CN109568169B - Active polypeptide with hair growth effect - Google Patents
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- CN109568169B CN109568169B CN201910057508.8A CN201910057508A CN109568169B CN 109568169 B CN109568169 B CN 109568169B CN 201910057508 A CN201910057508 A CN 201910057508A CN 109568169 B CN109568169 B CN 109568169B
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
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- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
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- A—HUMAN NECESSITIES
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- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
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- A61K2800/00—Properties of cosmetic compositions or active ingredients thereof or formulation aids used therein and process related aspects
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Abstract
The invention discloses an active polypeptide with hair growth effect, wherein the polypeptide is wrapped in a nano-inclusion by a lipid component, the nano-inclusion also comprises a polyalcohol, a surfactant and a pH buffering agent, the mass percentage concentration of the active polypeptide with hair growth effect is 0.0001-5%, the mass percentage concentration of the lipid component is 2-20%, the mass percentage concentration of the polyalcohol is 2-20%, the mass percentage concentration of the surfactant is 0.1-10%, and the mass percentage concentration of the pH buffering agent is 0.001-1%. The active polypeptide with the hair growth effect has good stability and high safety; easy transdermal absorption; compared with the active polypeptide before nano-encapsulation, the active polypeptide after nano-encapsulation has higher accumulation amount in the skin, and the same feeding amount can achieve more excellent effect of promoting hair growth.
Description
Technical Field
The invention belongs to the field of skin-care external medicines, and relates to an active polypeptide with a hair growth effect.
Background
The hair is mainly divided into two parts of hair shaft and hair root. The hair exposed on the surface of the skin is called a hair shaft, and the part buried in the skin and in the hair follicle is called a hair root, and the part protruding into the hair bulb from the bottom of the hair follicle is a hair papilla. The papilla is connected to the capillaries in the dermis to deliver nutrients to the hair through blood circulation. The collagen is an important nutrient component, controls the thickness, elasticity and wettability of the hair, is sufficient at the root of the hair, is beneficial to promoting the growth of the hair, and can make the hair strong and firm.
The hair matrix cells at the ends of the hair follicle have a decisive effect on the growth of the hair. Hair matrix cells have stem cell characteristics, and grow by proliferating through continuous division, moving upward, gradually degenerating and dying in the process, and accumulating a large amount of hard keratin in the cells to form various parts of the hair.
The growth cycle of hair is divided into three phases: growth stage, regression stage, resting stage. The growth cycles of the hair, eyebrows, and eyelashes are similar, except that the respective growth cycle times are different. The hair had an initial life of about 2-6 years, followed by a resting period and shedding, with a daily growth length of about 0.3 mm. The eyebrow growth period is about 2 months, and the eyebrow grows about 0.2mm per day. Eyelash growth cycle is about 5-6 months, wherein the duration of growth period is (34 + -9) days on average, and the average growth rate is (0.12 + -0.05) mm/day.
Alopecia is a common disease, which not only affects the appearance of a person, but also causes a heavy psychological burden on the patient. Androgenic alopecia is the most common type of hair loss. With increasingly prominent social pressure, environmental pollution and other problems, the incidence of alopecia diseases is increasing year by year and the alopecia diseases are in a trend of being younger.
The treatment regimen used varies according to severity in response to the problem of missing or thinning hair, eyebrows, and eyelashes. Minoxidil is a medicament which is commonly used for treating alopecia in clinic at present, and 2 percent of minoxidil can make most male and female patients with male hereditary alopecia grow visible hair/fine hair, but side effects such as scalp inflammation and facial hair increase can occur after administration. In addition, the treatment medicine is bimatoprost, which can promote the growth of eyelashes, but the excessive use of the medicine can cause damage to optic nerves, possibly cause vision loss and even blindness, and can also cause symptoms such as retina swelling and eye inflammation. In recent years, the application of active polypeptides to solve hair loss and promote hair regeneration has been receiving attention from dermatologists and cosmetology experts, and these active polypeptides are used as raw materials in hair growth and care products to achieve good hair growth promoting effects.
