Composition for promoting hair regeneration and application

Abstract

The invention provides a composition for promoting hair regeneration and application. The composition comprises an exosome and dipeptide diaminobutyrylbenzylamide diacetate. In the composition, the dipeptide diaminobutyrylbenzylamide diacetate can cooperate with the exosome to promote hair regeneration and prevent alopecia, and the composition has excellent alopecia preventing and hair growing effects. And the effect of the composition is obviously superior to the effect of the exosome and the dipeptide diaminobutyrylbenzylamide diacetate which are used independently and the effect of the exosome and the dipeptide diaminobutyrylbenzylamide diacetate which are superposed.

Description

Technical Field

[0001] The present invention belongs to the technical field of exosome application, and relates to a composition for promoting hair regeneration and its application. Background Art

[0002] Hair loss is the abnormal loss of hair caused by factors such as genetics, immunity, stress, taking certain medications, endocrine disorders, etc., which causes sparse hair or the formation of bald spots, causing great mental distress to the person concerned.

[0003] In medicine, minoxidil and finasteride are commonly used to treat hair loss. Minoxidil needs to be used regularly once or twice a day to prevent hair loss, but it is prone to side effects such as itching and allergic dermatitis. Finasteride tablets are a 5α-reductase inhibitor that can inhibit the conversion of testosterone to dihydrotestosterone in the body, thereby inhibiting the shrinkage of hair follicles and reversing hair loss. The probability of impotence, loss of libido and decreased semen volume caused by the use of finasteride is as high as 8.1%, 6.4% and 3.7% respectively. Faced with such a high failure rate, safe and effective hair loss prevention methods are necessary.

[0004] At present, exosomes are gradually being used to treat hair loss. For example, patent CN 111073881 A discloses a method for preparing exosomes with hair regeneration effects. The exosomes prepared by this method have high permeability and do not have the side effects that occur when synthetic drugs are used. U.S. Patent US 11,185,481 B2 describes that exosomes derived from stem cells promote or enhance wound healing and hair growth. However, there is still a problem that exosomes are not effective in treating hair loss. Summary of the invention

[0005] The purpose of the present invention is to provide a composition for promoting hair regeneration and its application, so as to solve the problem that the existing exosomes are not effective in treating hair loss.

[0006] To achieve the above object, the present invention adopts the following technical solutions: The present study found that dipeptide diaminobutyryl benzylamide diacetate can cooperate with exosomes to promote hair regeneration and prevent hair loss.

[0007] Dipeptide diaminobutyryl benzylamide diacetate is a peptide that simulates the activity of the snake venom toxin Waglerin I, with the sequence: H-β-Ala-Pro-Dab-NH-Bzl, which acts on the postsynaptic membrane and is a reversible antagonist of muscle nicotinic acetylcholine receptors (nmAChR). Dipeptide diaminobutyryl benzylamide diacetate can reduce the formation of wrinkles by inhibiting muscle contraction and has excellent wrinkle removal performance. In the present application, dipeptide diaminobutyryl benzylamide diacetate can cooperate with exosomes to promote hair regeneration and prevent hair loss, and has excellent anti-hair loss and hair growth effects. This helps to achieve excellent anti-hair loss effects even when using a small amount of exosomes.

[0008] Based on the above application, the present application provides a composition for promoting hair regeneration, which comprises exosomes and dipeptide diaminobutyryl benzylamide diacetate. The mass ratio of exosomes to dipeptide diaminobutyryl benzylamide diacetate is 1: (0.01-100). Preferably, the mass ratio of exosomes to dipeptide diaminobutyryl benzylamide diacetate is 1: (0.1-10).

[0009] In the present application, the exosomes are selected from one or more of cell culture supernatant, plasma, serum, saliva, urine, amniotic fluid, malignant ascites and other biological fluids.

[0010] The present application provides a method for culturing exosomes through tissue, the method comprising: (1) Disinfecting, cleaning, decomposing and digesting the skin tissue to obtain separated skin tissue; (2) Add 0.05% pancreatic digestive enzyme to the separated skin tissue to digest until the cells on the skin tissue fall off; obtain a single cell suspension by repeated blowing. The single cell suspension is filtered, centrifuged, washed, and precipitated to obtain primary epidermal stem cells.

[0011] (3) Primary epidermal stem cells were cultured in culture medium. When the cell confluence reached about 80%, the cells were subcultured. During the subculture process, the culture supernatant was collected.

