A hair follicle targeting liposome, a preparation method and application thereof in medicines and cosmetics

Abstract

The application provides a hair follicle targeting liposome, a preparation method and application thereof in medicines and cosmetics, relates to the technical field of biological medicine, and is composed of 20-80 parts by weight of lecithin and 5-15 parts by weight of notoginseng saponins. The lecithin is used as a skeleton, the surface of the liposome presents a negative Zeta potential, the electrostatic repulsion between the liposome and the cuticle of the skin is enhanced, non-specific adsorption is reduced, the particle is beneficial to migration and retention along the hair follicle path, and skin retention and hair follicle targeting are improved. Meanwhile, the notoginseng saponins are used as a film material, inflammation caused by androgenetic alopecia is reduced, hair follicle generation is promoted, collagen deposition is improved, the lecithin and the notoginseng saponins are synergistically used, the film material has the characteristics of film forming, high encapsulation rate, hair follicle targeting and retention. The technical problems of low encapsulation rate of the liposome and single function of the film material in the prior art are solved, and the hair follicle targeting and retention are improved.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a hair follicle-targeting liposome, its preparation method, and its application in pharmaceuticals and cosmetics. Background Technology

[0002] Minoxidil (MND), an FDA-approved topical treatment for hair loss, works by dilating blood vessels, promoting microcirculation around hair follicles, and prolonging the growth phase of hair follicles. However, its clinical application still faces the following bottlenecks: (1) Low transdermal efficiency: Minoxidil has poor lipophilicity (Log P=-0.41), resulting in low transdermal efficiency, making it difficult to penetrate the stratum corneum barrier and leading to low bioavailability; (2) Lack of targeting: Traditional solutions or tinctures cannot effectively accumulate in hair follicles, causing a large amount of the drug to enter the systemic circulation, leading to side effects such as hirsutism and contact dermatitis; (3) Poor stability: Minoxidil is easily oxidized and degraded, and penetration enhancers such as propylene glycol in the formulation further aggravate skin irritation. On the other hand, its clinical efficacy is limited by the conversion efficiency of sulfonate transferase (SLUT) in the outer root sheath of hair follicles. Studies have shown that only about 1-2% of topical minoxidil can be effectively converted into the active form minoxidil sulfate to exert its hair growth-promoting effect.

[0003] Liposomes, as nanoscale drug carriers, can improve drug permeability by controlling particle size, surface charge, and membrane composition. Studies have shown that liposomes with a particle size of 100–300 nm can selectively accumulate at hair follicle openings. Lipid bilayers can encapsulate water-soluble or lipid-soluble drugs, delaying degradation and reducing irritation. However, traditional liposomes (such as phosphatidylcholine liposomes) still have the following drawbacks: low encapsulation efficiency and limited membrane functionality; existing membranes only serve as physical barriers and lack synergistic therapeutic activity.

[0004] Therefore, there is an urgent need to screen a new, safe, and efficient preparation method to promote skin retention and hair follicle targeting of MND and improve the bioavailability of MND.

[0005] In view of this, the present invention is hereby proposed. Summary of the Invention

[0006] One of the objectives of this invention is to provide a hair follicle-targeting liposome to solve the technical problems of low liposome encapsulation rate and poor hair follicle targeting and retention caused by the single functionality of membrane materials in the prior art.

[0007] The second objective of this invention is to provide a method for preparing the above-mentioned hair follicle-targeting liposomes.

[0008] A third objective of this invention is to provide the application of the above-mentioned hair follicle-targeting liposomes or hair follicle-targeting liposomes prepared by the above-mentioned preparation method in the preparation of hair follicle-targeting products.

[0009] The fourth objective of this invention is to provide a product.

[0010] The fifth objective of this invention is to provide a method for preparing the above-mentioned product.

[0011] In order to achieve the above-mentioned objectives of the present invention, the following technical solution is adopted:

[0012] In a first aspect, the present invention provides a hair follicle-targeting liposome, the liposome being composed of 20-80 parts by weight of lecithin and 5-15 parts by weight of Panax notoginseng saponins.

[0013] The structural formula of the notoginsenosides is shown below:

[0014] or

[0015] .