However, polypeptide components are often directly added into the prescription in the existing products, which brings about the problems that the stability of the polypeptide in the product is poor, so that the hair growth polypeptide which originally has high-efficiency activity cannot fully exert the due effect, and the degradation products generated by the degradation of the polypeptide in the product may possibly generate potential harm to human bodies. In addition, due to the existence of the skin barrier, the communication of substances inside and outside the body is limited, and the polypeptide in the product cannot be smoothly absorbed through the skin barrier, so that the effect of promoting hair growth is difficult to be fully exerted. In order to increase the transdermal absorption and achieve the desired effect, the amount of the feed needs to be increased, which leads to an increase in cost.
In conclusion, there is a need in the art for a skin care product or a medical product for external use for skin, which has good stability, high safety, easy transdermal absorption, small dosage and excellent effect and can make up for the defects of the prior art.
Disclosure of Invention
The technical problem to be solved by the invention is to provide the active polypeptide with the hair growth effect, which has the advantages of good stability, high safety, easy transdermal absorption, small dosage and excellent effect.
Therefore, the invention provides an active polypeptide with hair growth effect, which is wrapped in a nano-inclusion body.
The active polypeptide with hair growth effect is biotin tripeptide-1, myristoyl pentapeptide-17, myristoyl hexapeptide-16, tripeptide-1 copper and acetyl tetrapeptide-3, and the mass percentage concentration of each component is 0.0001-5%.
The coating of the present invention is a coating with a lipid component.
The lipid component is one or a combination of more of soybean lecithin, egg yolk lecithin, phosphatidylethanolamine, lysophosphatidylcholine, cholesterol, lauric acid, myristic acid, palmitic acid, stearic acid, isopropyl myristate, isopropyl palmitate, oleic acid, linoleic acid glyceride and vitamin E, and the mass percentage concentration of each component is 2-20%.
The nano inclusion also comprises the following components: polyol, surfactant, pH buffer.
The polyhydric alcohol is one or a combination of more of propylene glycol, glycerol, 1, 2-hexanediol and 1, 3-butanediol, and the mass percentage concentration of each component is 2-20%.
The surfactant is tween 20, tween 40, tween 60, tween 80, poloxamer and tyloxapol, and the mass percentage concentration of each component is 0.1-10%.
The pH buffering agent is a combination of disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate or dipotassium hydrogen phosphate, and the mass percentage concentration of each component is 0.001-1%.
The active polypeptide product form with hair growth effect of the present invention includes but is not limited to hair care essence, shampoo, lotion, mascara, eyeliner and eyebrow pencil.
The hair of the present invention is hair, eyebrows, or eyelashes.
To further facilitate understanding of the present invention, the mechanism of action of the above active polypeptide having hair growth effect is described as follows:
biotin Tripeptide-1 (CAS number: 299157-54-3) can promote the synthesis of collagen IV and laminin 5, delay hair follicle aging, promote hair growth, improve hair follicle structure, fix hair in dermal hair follicle, and prevent alopecia.
Myristoyl Pentapeptide-17 (Myristoyl Pentapeptide-17) stimulates the expression of keratin gene and promotes the production of keratin, thereby promoting hair growth.
Myristoyl Hexapeptide-16 (Myristoyl Hexapeptide-16) has the same action mechanism as Myristoyl pentapeptide-17, and stimulates the expression of keratin gene, thereby achieving the purpose of promoting hair growth.
Tripeptide-1 Copper (Copper Peptide) can expand hair follicles, accelerate hair growth, inhibit hair loss, prevent alopecia and promote hair growth in addition to the anti-wrinkle and anti-aging effects.
Acetyl Tetrapeptide-3 (CAS number: 827306-88-7) stimulates dermal hair emulsion or perifollicular fiber cells to generate collagen type III, laminin and other matrix proteins and collagen type VII, so that the hair follicle is more full and healthy and is beneficial to hair growth.
Compared with the prior art, the invention has the advantages that:
(1) the active polypeptide with hair growth effect is subjected to nano-encapsulation, so that the stability and the use safety of the polypeptide are improved.