[0012] (4) The culture supernatant is centrifuged to remove cell debris in the supernatant; the supernatant after centrifugation is transferred to a concentration tube to concentrate cytokines to obtain an exosome concentrate.

[0013] In addition, the composition of the present application further comprises excipients and / or active ingredients, wherein the excipients comprise at least one of a surfactant, a thickener, a pH regulator, a preservative and a fragrance, and the active ingredient comprises at least one of an extract of Platycladus orientalis leaves, adenosine, niacinamide and panthenol.

[0014] In the present application, the excipients are the excipients commonly used by those skilled in the art, such as surfactants including at least one of sodium laureth sulfate, cocamidopropyl betaine and decyl glucoside, thickeners including at least one of carbomer, xanthan gum and cellulose, pH adjusters including at least one of sodium hydroxide, citric acid, arginine and triethanolamine, preservatives including at least one of phenoxyethanol, sodium dehydroacetate, sodium benzoate and potassium sorbate, and fragrances including lavender essential oil or rose water.

[0015] In the present application, the types and contents of excipients and active ingredients in the composition are set according to actual needs, and the present application does not impose any restrictions.

[0016] The preparation method of the composition provided in the present application comprises: dissolving exosomes and dipeptide diaminobutyryl benzylamide diacetate in water and / or alcohol and mixing them to obtain the composition, wherein the alcohol is anhydrous ethanol, etc. Of course, the preparation method of the composition can also be prepared in other ways.

[0017] In addition, the present application also provides a cosmetic, which comprises the above composition. The cosmetic in the present application is used in the form of, but not limited to, essence, gel, cream or spray.

[0018] The present invention has the following beneficial effects: The dipeptide diaminobutyryl benzylamide diacetate in the present application can cooperate with exosomes to promote hair regeneration and prevent hair loss, and has excellent anti-hair loss and hair growth effects. And the effect of this combination is significantly better than the effect of exosomes and dipeptide diaminobutyryl benzylamide diacetate when used alone and the superposition of the two. DETAILED DESCRIPTION

[0019] The exosomes in the embodiments of the present application may be commercially available concentrated solutions or freeze-dried powders, or may be obtained through tissue culture. The technical solution of the present invention is further explained and illustrated by specific embodiments below.

[0020] Example 1 An embodiment of the present application provides a composition for promoting hair regeneration, which comprises exosomes and dipeptide diaminobutyrylbenzylamide diacetate in a mass ratio of 1:1.

[0021] The preparation method of the composition comprises the steps of dissolving exosomes and dipeptide diaminobutyryl benzylamide diacetate in water respectively, and then mixing the mixture evenly to form the composition.

[0022] Example 2 An embodiment of the present application provides a composition for promoting hair regeneration, which comprises exosomes and dipeptide diaminobutyryl benzylamide diacetate in a mass ratio of 1:0.1.

[0023] The preparation method of the composition is the same as that of Example 1.

[0024] Example 3 An embodiment of the present application provides a composition for promoting hair regeneration, which comprises exosomes and dipeptide diaminobutyryl benzylamide diacetate in a mass ratio of 1:10.

[0025] The preparation method of the composition is the same as that of Example 1.

[0026] Example 4 An embodiment of the present application provides a composition for promoting hair regeneration, which comprises exosomes and dipeptide diaminobutyryl benzylamide diacetate in a mass ratio of 1:0.01.

[0027] The preparation method of the composition is the same as that of Example 1.

[0028] Example 5 An embodiment of the present application provides a composition for promoting hair regeneration, which comprises exosomes and dipeptide diaminobutyryl benzylamide diacetate in a mass ratio of 1:100.

[0029] The preparation method of the composition is the same as that of Example 1.

[0030] Comparative Example 1 The composition of this comparative example includes dipeptide diaminobutyryl benzylamide diacetate at a concentration of 0.04%, and the remainder is water.

[0031] Comparative Example 2 The composition of this comparative example includes exosomes at a concentration of 0.04%, and the remainder is water.

[0032] In order to determine whether the composition provided in this application has the effect of preventing hair loss and promoting hair regeneration, this application adopts a human efficacy evaluation experiment for specific determination, and the specific contents are as follows: 1. Subject inclusion criteria 18-60 years old, healthy male or female; hair length between 5-40cm; those who have hair loss and mild hair thinning, and the hair loss count is greater than 10 according to the 60-combing method, and still greater than 10 after a 2-week washing period; those who have not undergone special hair treatments such as hair dyeing, perming, and styling in the past month; those who can cooperate with the staff's arrangements on site; those who can read and understand all the contents of the informed consent form and voluntarily sign the informed consent form.

