CN114642733B - Composition for treating androgenetic alopecia and preparation method thereof - Google Patents
Composition for treating androgenetic alopecia and preparation method thereof Download PDFInfo
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- CN114642733B CN114642733B CN202210201304.9A CN202210201304A CN114642733B CN 114642733 B CN114642733 B CN 114642733B CN 202210201304 A CN202210201304 A CN 202210201304A CN 114642733 B CN114642733 B CN 114642733B
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- ZRKFYGHZFMAOKI-QMGMOQQFSA-N tgfbeta Chemical compound C([C@H](NC(=O)[C@H](C(C)C)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CC(N)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H]([C@@H](C)O)NC(=O)[C@H](CC(C)C)NC(=O)CNC(=O)[C@H](C)NC(=O)[C@H](CO)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@@H](NC(=O)[C@H](CC(C)C)NC(=O)[C@@H](N)CCSC)C(C)C)[C@@H](C)CC)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](C)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H]([C@@H](C)O)C(=O)N[C@@H](CC(N)=O)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C=CC=CC=1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](C)C(=O)N[C@@H](CC(C)C)C(=O)N1[C@@H](CCC1)C(=O)N1[C@@H](CCC1)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CCC(O)=O)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CO)C(=O)N[C@@H](CCCNC(N)=N)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CC(C)C)C(O)=O)C1=CC=C(O)C=C1 ZRKFYGHZFMAOKI-QMGMOQQFSA-N 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- OUYCCCASQSFEME-UHFFFAOYSA-N tyrosine Natural products OC(=O)C(N)CC1=CC=C(O)C=C1 OUYCCCASQSFEME-UHFFFAOYSA-N 0.000 description 1
- 235000002374 tyrosine Nutrition 0.000 description 1
- 238000002137 ultrasound extraction Methods 0.000 description 1
- 230000003827 upregulation Effects 0.000 description 1
- 238000001291 vacuum drying Methods 0.000 description 1
- WKOLLVMJNQIZCI-UHFFFAOYSA-N vanillic acid Chemical compound COC1=CC(C(O)=O)=CC=C1O WKOLLVMJNQIZCI-UHFFFAOYSA-N 0.000 description 1
- TUUBOHWZSQXCSW-UHFFFAOYSA-N vanillic acid Natural products COC1=CC(O)=CC(C(O)=O)=C1 TUUBOHWZSQXCSW-UHFFFAOYSA-N 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
Classifications
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- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A61K31/185—Acids; Anhydrides, halides or salts thereof, e.g. sulfur acids, imidic, hydrazonic or hydroximic acids
- A61K31/19—Carboxylic acids, e.g. valproic acid
- A61K31/192—Carboxylic acids, e.g. valproic acid having aromatic groups, e.g. sulindac, 2-aryl-propionic acids, ethacrynic acid
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- A61K31/33—Heterocyclic compounds
- A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
- A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
- A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
- A61K31/506—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim not condensed and containing further heterocyclic rings
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- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/565—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol
- A61K31/567—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids not substituted in position 17 beta by a carbon atom, e.g. estrane, estradiol substituted in position 17 alpha, e.g. mestranol, norethandrolone
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- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
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- A61K31/56—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids
- A61K31/58—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin
- A61K31/585—Compounds containing cyclopenta[a]hydrophenanthrene ring systems; Derivatives thereof, e.g. steroids containing heterocyclic rings, e.g. danazol, stanozolol, pancuronium or digitogenin containing lactone rings, e.g. oxandrolone, bufalin
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- A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/08—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing oxygen, e.g. ethers, acetals, ketones, quinones, aldehydes, peroxides
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- A61K47/06—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite
- A61K47/16—Organic compounds, e.g. natural or synthetic hydrocarbons, polyolefins, mineral oil, petrolatum or ozokerite containing nitrogen, e.g. nitro-, nitroso-, azo-compounds, nitriles, cyanates
- A61K47/18—Amines; Amides; Ureas; Quaternary ammonium compounds; Amino acids; Oligopeptides having up to five amino acids
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- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The invention provides a composition for treating androgenetic alopecia and a preparation method thereof. The composition comprises an antiandrogen main drug and a choline-ferulic acid ionic liquid. The composition can be directly smeared on skin for transdermal administration, can reduce adverse reaction of the whole body, enhances the compliance of patients, has the advantages of simple and beneficial components, high skin permeation and skin retention of medicines, hair follicle targeting, good treatment effect, convenient administration and the like, and provides a new thought for the design of external preparations for treating androgenetic alopecia.
Description
Technical Field
The invention belongs to the technical field of medicines, and particularly relates to a composition for treating androgenetic alopecia and a preparation method thereof.
Background
Androgenic alopecia is often associated with hypersecretion of androgens and is often manifested as excessive spilling of scalp fat. The course of androgenetic alopecia is as long as several years, and has influence on the quality of life and physical and mental health of patients. Hair growth is closely related to hormones, whereas hair in different areas is sensitive to androgens to a different degree. For hair loss patients, there is often more androgen receptor expression in the scalp hair follicles, enhancing the sensitivity of hair papilla cells to androgens. After the in vivo androgenic testosterone acts on androgen receptor in hair follicle, the dihydrotestosterone with higher activity is formed under the action of 5 alpha-reductase in cytoplasm, and the dihydrotestosterone enters cells to check metabolism to influence, so that toxic effect can be generated on hair follicle, hair follicle atrophy and hair enters telogen and then alopecia. Hair papilla cells play an important core role in the hair follicle growth and development process, androgen receptors are specific markers of human hair papilla cells and are sensitive to androgens, and the hair papilla cells can induce various signal paths and secrete various growth factors to influence the hair follicle growth and development, such as Wnt/beta-catenin signal paths, insulin-like growth factors (IGF-1), transforming Growth Factors (TGF) and the like. Therefore, hair loss treatment is more focused on targeted delivery of drugs in hair follicles to achieve maximization of drug efficacy.