(2) The polypeptide is coated by nanometer, and the transdermal absorption is increased.
(3) The nano-encapsulated active polypeptide with hair growth effect has higher accumulation amount in skin, and the same feeding amount can achieve better effect of promoting hair growth.
Drawings
FIG. 1 in vitro cumulative transdermal and cumulative skin Retention of the polypeptide (24h)
FIG. 2 average Density of anagen Hair
FIG. 3 average Density of telogen hairs
FIG. 4 percentage of eyebrow growth (30 days)
Figure 5% eyelash growth
FIG. 6 eyelash growth length
Detailed Description
For a better understanding of the present invention, the following detailed description is given in conjunction with the following examples and drawings, but is not limited to the following examples.
Example 1
Prescription
The preparation method comprises the following steps:
1. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
2. taking biotin tripeptide-1, tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 52.1 nm.
Example 2
Prescription
The preparation method comprises the following steps:
1. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
2. taking the myristoyl pentapeptide-17, the Tween 20, the disodium hydrogen phosphate and the sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), and stirring for dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 55.8 nm.
Example 3
Prescription
The preparation method comprises the following steps:
1. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
2. taking myristoyl hexapeptide-16, tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 56.4 nm.
Example 4
Prescription
The preparation method comprises the following steps:
1. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
2. taking tripeptide-1 copper, Tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 41.3 nm.
Example 5
Prescription
The preparation method comprises the following steps:
1. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
2. taking acetyl tetrapeptide-3, Tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), and stirring for dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 47.9 nm.
Example 6
Prescription
The preparation method comprises the following steps:
1. taking phosphatidylethanolamine, isopropyl myristate, oleic acid, propylene glycol and 1, 3-butanediol in the phase A according to the proportion of the formula, and stirring and dissolving the mixture at the heating temperature of 40 ℃ for later use;
2. taking biotin tripeptide-1, tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 32.5 nm.
Example 7
Prescription
The preparation method comprises the following steps:
1. taking phosphatidylethanolamine, isopropyl myristate, oleic acid, propylene glycol and 1, 3-butanediol in the phase A according to the proportion of the formula, and stirring and dissolving the mixture at the heating temperature of 40 ℃ for later use;
2. taking the myristoyl pentapeptide-17, the Tween 20, the disodium hydrogen phosphate and the sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), and stirring for dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 35.8 nm.
Example 8
Prescription
The preparation method comprises the following steps:
1. taking phosphatidylethanolamine, isopropyl myristate, oleic acid, propylene glycol and 1, 3-butanediol in the phase A according to the proportion of the formula, and stirring and dissolving the mixture at the heating temperature of 40 ℃ for later use;
2. taking myristoyl hexapeptide-16, tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 37.6 nm.
Example 9
Prescription
The preparation method comprises the following steps:
1. taking phosphatidylethanolamine, isopropyl myristate, oleic acid, propylene glycol and 1, 3-butanediol in the phase A according to the proportion of the formula, and stirring and dissolving the mixture at the heating temperature of 40 ℃ for later use;
2. taking tripeptide-1 copper, Tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 30.5 nm.
Example 10
Prescription
The preparation method comprises the following steps:
1. taking phosphatidylethanolamine, isopropyl myristate, oleic acid, propylene glycol and 1, 3-butanediol in the phase A according to the proportion of the formula, and stirring and dissolving the mixture at the heating temperature of 40 ℃ for later use;
2. taking acetyl tetrapeptide-3, Tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), and stirring for dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 33.2 nm.
Example 11
Prescription
The preparation method comprises the following steps:
1. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
2. taking biotin tripeptide-1, tween 20, disodium hydrogen phosphate and sodium dihydrogen phosphate in the phase B according to the proportion of the formula, adding warm water (20-35 ℃), stirring and dissolving for later use;
3. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
4. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
5. Detecting the particle size of the polypeptide nano inclusion with hair growth activity to obtain the inclusion particle size of 53.4 nm.
