CN113425845B

CN113425845B - Carrier preparation for preventing and/or treating alopecia and preparation method and application thereof

Info

Publication number: CN113425845B
Application number: CN202110915726.8A
Authority: CN
Inventors: 黄丹; 曾洋; 阳秀平; 梁臻; 黄浩喜; 苏忠海
Original assignee: Chengdu Beite Pharmaceutical Co ltd
Current assignee: Chengdu Beite Pharmaceutical Co ltd
Priority date: 2021-08-10
Filing date: 2021-08-10
Publication date: 2022-12-02
Anticipated expiration: 2041-08-10
Also published as: CN113425845A

Abstract

The invention provides a carrier preparation for preventing and/or treating alopecia, a preparation method and application thereof, and belongs to the field of pharmaceutical preparations. The experimental result shows that the carrier preparation provided by the invention has good transdermal performance and high adsorption capacity on scalp, can deeply permeate into hair follicle and be retained in the hair follicle for a long time, can maintain the effective concentration of the medicament for a long time, improves the bioavailability, reduces the using amount of active ingredients, and ensures that the low-content active ingredients can also play an excellent anti-hair loss effect. The transfersome preparation of the invention has simple preparation method and mild condition, and has good application prospect in preparing the medicine or the cosmetics for preventing and/or treating alopecia (especially androgenetic alopecia).

Description

Carrier preparation for preventing and/or treating alopecia and preparation method and application thereof

Technical Field

The invention belongs to the field of pharmaceutical preparations, and particularly relates to a carrier preparation for preventing and/or treating alopecia, and a preparation method and application thereof.

Background

With the increasingly fierce social competition, the working and living pressure of people is increased, the proportion of people with alopecia and poliosis in modern society is increased due to mental stress, night stay and the like, and the trend of the youthfulness is more and more obvious. The alopecia comprises androgen-induced alopecia, telogen alopecia, alopecia areata, lupus alopecia, anagen hair loss syndrome, etc.

Androgenetic alopecia (AGA for short), also called androgenetic alopecia or seborrheic alopecia, is the most common type of alopecia in clinical practice. There is a large body of evidence that AGA has a clear genetic predisposition between parents and progeny, and occurs in both male and female sexes. Research shows that the growth phase of the AGA is greatly shortened, and the AGA enters the catagen phase in advance when the hair follicle does not finish growth and growth in the growth phase, so that the hair follicle is miniaturized and becomes thin; meanwhile, the hair follicles enter the catagen phase in advance, resulting in a decrease in the number of hairs in the anagen phase, while the number of hairs in the catagen and telogen phases is increased, resulting in a decrease in the total number of hairs. AGA is manifested by marked hair loss, accompanied by increased scalp lipid secretion and increased head lipids. At present, minoxidil, finasteride and the like are commonly used chemical drugs for treating androgenetic alopecia, but the drugs have obvious side effects, the minoxidil can cause scalp dryness, dandruff, scalp erythema, inflammation and irritation and can influence blood pressure, and the finasteride has the effect of inhibiting the activity of male hormones.

Corticosteroid-17 α -propionate (also known as 17 α -propionyloxy-21-hydroxy-pregn-4-ene-3,20-dione) is a known androgen antagonist. It has been found that cortexolone-17 α -propionate can be used to treat acne, alopecia, and other skin and skin accessory disorders.

Chinese patent application No. CN201680036665.5 discloses a high concentration formulation of cortexolone-17 α -propionate for the treatment of alopecia, which comprises cortexolone-17 α -propionate at a concentration of 2.5% to 15% by weight and one or more pharmaceutically acceptable solvents, comprising less than 5% by weight water. However, the preparation described in this application is a solution preparation, which has limited permeability to the skin, resulting in that the cortexolone-17 α -propionate in the preparation is difficult to penetrate the stratum corneum of the skin and is unable to act on the deep structures of hair follicles, limiting the therapeutic effect thereof on alopecia. Therefore, it is of great significance to develop a pharmaceutical preparation which has good transdermal properties, can deeply permeate into hair follicles, can be retained in the hair follicles for a long time, and can exert excellent alopecia preventing and hair growing effects.