The treatment mode of androgenetic alopecia is mainly drug treatment, and currently, the drugs approved by the FDA for treating androgenetic alopecia are minoxidil and finasteride, and in addition, the drugs for treating female androgenetic alopecia are estrol cyproterone, drospirenone, ethinyl estradiol, spironolactone and the like. Wherein, finasteride is an inhibitor of 5 alpha-reductase, can effectively reduce the conversion of testosterone in blood and scalp to dihydrotestosterone, reduce the injury of dihydrotestosterone to hair follicle, inhibit the hair follicle of scalp from becoming smaller, and reverse the alopecia process. The oral dosage of finasteride is 1 mg/time/day. However, when finasteride is orally taken, adverse reactions such as hyposexuality and impotence occur in male patients suffering from alopecia, so that the finasteride is prepared into an external preparation to reduce systemic adverse reactions, and has important practical significance.
Disclosure of Invention
The present invention aims to solve at least one of the technical problems in the prior art described above. Therefore, the first aspect of the invention provides a composition for treating androgenetic alopecia, which has good transdermal effect, high skin retention and strong hair follicle targeting effect.
In a second aspect, the present invention provides a method for preparing the above composition for treating androgenetic alopecia.
In a third aspect, the invention provides the use of the above composition in the manufacture of a medicament for the treatment of androgenetic alopecia.
According to a first aspect of the present invention, there is provided a composition for treating androgenic alopecia, the composition comprising an anti-androgenic alopecia drug and a choline-ferulic acid ionic liquid, the choline-ferulic acid ionic liquid carrying the anti-androgenic alopecia drug.
In the invention, in order to promote the targeting and detention effect of the anti-androgenetic alopecia medicine in the skin and further enhance the effect of the anti-androgenetic alopecia medicine on treating androgenetic alopecia, the anti-androgenetic alopecia medicine is loaded into the choline-ferulic acid ionic liquid. The inventor finds that the drug-assisted effect of acid sources such as tartaric acid, benzoic acid, L-malic acid, cysteine, L-arginine, citric acid, vitamin C, tyrosine, phenylalanine, cinnamic acid, vanillic acid, folic acid, glycine and the like is not obvious, and a great amount of experiments show that ferulic acid has a stronger drug-assisted effect. The choline-ferulic acid ionic liquid disclosed by the invention has good safety, can obviously improve the saturated solubility of the anti-androgenetic alopecia medicine, promotes the percutaneous permeation and retention of the anti-androgenetic alopecia medicine, especially the retention at the hair follicle part, and has an excellent synergistic effect on treating androgenetic alopecia by the anti-androgenetic alopecia medicine.
In particular, in the invention, the choline-ferulic acid ionic liquid has excellent dissolution property and can be used as a carrier for drug delivery. Choline-ferulic acid ionic liquid can be used as a novel penetration enhancer for promoting transdermal drug delivery. The mechanism of ionic liquid permeation promotion comprises the following two types: firstly, the ionic liquid can open tight connection in the stratum corneum and promote the paracellular transport of the medicine; secondly, the ionic liquid can extract lipid components of cell membranes and promote the transcellular transport of medicines.
In the present invention, choline is a nutrient necessary for human body, and a great deal of research has shown that ionic liquids prepared using choline as a cation have biodegradability and low toxicity.
In the present invention, ferulic acid has a proliferation promoting effect on human hair papilla cells cultured in vitro, and can regulate the expression of IGF-1 in human hair papilla cells to thereby affect hair growth.
In some embodiments of the invention, the mass concentration of the anti-androgenic alopecia drug in the composition is (0.1-3%).
In some embodiments of the invention, the molar ratio of choline to ferulic acid in the choline-ferulic acid ionic liquid is 1 (1-2).
In some preferred embodiments of the present invention, the molar ratio of choline to ferulic acid in the choline-ferulic acid ionic liquid is 1 (1-1.8).
In some more preferred embodiments of the present invention, the molar ratio of choline to ferulic acid in the choline-ferulic acid ionic liquid is 1 (1 to 1.5).
In some more preferred embodiments of the present invention, the choline is selected from at least one of choline hydroxide, choline bicarbonate.
In some more preferred embodiments of the present invention, the anti-androgenic alopecia drug is selected from at least one of finasteride, minoxidil, estrol cyproterone, drospirenone, ethinyl estradiol, and spironolactone.
Still further, the anti-androgenic alopecia drug is finasteride.
According to a second aspect of the present invention, there is provided a method of preparing a composition for treating androgenetic alopecia, comprising:
s1: mixing the ferulic acid dispersion liquid with a choline solution, stirring, and drying to obtain the choline-ferulic acid ionic liquid;
s2: dissolving the anti-androgenetic alopecia medicine in the choline-ferulic acid ionic liquid prepared in the step S1 to obtain the composition for treating androgenetic alopecia.
In some embodiments of the invention, the ferulic acid dispersion of S1 is a ferulic acid absolute ethanol dispersion.
In some embodiments of the invention, the stirring time of S1 is 3h to 5h.
In some embodiments of the invention, the drying temperature of S1 is from 35 ℃ to 50 ℃.
In some embodiments of the invention, the drying time of S1 is 60h to 75h.
In some preferred embodiments of the invention, the choline is present at a concentration of 75% to 85% by mass.
In some embodiments of the invention, the dissolving of S2 comprises ultrasonic dissolving and shaking dissolving at constant temperature.
According to a third aspect of the present invention, there is provided the use of a composition as described above in the manufacture of a medicament for the treatment of androgenetic alopecia.
The beneficial effects of the invention are as follows:
1. the composition prepared by the invention has the advantages of simple and beneficial components, high skin permeation and skin retention of the medicine, hair follicle targeting, good treatment effect, convenient administration and the like.