Comparative example 1
Prescription (1 kilogram essence)
The preparation method comprises the following steps:
adding the sodium hyaluronate into water according to the formula ratio, stirring to uniformly mix, heating to 80-85 ℃, and keeping the temperature and stirring to uniformly disperse the sodium hyaluronate. Cooling to below 40 deg.C, adding glycerol, aloe gel, biotin tripeptide-1, ceramide 3, octyl glycol and 1, 2-hexanediol, and stirring. The pH of the solution was adjusted to about 5.5 with 15% triethanolamine.
Comparative example 2
According to the formula and the preparation method of the comparative example 1, a blank essence is prepared. Mixing the example 11 and the blank essence according to a mass ratio of 1: 1, and evenly mixing to obtain the nano-encapsulated biotin tripeptide-1 essence containing about 0.1 percent. Example 12 stability test of polypeptide nanocapsules having Hair growth Activity
The polypeptide nanocapsules with hair growth activity obtained in examples 1 to 11 were placed in a closed container at room temperature for 30 days, and the properties and particle size of the samples were measured, and the results are shown in table 1.
TABLE 1 stability test results of polypeptide nanocapsules having hair growth activity
As can be seen from the results in table 1, the polypeptide nanocapsules with hair growth activity in examples 1 to 11 were not aggregated, precipitated, and delaminated after being placed for 30 days, and had no significant change in particle size, and had good stability, and were capable of nanocapsuling active polypeptide.
Example 13 Nano-encapsulated polypeptide having Hair growth Activity and Compound stability test thereof
13.1 Instrument
Constant temperature and humidity chamber, High Performance Liquid Chromatograph (HPLC)
13.2 test specimens
Example 1 nano-encapsulated biotin tripeptide-1, comparative example 1 ordinary biotin tripeptide-1 essence, and comparative example 2 nano-encapsulated biotin tripeptide-1 essence.
13.3 test basis
9001 raw material medicine and preparation stability test guiding principle of the four-part general rule of the year 2015 pharmacopoeia
13.4 test conditions and test items
And (3) accelerated test: the constant temperature and humidity box is 40 +/-2 ℃, RH 75% +/-5%, and the content of the polypeptide in each sample is detected by HPLC at 1,2, 3 and 6 months respectively to evaluate the stability of the sample.
And (3) long-term test: the stability of the samples was evaluated by measuring the polypeptide content in each sample by HPLC at months 3, 6, 9, 12, 18, 24 and 36 in a constant temperature and humidity chamber at 25 ℃. + -. 2 ℃ and RH 60%. + -. 10%.
13.5 stability test results
Example 1 stability data of samples of nano-encapsulated biotin tripeptide-1, the ordinary biotin tripeptide-1 essence of comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence of comparative example 2 after being placed under accelerated test conditions for 6 months are shown in table 2 below:
TABLE 2 stability test data for 6 months accelerated
Example 1 stability data of samples of nano-encapsulated biotin tripeptide-1, the ordinary biotin tripeptide-1 essence of comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence of comparative example 2 after being placed under long-term test conditions for 6 months are shown in table 3 below:
TABLE 3 Long term 6 month stability test data
As can be seen from the results in tables 2 and 3, after the accelerated test and the long-term test of the essence of the nano-encapsulated biotin tripeptide-1 in example 1 and the essence of the nano-encapsulated biotin tripeptide-1 in comparative example 2 for 6 months, the content of the biotin tripeptide-1 in the product does not change significantly, and no oil-water separation occurs, which indicates that the polypeptide has good stability after being nano-encapsulated and after being compounded with a matrix. In contrast, in the case of the common biotin tripeptide-1 essence in comparative example 1, under the conditions of being accelerated for 6 months and being prolonged for 6 months, the content of biotin tripeptide-1 in the product is reduced to different degrees, and the reduction of the content of polypeptide inevitably causes the reduction of the efficacy of the product, and even harmful degradation products can be generated, thus having potential harm to human bodies. Therefore, after the active polypeptide is nano-coated, the stability and the safety of the active polypeptide can be improved, and a more excellent hair growth promoting effect can be obtained under the condition of the same feeding amount.