In 1996, cevc et al, the physical system of medical university of munich, germany, first proposed a systematic study report on carriers. The transfersome is improved from the conventional liposome by a prescription, and researches show that the addition of different additives into the phospholipid component of the liposome can obviously influence the properties and functions of the liposome, so that the lipid membrane of the liposome has high deformability, and small-molecule and large-molecule medicines can successfully enter systemic circulation. Therefore, the transfersome is defined as a lipoid aggregate which has high deformation capacity, takes the hydration pressure of skin as the power and can efficiently penetrate through pores which are several times smaller than the lipoid aggregate. However, it is well known in the art that the composition of the carrier has a significant effect on its transdermal properties, as well as on the time that the drug being carried is retained in the hair follicle. At present, no report is found on a carrier which has good transdermal performance, can deeply permeate into hair follicles, and can enable cortexolone-17 alpha-propionate to be retained in the hair follicles for a long time so as to exert an excellent anti-alopecia effect.

Disclosure of Invention

The invention aims to provide a carrier preparation for preventing and/or treating alopecia, a preparation method and application thereof.

The invention provides a carrier preparation for preventing and/or treating alopecia, which is prepared from the following raw materials in parts by weight: 0.5 to 2.0 portions of medicine for preventing and/or treating alopecia, 2.0 to 5.0 portions of lipid and 0.1 to 3.0 portions of surfactant.

Further, the material is prepared from the following raw materials in parts by weight: 1 part of medicine for preventing and/or treating alopecia, 2.6 to 4.6 parts of lipid and 0.2 to 2.2 parts of surfactant;

and/or the medicament for preventing and/or treating alopecia is an androgen antagonist;

and/or, the lipid is a phospholipid;

and/or the surfactant is an edge activator.

Further, the material is prepared from the following raw materials in parts by weight: 1 part of medicine for preventing and/or treating alopecia, 2.6 parts of lipid and 2.2 parts of edge activator;

and/or, the androgen antagonist is cortexolone-17 alpha-propionate;

and/or the phospholipids are selected from egg yolk lecithin, dimyristoyl phosphatidylcholine, soybean phosphatidylcholine, one or more of phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidic acid, phosphatidylethanolamine, sphingomyelin, lysophosphatidylcholine, preferably egg yolk lecithin;

and/or the edge activating agent is selected from one or more of sodium cholate, sodium deoxycholate, tween and span, and is preferably sodium cholate.

Further, the tween is tween 20, tween 60 or tween80, and the span is span 60, span 65 or span 80.

Further, the transfer agent preparation is an external preparation, preferably a transdermal administration preparation.

Further, the transdermal administration preparation is a liquid or semisolid preparation, and the semisolid preparation is a suspension, an emulsion, a microemulsion, a cream, a gel, a foam or an ointment.

The present invention also provides a method for preparing the above-mentioned delivery agent preparation for preventing and/or treating alopecia, comprising the steps of:

(1) Dissolving a drug for preventing and/or treating alopecia, a lipid and a surfactant in an organic solvent to obtain a mixture;

(2) Volatilizing the organic solvent in the mixture to obtain a mixture film;

(3) Adding a hydration medium into the mixture film for hydration and annealing to obtain primary liquid medicine;

(4) Homogenizing the primary medicinal liquid to obtain the final product.

Further, in the step (1), the organic solvent is a polyol; the concentration of the drug for preventing and/or treating alopecia in the preparation for delivery is 5 to 10% by weight;

in the step (3), the hydration medium is water or water phase solution, the hydration mode is ultrasonic treatment, and the annealing mode is standing at room temperature;

in the step (4), the homogenizing treatment mode is high-pressure homogenizing, high-shear emulsification or high-pressure micro-jet treatment.

Further, in the step (1), the polyalcohol is selected from one or more of ethanol, propylene glycol, butanediol and pentanediol, and is preferably ethanol;

in the step (3), the aqueous phase solution is phosphate buffer solution.

In addition to the above-mentioned methods, the carrier preparation for preventing and/or treating alopecia according to the present invention can be prepared according to other conventional methods for preparing carrier preparations in the art.

The invention also provides the application of the transfersome in carrying cortexolone-17 alpha-propionate, wherein the transfersome is prepared from the following raw materials in parts by weight: 2.0 to 5.0 portions of lipid and 0.1 to 3.0 portions of surfactant.

Further, the transfersome is prepared from the following raw materials in parts by weight: 2.6 to 4.6 portions of lipid and 0.2 to 2.2 portions of surfactant;

and/or, the lipid is a phospholipid;

and/or the surfactant is an edge activator.

