Medicinal preparation for promoting hair growth and preparation method thereof

Abstract

The invention relates to the technical field of external medicinal preparations for hair, in particular to a medicinal preparation for promoting hair growth and a preparation method thereof. The pharmaceutical preparation is prepared from the following components in parts by mass: 0.1 to 0.5 part of exosome, 0.2 to 0.7 part of collagen and 2.5 to 5 parts of natural plant extract. The pharmaceutical preparation disclosed by the invention can promote and stimulate the growth of hair, improve the quality of the hair and thicken the hair; the hair follicle resuscitation can be stimulated, so that the hair follicle is regenerated, and the number of hair follicles is increased.

Description

Technical Field

[0001] The invention relates to the technical field of hair external medicinal preparations, and in particular to a medicinal preparation for promoting hair growth and a preparation method thereof. Background Art

[0002] Hair loss refers to the phenomenon of hair falling out. Normally, the hair that falls out is in the regression phase and the resting phase. Since the hair entering the regression phase and the hair entering the growth phase are constantly in a dynamic balance, the normal amount of hair can be maintained. Hair loss has a great impact on people's psychology and daily life.

[0003] Exosomes secreted by cells contain a variety of growth factors, such as stem cell factor (SCF), insulin-like growth factor 1 (IGF-1), vascular endothelial growth factor (VEGF), epidermal growth factor (EGF) and keratinocyte growth factor (KGF). After entering the target cells, exosomes can interact with cells and mediate intercellular signal transduction. These factors can not only repair damaged hair follicles and activate the growth of hair follicle stem cells, but also promote hair growth and improve hair quality.

[0004] There are more and more reports on the use of plant extracts such as traditional Chinese medicine to treat hair loss. These plant-derived ingredients are relatively safe, effective and easy to use. Chinese herbal plant extracts have the advantages of easy access, simple production process and low price. They are also compatible with many cosmetic raw materials. These plant extracts often have the effects of promoting hair growth, improving hair quality, stimulating hair follicle regeneration, improving hair follicle vitality, and anti-oxidation, anti-inflammatory and antibacterial.

[0005] Collagen has a strong affinity for protein molecules on the surface of hair and can bind to the surface of hair through physical adsorption. Collagen is resistant to rinsing, and its affinity increases with the increase of its molecular weight. Collagen with a small molecular weight can penetrate the cortex of the hair to achieve the effect of nourishing the hair. Supplementing collagen for hair can promote the smoothness and luster of hair. Collagen is an important component for maintaining healthy skin and hair, and helps to enhance the elasticity and toughness of hair. At the same time, collagen can promote blood circulation and improve the nutritional supply of the scalp, thereby reducing the occurrence of hair loss. In addition, collagen can also stimulate the growth of hair follicles and promote hair growth.

[0006] However, in existing studies, there are few reports on the combined use of exosomes with collagen and natural plant extracts in hair topical pharmaceutical preparations; therefore, providing a method for combining exosomes with collagen and natural plant extracts for preparing hair growth pharmaceutical preparations with excellent effects is one of the technical problems that need to be solved at present. Summary of the invention

[0007] The purpose of the invention is to prepare a hair external medicinal preparation, more specifically a medicinal preparation for promoting hair growth, which can promote the generation of hair follicle cells and stimulate hair follicles to enter the growth phase, thereby significantly improving the problem of hair loss; helping hair loss patients, using a smear auxiliary material treatment method, combining prevention with treatment, reducing the pain of patients during the treatment process, and shortening the treatment time.

[0008] On the one hand, the pharmaceutical preparation for promoting hair growth of the present invention involves three raw materials: the first raw material is exosomes produced by cell culture; the second component is collagen; and the third component is natural plant extracts.

[0009] Furthermore, the pharmaceutical preparation comprises the following raw materials in parts by mass: 0.1-0.5 parts of exosomes, 0.2-0.7 parts of collagen and 2.5-5 parts of natural plant extracts.

[0010] Furthermore, the natural plant extracts are extracts of Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberry.