2. The composition for treating androgenetic alopecia prepared by the invention can be directly smeared on skin for transdermal administration, and compared with the oral tablet of the anti-androgenetic alopecia medicament on the market, the composition can reduce adverse reaction of the whole body, enhance the compliance of patients and provide a new thought for the design of external preparations for treating androgenetic alopecia.
3. The invention has simple and convenient instrument and equipment, and the preparation process is efficient, simple and controllable, and is suitable for industrialized mass production.
Drawings
The invention is further described with reference to the accompanying drawings and examples, in which:
FIG. 1 is a cytotoxicity evaluation of choline-ferulic acid ion liquid prepared in example 1 of the present invention in human immortalized keratinocytes.
FIG. 2 shows cytotoxicity evaluation of choline-ferulic acid ion liquid prepared in example 1 of the present invention on human hair papilla cells.
FIG. 3 shows the damage and recovery of skin after the choline-ferulic acid ion liquid prepared in example 1 of the present invention is applied.
FIG. 4 shows the cumulative release of finasteride from finasteride-loaded choline-ferulic acid ion solution prepared in example 2 of the present invention.
Figure 5 shows uptake of coumarin 6 by human papilla cells in different formulations.
Fig. 6 shows the distribution of coumarin 6 after transdermal observation in different prescriptions by a confocal laser microscope.
Fig. 7 shows changes in hair growth after administration to each experimental group of androgenic alopecia model mice.
FIG. 8 is a graph showing hair growth scores after administration to each experimental group of androgenetic alopecia model mice.
Fig. 9 is a graph showing regenerative hair coverage after administration to each experimental group of androgenetic alopecia model mice.
Fig. 10 is a graph showing hematoxylin-eosin staining results of transection of the back skin hair follicle of mice after administration of each experimental group of androgenic alopecia model mice.
Fig. 11 is a relative number of hair follicles in hematoxylin-eosin stained sections of the back skin hair follicle transection of each experimental group of mice.
Fig. 12 is the relative thickness of skin in hematoxylin-eosin stained sections of the back skin hair follicle transection of mice in each experimental group.
FIG. 13 shows hematoxylin-eosin staining of hair follicles of the back skin of mice after administration of each experimental group of androgen-derived alopecia model mice.
Fig. 14 is a graph of relative depth quantification of hair follicles in hematoxylin-eosin stained sections of back skin hair follicles of mice of each experimental group.
FIG. 15 shows the relative expression levels of 5α -reductase mRNA in skin of mice model of androgenic alopecia after administration of each experimental group.
FIG. 16 shows the relative expression levels of IGF-1mRNA in skin of mice with androgenic alopecia models after each experimental group of mice was dosed.
FIG. 17 shows the relative expression levels of TGF-. Beta.1 mRNA in skin after administration of each experimental group of mice with androgenic alopecia model.
FIG. 18 shows the relative expression levels of VEGF mRNA in skin after administration of each experimental group of mice with androgenic alopecia model.
Detailed Description
The conception and the technical effects produced by the present invention will be clearly and completely described in conjunction with the embodiments below to fully understand the objects, features and effects of the present invention. It is apparent that the described embodiments are only some embodiments of the present invention, but not all embodiments, and that other embodiments obtained by those skilled in the art without inventive effort are within the scope of the present invention based on the embodiments of the present invention.
Example 1
The choline-ferulic acid ionic liquid is prepared in the embodiment and comprises the following raw materials:
choline bicarbonate solution (80%, w/w) 5.32g
Ferulic acid (99%) 5.05g
Absolute ethyl alcohol 4mL
The specific process is as follows:
adding ferulic acid into absolute ethyl alcohol, dispersing uniformly, then slowly dripping choline bicarbonate solution, stirring for 4 hours at room temperature, rotationally evaporating most of water and ethanol at 40 ℃, and vacuum drying the obtained liquid at 40 ℃ for 72 hours to obtain choline-ferulic acid ionic liquid. The liquid was an orange-yellow viscous liquid.
Example 2
The present example prepared a composition for treating androgenetic alopecia, comprising the following raw materials:
finasteride 0.03g
Choline-ferulic acid ionic liquid 9.97g
The specific process is as follows:
adding finasteride into choline-ferulic acid ionic liquid, performing ultrasonic treatment for 0.5h to accelerate dissolution, and continuously placing in a constant temperature shaking table for shaking for 48 hours at a speed of 100r/min to completely dissolve.
Example 3
The present example prepared a composition for treating androgenetic alopecia, comprising the following raw materials:
minoxidil 0.2g
Choline-ferulic acid ionic liquid 9.5g
The specific process is as follows:
adding minoxidil into choline-ferulic acid ionic liquid, performing ultrasonic treatment for 0.5h to accelerate dissolution, and continuously placing in a constant temperature shaking table for shaking for 48 hours at a speed of 100r/min to completely dissolve.
Example 4
The present example prepared a composition for treating androgenetic alopecia, comprising the following raw materials:
estrol cyproterone 0.1g
Choline-ferulic acid ionic liquid 9.9g
The specific process is as follows:
adding estrol cyproterone into choline-ferulic acid ionic liquid, performing ultrasonic treatment for 0.5h to accelerate dissolution, and continuously placing in a constant temperature shaking table for shaking for 48 hours at a speed of 100r/min to completely dissolve.
Example 5
The present example prepared a composition for treating androgenetic alopecia, comprising the following raw materials:
drospirenone 0.1g
Choline-ferulic acid ionic liquid 9.9g
The specific process is as follows:
adding drospirenone into choline-ferulic acid ionic liquid, performing ultrasonic treatment for 0.5h to accelerate dissolution, and continuously placing in a constant temperature shaking table for shaking for 48 hours at a speed of 100r/min to completely dissolve.
Example 6
The present example prepared a composition for treating androgenetic alopecia, comprising the following raw materials:
ethinyl estradiol 0.1g
Choline-ferulic acid ionic liquid 9.9g
The specific process is as follows:
adding ethinyl estradiol into choline-ferulic acid ionic liquid, performing ultrasonic treatment for 0.5h to accelerate dissolution, and continuously placing in a constant temperature shaking table for shaking for 48 hours at a speed of 100r/min to completely dissolve.