Further, the transfersome is prepared from the following raw materials in parts by weight: 2.6 parts of lipid and 2.2 parts of edge activator;

and/or, the phospholipid is selected from one or more of egg yolk lecithin, dimyristoyl phosphatidylcholine, soybean phosphatidylcholine, egg phosphatidylcholine, phosphatidylserine, phosphatidylglycerol, phosphatidylinositol, phosphatidic acid, phosphatidylethanolamine, sphingomyelin and lysophosphatidylcholine, and preferably egg yolk lecithin;

The edge activator is edge activator, EA for short, and is a surfactant.

The transfersome for carrying cortexolone-17 alpha-propionate according to the present invention as described above may be prepared according to a conventional method for preparing transfersome in the art, such as a thin film hydration method.

The invention also provides a medicament or daily chemical product for preventing and/or treating alopecia, which is prepared from the transfersome preparation.

The invention also provides application of the transfersome preparation in preparing a medicament or a daily chemical product for preventing and/or treating alopecia.

Further, the alopecia is androgenetic alopecia, telogen effluvium or anagen-phase hair loss syndrome, and is preferably androgenetic alopecia.

The components and the proportion of the external multi-sac particle system composition provided by the invention are closely related to the effects of promoting the active ingredients to penetrate through the stratum corneum of the skin, penetrate into hair follicles, improve the retention performance in the hair follicles and improve the stability of the composition. The concrete expression is as follows:

(1) The lipid bilayer structure composed of the lipid and the edge active agent in the composition can be assembled into a microcapsule, and compared with a single-phase solvent system, the microcapsule is easier to penetrate through the surface layer of the skin, smoothly enters the inside of skin stratum corneum cells, interacts with keratin, reduces the compactness of the stratum corneum cells, forms a lipid channel and promotes the active substance to penetrate through the stratum corneum;

(2) The addition of a proper amount of the edge active agent can stabilize the bilayer structure of the microcapsule, increase the membrane strength, facilitate the wrapping of substances, and play a role in stabilizing the lipid membrane and reducing leakage;

(3) The addition of the edge activators such as sodium cholate, sodium deoxycholate, tween, span and the like enables the lipid membrane to have high deformability, and can efficiently penetrate through a pore canal which is a plurality of times smaller than the lipid membrane by taking the skin hydration pressure as power, and deeply penetrate into hair follicles, so that the sodium cholate and other nonionic activators have good safety and no stimulation to skin;

(4) The single microcapsule system is provided with positive charge components, so that the liposome with positive charge is easier to adsorb on human hair and scalp with negative charge, has stronger adhesion with a cell biomembrane, is easier to deeply permeate into hair follicles, can fully exert the effect of active ingredients, greatly improve the bioavailability, reduce the using amount of the active ingredients and ensure that low-content active substances can also exert excellent anti-dropping effect;

in conclusion, the multi-sac particle system external composition has good transdermal performance, increases the adsorption capacity of the preparation on the scalp, deeply permeates into hair follicles, can be retained in the hair follicles for a long time, can maintain effective concentration for a long time, improves bioavailability, reduces the using amount of active ingredients, and ensures that low-content active substances can also play an excellent anti-hair loss effect.

Compared with the prior art, the transfer agent preparation provided by the invention has the following beneficial effects:

(1) Most anti-hair loss product preparations sold in the market at present are common, the anti-hair loss mechanism is single, and the anti-hair loss effect is poor. The invention adopts specific androgen sites to combine with the medicament corticosteroid ketone-17 alpha-propionate, and prepares the medicament into a transfersome preparation which can penetrate into hair follicle glands in a targeted and positioned manner, plays the effects of preventing hair loss, growing hair and fixing hair and can effectively reduce the side effect brought by the medicament;

(2) Because the skin has a barrier, the anti-hair loss active substance is difficult to permeate into hair follicles and cannot directly act on a target site, and the lipid bilayer structure of the transfer agent can enter the inside of skin stratum corneum cells to promote the active substance to permeate the stratum corneum; but also has high deformability and can deeply penetrate into hair follicles; the liposome with positive charges can increase the adsorption of the preparation on the scalp, greatly improve the bioavailability, reduce the using amount of active ingredients and play an excellent anti-alopecia and hair-growing effect;

