Description
The invention discloses a traditional Chinese medicine composition, a preparation method and application thereof.
It should be understood that the expression "one or more of … …" individually includes each stated object after the expression and various combinations of two or more of the stated objects unless otherwise understood from the context and usage. The expression "and/or" in combination with three or more recited objects should be understood as having the same meaning unless otherwise understood from the context.
The use of the terms "comprising," "having," or "containing," including grammatical equivalents thereof, should generally be construed as open-ended and non-limiting, e.g., not to exclude other unrecited elements or steps, unless specifically stated otherwise or otherwise understood from the context.
It should be understood that the order of steps or order of performing certain actions is not important so long as the invention remains operable. Furthermore, two or more steps or actions may be performed simultaneously.
The use of any and all examples, or exemplary language, such as "e.g." or "comprising" herein is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.
Furthermore, the numerical ranges and parameters setting forth the present invention are approximations that may vary as precisely as possible in the exemplary embodiments. However, any numerical value inherently contains certain standard deviations found in their respective testing measurements. Accordingly, unless explicitly stated otherwise, it is to be understood that all ranges, amounts, values and percentages used in this disclosure are modified by "about". As used herein, "about" generally means that the actual value is within plus or minus 10%, 5%, 1% or 0.5% of a particular value or range.
The invention provides a traditional Chinese medicine formula, wherein the extract of the traditional Chinese medicine formula can reverse the action of androgens to a certain extent in an androgenic alopecia animal model, and promote the hair growth of C57BL/6 mice; can regulate and control gene expression of key cell factor DKK-1 in DPC cell, so as to exert the effect of promoting hair growth.
The invention provides a medicine for preparing hair growth and development, which is prepared from the following components: 0.01 to 15 parts of black sesame, 2.5 to 100 parts of vine of multiflower knotweed, 2.5 to 100 parts of eclipta, 2.5 to 100 parts of cacumen biotae, 0.4 to 10 parts of herba lycopi, 2.5 to 100 parts of rhizoma dioscoreae, 1.5 to 60 parts of radix scutellariae, 1.5 to 60 parts of cortex phellodendri, 1.5 to 50 parts of cassia twig, 0.75 to 30 parts of pseudo-ginseng, 0.75 to 30 parts of rosewood and 0.075 to 3 parts of storax. The traditional Chinese medicine formulas with different proportions have the effect of growing and developing hair, and the optimal traditional Chinese medicine formula for growing and developing hair is obtained.
The invention also provides a preparation method of the traditional Chinese medicine formula extract, which comprises the steps of crushing the single medicines, extracting with 90% ethanol, concentrating, freeze-drying (at-65 ℃ for 24 hours and under vacuum degree of 1 Pa), and dissolving.
The invention also provides a preparation method of the traditional Chinese medicine composition extract, which comprises the steps of taking the formula amount of pseudo-ginseng, black sesame, rosewood heart wood, herba lycopi, rhizoma dioscoreae, radix scutellariae, cortex phellodendri, cacumen biotae, caulis polygoni multiflori, eclipta, cassia twig and storax, adding into an ethanol aqueous solution with the volume fraction of 50% by weight for soaking for at least 72 hours, obtaining a soaked traditional Chinese medicine solid-liquid mixture, filtering and separating, adding into a glycerol aqueous solution with the volume fraction of 50% by weight of 2.5% of 1, 3-butanediol, and stirring uniformly to obtain the traditional Chinese medicine composition extract.
The traditional Chinese medicine formula extract provided by the invention can reverse the action of androgens to a certain extent, promote the hair growth of C57BL/6 mice, regulate and control the gene expression of a key cell factor DKK-1 in DPC cells, which participates in the occurrence process of androgens alopecia, so as to exert the efficacy of hair growth and development.
The traditional Chinese medicine formula provided by the invention is applied to preparing medicines for growing and developing hair on the other hand, and comprises different oral medicines such as decoction, pills and the like and different external medicines such as leaching, spraying and the like.
In examples 1 to 10, comparative examples 1 to 4 and verification examples 1 to 3 of the present invention, all the raw materials and reagents used were commercially available.
The invention is further illustrated by the following examples:
Example 1
3.75 Parts of black sesame, 2.5 parts of vine of multiflower knotweed, 2.5 parts of eclipta, 2.5 parts of cacumen biotae, 2.5 parts of herba lycopi, 2.5 parts of rhizoma dioscoreae, 1.5 parts of radix scutellariae, 1.5 parts of cortex phellodendri, 1.5 parts of cassia twig, 0.75 part of pseudo-ginseng, 0.75 part of rosewood and 0.075 part of storax. The use concentration (crude drug amount) is that of human: 0.11g/kg; rats: 0.7g/kg; mice: 1g/kg.