Example 7
The present example prepared a composition for treating androgenetic alopecia, comprising the following raw materials:
spirolactone 0.3g
Choline-ferulic acid ionic liquid 9.7g
The specific process is as follows:
adding spirolactone into choline-ferulic acid ionic liquid, performing ultrasonic treatment for 0.5h to accelerate dissolution, and continuously placing in a constant temperature shaking table for shaking for 48 hours at a speed of 100r/min to completely dissolve.
Comparative example
This comparative example produced a finasteride suspension, differing from example 2 in the use of water instead of choline-ferulic acid ionic liquid, comprising the following raw materials:
finasteride 0.03g
9.97g of water
The specific process is as follows:
adding finasteride into water, performing ultrasonic treatment for 0.5h, continuously placing in a constant temperature shaking table for shaking for 48 hours at 100r/min, and uniformly mixing to obtain the finasteride.
Test example 1 evaluation of safety of Choline-Ferulic acid Ionic liquid
1. Cytotoxicity test
Considering the safety problem of ionic liquids for treating androgenic alopecia, cytotoxicity experiments were used to evaluate the safety of blank ionic liquids on human immortalized keratinocytes (HaCaT) and human hair papilla cells (HDPC). Taking HaCaT in logarithmic phase, digesting with pancreatin and resuspension with complete medium, adding 10 μl into blood cell counting plate, counting cells under microscope, diluting with DMEM complete medium to obtain final cell number concentration of about 1×10 5 And each mL. The 96-well plate was removed and 100. Mu.L of cell solution was added to each well in the middle region of the well plate so that the cell concentration in each well was about 1X 10 4 And/or holes. The 96-well plate was placed in a 37 ℃ incubator overnight to allow the cells to adhere. The ionic liquid prepared in example 1 was diluted to final concentrations of 25, 50, 100, 125 μg/mL, respectively, using DMEM medium in a clean bench for use. Taking out 96-well plate from incubator, observing cell growth state under microscope, removing culture medium in each well after cell is completely adhered, adding 100 μl of the above ion with concentration of 25, 50, 100, 125 μg/mLLiquid solution, the 96-well plate was placed in an incubator for further culture for 24 hours, and the ionic liquid solutions prepared in example 1 with different concentrations were examined for toxicity to cells. MTT with mother liquor concentration of 5mg/mL was diluted with DMEM medium at a ratio of 1:5, the 96-well plate was removed from the incubator, 120. Mu.L of MTT dilution was added to each well, the 96-well plate was wrapped with tinfoil, and the plate was placed in the incubator for 4 hours in a dark place. The medium in the wells was removed, 150. Mu.L of DMSO was added to each well and the crystals were allowed to develop well by shaking on a shaker for 15-20 min. The whole process keeps the light-proof operation. Absorbance was measured for each well in a 96-well plate at 490nm using an enzyme-labeled instrument. Cell viability was calculated as follows reflecting cytotoxicity.
Wherein, the administration group is a group which is not administered with cells, the control group is a group which is not administered with cells, and the zeroing group is a group which is not administered with cells.
HDPC cells were treated in the same way.
As shown in the experimental results in figures 1 and 2, the choline-ferulic acid ionic liquid prepared in the example 1 is diluted to 25, 50, 100 and 125 mug/mL and then acts on HaCaT and HDPC cells, the survival rate of the cells reaches more than 80%, which shows that the ionic liquid prepared in the example 1 with the concentration of 25, 50, 100 and 125 mug/mL has good safety on the two cells.
2. Skin recovery experiment after choline-ferulic acid ion liquid action
The effect of the choline-ferulic acid ionic liquid prepared in example 1 on the skin barrier function can be examined by evaluating the change in the percutaneous moisture loss (TEWL) value. BALB/c mice were taken for about 4 weeks, and the back hair of the mice was removed with a razor. The TEWL value of the normal skin on the back of the mice was measured using a percutaneous moisture meter, and then divided into two groups on average, one group was not treated at all, and one group was coated with choline-ferulic acid ionic liquid, and the TEWL value of the skin on the back of the mice was measured using a percutaneous moisture meter after 5h, 10h, 1d, 2d, 3d, respectively, and recorded.
As a result, as shown in FIG. 3, the TEWL value of the mice before the back skin treatment was measured to be about 6.83g/m 2 h, after the choline-ferulic acid ionic liquid is smeared on the skin for 10h, the TEWL value of the skin reaches the maximum value of 11.18g/m 2 h, whereas the skin TEWL value of the untreated group is only 6.96g/m 2 h, showing that the choline-ferulic acid ion liquid can significantly improve the TEWL value of skin, and that the larger the TEWL value is, the more water is scattered from the skin, and showing that the skin barrier is damaged. Thus, the ionic liquid can reduce the barrier function of the skin after being applied to the skin, which is advantageous for transdermal drug delivery. Thereafter, the TEWL value of the skin applied to the choline-ferulic acid ion liquid group gradually decreased to reach a normal level consistent with the untreated group by about day 3, indicating that the barrier function of the skin had been restored. This suggests that the initial stage of administration of the choline-ferulic acid ion liquid can disrupt the skin barrier function, facilitating drug delivery, while the skin can return to normal after a period of administration, indicating that the effect of the choline-ferulic acid ion liquid on the skin barrier function is reversible. P in the figure<0.001, which is obtained as compared with the 0.9% physiological saline group.
Test example 2 finasteride saturation determination
Preparation of ionic liquid-carried finasteride (saturated): taking the ionic liquid obtained in the example 1, diluting the ionic liquid with water to the concentration of 20%, 40%, 60%, 80% and 100% by mass, taking 4g of the ionic liquid with the five concentrations obtained by the preparation, respectively adding 500mg of finasteride (excessive) into 3 parts of each sample, carrying out ultrasonic treatment for 0.5 hour to accelerate dissolution, and continuously placing the mixture in a shaking table at a constant temperature of 37 ℃ for 100r/min to shake for 48 hours.