[0016] Furthermore, the surface of the liposomes is modified with vitamin A molecules, wherein the amount of vitamin A modification is less than 2 parts by weight.

[0017] Furthermore, the notoginseng saponins are selected from notoginseng saponins R1, notoginseng saponins R2, notoginseng saponins ST-4, notoginseng saponins R4, notoginseng saponins Fc, notoginseng saponins Fe, notoginseng saponins Ft1, notoginseng saponins K or notoginseng saponins Fd.

[0018] Furthermore, the liposomes satisfy at least one of conditions A1 to A3:

[0019] A1. The particle size of the liposomes is 150~250 nm;

[0020] A2. The PDI of the liposomes is 0.25~0.28;

[0021] A3. The Zeta point of the negatively charged liposome is -33.0 to -36.0 mV.

[0022] Secondly, the present invention provides a method for preparing the above-mentioned hair follicle-targeting liposomes, the method comprising dissolving lecithin and notoginsenoside in an organic solvent according to the formula amount, rotary evaporating to form a dispersion film, injecting water, and incubating with ultrasound to obtain hair follicle-targeting liposomes.

[0023] Furthermore, after injecting water, vitamin A is added, followed by ultrasonic incubation.

[0024] The organic solvent comprises 1,000 to 3,000 parts by weight of dichloromethane and 600 to 2,000 parts by weight of anhydrous ethanol.

[0025] Thirdly, the present invention provides the application of the above-mentioned hair follicle-targeting liposomes or hair follicle-targeting liposomes prepared by the above-mentioned preparation method in the preparation of hair follicle-targeting products.

[0026] Fourthly, the present invention provides a product comprising an active ingredient and a carrier, wherein the carrier is the hair follicle-targeting liposome described above or a hair follicle-targeting liposome prepared by the above preparation method.

[0027] Furthermore, the active ingredient loading of the hair follicle-targeting liposome is 5%~20% w / w;

[0028] The active ingredients include at least one of the following: hair loss treatment drugs, whitening functional ingredients, antioxidant functional ingredients, anti-aging functional ingredients, or soothing and antipruritic functional ingredients.

[0029] Fifthly, the present invention provides a method for preparing the above-mentioned product, the method comprising dissolving the active ingredient together with lecithin and notoginsenoside in an organic solvent according to the above-mentioned preparation method;

[0030] The amount of the active ingredient added is 5 to 15 parts by weight.

[0031] This invention provides a hair follicle-targeting liposome, using lecithin as a framework to create a negative zeta potential on the liposome surface. This enhances the electrostatic repulsion between the liposome and the stratum corneum of the skin, reduces non-specific adsorption, and facilitates particle migration and retention along the hair follicle pathway, improving skin retention and hair follicle targeting. Simultaneously, it uses notoginseng saponins as a membrane material to reduce inflammation caused by androgenetic alopecia, promote hair follicle formation, and improve collagen deposition. Working synergistically with lecithin, it possesses film-forming properties, high encapsulation efficiency, hair follicle targeting, and retention. This solves the technical problems of low liposome encapsulation efficiency and limited membrane material functionality in existing technologies, leading to poor hair follicle targeting and retention. Attached Figure Description

[0032] To more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the specific embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are some embodiments of the present invention. For those skilled in the art, other drawings can be obtained from these drawings without creative effort.

[0033] Figure 1 The particle size distribution of the liposome sample with a weight ratio of 50:15:5 of lecithin, notoginsenoside R1 and minoxidil provided in Example 2 of the present invention in pure water.

[0034] Figure 2The particle size distribution of the liposome sample with a weight ratio of lecithin, notoginsenoside R1 and minoxidil of 50:15:10 in pure water in Example 2.

[0035] Figure 3 The particle size distribution of the liposome sample in pure water with a weight ratio of lecithin, notoginsenoside R1 and minoxidil of 50:15:15 in Example 2 is shown.

[0036] Figure 4 The particle size distribution of the liposome sample in pure water with a weight ratio of lecithin, ginsenoside Rh2 and minoxidil of 50:15:5 in Comparative Example 2.