(3) The transfersome preparation can improve the stability of the active ingredients and reduce the influence of the use environment on the active ingredients;

(4) The accelerated storage test result shows that the characteristics, the particle size distribution, the Zeta potential and the active matter content of the carrier preparation provided by the invention do not change significantly after the carrier preparation is placed for 6 months at normal temperature, which indicates that the carrier preparation has good stability;

(5) The skin irritation test result shows that the transfersome preparation provided by the invention has no irritation to scalp temperature and has high safety;

(6) The external phase of the carrier preparation provided by the invention is a water-soluble system, the internal phase is an active ingredient, the solubility of the active ingredient in the system can be effectively ensured, the carrier preparation is convenient to use and not greasy, no additional alcohol solvent is needed, and the product experience feeling is good.

In conclusion, the carrier preparation has good transdermal performance and high adsorption capacity on the scalp, can deeply permeate into hair follicles and stay in the hair follicles for a long time, can maintain the effective concentration of the medicament for a long time, improves the bioavailability, reduces the using amount of active ingredients, and ensures that the low-content active ingredients can also play an excellent anti-hair loss effect. The transfersome preparation of the invention has simple preparation method and mild condition, and has good application prospect in preparing drugs or cosmetics for preventing and/or treating alopecia (especially androgenetic alopecia).

Obviously, many modifications, substitutions, and variations are possible in light of the above teachings of the invention, without departing from the basic technical spirit of the invention, as defined by the following claims.

The present invention will be described in further detail with reference to the following examples. This should not be understood as limiting the scope of the above-described subject matter of the present invention to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention.

Drawings

Figure 1 is a schematic of the structure of a delivery agent formulation of the present invention.

FIG. 2 is a schematic of the skin permeation mechanism of a carrier formulation of the present invention.

Figure 3 is a schematic representation of the process for making a delivery body preparation of the present invention.

FIG. 4 shows the results of the test of the cumulative skin permeation per unit area (A) and the cumulative skin retention per unit area (B) for each group of drugs.

FIG. 5 shows the results of the hair follicle growth rate test for each group.

Detailed Description

The raw materials and equipment used in the examples of the present invention were all known products, and were obtained by purchasing commercially available products.

Wherein, the yolk lecithin is EPC for short, the dimyristoyl phosphatidylcholine is DMPC for short, span80 is Span80 for short, tween80 is Tween80 for short, the sodium cholate is SDC for short, and the pH value of the phosphate buffer solution is 7.4.

EXAMPLE 1 preparation of a Carrier formulation (F1) according to the invention

The preparation method (thin film hydration method) is as follows:

(1) Adding cortexolone-17 alpha-propionate, span80 and EPC into 12mL of ethanol, and performing ultrasonic treatment for 5 minutes (ultrasonic power: 40 kHz) by using a water bath ultrasonic instrument to ensure that all the components are completely dissolved to obtain a mixture; 12mL of the mixture, the concentration of Span80 was 5% mg/mL, the concentration of EPC was 95% mg/mL;

(2) Evaporating the mixture with a rotary evaporator (rotating speed 250 rpm) at 60 deg.C under reduced pressure to obtain continuous lipid film after the ethanol is completely evaporated;

(3) Adding phosphate buffer (pH7.4) into the lipid film, performing ultrasonic treatment for 10 min to obtain hydrated preparation, and standing the hydrated preparation at room temperature for 30 min to obtain primary medicinal liquid;

(4) And (3) circulating the primary liquid medicine in a high-pressure homogenizer at 45 ℃ under 10000bar pressure for 5 times for high-pressure homogenization, cooling to room temperature, filtering by a filter (the aperture of a filtering membrane is 200 nm), and adding deionized water to make the volume of the solution constant to 50mL to obtain the transfersome preparation, namely F1 for short.

The charge concentration of cortexolone-17 α -propionate was 5% by weight based on 50mL of the carrier preparation.

Example 2 preparation of a Carrier formulation (F2) according to the invention

The preparation method comprises the following steps: the carrier preparation of the present invention, abbreviated as F2, was prepared by the preparation method of example 1, except that the lipid and the type of the edge-activating agent were replaced according to table 1, and the feeding concentrations of cortexolone-17 α -propionate, lipid and edge-activating agent were controlled according to table 1.