[0033] 2. Exclusion criteria for subjects Pregnant or breastfeeding women, or those who are planning to get pregnant in the near future; those with psychiatric or psychological diseases; or those with long-term sleep or emotion control disorders; those who have used cosmetics with anti-hair loss effects or other samples with such effects or hair growth effects in the past 3 months; those who have taken or topically applied any drugs that affect hair growth in the past 6 months; those who have received hair transplant treatment; those with curly hair; those with highly sensitive constitutions; those who have participated in other clinical trials in the past 2 months; those who are considered unsuitable for participation in the trial by clinical evaluation.

[0034] 3. Subject Restrictions Subjects are not allowed to wash their hair within 48±4 hours before screening and each visit during the trial, and the time without washing hair before each visit is basically the same. They are not allowed to comb their hair on the day of the visit; they are not allowed to have their hair cut within 2 weeks before each visit evaluation during the trial; they are not allowed to perform any hair care and hairdressing measures during the trial, and are not allowed to receive any anti-hair loss or hair growth treatments; they need to maintain their original lifestyle habits during the trial and avoid large mood swings.

[0035] 4. Testing process 4.1 Preliminary experiment (1) Volunteer subjects were recruited according to the requirements and signed informed consent. Before enrollment, subjects were asked a series of questions about their medical history, health status, etc. according to the inclusion and exclusion criteria. The number of hair loss was counted using the 60-combing method and T was recorded. -2W , those with hair loss count greater than 10 can be included in the group; (2) Qualified subjects underwent a 2-week washout period and used washout samples; after the washout period (T 0W ) The subjects were required to perform the combing method 60 times again. Only those whose hair loss count was still greater than 10 could enter the formal test.

[0036] 4.2 Formal Experiment (1) The experiment officially started at week 0. After the subjects adapted to the required environment for 30 minutes, the examiner used the 60-comb method to count the number of hair loss and the corresponding instruments to measure the overall hair density (visual assessment or image assessment), local hair density, growth phase to rest phase ratio, local hair number, and hair diameter, recorded as T 0W ; (2) The subjects used Example 1, Comparative Examples 1 and 2 for 8 consecutive weeks, and were revisited at the 4th and 8th weeks, respectively. In Example 1, the amount of exosomes added was 0.04%, the amount of dipeptide diaminobutyryl benzylamide diacetate added was 0.04%, and the rest was water. During the revisit, after adapting to the required environment for 30 minutes, the inspector used the 60-comb method to count and the corresponding instruments to measure the number of hair loss, overall hair density (visual assessment or image assessment), local hair density, growth phase to rest phase ratio, local hair number, and hair diameter, which were recorded as T 4W , T8W , and get Table 1.

[0037] 4.3 Test parameters (1) Hair loss count During each visit, trained staff combed the subject's hair 60 times, counted the number of hairs that fell out, and recorded the number. Test time: T -2W , T 0W , T 4W , T 8W There was no significant reduction in hair loss counts, indicating that the test product had an effect in improving hair loss.

[0038] (2) Overall hair density The subject's hair on the top of the head should be combed symmetrically and smoothly on both sides. The same hairstyle must be maintained for each visit (refer to the full head hair photo taken for the first time). The hair density in the top area of ​​the head is assessed. The overall hair density assessment includes clinical assessment or image assessment, and the average of the two is calculated as the overall hair density. Test time point: T 0W , T 4W , T 8W There was no significant reduction in overall hair density, indicating that the test product had an effect in improving hair loss.

[0039] Visual assessment: Hair density was assessed on-site by a dermatologist at each visit and the score was recorded.

[0040] Image evaluation: Take a full-head hair photo at each visit. Specific steps: The subject wears a black non-reflective scarf and combs the hair smoothly (the hairstyle remains the same each time, and the follow-up photo refers to the first photo), places the chin on the shooting stand, fixes the shooting parameters of the digital camera, and takes a full-head hair photo with the top of the head as the center. Each time a photo is taken, a label needs to be attached, including the trial number, subject number and visit time, and a standard color palette (containing at least black, white and gray) is set. After all visits are completed, the dermatologist will evaluate the hair density of the photos taken and record the rating.