[0037] Figure 5 The particle size distribution of the liposome sample in Comparative Example 2 with a weight ratio of lecithin, ginsenoside Rh2 and minoxidil of 50:15:10 in pure water is shown.

[0038] Figure 6 The particle size distribution of the liposome sample in Comparative Example 2 with a weight ratio of lecithin, ginsenoside Rh2 and minoxidil of 50:15:15 in pure water is shown.

[0039] Figure 7 The particle size distribution of the liposome sample in pure water with a weight ratio of 50:15:15:1 in lecithin, notoginsenoside R1, minoxidil and vitamin A in Example 3 is shown.

[0040] Figure 8 The results of the proliferation-promoting effects of different liposomes on human dermal papilla cells in Example 4;

[0041] Figure 9 The results of the effects of different liposomes on the migration of human dermal papilla cells in Example 5;

[0042] Figure 10 Example 6 shows the cumulative permeation of minoxidil in porcine skin by different liposomes through an in vitro transdermal experiment.

[0043] Figure 11 Example 6: The retention of minoxidil in porcine skin at 24 h by different liposomes in an in vitro transdermal experiment;

[0044] Figure 12 This describes the permeation and distribution of the notoginsenoside R1 liposome loaded with rhodamine B in the skin in Example 7.

[0045] Figure 13 This is a comparison of hair growth in mice under different liposome treatments in Example 8. Detailed Implementation

[0046] Unless otherwise defined herein, the scientific and technical terms used in conjunction with this invention shall have the meanings commonly understood by one of ordinary skill in the art. The meaning and scope of terms shall be clear; however, in any case of potential ambiguity, the definitions provided herein shall prevail over any dictionary or foreign definitions. In this application, unless otherwise stated, the use of "or" means "and / or". Furthermore, the use of the term "comprising" and other forms is non-limiting.

[0047] Generally, the nomenclature and techniques used in cell and tissue culture, molecular biology, immunology, microbiology, genetics, and protein and nucleic acid chemistry and hybridization, together with those described herein, are those well-known and commonly used in the art. Unless otherwise stated, the methods and techniques of the present invention are generally carried out according to conventional methods well-known in the art and described in various general and more specific references, which are cited and discussed throughout this specification. Enzymatic reactions and purification techniques are carried out according to the manufacturer's instructions, as commonly practiced in the art, or as described herein. The nomenclature, laboratory procedures, and techniques used in analytical chemistry, synthetic organic chemistry, and medical and medicinal chemistry, together with those described herein, are those well-known and commonly used in the art.

[0048] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0049] The present invention provides a hair follicle-targeting liposome, wherein the liposome is composed of 20-80 parts by weight of lecithin and 5-15 parts by weight of Panax notoginseng saponins;

[0050] The structural formula of the notoginsenosides is shown below:

[0051] or .

[0052] Using lecithin as a framework, the liposome surface exhibits a negative zeta potential, enhancing the electrostatic repulsion between it and the stratum corneum of the skin, reducing non-specific adsorption, and facilitating particle migration and retention along the hair follicle pathway, thus improving skin retention and hair follicle targeting. Simultaneously, using notoginseng saponins as the membrane material reduces inflammation caused by androgenetic alopecia, promotes hair follicle generation, and improves collagen deposition. Working synergistically with lecithin, this method possesses film-forming properties, high encapsulation efficiency, hair follicle targeting, and retention. It solves the technical problems of low liposome encapsulation efficiency and limited membrane material functionality in existing technologies, which lead to poor hair follicle targeting and retention.

[0053] In some specific embodiments, the surface of the liposomes is modified with vitamin A molecules, wherein the amount of vitamin A modification is <2 parts by weight. Vitamin A modification further improves the retention of the liposomes.

[0054] In some specific embodiments, the notoginsenosides represented by structural formula A are selected from notoginsenoside R1, notoginsenoside R2 or notoginsenoside ST-4; the notoginsenosides represented by structural formula B are selected from notoginsenoside R4, notoginsenoside Fc, notoginsenoside Fe, notoginsenoside Ft1, notoginsenoside K or notoginsenoside Fd.