Example 14 in vitro cumulative transdermal and cumulative skin Retention assays
14.1 instruments
Intelligent medicine transdermal diffusion tester, High Performance Liquid Chromatograph (HPLC)
14.2 test specimens
Example 1 nano-encapsulated biotin tripeptide-1, comparative example 1 ordinary biotin tripeptide-1 essence, and comparative example 2 nano-encapsulated biotin tripeptide-1 essence.
14.3 test methods
The transdermal properties of the samples were evaluated using a vertical Franz cell method. Fixing the in vitro skin of SD rat abdomen between the diffusion cell receiving chamber and the supply chamber, taking 1g sample on the skin surface of the supply chamber, and having effective diffusion area of 3.14cm2Adding normal saline as receiving liquid into the receiving pool, discharging air bubbles to make one side of dermis completely contact with the receiving liquid, stirring and diffusing at 32 ℃ at 300 r/min. 0.5mL of receiving solution is taken in 4h, 8h, 12h, 16h, 20h and 24h respectively, and equal amount of constant temperature blank receiving solution is supplemented in time. Measuring the concentration of the polypeptide in the receiving solution by HPLC, and calculating the cumulative transdermal quantity of the polypeptide per unit area at different times according to the following formula:
wherein: qnCumulative transdermal mass; cnIs the concentration of the polypeptide in the receiving fluid at the time of the sampling; v is the volume of the physiological saline in the receiving pool; ciThe concentration of the polypeptide in the receiving solution from the 1 st to the last sampling; viIs the volume sampled each time; and a is the effective diffusion area.
And after 24h, taking down the skin, washing out residual liquid of the sample by ultrapure water, shearing, adding ultrapure water for homogenization treatment, performing ultrasonic treatment for 5min, centrifuging at 10000r/min for 10min, taking the supernatant, detecting by an HPLC method, and calculating the skin retention of the polypeptide in unit area according to the following formula:
Qs＝Cs×V/A
wherein Q issIs the cumulative hold up; csThe mass concentration of the polypeptide in the skin sample fluid measured at the sampling time point; v is the volume of the supernatant; and a is the effective diffusion area.
14.4 test results
After 24h transdermal diffusion test of the nano-encapsulated biotin tripeptide-1 in example 1, the ordinary biotin tripeptide-1 essence in comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence in comparative example 2, the in vitro accumulated transdermal amount and the accumulated skin retention amount of the polypeptides in the sample are shown in fig. 1.
FIG. 1 shows the results of example 1, that the cumulative transdermal amount of nano-encapsulated biotin tripeptide-1 over 24 hours is 57.36. mu.g/cm2The cumulative skin retention was 43.89. mu.g/cm2Comparative example 1 the amount of accumulated skin of the common biotin tripeptide-1 essence for 24 hours was 26.31. mu.g/cm2The cumulative skin retention was 3.56. mu.g/cm2Comparative example 2 cumulative transdermal amount of nano-encapsulated biotin tripeptide-1 essence is 57.50 mug/cm2The cumulative skin retention was 42.73. mu.g/cm2. Therefore, the transdermal quantity and the skin retention quantity of the polypeptide are lower due to the limitation of a skin barrier before nano-coating, and after nano-coating, the transdermal quantity and the skin retention quantity of the polypeptide are obviously improved, particularly the skin retention quantity is obviously improved, which shows that the polypeptide can enhance transdermal absorption after nano-coating, and is accumulated in the skin, so that the effect of promoting hair growth in the skin is more effectively exerted. Nanocapsulated polypeptides andthe matrix compound does not affect the transdermal absorption and skin retention of the polypeptide in the product, still has larger accumulated skin retention amount, and is beneficial to enhancing the effect of promoting hair growth.
Example 15 testing of the Effect on Hair growth in an in vitro culture model
15.1 sample Condition
Scalp tissue of healthy volunteers is taken, and the volunteers have no problems of hair dyeing, head skin diseases and the like and are between the ages of 20 and 40 years old. Scalp tissue was separated under a microscope and anagen hair follicle tissue was collected.