The sample obtained above and comparative example 1 were respectively taken and filtered through a microporous filter membrane, diluted with a mobile phase, and the saturated solubility of finasteride in different systems was measured by a high performance liquid phase method.
Chromatographic conditions: the chromatographic column is XB-C18 column250X 4.6mm,5 μm); the mobile phase is water: acetonitrile=50: 50 (v/v); flow rate: 1mL/min; detection wavelength: 210nm;sample injection amount: 20. Mu.L. The experimental results are shown in Table 1.
TABLE 1 saturation solubility of finasteride
From the table above, the saturation solubility of finasteride in choline-ferulic acid ionic liquid can reach 1088.32 times of that in water, and the solubility of finasteride gradually increases along with the increase of the concentration of the ionic liquid, which indicates that the ionic liquid is favorable for solubilizing the finasteride.
Test example 3 in vitro Release test of finasteride-loaded Ionic liquid
The composition for treating androgenic alopecia prepared in example 2 was taken and examined for the release characteristics of finasteride from choline-ferulic acid ionic liquid.
(1) Screening of the receiving solution: the saturated solubility of finasteride in various media was determined to be 26.72.+ -. 0.49. Mu.g/mL in water and 464.96.+ -. 25.00. Mu.g/mL in 30% (v/v) PEG400 in physiological saline. In this experiment, 300mg of example 2 was administered to the supply tank, i.e., 900. Mu.g of finasteride was administered, and the volume required for total dissolution of finasteride in 30% (v/v) PEG400 in physiological saline was 900. Mu.g/464.96. Mu.g/mL. Apprxeq.1.94 mL. And the volume of the receiving tank is 7.5mL, thereby meeting the condition of groove leakage.
The experiment adopts a 0.45 mu m organic nylon filter membrane as a support, is fixed between a supply tank and a receiving tank of a Franz diffusion tank, and adds medicine-carrying ionic liquid into the supply tank. Franz diffusion cell effective release area is 3.14cm 2 The receiving well volume was 7.5mL and 30% (v/v) PEG400 saline was used as the release medium. Injecting release medium into a receiving pond, placing the diffusion pond in a diffusion instrument, placing the diffusion pond in a constant-temperature water bath at 32 ℃, taking 1mL of release medium at the rotation speed of 250rpm for 5min, 10min, 20min, 30min, 45min, 1h, 1.5h, 2h, 3h, 4h, 6h, 9h and 12h respectively, and supplementing the blank release medium with the same volume; filtering the obtained sample with 0.22 μm microporous membrane, and measuring finasteride content by high performance liquid chromatography, and calculating cumulative release according to the following formulaAnd calculating the cumulative release (%) according to the ratio of the cumulative release and the total drug loading of the ionic liquid.
Where Cn represents the drug concentration measured in the nth sample, ci is the drug concentration measured in each sample, V represents the volume of the release medium, vi represents the volume of each sample, and Qn (μg) is the cumulative release amount of the drug.
Chromatographic conditions: the chromatographic column is XB-C18 column250X 4.6mm,5 μm); the mobile phase is water: acetonitrile=50: 50 (v/v); flow rate: 1mL/min; detection wavelength: 210nm; sample injection amount: 20. Mu.L.
As shown in FIG. 4, the release rate of finasteride in the ionic liquid is 9.94+/-4.17% in 5min, the release rate is faster, and the release rate reaches 97.69+/-9.40% in 12h, so that the finasteride in the ionic liquid is completely released. In vitro release of finasteride-loaded ionic liquids presents a slow release trend.
Test example 4 transdermal test of compositions for treating androgenetic alopecia
1. In vitro transdermal experiments
(1) Skin treatment: about 200g SD rats were anesthetized with 20% (w/v) Ulatan, sacrificed by cervical dislocation, abdominal hair was removed from the rats, abdominal skin was separated with a scalpel, and subcutaneous tissues, blood vessels, and fat layers were removed. The skin was thoroughly cleaned with physiological saline and the filter paper was blotted to remove excess water from the skin surface. Finally, the skin was encapsulated with aluminum foil paper and stored at-20 ℃. The skin is carefully inspected to ensure its integrity prior to transdermal experiments using the skin.
(2) In vitro transdermal experiments: a piece of skin is placed in normal saline, thawed at room temperature, and then the moisture on the skin surface is sucked up by filter paper, and the skin is mounted between a supply tank and a receiving tank of a diffusion tank, and the skin horny layer faces the side of the supply tank. Diffusion used in this experimentThe effective permeation area of the pool is 3.14cm 2 The receiving well volume was 7.5mL. Firstly, the receiving pool is filled with 7.5mL of physiological saline of 30% (v/v) PEG400, and the temperature is kept at constant temperature of 32 ℃ and is kept in balance with the skin for more than 30 minutes. Then, 300mg of the liquids obtained in example 2 and comparative example were added to the supply tank, respectively, and the upper side of the supply tank was sealed with a sealing film to prevent volatilization of the drugs during the experiment. The stirring speed and the water bath temperature of the diffusion cell were set to 250rpm and 32.+ -. 1 ℃ respectively. In the transdermal test, 1mL of the receiving solution was withdrawn from the receiving well at predetermined times (6 h, 9h, 12h, 24 h) using a sampling needle, and an equal volume of blank receiving solution was immediately replenished to keep the volume of the solution in the receiving well unchanged. The content of finasteride in the sample is measured by high performance liquid chromatography after the sample is filtered by a 0.22 mu m microporous filter membrane, and the accumulated transmission amount of finasteride per unit area is calculated according to the following formula.
Wherein C is n Represents the measured concentration at the nth sampling point, C i Represents the measured concentration at the ith sampling point (i=6, 9, 12, 24), V 0 Represents the volume of the receiving reservoir, V represents the volume of each sample, A represents the effective permeation area, qn (μg/cm 2 ) The transmission amount is accumulated per unit area.