Example 2
7.5 Parts of black sesame, 5 parts of vine of multiflower knotweed, 5 parts of eclipta, 5 parts of cacumen biotae, 5 parts of herba lycopi, 5 parts of rhizoma dioscoreae, 3 parts of radix scutellariae, 3 parts of cortex phellodendri, 3 parts of cassia twig, 1.5 parts of radix notoginseng, 1.5 parts of rosewood and 0.15 part of storax. The use concentration (crude drug amount) is that of human: 0.22g/kg; rats: 1.4g/kg; mice: 2g/kg.
Example 3
15 Parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 10 parts of herba lycopi, 10 parts of rhizoma dioscoreae, 6 parts of radix scutellariae, 6 parts of cortex phellodendri, 6 parts of cassia twig, 3 parts of pseudo-ginseng, 3 parts of rosewood and 0.3 part of storax. The use concentration (crude drug amount) is that of human: 0.44g/kg; rats: 2.8g/kg; mice: 4g/kg.
Example 4
15 Parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 10 parts of herba lycopi, 10 parts of rhizoma dioscoreae, 6 parts of radix scutellariae, 6 parts of cortex phellodendri, 6 parts of cassia twig, 3 parts of pseudo-ginseng, 3 parts of rosewood and 0.3 part of storax. The concentration used was 50. Mu.g/mL.
Example 5
30 Parts of pseudo-ginseng, 0.01 part of black sesame, 30 parts of rosewood heart wood, 0.4 part of herba lycopi, 100 parts of rhizoma dioscoreae septemlobae, 60 parts of radix scutellariae, 60 parts of cortex phellodendri, 100 parts of cacumen biotae, 100 parts of caulis polygoni multiflori, 100 parts of eclipta, 50 parts of cassia twig and 3 parts of storax. The use concentration (crude drug amount) is that of human: 0.01g/kg; rats: 0.064g/kg; mice: 0.091g/kg.
Comparative example 1
15 Parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 0 part of herba lycopi, 0 part of rhizoma dioscoreae, 0 part of radix scutellariae, 0 part of cortex phellodendri, 0 part of cassia twig, 0 part of pseudo-ginseng, 0 part of rosewood and 0 part of storax. The concentration used was 50. Mu.g/mL.
Comparative example 2
15 Parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 10 parts of herba lycopi, 0 part of rhizoma dioscoreae, 6 parts of radix scutellariae, 6 parts of cortex phellodendri, 6 parts of cassia twig, 3 parts of pseudo-ginseng, 0 part of rosewood and 0 part of storax. The concentration used was 50. Mu.g/mL.
Example 6 preparation method of Chinese medicinal composition extract
The medicines in the formula of the example 1 are weighed according to 3.75 parts of black sesame, 2.5 parts of vine of multiflower knotweed, 2.5 parts of eclipta, 2.5 parts of cacumen biotae, 2.5 parts of herba lycopi, 2.5 parts of rhizoma dioscoreae, 1.5 parts of radix scutellariae, 1.5 parts of cortex phellodendri, 1.5 parts of cassia twig, 0.75 part of pseudo-ginseng, 0.75 part of rosewood and 0.075 part of storax, mixed and crushed, soaked in 1000mL of 90% ethanol for 30min, extracted for 30min by standard ultrasound for 3 times, and the supernatant is subjected to suction filtration by using a Buchner funnel, and the filtrate is collected. The filtrate was prepared into extract lyophilized powder by using a rotary evaporator, 2.79g of the lyophilized powder (crude drug amount 22.325g, extraction rate: 12.5%) was weighed, and dissolved in 223.25mL of 75% ethanol, and mixed uniformly for use, to obtain a Chinese medicinal composition extract solution. The application concentration (crude drug amount) is as follows: 0.11g/kg; rats: 0.7g/kg; mice: 1g/kg.