[0055] In some specific embodiments, the liposomes have a particle size of 150~250 nm; in some specific embodiments, the PDI of the liposomes is 0.25~0.28; in some specific embodiments, the Zeta point of the negatively charged liposomes is -33.0~-36.0 mV.

[0056] According to another aspect of the present invention, a method for preparing the above-mentioned hair follicle-targeting liposomes is also provided. The method includes dissolving lecithin and notoginseng saponins in an organic solvent according to the formula amount, rotary evaporating to form a dispersion film, injecting water, and incubating with ultrasound to obtain hair follicle-targeting liposomes.

[0057] In some specific embodiments, after water injection, vitamin A is added and the mixture is incubated with ultrasound. Vitamin A is added and incubated after lecithin and notoginsenosides form liposomes, allowing vitamin A to modify the liposome surface and thus improve liposome retention.

[0058] In some specific embodiments, the organic solvent includes 1,000 to 3,000 parts by weight of dichloromethane and 600 to 2,000 parts by weight of anhydrous ethanol.

[0059] According to another aspect of the present invention, the application of the above-described hair follicle-targeting liposomes or hair follicle-targeting liposomes prepared by the above-described preparation method in the preparation of hair follicle-targeting products is also provided.

[0060] According to another aspect of the present invention, a product is also provided, comprising an active ingredient and a carrier, wherein the carrier is the hair follicle-targeting liposome described above or a hair follicle-targeting liposome prepared by the above preparation method.

[0061] Using follicle-targeting liposomes as carriers to load active ingredients, the active ingredients are delivered to the area around the hair follicles by utilizing the carrier's high follicle targeting and retention properties. The active ingredients are maintained around the hair follicles to exert their effects, and the transdermal efficiency and stability of the active ingredients are improved.

[0062] When liposomes are modified with vitamin A, vitamin A also regulates the expression of various metabolic enzymes by activating the retinoic acid receptor (RAR), upregulates the transcription level of sulfotransferases (SULT1A1 / 2A1), and improves the conversion efficiency of active ingredients (such as minoxidil).

[0063] In some specific embodiments, the active ingredient loading of the hair follicle-targeting liposome is 5%~20% w / w.

[0064] In some specific embodiments, the active ingredient includes at least one of the following: hair loss treatment drugs, whitening functional ingredients, antioxidant functional ingredients, anti-aging functional ingredients, or soothing and antipruritic functional ingredients;

[0065] Among them, the hair loss treatment drugs include at least one of minoxidil, ninodil, unodil or rhodamine; the whitening functional ingredients include at least one of glycyrrhizin, 4-butylresorcinol or phenylethylresorcinol; the antioxidant functional ingredients include at least one of resveratrol, pterostilbene, coenzyme Q10, astaxanthin, rhodioloside, quercetin, curcumin, tetrahydrocurcumin or tocopherol E and its analogues; the anti-aging functional ingredients include at least one of psoralen, silymarin, pro-xylene, retinol and its analogues or peptides; and the soothing and antipruritic functional ingredients include at least one of paeonol, gentiopicrin, oleuropein, hydroxytyrosol or asiaticoside.

[0066] In some specific embodiments, the product possesses at least one of the following functions: skin retention, hair follicle targeting, promotion of human dermal papillary cell proliferation, or promotion of human dermal papillary cell migration.

[0067] According to another aspect of the present invention, a method for preparing the above-mentioned product is also provided, the method comprising dissolving the active ingredient together with lecithin and notoginsenoside in an organic solvent according to the above-mentioned preparation method; wherein the amount of the active ingredient added is 5 to 15 parts by weight.

[0068] The aforementioned products include pharmaceuticals or cosmetics.

[0069] The present invention will be further illustrated by the following examples. Unless otherwise specified, the materials in the examples are prepared according to existing methods or purchased directly from the market.

[0070] Example 1

[0071] A blank liposome using notoginsenoside R1 as the membrane material is prepared according to the following steps:

[0072] Lecithin and Panax notoginseng saponin R1 were dissolved in dichloromethane and anhydrous ethanol and sonicated for 30 min to fully dissolve. The organic reagents were recovered by rotary evaporation in a 50°C water bath for 10-15 min to form a dispersed film. 5 mL of pure water was injected and incubated at 40°C for 30 min to fully hydrate. The mixture was then sonicated for 10 min to obtain blank liposomes with Panax notoginseng saponin R1 as the membrane material.