Example 3 preparation of a Carrier formulation of the invention (F3)

The preparation method comprises the following steps: the carrier preparation of the present invention, abbreviated as F3, was prepared by the preparation method of example 1, except that the lipid and the species of the limbal activating agent were replaced according to table 1, and the feeding concentrations of cortexolone-17 α -propionate, lipid and limbal activating agent were controlled according to table 1.

Example 4 preparation of a Carrier formulation according to the invention (F4)

The preparation method comprises the following steps: the carrier preparation of the present invention, abbreviated as F4, was prepared by the preparation method of example 1, except that the lipid and the type of the edge-activating agent were replaced according to table 1, and the feeding concentrations of cortexolone-17 α -propionate, lipid and edge-activating agent were controlled according to table 1.

Example 5 preparation of a Carrier formulation of the invention (F5)

The preparation method comprises the following steps: the carrier preparation of the present invention, abbreviated as F5, was prepared by the preparation method of example 1, except that the lipid and the type of the edge-activating agent were replaced according to table 1, and the feeding concentrations of cortexolone-17 α -propionate, lipid and edge-activating agent were controlled according to table 1.

Example 6 preparation of a Carrier formulation of the invention (F6)

The preparation method comprises the following steps: the carrier preparation of the present invention, abbreviated as F6, was prepared by the preparation method of example 1, except that the lipid and the type of the edge-activating agent were replaced according to table 1, and the feeding concentrations of cortexolone-17 α -propionate, lipid and edge-activating agent were controlled according to table 1.

Table 1 examples 1-6 raw material cortexolone-17 a-propionate, lipids, marginating agent and dosing concentrations thereof for delivery agent formulations

Table 2 examples 1-6 raw material cortexolone-17 a-propionate, lipid, and edge-activator charge (mg) for delivery formulations

The following is the preparation of comparative example samples.

Comparative example 1

A single-phase solution of a non-aqueous solvent system was prepared by uniformly mixing 28wt.% of diethylene glycol monoethyl ether, 28wt.% of ethanol, 0.1wt.% of tween80, and the balance propylene glycol, and used as a sample of comparative example 1.

Comparative example 2

A single-phase solution of a non-aqueous solvent system was prepared by uniformly mixing 5wt.% of cortexolone-17 α -propionate, 30wt.% of diethylene glycol monoethyl ether, 30wt.% of ethanol, 0.1wt.% of tween80, and the balance propylene glycol, and used as a sample of comparative example 2.

The advantageous effects of the present invention are demonstrated by test examples below.

Test example 1 particle diameter, zeta potential and encapsulation efficiency test

1. Test method

The particle size zeta potential and the encapsulation efficiency (EE%) of the carrier preparations F1 to F6 obtained in examples 1 to 6 were examined, and the properties of the samples were observed.

Particle size, PDI, zeta potential test methods: the DLS method computer detection system of Malvern Zetasizer was used to determine vesicle particle size and particle size distribution (particulate distribution Index); the electrophoretic mobility technique of the Malvern Zetasizer was used to measure the zeta potential.

Envelope efficiency (EE%) calculation formula: EE% = (total drug addition-free drug amount)/total drug addition × 100%

The detection method of the amount of the free medicine comprises the following steps: after ultracentrifugation of the delivery vehicle formulation, the supernatant is removed and the precipitated vesicles are disrupted using a suitable solvent capable of dissolving the precipitate. The resulting solution was then diluted and filtered using a syringe and filter (0.22-0.45 μm) to remove precipitates and impurities. The amount of free drug was detected by using a validated analytical detection method (modified High Performance Liquid Chromatography (HPLC) or spectrophotometry).

2. Test results

The results are shown in Table 3.

TABLE 3 measurement results of particle diameter, PDI, zeta potential and encapsulation efficiency of the delivery agent preparations F1 to F6

As can be seen from Table 3, the transfersome preparations F1 to F6 prepared in examples 1 to 6 of the present invention are all nanoscale preparations having a positive surface charge. Among them, the carrier preparation F6 prepared in example 6 has a large particle size, and therefore F6 was not detected in the stability test below. The carrier preparations F1 to F5 prepared in the embodiments 1 to 5 have the particle size of 100 to 300nm, the Zeta potential of 0 to +30mV and the encapsulation rate of about 50 percent, thereby meeting the requirements of practical application.