[0011] Furthermore, the exosomes are obtained by umbilical cord crushing, primary culture, subculture and separation.

[0012] Furthermore, the method for preparing exosomes comprises the following steps:

[0013] Step (1), fragmentation: clean the umbilical cord, peel off the amniotic membrane and arteries and veins of the umbilical cord, remove Wharton's jelly, and cut it into pieces;

[0014] Step (2), primary culture: placing the obtained fragments in a vessel containing culture medium for culture;

[0015] Step (3), subculture: when the cell confluence in step (2) reaches 68-85%, discard the culture medium and rinse the cell growth surface; add a trypsin aqueous solution to obtain a mixture, then add the culture medium to form a single cell suspension, centrifuge, discard the supernatant, add the culture medium to resuspend the cells, and subculture;

[0016] Step (4), separation: collecting the culture supernatant when the cells are cultured to the third generation, and centrifuging to remove cell debris; filtering the supernatant, collecting the filtrate, and freeze-drying to obtain the exosomes.

[0017] Furthermore, the collagen is type I collagen. The collagen can be selected as type I collagen α1 (COL1a1) recombinant protein, Shanghai Bolson Biotechnology Co., Ltd.

[0018] Furthermore, the mass ratio of the exosomes to collagen is 1:1-1.5.

[0019] Furthermore, the preparation method of the natural plant extract comprises the following steps:

[0020] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them respectively, and mix them according to the following weight proportions: 1-3 parts of Polygonum multiflorum, 0.5-2.5 parts of ginger, 1-2.5 parts of Platycladus orientalis leaves and 1.2-3.5 parts of mulberries to obtain a mixed powder;

[0021] Step 2: adding water to the mixed powder of step 1, decocting to obtain a decocted material;

[0022] Step 3: cooling the decoction, adding ethanol to obtain a mixed solution, adjusting the ethanol concentration in the mixed solution to 45-65wt%, heating and stirring, cooling, filtering, and concentrating to obtain a concentrate;

[0023] Step 4: Add the concentrate to an extractant consisting of an organic phase and an aqueous phase for extraction, then pour out the aqueous phase, concentrate the extracted organic phase, remove the organic solvent, and obtain a concentrate, which is the natural plant extract.

[0024] Furthermore, in step 1, the particle size selected after crushing is 0.05-1 mm.

[0025] Furthermore, in step 2, the mass ratio of water to mixed powder is 10-28:1.

[0026] Furthermore, in step 2, the decoction temperature is 75-90°C.

[0027] Furthermore, in step 2, the decoction time is 5-10 hours.

[0028] Furthermore, in step 2, the decoction method is to add water to the volume before decoction every 30 minutes.

[0029] Furthermore, in step 3, the decoction is cooled to 15-30°C.

[0030] Furthermore, in step 3, the temperature of heating and stirring is 45-55°C.

[0031] Furthermore, in step 3, the heating stirring speed is 60-500 rpm.

[0032] Furthermore, in step 3, the heating and stirring time is 10-30 hours.

[0033] Furthermore, in step 3, filtration is performed using a filter membrane with a pore size of 0.1-2 microns.

[0034] Furthermore, in step 3, the concentration is concentrated under reduced pressure.

[0035] Furthermore, in step 3, the concentration temperature is 55-60°C

[0036] Furthermore, in step 3, the mixture is concentrated to 0.3-0.8 times the mass of the powder.

[0037] Furthermore, in step 4, the mass ratio of the extractant to the concentrate is 6-20:1.

[0038] Furthermore, in step 4, the volume ratio of the organic phase to the aqueous phase in the extractant is 2.5-4:1.

[0039] Furthermore, in step 4, the organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 4-6:1.

[0040] Furthermore, in step 4, the aqueous phase is a 0.2-0.55 wt % calcium chloride aqueous solution.

[0041] Furthermore, in step 4, the extraction temperature is 35-45°C.

[0042] Furthermore, in step 4, the extraction time is 4-7 hours.