[0073] The liposomes contained lecithin and notoginsenoside R1 in a weight ratio of 50:15. The weights of dichloromethane were 3000 parts and anhydrous ethanol was 2000 parts.

[0074] Comparative Example 1

[0075] Unlike Example 1, ginsenoside Rh2 was used instead of notoginsenoside R1.

[0076] Example 2

[0077] Based on Example 1, minoxidil was loaded and prepared according to the following steps:

[0078] Lecithin, Panax notoginseng saponin R1, and minoxidil were dissolved in dichloromethane and anhydrous ethanol and sonicated for 30 min to fully dissolve. The organic reagents were recovered by rotary evaporation in a 50°C water bath for 10-15 min to form a dispersed film. 5 mL of pure water was injected and incubated at 40°C for 30 min to fully hydrate. The mixture was then sonicated for 10 min to obtain minoxidil-loaded Panax notoginseng saponin R1 liposomes.

[0079] The weight ratios of lecithin, notoginsenoside R1, and minoxidil were 50:15:5, 50:15:10, and 50:15:15, respectively; the weight ratios of dichloromethane and anhydrous ethanol were 3000 parts and 2000 parts, respectively.

[0080] The prepared liposomes showed no significant precipitation after centrifugation, indicating that minoxidil can be completely encapsulated by the liposomes of Panax notoginseng saponin R1.

[0081] Comparative Example 2

[0082] Unlike Example 2, minoxidil was loaded onto Comparative Example 1.

[0083] The prepared liposomes showed no significant precipitation after centrifugation, indicating that minoxidil can be completely encapsulated by ginsenoside Rh2 liposomes.

[0084] Experiment 1

[0085] 1. Determine the particle size and potential of the liposomes prepared in Example 2 and Comparative Example 2.

[0086] The prepared drug liposomes were analyzed in pure water using a high-sensitivity Zetasizer Nano s90 nanoparticle size analyzer. The results are as follows: Figures 1-6 As shown in Table 1, the average particle size, dispersion index (PDI), and zeta potential were measured.

[0087] 2. Calculation of drug loading

[0088] The prepared liposome solution was centrifuged at 15000 rpm for 20 min to observe for precipitation and calculate its drug loading. The results are shown in Table 1. The drug loading was calculated according to the following formula:

[0089] ;

[0090] Table 1. Average particle size, PDI, and Zeta potential

[0091]

[0092] The weight ratio in the table above is lecithin:saponin:minoxidil.

[0093] Data shows that liposomes with a weight ratio of 50:15:15 exhibited the highest drug loading rate, along with suitable particle size, high uniformity, and high stability. At the same weight ratio, compared to liposomes using notoginsenoside R1 as the membrane material, liposomes using ginsenoside Rh2 as the membrane material showed an excessively high particle size distribution (PDI), indicating poor particle size distribution uniformity and affecting the consistency of drug delivery. Therefore, in Example 1, the blank liposomes using notoginsenoside R1 as the membrane material demonstrated a better loading effect on minoxidil than those using ginsenoside Rh2 as the membrane material in Comparative Example 1.

[0094] Example 3

[0095] A vitamin A-modified notoginsenoside R1 liposome loaded with minoxidil was prepared according to the following steps:

[0096] Lecithin, notoginsenoside R1, and minoxidil were dissolved in dichloromethane and anhydrous ethanol, and sonicated for 30 min to ensure complete dissolution. The organic reagents were then recovered by rotary evaporation in a 50°C water bath for 10–15 min, forming a dispersed thin film. 5 mL of pure water was added, and the mixture was incubated at 40°C for 30 min. Vitamin A was then added, and the mixture was incubated in a 40°C water bath with magnetic stirring for 30 min to ensure complete hydration. The mixture was then sonicated for 10 min to obtain vitamin A-modified notoginsenoside R1 liposomes.