Test example 2 stability test

1. Test method

After the carrier formulations F1 to F5 prepared in examples 1 to 5 were left in a closed High Density Polyethylene (HDPE) bottle at room temperature for 1,2, 3, and 6 months, the particle size, zeta potential, and encapsulation efficiency (EE%) of the samples were measured, and the properties of the samples were observed.

The particle diameter and zeta potential were measured in the same manner as in test example 1, and the encapsulation efficiency was calculated in the same manner as in test example 1.

2. Test results

The results of the measurements are shown in tables 4 to 6.

Table 4 particle size stability test results for each delivery vehicle formulation

Table 5 zeta potential stability test results for each of the delivery agent formulations

Table 6 encapsulation efficiency stability test results for each delivery vehicle formulation

Test results show that the sample does not have agglomeration, discoloration and layering after being placed for 6 months, the active ingredient does not have crystallization in the transfer agent preparation, and the particle size, zeta potential and encapsulation rate of the sample do not have significant changes, so that the practical application requirements are still met.

The experimental results show that the transfer agent preparation provided by the invention has good stability.

Test example 3 irritation test

1. Test method

Test samples: the carrier formulations F1 to F5 obtained in examples 1 to 5 and the sample of comparative example 1 obtained in comparative example 1.

The experimental steps are as follows: taking 42 healthy rabbits with the weight of 2.0 +/-0.2 kg, randomly dividing the rabbits into 7 groups, removing hairs on two sides of the skin on the back of the rabbits 24 hours before the experiment, checking whether the removed hairs are injured 24 hours after the hairs are removed, and not suitable for skin irritation test of the injured skin. The experimental groups were applied with the transfersome preparations F1 to F52 times (1 time each in the morning and evening, 1mL film each time) each day obtained in examples 1 to 5 for 7 days; the same volume of the sample of comparative example 1 applied the same number of times was used as a control. The skin condition of the test sites of each group of mice was observed, and the test results are shown in table 7.

2. Test results

TABLE 7 summary of skin irritation observations for each sample

Remarking: "-" indicates no obvious hyperemia, redness and swelling; "+" indicates visible congestion to the naked eye; "+ + +" indicates that there was congestion as well as a red swelling visible to the naked eye.

As can be seen from the test results in Table 7, no hyperemia or redness occurred in the skin of the rabbits within 3 days using the samples of comparative example 1; however, the high doses of propylene glycol and ethanol in the samples of comparative example 1 produced some irritation to the rabbit skin from day 4 onwards.

After the carrier preparations F1 to F5 prepared in the embodiments 1 to 5 of the invention are used, the phenomena of congestion and red swelling do not appear on the skin of the rabbits, which shows that the carrier preparation of the invention has no irritation on the skin of the rabbits.

Test example 4 in vitro transdermal test

1. Test method

Test samples: carrier formulation F3 from example 3, comparative example 2 from comparative example 2.

The experimental steps are as follows: the transdermal experiment of the in vitro Bama miniature pig skin is carried out by adopting a vertical Franz diffusion cell method. The abdominal skin of the Bama miniature pig is fixed between a receiving chamber and a supplying chamber, 1g of each of the sample in the comparative example 2 and the carrier preparation F3 prepared in the example 3 is taken in the supplying chamber, 50% of ethanol and 50% of phosphate buffer solution are taken as receiving liquid by mass percentage, stirring and diffusion are carried out under the condition of 32 +/-1 ℃, and the stirring speed is 600rpm. 0.5mL of the receiving solution was taken at 1,2,4,6,8, 10, 12h and 24h, and an equal amount of a constant temperature fresh receiving solution was immediately replenished. And (4) performing HPLC analysis, and calculating the cumulative skin permeation amount of a specific drug per unit area at different times. And after 24h, taking down the skin, cleaning, shearing, grinding into homogenate, adding a proper amount of receiving liquid, centrifuging, taking supernatant, analyzing by HPLC, and calculating the cumulative retention of the specific medicine on the skin per unit area. The specific drug tested in this experiment was cortexolone-17 α -propionate.

2. Test results

The results of the experiment are shown in FIG. 4.As can be seen from the test results of FIG. 4A, the cumulative skin penetration per unit area of group 2 cortexolone-17 α -propionate was 38.1 μ g/cm ² Whereas the transfersome formulation F3 group cortexolone-17 alpha-propionate had a cumulative skin permeation per unit area of 34.2. Mu.g/cm ² It is shown that the cumulative transdermal flux per unit area of drug is comparable for carrier formulation F3 to the sample of comparative example 2.