[0043] Furthermore, in step 4, the concentration is concentrated under reduced pressure.

[0044] Furthermore, in step 4, the concentration temperature is 46-52°C.

[0045] Furthermore, in step 4, when no organic solvent component of the organic phase is detected in the concentrate, concentration is stopped to obtain a concentrate, which is a natural plant extract.

[0046] Furthermore, the preparation method of the natural plant extract comprises the following steps:

[0047] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them respectively, and mix them according to the following weight proportions: 1-3 parts of Polygonum multiflorum, 0.5-2.5 parts of ginger, 1-2.5 parts of Platycladus orientalis leaves and 1.2-3.5 parts of mulberries to obtain a mixed powder;

[0048] Step 2: Add 10-28 times the mass of water to the mixed powder of step 1, and boil at 75-90° C. for 5-10 hours to obtain a boiled material;

[0049] Step 3: Cool the decoction, add ethanol to obtain a mixed solution, adjust the ethanol concentration in the mixed solution to 45-65wt%, heat and stir for 10-30 hours, cool, filter, and concentrate under reduced pressure at 55-60°C to 0.3-0.8 times the mass of the mixture powder to obtain a concentrate;

[0050] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 2.5-4:1, which is 6-20 times the mass, and extracting at 35-45° C. for 4-7 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 46-52° C. to remove the organic solvent. When the organic solvent component of the organic phase cannot be detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0051] The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 4-6:1, and the aqueous phase is a 0.2-0.55wt% calcium chloride aqueous solution.

[0052] In another aspect, the present invention provides a method for preparing the pharmaceutical preparation, comprising the step of mixing the raw materials.

[0053] On the other hand, the present invention also provides preparing the drug preparation into a dressing external-use drug product.

[0054] On the other hand, the present invention also provides preparing the pharmaceutical preparation into a gel-like external pharmaceutical product, wherein the gel-like external pharmaceutical product is composed of: 4-10% of the above-mentioned pharmaceutical preparation, 0.4-1% carbomer 940, 2-7% glycerol, 1.5-5% butylene glycol, 0.02-0.1% EDTA-2Na, 0.4-0.8% triethanolamine and the remainder water in terms of mass percentage.

[0055] Beneficial effects:

[0056] The pharmaceutical preparation prepared by combining exosomes with collagen and natural plant extracts can effectively stimulate hair growth, improve hair quality, and make hair thicker; and can promote hair follicle recovery, regenerate hair follicles, and increase the number of hair follicles.

[0057] After the drug preparation of the present invention is diluted to 10wt%, the hair length of the mouse test area reaches 6.14±0.67mm, the number of hair follicles reaches 46.2±4.9, and the hair diameter reaches 12.04±1.34μm after 20 days.

[0058] The use of exosomes and collagen in the pharmaceutical preparation of the present invention can give hair a better growth effect and enhance the vitality of hair follicles; the exosomes and collagen used in the pharmaceutical preparation of the present invention can mutually promote each other to increase the hair growth rate, improve the hair quality and enhance the vitality of hair follicles. When the mass ratio of exosomes to collagen in the pharmaceutical preparation of the present invention is 1:1-1.5, the effect of the obtained preparation is better.

[0059] The invention uses natural plant extracts to further enhance the hair growth effect of the preparation; and when extracting the plants, Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries are boiled in water, and then ethanol is added to adjust the ethanol concentration to 45-65wt%, and the mixture is heated and stirred for extraction, concentrated, and then extracted with an extractant obtained by combining an oil phase composed of ethyl acetate and cyclohexane with a water phase composed of a calcium chloride aqueous solution; the extract obtained by the extraction method of the invention can significantly enhance the effects of the preparation on promoting hair growth, improving hair quality and activating hair follicle vitality.

[0060] The present invention uses 45wt%-65wt% of the plant extract obtained by extraction, so that the preparation has a better hair growth effect.