(3) Ex vivo skin hold-up experiment: after the transdermal test, the skin mounted in the middle of the receiving well and the supply well was carefully removed, and the skin was scrubbed three times with methanol and distilled water, respectively, to remove the excessive medicinal liquid remaining on the skin surface. The skin was cut into small pieces and 1mL of methanol was added. Ultrasonic extraction is carried out for 30min, and standing is carried out for 2h at room temperature. Filtering the supernatant with 0.22 μm microporous membrane, and measuring finasteride content in the filtrate by high performance liquid chromatography to obtain skin retention.
Chromatographic conditions: the chromatographic column is XB-C18 column250X 4.6mm,5 μm); the mobile phase is water: acetonitrile=50:50 (v/v); flow rate:1mL/min; detection wavelength: 210nm; sample injection amount: 20. Mu.L. The experimental results are shown in Table 2.
TABLE 2 in vitro transdermal results
Note that: * Is statistically different from the comparative examples (P < 0.05).
The test uses the aqueous suspension of finasteride of comparative example for comparison, and the result shows that the choline-ferulic acid ionic liquid can well enhance the skin permeation and skin retention condition of finasteride, and has the treatment of Li Yuxiong hormone-induced alopecia.
2. In vivo transdermal experiments
About 200g SD rats were anesthetized with 20% (w/v) Ulatan, and after shaving the abdominal hair with a shaver, depilation was performed using a depilatory cream, and after skin recovery was performed for 24 hours, a subsequent experiment was performed. Rats were anesthetized with 20% (w/v) uratam and fixed to the rat plates, and the supply cells of the diffusion cells (administration area 3.14 cm) were filled with 502 glue 2 ) The solution was applied to the abdominal skin of a rat, 300mg of each of example 2 and comparative example 1 was applied to the supply tank, the upper end of the supply tank was sealed with a sealing film, and an in vivo transdermal test was performed for 24 hours. After 24 hours, the rats were sacrificed by cervical dislocation, and the skin at the administration site was rubbed 3 times with cotton balls stained with water, methanol, and water, respectively, and the skin was removed rapidly. Cutting in an EP tube, adding 1mL of methanol, performing ultrasonic treatment for 30min to extract the medicine, and standing at room temperature for 2h. The supernatant was filtered through a 0.22 μm microporous membrane and the drug content was measured by HPLC.
Chromatographic conditions: the chromatographic column is XB-C18 column250X 4.6mm,5 μm); the mobile phase is water: acetonitrile=50: 50 (v/v); flow rate: 1mL/min; detection wavelength: 210nm; sample injection amount: 20. Mu.L. The experimental results are shown in Table 3.
TABLE 3 in vivo transdermal results
Note that: * Is statistically different from the comparative examples (P < 0.05).
The test uses the aqueous suspension of finasteride of comparative example for comparison, and the result shows that the choline-ferulic acid ionic liquid can well enhance the skin permeation and skin retention condition of finasteride, and has the treatment of Li Yuxiong hormone-induced alopecia.
Test example 5 Choline-Ferulic acid ion liquid hair follicle targeting ability test carrying finasteride
1. Cellular uptake
In order to observe the uptake of the drug by the cells using a confocal laser microscope, human hair papilla cells, which have an important role in hair follicle formation and growth, were used as model cells, and the cell uptake intensities of different prescriptions were compared. Since finasteride is a fat-soluble drug, hydrophobic coumarin 6 (which shows green fluorescence) was used as a model drug. Culturing human hair papilla cells until they are in logarithmic phase, digesting the cells with pancreatin, centrifuging, adding culture medium, suspending, and counting to obtain 1×10 cells 5 Density of dish inoculated in a laser confocal dish, placed at 37℃in 5% CO 2 Culturing in an incubator for 24 hours to enable the incubator to be completely adhered. After 24h, the medium was discarded and washed three times with PBS. Then 1mL of serum-free DMEM was added as a blank and 1mL of diluted coumarin 6 suspension and coumarin 6-loaded ionic liquid (loaded with ionic liquid prepared in example 1) were placed in an incubator and incubated for 4h, respectively. Removing the culture medium containing the medicine, washing the cells with precooled PBS for three times, adding 4% paraformaldehyde solution to fix the cells, finally adding DAPI solution to dye for 25min to dye the cell nuclei into blue, and washing the cells with precooled PBS for three times. The whole operation process keeps light-proof. Finally, observing and photographing by using a laser confocal microscope.
The cell uptake results are shown in FIG. 5, and in the control group, the nuclei were seen to be blue-stained, and green fluorescence of coumarin 6 was not seen. The results of coumarin 6 suspension group were similar to those of the control group, indicating that the hair papilla cells did not ingest coumarin 6. In the coumarin 6 ionic liquid group, the cytoplasm has obvious green fluorescence distribution and less overlap with blue fluorescence, which indicates that the ionic liquid can increase the uptake of the hair papilla cells to the coumarin 6 and is mainly distributed in the cytoplasm. The result shows that the ionic liquid can enhance the uptake of the medicine by the hair papilla cells, and is more beneficial for the medicine to enter the cells to play a role.