Example 7 preparation method of Chinese medicinal composition extract
The preparation method comprises the steps of weighing 7.5 parts of black sesame, 5 parts of vine of multiflower knotweed, 5 parts of eclipta, 5 parts of cacumen biotae, 5 parts of herba lycopi, 5 parts of rhizoma dioscoreae, 3 parts of radix scutellariae, 3 parts of cortex phellodendri, 3 parts of cassia twig, 1.5 parts of radix notoginseng, 1.5 parts of rosewood and 0.15 part of storax, mixing and crushing, soaking in 1000mL of 90% ethanol for 30min, carrying out standard ultrasonic extraction for 3 times, and carrying out suction filtration on the supernatant by using a Buchner funnel to collect filtrate. Preparing the filtrate into extract lyophilized powder by using a rotary evaporator, weighing a certain mass of lyophilized powder, weighing 5.58g of lyophilized powder (crude drug amount 44.65g, extraction rate 12.5%), dissolving in 223.25mL of 75% ethanol, mixing uniformly, and obtaining the Chinese medicinal composition extract solution. The application concentration (crude drug amount) is as follows: 0.22g/kg; rats: 1.4g/kg; mice: 2g/kg.
Example 8 preparation method of Chinese medicinal composition extract
The preparation method comprises the steps of weighing 15 parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 10 parts of herba lycopi, 10 parts of rhizoma dioscoreae, 6 parts of radix scutellariae, 6 parts of cortex phellodendri, 6 parts of cassia twig, 3 parts of pseudo-ginseng, 3 parts of rosewood and 0.3 part of storax, mixing and crushing, soaking in 1000mL of 90% ethanol for 30min, carrying out standard ultrasonic extraction for 3 times, carrying out suction filtration on the supernatant by using a Buchner funnel, and collecting filtrate. Preparing the filtrate into extract lyophilized powder by using a rotary evaporator, weighing a certain mass of lyophilized powder, weighing 11.16g of lyophilized powder (crude drug amount 89.3g, extraction rate 12.5%), dissolving in 223.25mL of 75% ethanol, mixing uniformly, and obtaining the Chinese medicinal composition extract solution. The application concentration (crude drug amount) is as follows: 0.44g/kg; rats: 2.8g/kg; mice: 4g/kg.
Example 9 preparation method of Chinese medicinal composition extract
The preparation method comprises the steps of weighing 15 parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 10 parts of herba lycopi, 10 parts of rhizoma dioscoreae, 6 parts of radix scutellariae, 6 parts of cortex phellodendri, 6 parts of cassia twig, 3 parts of pseudo-ginseng, 3 parts of rosewood and 0.3 part of storax, mixing and crushing, soaking in 1000mL of 90% ethanol for 30min, carrying out standard ultrasonic extraction for 3 times, carrying out suction filtration on the supernatant by using a Buchner funnel, and collecting filtrate. Preparing the filtrate into extract freeze-dried powder by using a rotary evaporator, weighing a certain mass of freeze-dried powder, dissolving the powder in DMSO to prepare extract mother solution, and obtaining extract solution of the traditional Chinese medicine composition, wherein the extract solution is used for cell experiments and the use concentration is 50 mug/mL.
Example 10 preparation method of Chinese medicinal composition extract
(1) The preparation method comprises the steps of crushing each medicine in the formula of example 5, adding 30 parts of pseudo-ginseng, 0.01 part of black sesame, 30 parts of rosewood heart wood, 0.4 part of herba lycopi, 100 parts of rhizoma dioscoreae, 60 parts of radix scutellariae, 60 parts of cortex phellodendri, 100 parts of cacumen biotae, 100 parts of caulis polygoni multiflori, 100 parts of eclipta, 50 parts of cassia twig and 3 parts of storax, and then adding 3800.46mL of 50% ethanol aqueous solution with the volume fraction to soak for at least 72 hours to obtain a soaked Chinese medicine solid-liquid mixture.
(2) Filtering and separating the solid-liquid mixture of the traditional Chinese medicine soaked in the step (1), adding 100mL of 1, 3-butanediol and 100mL of glycerin water solution with the volume fraction of 50%, and uniformly stirring to obtain the traditional Chinese medicine composition extract. The application concentration (crude drug amount) is as follows: 0.01g/kg; rats: 0.064g/kg; mice: 0.091g/kg.
Comparative example 3
The preparation method comprises the steps of weighing 15 parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 0 part of herba lycopi, 0 part of rhizoma dioscoreae, 0 part of radix scutellariae, 0 part of cortex phellodendri, 0 part of cassia twig, 0 part of radix notoginseng, 0 part of rosewood heart and 0 part of storax, mixing and crushing, soaking in 1000mL of 90% ethanol for 30min, carrying out standard ultrasonic extraction for 30min, carrying out suction filtration on the supernatant by using a Buchner funnel for 3 times, and collecting filtrate. Preparing the filtrate into extract freeze-dried powder by using a rotary evaporator, weighing a certain mass of freeze-dried powder, dissolving the powder in DMSO to prepare extract mother solution, and obtaining extract solution of the traditional Chinese medicine composition, wherein the extract solution is used for cell experiments and the use concentration is 50 mug/mL.