The nano-encapsulated biotin tripeptide-1 of example 1, the ordinary biotin tripeptide-1 essence of comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence of comparative example 2 were used as test samples.
15.2 tissue culture
Hair follicle tissue was added to a medium containing 10ng/mL cortisol, L0. mu.g/mL insulin, 2mM L-glutamine, 100U/mL penicillin, and placed at 37 ℃ in 5% CO2The culture was continued for 14 days. Each test sample was added, and the other two groups were added with an equal volume of physiological saline as a blank control group and 2% minoxidil as a positive control group, respectively.
15.3 data determination
The hair length growth rate was measured at 0, 7 and 14 days of culture, respectively, and the results are shown in Table 4 below.
TABLE 4 Hair growth Rate
| Grouping | 7 days | 14 days |
| Blank control group | 4.8％ | 10.5％ |
| Minoxidil | 7.7％ | 13.2％ |
| Example 1 | 11.0％ | 19.3％ |
| Comparative example 1 | 8.5％ | 14.7％ |
| Comparative example 2 | 10.9％ | 19.1％ |
15.4 analysis of results
As can be seen from the results in Table 4, the hair growth rate of the minoxidil group was significantly greater than that of the blank control group, indicating that the culture was successful.
As can be seen from the data in the table, the hair growth rates of the nano-encapsulated biotin tripeptide-1 in example 1, the ordinary biotin tripeptide-1 essence in comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence in comparative example 2 are obviously higher than those of the blank control group, which indicates that the polypeptide in the sample has the effect of promoting hair growth. The hair growth rates of example 1 and comparative example 2 were greater than those of comparative example 1, indicating that the peptides were nano-encapsulated, and the promotion of hair growth was more significant due to good transdermal absorption and higher skin retention. After the nano-encapsulated active polypeptide with the hair growth effect is compounded with the matrix, the hair growth promoting effect of the nano-encapsulated active polypeptide is not influenced, is superior to 2 percent of minoxidil group, and is expected to replace minoxidil to treat alopecia and promote hair growth.
EXAMPLE 16 clinical trial for promoting Hair growth
16.1 volunteer situations
100 volunteers with androgenic alopecia were selected, the average age was 46 years, all volunteers had no iron deficiency anemia, thyroid gland and associated pathological features, and their head hair had not received any treatment. In this experimental area, there were at least 200 hairs, 70% of which were in anagen phase.
16.2 sample Condition
The nano-encapsulated biotin tripeptide-1 of example 1, the ordinary biotin tripeptide-1 essence of comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence of comparative example 2 were used as test samples, and a placebo group and a minoxidil group were also used, and the concentrations of the drugs in the groups were the same, so that 20 people in each group were treated.
16.3 test methods
Cutting hair in the head area (the area of the hair cutting area is about 1.8 cm)2) The picture of the haircut part was taken 3 consecutive days to evaluate the initial and final stage of hair growth. The medicine is applied once a day in the evening for 4 months. Data were recorded on day 0 and 4 months using a microscope and an automated data image acquisition analyzer.
Comparing day 0 with month 4, the average density of each group of hair at anagen and the average density of each group of hair at telogen are shown in fig. 2 and fig. 3, respectively.
16.4 test results
The test result shows that the polypeptide can effectively promote the hair growth, increase the hair in the growth period of the hair, simultaneously effectively reduce the hair in the telogen period and effectively prevent the hair from falling off. Compared with comparative example 1, the effects of example 1 and comparative example 2 on promoting hair growth and preventing hair loss are more significant, indicating that the polypeptide is nano-encapsulated and can exert more excellent effects due to better transdermal absorption and higher skin retention.
EXAMPLE 17 clinical trials for promoting eyebrow growth
17.1 volunteer situations
100 volunteers with obvious eyebrow loss were selected and the number of eyebrows was recorded separately.
17.2 sample Condition
The nano-encapsulated biotin tripeptide-1 of example 1, the ordinary biotin tripeptide-1 essence of comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence of comparative example 2 were used as test samples, and a placebo group and a bimatoprost group were also set, and the concentrations of the drugs in the groups were the same, and 20 people in each group were selected.