As can be seen from FIG. 4B, the skin retention per unit area of group of cortexolone-17 α -propionate of comparative example 2 was 38.5 μ g/cm ² And the skin retention per unit area of the carrier preparation F3 group cortexolone-17 alpha-propionate is 56.4 mu g/cm ² It is shown that the transfersome formulation F3 of the present invention significantly increased the retention of the drug cortexolone-17 α -propionate in the skin.

The experimental results show that the delivery preparation prepared by the invention can prolong the detention time of the medicament corticosteroid-17 alpha-propionate in hair follicles, slowly release and control the release, remarkably improve the bioavailability of the medicament and enhance the effects of preventing hair loss and growing hair.

Test example 5 in vitro test for promoting Hair follicle growth

1. Test method

Test samples: the carrier preparations F1 to F5 prepared in examples 1 to 5, positive control minoxidil (commercially available).

The experimental steps are as follows: isolation and extraction of mouse whisker hair follicles and in vitro culture, starting the day after the culture, 5% of CO ₂ Incubate in incubator for 14 days. And simultaneously setting a blank control group (without medicine) and a minoxidil group, wherein the concentration of minoxidil in a minoxidil group culture solution is 10 mu g/mL.

Whisker hair follicle length was photographed every 7 days and measured, 3 times each. Data are presented as mean ± standard deviation.

The hair follicle growth values after 14 days of incubation after each group dosing were calculated using the following formula:

hair follicle growth value (μm) = average length of hair follicle in each group D14-average length of hair follicle in each group D0

The relative growth rate of the follicles after 14 days post-drug incubation of each group was calculated using the following formula:

hair follicle relative growth rate (%) = (average length of hair follicle in each group D14-average length of hair follicle in each group D0)/(average length of hair follicle in blank control group D14-average length of hair follicle in blank control group D0) × 100%

Wherein the average hair follicle length D14 represents the average hair follicle length measured on the 14 th day of incubation after the drug administration, and the average hair follicle length D0 represents the average hair follicle length measured on the day before the drug administration.

2. Test results

TABLE 8 in vitro test results for promoting hair follicle growth

Group of	Hair follicle growth value (mum)	Relative growth rate of hair follicle
			Blank control group	50.1±4.3	100.0％±3.4
Minoxidil group	73.5±3.7	136.7％±2.8
			F1	82.3±5.2	159.7％±2.7
F2	86.4±4.6	162.4％±3.7
			F3	102.5±5.7	196.2％±3.2
F4	91.7±3.4	172.4％±5.4
			F5	93.2±6.1	178.4％±2.9

As can be seen from the results of Table 8 and FIG. 5, the hair follicle growth values of the groups F1 to F5 and the minoxidil group after 14 days administration were significantly increased and the relative growth rates of the hair follicles were significantly increased, as compared with the control group. Compared with the minoxidil group, the hair follicle growth values of the transfer agent groups F1 to F5 after 14 days of administration are obviously increased, and the relative growth rates of the hair follicles are obviously improved, wherein the hair follicle growth value of the transfer agent group F3 is the largest (as large as 102.5 +/-5.7 mu m), and the relative growth rate of the hair follicles is the highest (as high as 196.2% +/-3.2).

The experimental result shows that the transfer agent provided by the invention can obviously promote the growth of hair follicles, and the promoting effect of the transfer agent is even better than that of a positive control minoxidil. The delivery agent preparation provided by the invention can be used for preparing pharmaceutical preparations and cosmetics for storing hair growth.

Test example 6 Hair growth Effect animal test

1. Test method

A sample to be tested: transfersome formulation F3, comparative example 1 sample, minoxidil (commercially available).

After the acclimation period of the mice, the hair was shaved off by a skin preparation knife from the lower neck parallel to the back of each mouse in an area of about 2cm by 3cm as a depilation observation area. Depilatory creams were then applied separately according to the instructions to remove residual hair and confirm that the current mouse hair growth was at rest (skin pink and no breakage). The day of shaving was defined as day 0 of the experiment. After shaving for 24h, the hairs were randomly grouped according to body weight. The grouping situation is as follows:

group a (blank control group): physiological saline;

group B (model control group): testosterone propionate;

group C: testosterone propionate + comparative example 1 sample;

group D: testosterone propionate + carrier preparation F3;

group E: testosterone propionate + minoxidil 5% solution.