[0061] When extracting plant active ingredients, the present invention uses ethyl acetate and cyclohexane to efficiently dissolve and extract hair growth active ingredients in a synergistic manner, giving the preparation a better hair growth effect; and when the ratio of ethyl acetate to cyclohexane in the organic phase is 4-6:1, the effect of the preparation is better. When other organic phases are used to replace the organic phase of the present invention for extraction to obtain plant extracts, the hair growth effect of the prepared preparation is worse than that of the preparation of the present invention.

[0062] The hair growth promoting pharmaceutical preparation of the present invention can promote the generation of hair follicle cells and stimulate hair follicles to enter the growth phase, thereby significantly improving the problem of hair loss; it helps hair loss patients to combine prevention with treatment by using a smear auxiliary material treatment method, thereby alleviating the pain of the patient during the treatment process and shortening the treatment time. DETAILED DESCRIPTION

[0063] The present invention will be described below in conjunction with specific embodiments, and various effects of the present invention will be more clearly presented. It should be understood by those skilled in the art that these specific embodiments are used to illustrate the present invention, rather than to limit the present invention.

[0064] Unless otherwise specified, various raw materials, reagents, instruments and equipment used in the present invention can be purchased from the market or prepared by existing methods.

[0065] 1. Preparation of exosomes: Step (1), fragmentation: The umbilical cord of a healthy full-term caesarean section baby was washed with physiological saline, the amniotic membrane and arteries and veins of the umbilical cord were peeled off, Wharton's jelly was removed, and the fragments were cut into pieces of about 1 mm in size;

[0066] Step (2), primary culture: placing the obtained fragments in a vessel containing DMEM culture medium and culturing at 37°C and 5% CO2;

[0067] Step (3), subculture: When the cell fusion degree of the primary culture in step (2) reaches 70%, the DMEM culture medium is discarded, PBS buffer is added to rinse the cell growth surface, and the washing solution is discarded; then a 0.25wt% trypsin aqueous solution is added, and the trypsin is shaken so that the trypsin evenly covers the bottom of the vessel to obtain a mixture, 10 times the volume of the mixture is added to the DMEM culture medium, and the mixture is repeatedly blown to form a single cell suspension, centrifuged at 1500rpm for 8 minutes, the supernatant is discarded, and the cells are resuspended in DMEM culture medium at a density of 1.5×10 5 cels were subcultured at 37°C and 5% CO2;

[0068] Step (4), separation: collecting the culture supernatant when the cells were cultured to the third generation, centrifuging at 4°C and 2000 g for 35 min to remove cell debris; filtering the supernatant after centrifugation with a 0.22 μm filter pore, collecting the filtrate, and freeze-drying to obtain the exosomes.

[0069] 2. Preparation of natural plant extracts

[0070] 1. Extract 1: Prepare as follows:

[0071] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0072] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0073] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 52.5wt%, heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered with a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0074] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0075] The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 5:1, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0076] Extract 2: Prepare as follows:

[0077] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 1.8 parts of Polygonum multiflorum, 1.2 parts of ginger, 1.7 parts of Platycladus orientalis leaves and 3.5 parts of mulberries by weight to obtain a mixed powder;

[0078] Step 2: Add 20 times the mass of water to the mixed powder of step 1, boil at 82° C. for 5.5 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0079] Step 3: The decoction is cooled to 25° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 45wt%, the mixture is heated to 50° C., stirred at 80 rpm for 15 hours, cooled to 26° C., filtered through a 0.22 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.4 times the mass of the mixture powder to obtain a concentrate;

[0080] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3:1 with a mass of 15 times, extracting at 42° C. for 5.5 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, stopping the concentration to obtain a concentrate, which is the natural plant extract;

[0081] The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 4:1, and the aqueous phase is a 0.25 wt % calcium chloride aqueous solution.