Comparative example 4
The preparation method comprises the steps of weighing 15 parts of black sesame, 10 parts of vine of multiflower knotweed, 10 parts of eclipta, 10 parts of cacumen biotae, 10 parts of herba lycopi, 0 part of rhizoma dioscoreae, 6 parts of radix scutellariae, 6 parts of cortex phellodendri, 6 parts of cassia twig, 3 parts of pseudo-ginseng, 0 part of rosewood and 0 part of storax, mixing and crushing, soaking in 1000mL of 90% ethanol for 30min, carrying out standard ultrasonic extraction for 30min, carrying out suction filtration on the supernatant by using a Buchner funnel for 3 times, and collecting filtrate. Preparing the filtrate into extract freeze-dried powder by using a rotary evaporator, weighing a certain mass of freeze-dried powder, dissolving the powder in DMSO to prepare extract mother solution, and obtaining extract solution of the traditional Chinese medicine composition, wherein the extract solution is used for cell experiments and the use concentration is 50 mug/mL.
Verification example 1
An animal model of DHT-induced alopecia is constructed, in which the influence of low, medium and high different concentrations of the extract of the traditional Chinese medicine composition (example 6, example 7 and example 8) on the hair growth of mice is observed, and the specific experimental process is as follows:
(1) C57BL/6 mice (7 weeks old) were adapted to dehairing after 5 days of feeding, and control, model, minoxidil, example 6, example 7, and example 8 groups were set. The model group, minoxidil group and example group are used for constructing an androgenetic alopecia mouse model by subcutaneous injection of DHT induction on the 1 st day after dehairing, the DHT concentration is 7mg/kg in the first 5 days, the DHT concentration is 3.5mg/kg in the second 16 days, and the control group is injected with corresponding volumes of physiological saline without DHT.
(2) The prescription is used for drug administration: the animals were initially coated with 200 μl of the corresponding drug 2cm x 4cm on their dehairing backs for 15d daily 5 days after DHT or control solvent injection. Wherein the control group and the model group are coated with 75% ethanol solution, the minoxidil group is coated with 5% minoxidil solution, and the examples 6, 7 and 8 are coated with corresponding prescription solutions.
(3) The skin color change and hair growth on the back of the mice were recorded, and skin photographs of the mice were taken for a total of 3 recordings. (the molding day was recorded once every 10 days for day D0).
(4) The growth status of the new wool in the dehairing zone of the mice is scored every 10 days, and the scoring standard is as follows: the hair-free growth is 1 minute when the hair is shallow and full of the dehairing area; the length and density of the new hair are about half of the unhairing area and are 2 minutes; the no difference between the new hair growth and the unhairing area is 3 minutes; mice with irregular hair growth were calculated as area ratio. The average score for each group was calculated and plotted.
(5) And (3) data processing: experimental data were analyzed, counted and plotted using GRAPHPAD PRISM 5.0.0, SPSS20.0 software.
(6) The results were as follows:
As shown in fig. 1: after depilation, the skin at the back of each group of mice showed pink color at the depilation area. Over time, on day 10 post-dehairing, the control, minoxidil, example 6, example 7, example 8 mice dehaired zone skin darkened significantly and had begun to grow hair, with the example 8 mice dehaired zone hair being more concentrated, hair length longer, and the model mice dehaired zone skin only began to darken. On day 20, the control, example 6, example 7, example 8 mice were essentially full of hair but the model mice were not fully full of hair. It is apparent that the hair growth rate in the back dehairing zone was slower in the model group than in the other groups, and that the hair growth rate was the fastest in the mice of example 8 group.
Table 1 neonatal hair growth status score for each group of mice
Wherein table 1 corresponds to the data of fig. 2.
As shown in fig. 2 and table 1, the control group, minoxidil group, example 6 group, example 7 group, example 8 group mice showed significant new hair growth starting on day 10 after depilation, the score value increased day by day, and the example 8 group mice showed the fastest hair growth, suggesting that the formulation of example 8 group had significant hair growth promoting effect; on day 20, the back dehairing area of the model group mice was not fully overgrown, but the control group and the examples each group mice had no significant difference in the new hair and unhairing area. ** P <0.01 compared to control group and #P<0.05,# P <0.01 compared to model group.