[0097] The weight ratio of lecithin: Panax notoginseng saponin R1: minoxidil: vitamin A is 50:15:15:1 and 50:15:15:2. Dichloromethane is 3000 parts by weight, and anhydrous ethanol is 2000 parts by weight.

[0098] The results showed that liposomes with a weight ratio of lecithin: Panax notoginseng saponin R1: minoxidil: vitamin A of 50:15:15:2 had obvious solid precipitation and could not be modified.

[0099] Comparative Example 3

[0100] Unlike Example 3, vitamin D3 was used instead of vitamin A.

[0101] Comparative Example 4

[0102] Unlike Example 3, retinoic acid was used instead of vitamin A.

[0103] The results showed that vitamin D3 in Comparative Example 2 and retinoic acid-modified liposomes in Comparative Example 3 both showed significant precipitation and could not be modified.

[0104] Experiment 2

[0105] 1. Determine the particle size and potential of liposomes with a weight ratio of lecithin: Panax notoginseng saponin R1: minoxidil: vitamin A of 50:15:15:1 in Example 3.

[0106] The prepared drug liposomes were analyzed in pure water using a high-sensitivity Zetasizer Nano s90 nanoparticle size analyzer. The results are as follows: Figure 7 As shown in Table 2, the average particle size, dispersion index (PDI), and zeta potential were measured.

[0107] (4) Calculation of drug loading

[0108] Centrifuge the prepared liposome solution at 15000 rpm for 20 min, observe for precipitation, and calculate the drug loading capacity using the following formula:

[0109] ;

[0110] Table 2. Average particle size, PDI, and Zeta potential

[0111]

[0112] The weight ratio in the table above is lecithin: Panax notoginseng saponin R1: minoxidil: vitamin A.

[0113] Data shows that when vitamin A was selected, the drug loading of liposomes with a weight ratio of 50:15:15:1 was no different from that of unmodified liposomes, and the particle size was suitable, the uniformity was high, and the stability was high.

[0114] Example 4: Proliferative effect of liposomes on human dermal papillary cells

[0115] 1. Liposomes: R1 blank liposomes prepared in Example 1, R1 / MND liposomes prepared in Example 2, and VA / R1 / MND liposomes prepared in Example 3 were selected for preparation.

[0116] 2. Cell Culture

[0117] HFDPC cells (human dermal papillary cells) were passaged in DMEM-F12 medium containing 10% fetal bovine serum, penicillin (100 U / mL), and streptomycin (100 U / mL) at 37°C and 5% CO2. All cells used in the experiment were in the logarithmic growth phase.

[0118] 3. Effects of different liposomes on the proliferation of HFDPC cells

[0119] Cells were seeded at 50,000 cells / mL, 100 μL per well in 96-well plates and cultured in a 37°C, 5% CO2 incubator. After cell adhesion, four groups were set up: negative control group (NC), MND solution group, R1 blank liposome group, R1 / MND liposome group and VA / R1 / MND liposome group, and drug intervention was given for 24 h.

[0120] Cell viability was determined using the CCK-8 assay: The drug-containing culture medium was removed, the cells were washed twice with PBS, fresh culture medium was added, and CCK-8 reagent (10 μL / well) was added to each well. The reaction was carried out at 37°C for 2 h. Absorbance was measured at 450 nm to calculate cell viability.

[0121] .

[0122] The results are as follows Figure 8 As shown, compared with the MND solution and R1 blank liposome group, both the R1 / MND and VA / R1 / MND liposome groups exhibited significantly higher HFDPC cell proliferation activity. Specifically, the VA / R1 / MND liposome showed higher cell proliferation-promoting activity than the R1 / MND group, indicating that VA modification positively promoted the cell proliferation-promoting ability of liposomes. This suggests that VA / R1 / MND and R1 / MND liposomes have a significant pro-proliferative effect on human dermal papillary cells, promoting hair follicle cell proliferation and thus hair growth.

[0123] Example 5: The migration-promoting effect of liposomes in human dermal papillary cells

[0124] 1. Liposomes: R1 / MND prepared in Example 2 and VA / R1 / MND prepared in Example 3 were selected for preparation.