[0082] Extract 3: Prepare as follows:

[0083] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0084] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0085] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 20wt%, the mixture is heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered through a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0086] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0087] The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 5:1, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0088] Extract 4: Prepare as follows:

[0089] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0090] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0091] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 52.5wt%, heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered with a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0092] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0093] The organic phase is an organic solvent ethyl acetate, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0094] Extract 5: Prepare as follows:

[0095] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0096] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0097] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 52.5wt%, heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered with a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0098] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0099] The organic phase is an organic solvent cyclohexane, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0100] Extract 6: Prepare as follows:

[0101] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0102] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0103] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 52.5wt%, heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered with a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0104] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0105] The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 2:1, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0106] Extract 7: Prepare as follows:

[0107] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0108] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0109] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 52.5wt%, heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered with a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0110] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0111] The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 8:1, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0112] Extract 8: Prepare as follows:

[0113] Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them separately, and select a particle size between 0.05 and 1 mm after crushing. Mix 2.5 parts of Polygonum multiflorum, 1.8 parts of ginger, 2.2 parts of Platycladus orientalis leaves and 2.5 parts of mulberries by weight to obtain a mixed powder;

[0114] Step 2: Add 18 times the mass of water to the mixed powder of step 1, boil at 80° C. for 6 hours, and add water to the volume before boiling every 30 minutes to obtain a boiled material;

[0115] Step 3: the decoction is cooled to 28° C., ethanol is added to obtain a mixed solution, the ethanol concentration in the mixed solution is adjusted to 52.5wt%, heated to 52° C., stirred at 100 rpm for 12 hours, cooled to 28° C., filtered with a 0.5 μm filter membrane, and concentrated under reduced pressure at 58° C. to 0.45 times the mass of the mixture powder to obtain a concentrate;

[0116] Step 4: adding the concentrate to an extractant composed of an organic phase and an aqueous phase in a volume ratio of 3.5:1 with a mass of 12 times, extracting at 40° C. for 5.2 hours, then pouring out the aqueous phase, and concentrating the extracted organic phase under reduced pressure at 50° C. to remove the organic solvent. When the organic solvent component of the organic phase is no longer detected in the concentrate, the concentration is stopped to obtain a concentrate, which is the natural plant extract;

[0117] The organic phase is an organic solvent chloroform, and the aqueous phase is a 0.38 wt % calcium chloride aqueous solution.

[0118] 3. Drug preparations that promote hair growth

[0119] The composition of each preparation is shown in Table 1 below, and the raw materials are measured by mass. Among them, the exosomes are the exosomes prepared above; the collagen is type I collagen α1 (COL1 a1) recombinant protein, product number: BES20423RP, Shanghai Bolsen Biotechnology Co., Ltd.; the natural plant extracts are the above extracts 1-8. The mass ratio of exosomes to collagen is marked as K value.

[0120] Table 1: Raw material composition of each preparation

[0121] Serial number / copy Preparation 1 Preparation 2 Preparation 3 Preparation 4 Preparation 5 Preparation 6 exosomes 0.45 0.40 0 1.07 0.22 0.75 Collagen 0.62 0.50 1.07 0 0.85 0.32 Extract 1 3.43 3.15 3.43 3.43 3.43 3.43 K Value 1:1.38 1:1.25 / / 1:3.86 1:0.43

[0122] The extract used in preparations 1-6 is extract 1; then extracts 2-8 are used to replace extract 1 in preparation 1, and the resulting preparations are named preparations 7-13. Then the above preparations 1-13 are subjected to subsequent performance tests.

[0123] 4. Performance Testing

[0124] The above-mentioned preparation was diluted with an ethylene glycol aqueous solution to a dilution with a mass concentration of 10%, and then a performance test was performed; wherein the mass concentration of the ethylene glycol aqueous solution was 25%.