17.3 test methods
Each group of drugs was applied once a day, morning and evening, and the percentage of eyebrow growth was recorded 30 days after application.
17.4 test results
The test results are shown in FIG. 4. As can be seen from the results, the polypeptide significantly promoted the amount of eyebrow growth. Compared with comparative example 1, the effect of promoting the growth of eyebrows is more remarkable in example 1 and comparative example 2, and the fact that the polypeptide is nano-coated has better transdermal absorption and higher skin retention, so that the polypeptide can exert more excellent effect, is superior to bimatoprost, and is expected to replace bimatoprost in the aspect of the growth of the number of eyebrows.
EXAMPLE 18 clinical trial for promoting eyelash growth
18.1 volunteer situations
100 volunteers were selected and the number and length of upper and lower eyelash lashes were recorded separately.
18.2 sample Condition
The nano-encapsulated biotin tripeptide-1 of example 1, the ordinary biotin tripeptide-1 essence of comparative example 1 and the nano-encapsulated biotin tripeptide-1 essence of comparative example 2 were used as test samples, and a placebo group and a bimatoprost group were also set, and the concentrations of the drugs in the groups were the same, and 20 people in each group were selected.
18.3 test method
Each group of drugs was applied for 4 weeks, 8 weeks, once in the morning and evening, and the number and length of upper/lower eyelash was recorded.
18.4 test results
The percentage of eyelash growth and length of growth for 4 and 8 weeks are shown in fig. 5 and 6, respectively.
As can be seen from fig. 5, the percentage of eyelash growth was significantly greater in each of the polypeptide groups than in the placebo group. The percentage of eyelash growth is higher in example 1 and comparative example 2 than in comparative example 1, which is more advantageous to eyelash growth promotion, indicating that the polypeptide is nano-coated, and has better transdermal absorption and higher skin retention, so that more excellent effect can be exerted, and the effect of eyelash growth promotion is superior to that of bimatoprost, and is expected to replace bimatoprost.
As can be seen from fig. 6, each polypeptide group was able to significantly promote eyelash length. The length of eyelash growth is more remarkable in example 1 and comparative example 2 compared to comparative example 1, indicating that the polypeptide is nano-coated, and thus more excellent effect can be exerted due to better transdermal absorption and higher skin retention, and the effect of promoting eyelash length is superior to that of bimatoprost, and is expected to replace bimatoprost.
The foregoing is a more detailed description of the present invention in connection with specific preferred embodiments thereof, and is not intended to limit the invention to the particular forms disclosed. For those skilled in the art to which the invention pertains, several simple deductions or substitutions can be made without departing from the spirit of the invention, and all shall be considered as belonging to the protection scope of the invention.
Claims (5)
1. The active polypeptide with hair growth effect is the only active ingredient wrapped in a nano inclusion body, and the active polypeptide with hair growth effect is biotin tripeptide-1 with the mass percentage concentration of 0.0001-5%;
the coating is carried out by using lipid components, wherein the lipid components are soybean lecithin and cholesterol, and the mass percentage concentration of each component is 2-20%;
the nano inclusion also comprises the following components: the composition comprises polyhydric alcohol, a surfactant and a pH buffering agent, wherein the polyhydric alcohol is propylene glycol and 1, 2-hexanediol, and the mass percentage concentration of each component is 2% -20%;
the lipid component and the polyol constitute phase A, and the active polypeptide, the surfactant and the pH buffer constitute phase B;
the preparation method comprises the following steps:
a. taking the soybean lecithin, cholesterol, propylene glycol and 1, 2-hexanediol in the phase A according to the proportion of the formula, and stirring and dissolving the materials under the heating condition of 40 ℃ for later use;
b. taking biotin tripeptide-1, a surfactant and a pH buffering agent in the B phase according to the proportion of a formula, adding warm water with the temperature of 20-35 ℃, and stirring for dissolving for later use;
c. mixing the phase A and the phase B, and pre-emulsifying under the condition of 20000rpm by high-speed shearing;
d. homogenizing the mixture at 22500psi under high pressure, and circulating for 5 times to obtain nanometer-coated active polypeptide with hair growth promoting effect.