2. Laser confocal microscope observation of coumarin 6 distribution in skin
SD rats were anesthetized with 20% uratam, after shaving the abdominal hair with an electric shaver, depilatory was performed using a depilatory cream, and after skin repair for 24 hours, a subsequent experiment was performed. Rats were anesthetized with 20% uratam, fixed on a rat plate, and then a supply well of a diffusion cell (administration area of 3.14cm was prepared with 502 glue 2 ) Is stuck on the abdominal skin of a rat, 300mg of coumarin 6 suspension and coumarin 6-loaded ionic liquid are respectively administered in a supply tank in a dark place, and are uniformly distributed in the supply tank. The upper end of the supply tank is sealed by a sealing film and tinfoil to prevent the evaporation of water. In vivo transdermal behaviour was investigated for 24 hours. After various times of percutaneous permeation, rats were sacrificed by cervical dislocation, and the skin was rapidly removed by wiping 3 times with cotton balls stained with water and methanol, respectively. The skin at the administration site was cut into small pieces of about 2mm×5mm with surgical scissors, and after embedding with OCT frozen section embedding medium, the skin was slit on a frozen microtome to a section thickness of 20 μm, and the resulting skin section sample was adhered to a slide glass. Frozen skin slice samples were placed under a laser confocal microscope to observe the distribution of coumarin 6 in the skin, and the results are shown in fig. 6.
The results in fig. 6 show that coumarin 6 suspension is distributed almost exclusively in the stratum corneum during 24h transdermal process, and does not reach the deeper layers of the skin. The coumarin 6 ionic liquid group can reach deeper layers of skin, and more coumarin 6 distribution is visible in hair follicles (indicated by arrows in fig. 6), so that a better hair follicle detention effect is shown, and the method has important significance for treating alopecia.
3. Pigskin in vitro transdermal test
In order to quantitatively explore the effect of the finasteride-loaded choline-ferulic acid ionic liquid on hair follicle retention, pigskin is selected for in vitroTransdermal experiments. After the miniature Bama pigs were sacrificed, the abdominal hair was shaved off, the abdominal skin was peeled off, subcutaneous fat was removed, and the pigs were washed clean with physiological saline and stored at-20 ℃. Before use, thawing with physiological saline, sucking surface water with filter paper, and fixing between the supply tank and the receiving tank with horny layer facing the supply tank. The effective area of the diffusion cell was 3.14cm 2 The volume of the receiving solution was 7.5mL, the temperature of the diffuser water bath was adjusted to 32.+ -. 2 ℃ and the rotational speed was 250rpm. 300mg of the finasteride-carrying choline-ferulic acid ionic liquid prepared in example 2 and the finasteride suspension prepared in comparative example were respectively administered into a supply tank, 30% (v/v) PEG400 physiological saline as a receiving solution, 24h transdermal experiments were performed, 1mL of the receiving solution was withdrawn at 9, 12 and 24h, and the same volume of blank receiving solution was replenished. After the sample was filtered with a 0.22 μm microporous filter membrane, the finasteride content was measured by high performance liquid chromatography, and the cumulative permeation per unit area was calculated according to the following formula.
Wherein Cn represents the concentration measured at the nth sampling point, ci represents the concentration measured at the ith sampling point, V 0 Represents the volume of the receiving reservoir, V represents the volume of each sample, A represents the effective transdermal area, qn (μg/cm) 2 ) The transmission amount is accumulated per unit area. After the transdermal test is finished, the pig skin is taken out of the diffusion tank, the surface is cleaned by water, the residual medicine is removed by methanol, the skin containing hair follicle area and the skin not containing hair follicle area are separated by a surgical knife, 1mL of methanol is added, the medicine is extracted by ultrasonic for 30min, the supernatant is filtered by a microporous filter membrane with the thickness of 0.22 mu m, the subsequent filtrate is taken, and the content of finasteride in the filtrate is measured by high performance liquid chromatography, thus obtaining the skin hold-up.
Chromatographic conditions: the chromatographic column is XB-C18 column250X 4.6mm,5 μm); the mobile phase is water: acetonitrile=50:50 (v/v); flow rate: 1mL/min; detection wavelength: 210nm; sample injection amount: 20. Mu.L.
The experimental results are shown in Table 4.
Table 4 pigskin ex-vivo transdermal experiments
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Remarks: * Is statistically different from the comparative examples (P < 0.05).
As can be seen from table 4, there was a statistical difference between the cumulative permeation amounts of the finasteride suspension of comparative example 1 and the finasteride ionic liquid of example 2 for a 24h ex vivo transdermal experiment with pigskin; for skin retention in isolated non-follicular regions, there was no statistical difference between the two groups, while for skin retention of drug in follicular regions, the retention of the finasteride ionic liquid group was significantly higher than that of the finasteride aqueous suspension group, indicating that ionic liquid could promote drug retention in the skin by promoting retention of finasteride in follicular regions.
Test example 6 therapeutic Effect of finasteride-loaded Choline-Ferulic acid Ionic liquid on androgenic alopecia model mice
Establishment of androgenic alopecia model: the experiment uses C57BL/6 mice (7 weeks) as model animals, because the skin color of the C57BL/6 mice can be changed along with the hair growth period to change from pink to gray to black, and the observation in the experiment process is facilitated. The mice were shaved back hair and dehaired, and model animals were divided into five groups: blank control group, model group, blank ionic liquid group (example 1), finasteride-loaded ionic liquid group (example 2), minoxidil group. In addition to the placebo group, the remaining mice were injected subcutaneously with 0.1mL of 5mg/mL testosterone solution daily on the back, once daily with daily dosing for a total of 14 days. Animals were photographed and scored every 7 days after dosing began. The scoring criteria were: the initial state is 0 minutes; gray black coloration is 1 minute; the visible short hair was 2 points; sparse long hairs are 3 minutes; the dense long hair is 4 minutes; hair full growth was 5 points (final score for each component was the value of the present component minus the value of the model component). Animals were sacrificed on day 14 and hair regrowth coverage was recorded. After the experiment was completed, the mice were sacrificed by cervical dislocation, the back skin was removed, a portion of the skin was fixed in 4% paraformaldehyde, hematoxylin-eosin staining was performed after paraffin section, and another portion of the skin was preserved for a PCR experiment that measured the expression of mRNA of 5α -reductase, IGF-1, TGF- β1, and VEGF, which are closely related to hair growth. In the results of this part of the experiment, the expression is statistically different from the control group, # is statistically different from the model group, & expression is statistically different from the finasteride-loaded ionic liquid group, and P <0.05 is statistically different.