Verification example 2
By utilizing qPCR technology, the expression influence of the traditional Chinese medicine composition extracts (example 9, comparative example 3 and comparative example 4) on DKK-1 genes in a human hair papilla cell model is found, and the specific experimental process is as follows:
(1) DPC cells are spread in a 12-hole plate at 15 ten thousand per hole, after 24 hours of cell wall-attached growth, the traditional Chinese medicine composition extract with the final concentration of 50 mug/mL is added, and simultaneously, a DMSO solvent control group is arranged for treating the cells for 24 hours;
(2) RNA was extracted using FASTKING CDNA first strand synthesis kit (manufacturer: TIANGEN, lot: KR 116-02) and inverted to cDNA.
(3) QPCR reaction, procedure: 95 ℃ for 3min; (95 ℃,3s;60 ℃,30 s) 40 cycles.
TABLE 2 DKK-1 primer sequences
(4) And (3) data processing: experimental data were analyzed, counted and plotted using GRAPHPAD PRISM 5.0.0, SPSS20.0 software. *P<0.05,** P <0.01 compared to the control group and #P<0.05,## P <0.01 compared to the example 9 group.
(5) The results were as follows:
Table 3 qPCR results (n=4) of extracts of the herbal compositions (example 9, comparative example 3, comparative example 4) on DPC cells
Wherein table 3 corresponds to the data of fig. 3.
As shown in table 3 and fig. 3, DKK-1 genes were significantly reduced in each of the example 9 group, the comparative example 3 group and the comparative example 4 group, and the reduction was more significant in the example 9 group, compared to the control group; compared with the group of the example 9, the DKK-1 genes of the group of the comparative example 3 and the group of the comparative example 4 are obviously improved, and the results indicate that all the three groups of formulas have a certain alopecia preventing effect, and the effect of the group of the example 9 is more obvious. ** P <0.01 compared to the control group and #P<0.05,## P <0.01 compared to example 9.
Verification example 3
(1) The method for using the extract of the traditional Chinese medicine composition prepared in the embodiment 10 comprises the following steps: the Chinese medicinal composition is used twice a day, 2-4 mL of Chinese medicinal composition extract is smeared on the head (25 cm 2) each time, fifteen days is a treatment course, and administration is stopped after two to four treatment courses, so that a great amount of hair can be obviously grown.
(2) Clinical trial: the extract of the Chinese medicinal composition prepared in example 10 was used for promoting hair production, and 5 persons, male 4 persons, female 1 person, and male and female having an average age of 30 to 65 years, and all had long-term alopecia or about 100 average alopecia per day, wherein 4 men were long-term local hairless patients and 1 female were severe hairless patients, respectively corresponding to the extract of the Chinese medicinal composition prepared in example 10. About 2mL of the sample is taken and smeared on a alopecia part (25 cm 2), and the application is carried out 2 times a day for 1 month. The hair loss preventing effect was evaluated by evaluating the number of dropped hairs at each shampoo day, the hair growth promoting effect was counted by naked eyes, and the results are shown in Table 4. In case 3 of the verification example group, the alopecia area is not enlarged within 30 days, new stubbles or fuzz can grow on the affected part in the rest, the new hair can continuously grow, and serious hair loss is obviously reduced. Experiments prove that the traditional Chinese medicine composition extract prepared in the embodiment 10 has the effects of preventing alopecia and promoting hair growth for patients suffering from alopecia.
TABLE 4 effects of promoting hair production by the extracts of the Chinese medicinal compositions prepared in EXAMPLE 10
| Case of cases | Sex (sex) | Age of | Improving the situation |
| Case 1 | Man's body | 63 | New hair grows out from no hair place |
| Case 2 | Man's body | 58 | New hair grows out from no hair place |
| Case 3 | Man's body | 46 | The alopecia area is not enlarged |
| Case 4 | Man's body | 60 | New hair grows out from no hair place |
| Case 5 | Female | 37 | Fluff growing at hairless place |
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (7)
1. The traditional Chinese medicine composition for growing and/or nursing hair is characterized by comprising the following components in parts by weight:
0.01-15 parts of black sesame
2.5-100 Parts of tuber fleeceflower stem
2.5-100 Parts of eclipta
2.5-100 Parts of biota orientalis leaves
0.4-10 Parts of herba lycopi
2.5-100 Parts of rhizoma dioscoreae hypoglaucae
1.5-60 Parts of baikal skullcap root
1.5-60 Parts of phellodendron bark
1.5-50 Parts of cassia twig
0.75-30 Parts of pseudo-ginseng
0.75-30 Parts of rosewood heart wood
And 0.075-3 parts of storax.