[0125] SPF-grade C57BL / 6J male mice weighing about 23-25 ​​grams were randomly divided into blank control group, model control group, positive control group, and preparation 1-13 groups, with 8 mice in each group. Each group of mice was anesthetized with isoflurane, and the hair of the same position of the mice was removed, with an area of ​​3cm×3cm as the test area. A 12mg / mL dihydrotestosterone corn oil solution was prepared, and 0.1mL dihydrotestosterone corn oil solution was intraperitoneally injected into the mice of the other groups except the blank control group for 20 consecutive days to establish a seborrheic alopecia model. Then, in the positive control group, 0.5mg of ethylene glycol aqueous solution with a mass concentration of 6% minoxidil (ethylene glycol mass concentration in ethylene glycol aqueous solution is 30%) was applied to the depilatory area once in the morning and evening; the mice in the preparation 1-13 groups were applied with 0.5mg of the above dilution each time on the depilatory area, once in the morning and evening; the blank control group and the model control group were applied with an equal amount of normal saline in the same way. After 20 days, the hair growth of the test area of ​​each group of mice was observed and counted to obtain the average length and diameter of the new hair; then the mice were killed, the skin tissue of the test area was taken for HE staining, and the average number of hair follicles in the test area was calculated. The specific test results of the average length, diameter and number of hair follicles of the new hair are shown in Table 2.

[0126] Table 2: Average length, diameter and number of hair follicles

[0127]

[0128]

[0129] Based on the results of the above tests, the increase in the average length, diameter and number of hair follicles of each preparation group and positive control group compared with the model control group was calculated. The results are shown in Table 3. Among them, the increase in the average length of new hair / mm = the average length of new hair in the preparation group or positive control group - the average length of new hair in the model control group; the increase in the average number of new hair follicles / piece = the average number of new hair follicles in the preparation group or positive control group - the average number of new hair follicles in the model control group; the increase in the diameter of new hair / μm = the average diameter of new hair in the preparation group or positive control group - the average diameter of new hair in the model control group.

[0130] Table 3: The improvement of the preparation group compared with the model control group

[0131]

[0132]

[0133] From the test results in Table 2-3, it can be seen that the preparation prepared by combining exosomes with collagen and natural plant extracts in the present invention can effectively stimulate hair growth, improve hair quality, and make hair thicker; and can promote hair follicle recovery, regenerate hair follicles, and increase the number of hair follicles.

[0134] The test results of the preparations in Table 2-3 show that after the drug preparation of the present invention is diluted to 10wt%, the hair length of the mouse test area reaches 6.14±0.67mm after 20 days, the number of hair follicles reaches 46.2±4.9, and the hair diameter reaches 12.04±1.34μm.

[0135] The use of exosomes and collagen in the pharmaceutical preparation of the present invention can give hair a better growth effect and enhance the vitality of hair follicles; the test results of preparations 1, 3-4 in Table 2-3 show that the exosomes and collagen used in the pharmaceutical preparation of the present invention can mutually promote each other to increase the hair growth rate, improve the hair quality and enhance the vitality of hair follicles. By comparing preparations 5-6 with 1, 2, and 7, it can be seen that when the mass ratio of exosomes and collagen in the pharmaceutical preparation of the present invention is 1:1-1.5, the effect of the preparation is better.

[0136] The present invention uses natural plant extracts to further enhance the hair growth effect of the preparation; and when extracting the plants, Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries are boiled in water, and then ethanol is added to adjust the ethanol concentration to 45-65wt%, and the mixture is heated and stirred for extraction, concentrated, and then extracted with an extractant obtained by combining an oil phase composed of ethyl acetate and cyclohexane with an aqueous phase composed of a calcium chloride aqueous solution; the extract obtained by the extraction method of the present invention can significantly enhance the effect of the preparation on promoting hair growth, improving hair quality and activating hair follicle vitality, which can be specifically obtained by comparing preparations 1, 2, 7 with 8-13.

[0137] Comparison between preparations 1 and 8 shows that the present invention uses 45wt%-65wt% of the plant extract to make the preparation have a better hair growth effect.

[0138] The test results of preparations 1, 9-13 show that when extracting plant active ingredients, the present invention uses ethyl acetate and cyclohexane to efficiently dissolve and extract hair growth active ingredients in a synergistic manner, giving the preparation a better hair growth effect; and when the ratio of ethyl acetate to cyclohexane in the organic phase is 4-6:1, the effect of the preparation is better. When other organic phases are used to replace the organic phase of the present invention for extraction to obtain plant extracts, the hair growth effect of the prepared preparation is worse than that of the preparation of the present invention.