Figures 7 and 8 show the change in hair growth and scoring, respectively, of the groups after administration, and it can be seen that the hair of the control group can be regenerated without any treatment after shaving. Whereas if testosterone solution is injected subcutaneously after shaving and dehairing, the growth of the back hair of the model mice will be affected. After the blank ionic liquid, the finasteride-carrying ionic liquid and minoxidil are given, the hair regeneration of the model-making mice is facilitated, and the hair regeneration promoting effect is achieved: the finasteride-loaded ionic liquid group is larger than minoxidil group and larger than blank ionic liquid group.
Fig. 9 is a graph showing the coverage of regenerated hair from each group after dosing, showing that the coverage of hair from treated mice was in the finasteride-loaded ionic liquid group > minoxidil group > blank ionic liquid group, consistent with the previous scoring results.
Fig. 10 shows hematoxylin-eosin staining results of the transection of the skin hair follicles at the back of the mice after the end of the animal experiment, and fig. 11 and 12 show quantification results of fig. 10, which indicate that the number of hair follicles of the model building group is small, and is miniaturized, and the skin thickness is remarkably reduced in a stationary phase. The hair follicle density of the dosing group was significantly increased, with the finasteride-loaded ionic liquid group > minoxidil group > blank ionic liquid group. Fig. 13 shows hematoxylin-eosin staining results of longitudinal cutting of skin hair follicles on the back of mice after the end of animal experiments, and fig. 14 shows quantification results of fig. 13, which shows that the hair follicle depth of the blank ionic liquid group and the finasteride-loaded ionic liquid group is deeper than that of the model group, and the finasteride-loaded ionic liquid group is deeper than that of the blank ionic liquid group.
FIG. 15 shows that the expression of mRNA of 5α -reductase is increased after modeling compared to the control, meaning that the model group hair follicle has increased sensitivity to androgens, and that the blank ionic liquid group, the finasteride-loaded ionic liquid group, and the minoxidil group are statistically different from the model group, and that the effect of down-regulating the expression of mRNA of 5α -reductase is that the finasteride-loaded ionic liquid group is approximately equal to the minoxidil group > the blank ionic liquid group.
FIG. 16 shows that the expression of IGF-1mRNA was reduced after molding compared to the control, and that the blank and finasteride-loaded ionic liquid groups were statistically different from the model group, and that the upregulation effect on IGF-1mRNA expression was that in which finasteride-loaded ionic liquid group > blank ionic liquid group, and that minoxidil group was also statistically different from the model group. Indicating that the finasteride-loaded ionic liquid up-regulates the positive factor IGF-1 which promotes hair follicle growth.
FIG. 17 shows that TGF- β1mRNA expression is increased after modeling compared to control, and that there are statistical differences between the blank, finasteride-loaded and minoxidil-loaded groups and model groups, and that the effect of down-regulating TGF- β1mRNA expression is that the finasteride-loaded group is approximately equal to the minoxidil-loaded group > blank group. The finasteride-loaded ionic liquid is shown to down-regulate the factor TGF-beta 1 which inhibits hair follicle growth.
Fig. 18 shows that the expression of VEGF mRNA in the model group was similar to that in the control group, and that the blank ionic liquid group, finasteride-loaded ionic liquid group and minoxidil group were significantly higher than those in the model group, with statistical differences, but no statistical differences. The finasteride-loaded ionic liquid is shown to significantly promote angiogenesis of hair follicles and is beneficial to hair follicle growth.
While the embodiments of the present invention have been described in detail, the present invention is not limited to the above embodiments, and various changes can be made without departing from the spirit of the present invention within the knowledge of those skilled in the art. Furthermore, embodiments of the invention and features of the embodiments may be combined with each other without conflict.
Claims (8)
1. A composition for treating androgenetic alopecia comprising an anti-androgenetic alopecia drug and a choline-ferulic acid ionic liquid, wherein the choline-ferulic acid ionic liquid carries the anti-androgenetic alopecia drug; the anti-androgenetic alopecia medicine is finasteride.
2. The composition for treating androgenetic alopecia according to claim 1, wherein the mass concentration of the antiandrogenic alopecia drug in the composition is (0.1-3%).
3. The composition for treating androgenetic alopecia according to claim 1, wherein the molar ratio of choline to ferulic acid in the choline-ferulic acid ionic liquid is 1 (1-2).
4. A composition for the treatment of androgenic alopecia according to claim 3, wherein the choline is at least one selected from choline hydroxide and choline bicarbonate.
5. A method for preparing the composition for treating androgenetic alopecia according to any one of claims 1 to 4, comprising:
s1: mixing the ferulic acid dispersion liquid with a choline solution, stirring, and drying to obtain the choline-ferulic acid ionic liquid;
s2: dissolving the anti-androgenetic alopecia medicine in the choline-ferulic acid ionic liquid prepared in the step S1 to obtain the composition for treating androgenetic alopecia.
6. The method for preparing a composition for treating androgenetic alopecia according to claim 5, wherein the stirring time of S1 is 3-5 h.
7. The method for preparing a composition for treating androgenetic alopecia according to claim 5, wherein the drying temperature of S1 is 35-50 ℃.
8. The composition according to any one of claims 1 to 4 or the composition prepared by the preparation method according to any one of claims 5 to 7 for use in preparing a medicament for treating androgenetic alopecia.
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| CN101326275A (en) * | 2005-10-07 | 2008-12-17 | 阿拉巴马大学 | Multi-functional ionic liquid compositions |
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| CN101326275A (en) * | 2005-10-07 | 2008-12-17 | 阿拉巴马大学 | Multi-functional ionic liquid compositions |
| WO2020205409A1 (en) * | 2019-04-03 | 2020-10-08 | President And Fellows Of Harvard College | Ionic liquids for drug delivery |
| WO2021102084A1 (en) * | 2019-11-22 | 2021-05-27 | President And Fellows Of Harvard College | Ionic liquids for drug delivery |
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