[0125] 2. Cell Culture

[0126] HFDPC cells (human dermal papillary cells) were passaged in DMEM-F12 medium containing 10% fetal bovine serum, penicillin (100 U / mL), and streptomycin (100 U / mL) at 37°C and 5% CO2. All cells used in the experiment were in the logarithmic growth phase.

[0127] 3. Effects of R1 / MND and VA / R1 / MND liposomes on HFDPC cell scratching

[0128] Cells are arranged at 1×10 6 Each well was seeded into a 6-well plate and cultured at 37°C in a 5% CO2 incubator until cells adhered and formed a monolayer with a density >90%. Cells were then scratched, washed three times with PBS, and the scratched cells were removed. Medium containing 2% serum and drug-containing medium were added. Images were taken under a 4× microscope at 0 and 24 h. Results are shown below. Figure 9 As shown.

[0129] 4. The effect of R1 / MND and R1 / MND(+) liposomes on the migration of HFDPC cells

[0130] The results showed that, compared with the MND solution group, the liposome groups R1 / MND and VA / R1 / MND significantly reduced the distance of cell scratches and had a higher HFDPC cell migration effect. This indicates that R1 / MND and VA / R1 / MND liposomes have a significant pro-migration effect on human dermal papillary cells, promoting the migration of hair follicle cells and thus promoting hair growth.

[0131] Example 6: In vitro transdermal liposome experiment

[0132] 1. Experimental methods and materials

[0133] Experiments were conducted using pig skin of similar thickness. The Franz cell was used to investigate the differences in in vitro transdermal permeation of commercially available minoxidil solution (diluted to the same concentration as the prepared liposomes) and the prepared R1 / MND and VA / R1 / MND liposome solutions on pig skin. The effective diffusion area of ​​the Franz transdermal diffusion cell was 1.54 cm². 2 The volume of the liquid to be received is 14 mL.

[0134] Fresh skin samples were fixed between the supply and receiving cells, with the stratum corneum facing upwards. 0.3 mL of sample was placed in the supply cell and sealed with plastic wrap to prevent solution evaporation. A 30% 1,3-propanediol solution (v / v) was used as the diffusion medium in the receiving cell, and the medium was maintained at 32°C and stirred at 350 rpm throughout the experiment. Each sample was tested in triplicate. At predetermined time intervals (1 h, 2 h, 4 h, 8 h, 12 h, and 24 h), 1 mL of the receiving medium was withdrawn and replenished with the same volume of preheated fresh receiving solution. After 24 h, the permeation sites of the treated skin samples were wiped clean, cut into fragments, weighed, and then sonicated with methanol for 1 h. The drug concentration and skin retention of MND at each time point were determined by HPLC. Q1 (μg / cm³) 2 () represents the cumulative throughput within 24 hours.

[0135] 2. Permeability of the prepared minoxidil-loaded notoginsenoside R1 liposomes

[0136] The permeation of MND in MND solution and R1 / MND, VA / R1 / MND liposomes did not differ significantly from 0 to 4 h. The permeation increased over time in each experimental group. At 24 h, the cumulative permeation of MND in MND solution and R1 / MND, VA / R1 / MND liposomes was R1 / MND > VA / R1 / MND > MND solution, respectively. Figure 10 As shown, the results indicate that Panax notoginseng saponin R1 liposomes can significantly promote the transdermal utilization of MND.

[0137] 3. Skin retention of the prepared minoxidil-loaded notoginsenoside R1 liposomes

[0138] By measuring the MND content retained in pig skin of the MND solution group and the R1 / MND and VA / R1 / MND liposome groups, it was found that the MND retention in each experimental group was VA / R1 / MND > R1 / MND > MND solution, respectively. Figure 11 As shown, the retention of MND was significantly increased in the R1 / MND and VA / R1 / MND liposome groups compared with MND solution, indicating that the encapsulation of MND by Panax notoginseng saponin R1 liposomes and VA modification can significantly promote its retention in the skin, and the retention was further increased after VA modification.