2. The active polypeptide having hair growth effect of claim 1, wherein the surfactant is tween 20, tween 40, tween 60, tween 80, poloxamer, tyloxapol, and the mass percentage concentration of each component is 0.1% -10%.
3. The active polypeptide having hair growth effect according to claim 1, wherein the pH buffer is disodium hydrogen phosphate, sodium dihydrogen phosphate, potassium dihydrogen phosphate or a combination of dipotassium hydrogen phosphate, each at a concentration of 0.001% to 1% by mass.
4. The active hair-growth polypeptide product of any one of claims 1-3, wherein the active hair-growth polypeptide product is in the form of a hair conditioner, shampoo, lotion, mascara, eyeliner, eyebrow pencil, or the like.
5. The active polypeptide having hair growth effect according to claim 1, wherein the hair is hair, eyebrow or eyelash.
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| CN110075035A (en) * | 2019-06-10 | 2019-08-02 | 上海龙禧投资有限公司 | A kind of hair restorer of the part containing hair stroma cell activator |
| CN110123734A (en) * | 2019-06-10 | 2019-08-16 | 上海龙禧投资有限公司 | A kind of alopecia prevention and hair growth agent composition |
| CN110123656A (en) * | 2019-06-10 | 2019-08-16 | 上海龙禧投资有限公司 | One kind is for preventing hair loss and trichogenous composition |
| CN110200838A (en) * | 2019-06-21 | 2019-09-06 | 武汉百思凯瑞生物科技有限公司 | A kind of complex polypeptide nano vesicle and its preparation method and application |
| CN110538299B (en) * | 2019-09-09 | 2021-09-28 | 广州楚颜化妆品有限公司 | Hair-blacking anti-hair-loss oil-control anti-dandruff mite-removing external liquid medicine and preparation method thereof |
| CN111329779B (en) * | 2020-03-23 | 2022-09-13 | 烟台中科恩吉科创新产业园管理有限公司 | Active peptide composition for promoting hair growth and application thereof |
| CN111544360A (en) * | 2020-06-08 | 2020-08-18 | 广东悦生生物科技有限公司 | Hair nourishing and hair growing composition and preparation method thereof |
| CN112516006A (en) * | 2020-12-10 | 2021-03-19 | 武汉百思凯瑞生物科技有限公司 | Nanometer composition with hair loss prevention, hair growth promotion, hair fixing and hair blackening functions and preparation method and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106028A (en) * | 2015-08-28 | 2015-12-02 | 深圳市维琪医药研发有限公司 | Polypeptide composition for hair growth |
| CN105232358A (en) * | 2015-10-29 | 2016-01-13 | 中山卡丝生物科技有限公司 | Compound for promoting eyelash growth and preparation method thereof |
| CN106214509A (en) * | 2016-08-24 | 2016-12-14 | 上海波林丝生物科技有限公司 | A kind of compositions promoting eyelashes to grow |
| CN108721161A (en) * | 2018-05-16 | 2018-11-02 | 珀莱雅化妆品股份有限公司 | A kind of hair-growing composition and its method for preparing lipidosome |
-
2019
- 2019-01-22 CN CN201910057508.8A patent/CN109568169B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105106028A (en) * | 2015-08-28 | 2015-12-02 | 深圳市维琪医药研发有限公司 | Polypeptide composition for hair growth |
| CN105232358A (en) * | 2015-10-29 | 2016-01-13 | 中山卡丝生物科技有限公司 | Compound for promoting eyelash growth and preparation method thereof |
| CN106214509A (en) * | 2016-08-24 | 2016-12-14 | 上海波林丝生物科技有限公司 | A kind of compositions promoting eyelashes to grow |
| CN108721161A (en) * | 2018-05-16 | 2018-11-02 | 珀莱雅化妆品股份有限公司 | A kind of hair-growing composition and its method for preparing lipidosome |
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