[0139] 5. Application

[0140] Hair growth gel, mass percentage: 7.5% of the above preparation 1, 0.54% of carbomer 940, 3% of glycerol, 2% of 1,3-butylene glycol, 0.05% of EDTA-2Na, 0.54% of triethanolamine and the balance of water.

[0141] Preparation method: Carbomer 940 is mixed with water, stirred at 60 rpm for 0.3 hour, the above-mentioned preparation 1, glycerol, 1,3-butylene glycol, EDTA-2Na and triethanolamine are added, and stirring is continued for 0.5 hour to obtain the hair growth gel.

[0142] Volunteers with hair loss choose to use the hair growth gel prepared by the pharmaceutical preparation 1 of the present invention. The method of use is to apply it to the hair loss area and then apply it evenly. The usage amount is 1g / time, once in the morning and once in the evening every day, and it is used continuously for 3 months.

[0143] The hair growth promoting pharmaceutical preparation of the present invention can promote the generation of hair follicle cells and stimulate hair follicles to enter the growth phase, thereby significantly improving the problem of hair loss; it helps hair loss patients to combine prevention with treatment by using a smear auxiliary material treatment method, thereby alleviating the pain of the patient during the treatment process and shortening the treatment time.

[0144] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. A pharmaceutical preparation for promoting hair growth, characterized in that: The pharmaceutical preparation comprises the following raw materials in parts by mass: 0.1-0.5 parts of exosomes, 0.2-0.7 parts of collagen and 2.5-5 parts of natural plant extracts.

2. The pharmaceutical preparation according to claim 1, characterized in that The natural plant extracts are extracts comprising Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberry.

3. The pharmaceutical preparation according to claim 1, characterized in that The exosomes are obtained by umbilical cord crushing, primary culture, subculture and separation.

4. The pharmaceutical preparation according to claim 1, characterized in that The collagen is type I collagen.

5. The pharmaceutical preparation according to claim 1, characterized in that The mass ratio of the exosomes to collagen is 1:1-1.

5.

6. The pharmaceutical preparation according to claim 1, characterized in that The preparation method of the natural plant extract comprises the following steps: Step 1: Weigh dried Polygonum multiflorum, ginger, Platycladus orientalis leaves and mulberries, crush them respectively, and mix them according to the following weight proportions: 1-3 parts of Polygonum multiflorum, 0.5-2.5 parts of ginger, 1-2.5 parts of Platycladus orientalis leaves and 1.2-3.5 parts of mulberries to obtain a mixed powder; Step 2: adding water to the mixed powder of step 1, decocting to obtain a decocted material; Step 3: cooling the decoction, adding ethanol to obtain a mixed solution, adjusting the ethanol concentration in the mixed solution to 45-65wt%, heating and stirring, cooling, filtering, and concentrating to obtain a concentrate; Step 4: adding the concentrate to an extractant consisting of an organic phase and an aqueous phase for extraction, then pouring out the aqueous phase, concentrating the extracted organic phase, and removing the organic solvent to obtain a concentrate, which is the natural plant extract; The organic phase is composed of organic solvents ethyl acetate and cyclohexane in a volume ratio of 4-6:1, and the aqueous phase is a 0.2-0.55wt% calcium chloride aqueous solution.

7. The pharmaceutical preparation according to claim 6, characterized in that In step 2, the decocting time is 5-10 hours.

8. The pharmaceutical preparation according to claim 6, characterized in that In step 3, the heating and stirring time is 10-30 hours.

9. The pharmaceutical preparation according to claim 6, characterized in that In step 4, the mass ratio of the extractant to the concentrate is 6-20:

1.

10. A method for preparing the pharmaceutical preparation according to any one of claims 1 to 9, characterized in that: The method comprises the steps of mixing the raw materials.

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