The treatment method comprises the following steps: except for the blank control group injected with the normal saline, other groups of mice are injected subcutaneously at multiple points in the back epilation area according to the dose of 5mg/kg, and are continuously injected with testosterone propionate injection for 28 days to construct an androgen-induced alopecia mouse model. The corresponding drug to be tested is uniformly smeared on the unhairing area on the back of each group of mice 4h before and after testosterone propionate injection each time, 1 mL/time, 2 times a day, and continuously smeared for 30 days.

2. Test results

According to the experimental result, the hair in the depilated area on the back of the mice in the group A grows normally, and the hair does not grow in the mice in the group B, which indicates that the model of the mice with androgenetic alopecia is successfully modeled.

Compared with the blank control group (group A), the hair growth effect of the group D is obviously improved. After continuously applying the C, D, E group medicinal liquid for 30 days, the group C has no hair growth promoting effect basically, and the group D has obvious hair growth promoting effect. The carrier preparation prepared by the invention has obvious effects of preventing alopecia and growing hair on androgenetic alopecia.

Compared with group E, the hair growth effect of group D is obviously improved. Compared with the positive control drug minoxidil, the carrier preparation prepared by the invention has better anti-hair loss and hair growth effects on androgenetic alopecia.

In conclusion, the invention provides a carrier preparation for preventing and/or treating alopecia, a preparation method and application thereof. The carrier preparation has good transdermal performance and high adsorption capacity on scalp, can deeply permeate into hair follicles and be retained in the hair follicles for a long time, can maintain the effective concentration of the medicament for a long time, improves the bioavailability, reduces the using amount of active ingredients, and ensures that low-content active ingredients can also play an excellent anti-hair loss effect. The transfersome preparation of the invention has simple preparation method and mild condition, and has good application prospect in preparing the medicine or the cosmetics for preventing and/or treating alopecia (especially androgenetic alopecia).

Claims

1. A carrier preparation for preventing and/or treating alopecia, characterized in that: the composite material is prepared from the following raw materials in parts by weight: 1 part of cortexolone-17 alpha-propionate, 2.6 parts of egg yolk lecithin and 2.2 parts of sodium cholate.

2. The carrier formulation of claim 1, wherein: the transfer agent preparation is an external preparation.

3. The carrier formulation of claim 1, wherein: the transfer agent preparation is a transdermal administration preparation.

4. The delivery body preparation of claim 3, wherein: the transdermal administration preparation is a liquid or semisolid preparation, and the semisolid preparation is a suspension, an emulsion, a microemulsion, a cream, a gel, a foam or an ointment.

5. The method for producing the preparation for a delivery agent for preventing and/or treating alopecia according to any one of claims 1 to 4, wherein: the preparation method comprises the following steps:

(1) Dissolving cortexolone-17 alpha-propionate, egg yolk lecithin and sodium cholate in an organic solvent to obtain a mixture;

(2) Volatilizing the organic solvent in the mixture to obtain a mixture film;

(4) Homogenizing the primary medicinal liquid to obtain the final product.

6. The production method according to claim 5, characterized in that: in the step (1), the organic solvent is polyhydric alcohol; in the delivery preparation, the concentration of the corticosteroid-17 α -propionate is 5% to 10% by weight mg/mL; the dissolving mode is stirring or ultrasonic treatment;

7. The method of claim 6, wherein: in the step (1), the polyalcohol is selected from one or more of ethanol, propylene glycol, butanediol and pentanediol.

8. The method of claim 7, wherein: in the step (1), the polyhydric alcohol is ethanol.

9. A medicament or daily chemical product for preventing and/or treating alopecia, which is characterized in that: it is prepared from a delivery agent preparation according to any one of claims 1 to 4.

10. Use of the carrier preparation according to any one of claims 1 to 4 for the preparation of a medicament or a daily chemical product for the prevention and/or treatment of alopecia.

11. Use according to claim 10, characterized in that: the alopecia is androgen-induced alopecia, telogen alopecia or anagen-phase hair loss syndrome.

12. Use according to claim 11, characterized in that: the alopecia is androgenetic alopecia.