2. The traditional Chinese medicine composition according to claim 1, which is characterized by comprising the following components in parts by weight:
15 parts of black sesame
10 Parts of tuber fleeceflower stem
10 Parts of eclipta
10 Parts of biota orientalis
Herba lycopi 10 parts
10 Parts of rhizoma dioscoreae septemlobae
6 Parts of baikal skullcap root
6 Parts of phellodendron bark
Ramulus Cinnamomi 6 parts
Pseudo-ginseng 3 parts
Dalbergia wood 3 parts
0.3 Parts of storax;
Or (b)
Black sesame 3.75 parts
2.5 Parts of tuber fleeceflower stem
2.5 Parts of eclipta
2.5 Parts of biota orientalis
2.5 Parts of herba lycopi
2.5 Parts of rhizoma dioscoreae septemlobae
1.5 Parts of baikal skullcap root
1.5 Parts of phellodendron bark
1.5 Parts of cassia twig
Notoginseng 0.75 parts
0.75 Part of rosewood heart wood
0.075 Parts of storax;
Or (b)
7.5 Parts of black sesame
5 Parts of tuber fleeceflower stem
5 Parts of eclipta
5 Parts of biota orientalis
5 Parts of herba lycopi
5 Parts of rhizoma dioscoreae septemlobae
3 Parts of baikal skullcap root
Cortex phellodendri 3 parts
3 Parts of cassia twig
Notoginseng 1.5 parts
1.5 Parts of dalbergia wood
0.15 Parts of storax;
Or (b)
0.01 Part of black sesame
100 Parts of tuber fleeceflower stem
100 Parts of eclipta
100 Parts of biota orientalis
0.4 Part of herba lycopi
100 Parts of rhizoma dioscoreae septemlobae
60 Parts of baikal skullcap root
60 Parts of phellodendron bark
Ramulus Cinnamomi 50 parts
Pseudo-ginseng 30 parts
30 Parts of dalbergia wood
3 Parts of storax.
3. The method for preparing a Chinese medicinal composition according to claim 1 or 2, wherein the components in the formula amount are crushed, and the extract is collected by ethanol extraction with 50% ethanol aqueous solution or 90% ethanol aqueous solution.
4. A method of preparation according to claim 3, wherein the step of concentrating and/or freeze drying is further included after the collection of the extract;
the concentration temperature is 70 ℃, and the concentration ratio is 1:15; the freeze-drying pressure is 1 Pa, the temperature is-65 ℃, and the freezing time is 24: 24h.
5. The use of a Chinese medicinal composition according to claim 1 or 2 for the preparation of a medicament for promoting hair growth and/or hair growth.
6. The use of a Chinese medicinal composition according to claim 1 or 2 for the preparation of a medicament for the treatment and/or prophylaxis of androgenetic alopecia.
7. A medicament for growing and/or nourishing hair, comprising the Chinese medicinal composition according to claim 1 or 2 and pharmaceutically acceptable auxiliary materials.
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|---|---|---|---|---|
| CN102028759A (en) * | 2011-01-14 | 2011-04-27 | 霸王（广州）有限公司 | Chinese medicine hair-darkening empirical formula and preparation method thereof |
| CN106511924A (en) * | 2016-10-27 | 2017-03-22 | 哈尔滨乐泰药业有限公司 | Externally-applied pharmaceutical composition for treating alopecia and promoting hair growth and preparation method thereof |
| CN109419862A (en) * | 2017-08-28 | 2019-03-05 | 吴颖 | A kind of pharmaceutical formulation of hair growth |
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| CN102028759A (en) * | 2011-01-14 | 2011-04-27 | 霸王（广州）有限公司 | Chinese medicine hair-darkening empirical formula and preparation method thereof |
| CN106511924A (en) * | 2016-10-27 | 2017-03-22 | 哈尔滨乐泰药业有限公司 | Externally-applied pharmaceutical composition for treating alopecia and promoting hair growth and preparation method thereof |
| CN109419862A (en) * | 2017-08-28 | 2019-03-05 | 吴颖 | A kind of pharmaceutical formulation of hair growth |
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