[0041] (3) Local hair density Use a scalp mirror x60 (KC scalp diagnostic instrument from Korea) for measurement. A haircut area of ​​at least 1.5cm×1.5cm (temporal side of the top of the head) is fixed on the subject's head and positioned properly. During each visit, the hair cutting area is ensured to be consistent, and the hair is cut to a residual length of no more than 1mm. During the image acquisition process, the operator needs to put the subject in a comfortable position and place the dermatoscope in the center of the hair cutting area to take local hair images. When shooting, the dermatoscope lens is completely in contact with the scalp and kept vertical, and the clarity of the captured image is checked. Use image analysis software or manual counting methods to count the number and density of local hair and record them. Test time: T0W , T 4W , T 8W There was no significant reduction in local hair density, indicating that the test product had an effect in improving hair loss.

[0042] (4) Growth phase to rest phase ratio (i.e. the ratio of the number of hairs in the growth phase to the number of hairs in the rest phase) The measurement was performed using a scalp mirror x60 (KC Scalp Diagnostic Instrument from Korea). Hair growth is cyclical, which can be divided into the growth phase, regression phase, and resting phase. The number of hair in the growth phase and resting phase was obtained by manual counting. Test time: T 0W , T 4W , T 8W The higher the growth phase to telogen ratio, the better the test product is at improving hair growth.

[0043] (5) Local hair quantity Use scalp mirror x60 (KC scalp diagnostic instrument, Korea) to measure. Test time: T 0W , T 4W , T 8W There is no obvious reduction in the amount of hair in the captured image, indicating that the test product has an effect in improving hair loss.

[0044] (6) Hair diameter Use scalp mirror x150 (KC scalp diagnostic instrument, Korea) for measurement. Test time: T 0W , T 4W , T 8W The thicker the hair diameter, the better the test product's effect in improving hair growth.

[0045] Table 1: Human efficacy evaluation test data As can be seen from Table 1, the composition of Comparative Example 1 containing only dipeptide diaminobutyryl benzylamide diacetate cannot prevent hair loss or increase hair volume. The composition of Comparative Example 2 containing only exosomes can prevent hair loss, increase hair density and local hair quantity. Compared with Comparative Example 2, the composition of Example 1 containing both exosomes and dipeptide diaminobutyryl benzylamide diacetate can significantly prevent hair loss, increase hair density, local hair quantity and hair diameter, which indicates that dipeptide diaminobutyryl benzylamide diacetate can synergistically enhance the anti-hair loss and hair growth effects of exosomes.

[0046] The above description is only the preferred embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application shall be included in the protection scope of the present application.

Claims

1. Dipeptide diaminobutyryl benzylamide diacetate cooperates with exosomes to promote hair regeneration and prevent hair loss.

2. A composition for promoting hair regeneration, characterized in that: Includes exosomes and dipeptide diaminobutyrylbenzylamide diacetate.

3. The composition for promoting hair regeneration according to claim 2, characterized in that: The mass ratio of the exosomes to the dipeptide diaminobutyrylbenzylamide diacetate is 1:(0.01-100).

4. The composition for promoting hair regeneration according to claim 2, characterized in that: The mass ratio of the exosomes to the dipeptide diaminobutyrylbenzylamide diacetate is 1:(0.1-10).

5. The composition for promoting hair regeneration according to claim 2, characterized in that: The exosomes are selected from one or more of cell culture supernatant, plasma, serum, saliva, urine, amniotic fluid, malignant ascites and other biological fluids.

6. The composition for promoting hair regeneration according to claim 2, characterized in that: The composition further comprises auxiliary materials and / or active ingredients.

7. The composition for promoting hair regeneration according to claim 6, characterized in that: The auxiliary materials include at least one of a surfactant, a thickener, a pH regulator, a preservative and a fragrance, and the active ingredient includes at least one of a Platycladus orientalis leaf extract, adenosine, niacinamide and panthenol.

8. The composition for promoting hair regeneration according to claim 7, characterized in that: The surfactant includes at least one of sodium laureth sulfate, cocamidopropyl betaine and decyl glucoside, the thickener includes at least one of carbomer, xanthan gum and cellulose, the pH adjuster includes at least one of sodium hydroxide, citric acid, arginine and triethanolamine, the preservative includes at least one of phenoxyethanol, sodium dehydroacetate, sodium benzoate and potassium sorbate, and the fragrance includes lavender essential oil or rose water.

9. A method for preparing a composition, characterized in that: include: The exosomes according to any one of claims 2 to 8 and the dipeptide diaminobutyrylbenzylamide diacetate are dissolved in water and / or alcohol and mixed to obtain the mixture.

10. A cosmetic, characterized in that: A composition comprising any one of claims 2 to 8.

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