[0139] Example 7: Permeation and Distribution of Liposomes in the Skin

[0140] (1) Experimental methods and materials

[0141] Following the methods of Examples 2 and 4, R1 / RB and VA / R1 / RB liposomes were constructed by replacing minoxidil with rhodamine B (RB). Referring to the transdermal assay method in Example 6, samples from each experimental group were added to the sample pool. After 2 hours, skin samples were taken and embedded in OCT gel, cut into 15 μm sections, and observed under a fluorescence microscope.

[0142] (2) Permeation and distribution of RB liposomes in the skin

[0143] Depend on Figure 12 It can be seen that the low fluorescence intensity of the RB solution on the skin surface indicates that it cannot remain on the skin surface or even penetrate into the hair follicle. Compared with the RB solution, the VA / R1 / RB and R1 / RB liposomes significantly increased the retention of RB in the hair follicle, indicating that RB in the VA / R1 / RB and R1 / RB liposome groups is more likely to remain in the stratum corneum and epidermis to form a reservoir and penetrate into the hair follicle. The positive rate of R1 / RB and VA / R1 / RB liposomes in the hair follicle is significantly increased.

[0144] Example 8: The role of liposomes in hair growth in androgen-induced alopecia mice

[0145] (1) Experimental methods and materials

[0146] Six-week-old mice (C57BL / 6J) selected for this study were purchased from the Experimental Animal Center of Southern Medical University. Thirty mice were randomly divided into four groups: a blank control group (NC), an androgenic alopecia group (AGA), an androgenic alopecia R1 / MND liposome treatment group (AGA+R1 / MND), an androgenic alopecia VA / R1 / MND liposome treatment group (AGA+VA / R1 / MND), and an androgenic alopecia minoxidil treatment group (AGA+MND), with six mice in each group. Except for the blank control group, the remaining mice underwent androgenic alopecia modeling according to literature. After anesthetizing the mice with sodium pentobarbital, the hair on the back of the mice was shaved off in an area of ​​2cm × 3cm. Testosterone was slowly injected subcutaneously into the mice at a dose of 200 μL / mouse (testosterone (purity ≥95.0%) was dissolved at a dose of 5 mg / mL in a system of PEG300, Tween 80, and physiological saline). The blank control group received an equal volume of physiological saline. A liposome solution loaded with minoxidil was prepared, and the dosage was 100 μL / mouse. The dosage of minoxidil in the control group was 100 μL / mouse. The control group and the androgenic alopecia group were given an equal volume of physiological saline. The drug was applied to the back of the mice for 21 consecutive days.

[0147] The method for modeling androgenic alopecia refers to the published papers "Nitric oxide synergizes minoxidil delivered by transdermal hyaluronic acid liposomes for multimodalandrogenetic-alopecia therapy" and "Synergistic therapeutic effect of ginsenoside Rg3 modified minoxidil transfersomes (MXD-Rg3@TFs) on androgenic alopecia in C57BL / 6 mice".

[0148] (2) Obtaining skin tissue from experimental animals

[0149] After the last day of drug administration and recording, skin tissue samples were collected. All instruments were prepared and sterilized before sampling. Mice in each group were anesthetized by intraperitoneal injection of 50 mg / kg body weight of 1% sodium pentobarbital solution. After complete anesthesia, newly grown hair was shaved, and skin was harvested from the mouse's back using tissue scissors, carefully removing the fascia.

[0150] (3) Record of hair growth in mice

[0151] The growth of hair on the backs of mice was recorded by photographing on days 0, 7, 14, and 21. Figure 13 The study observed hair growth on the backs of mice after 21 days of continuous administration. Results showed that hair growth in the androgenetic alopecia model mice was slower than in the control group. However, the VA / R1 / MND and R1 / MND liposomes loaded with minoxidil, as well as the minoxidil (positive control) group, significantly promoted hair growth on the backs of the androgenetic alopecia model mice. Furthermore, the VA / R1 / MND and R1 / MND liposomes were more effective than the MND solution group.

[0152] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some or all of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A hair follicle-targeting liposome, characterized in that, The liposomes are composed of 20-80 parts by weight of lecithin and 5-15 parts by weight of Panax notoginseng saponins; The liposomes are modified with vitamin A molecules, wherein the amount of vitamin A modification is less than 2 parts by weight; The notoginseng saponins are selected from notoginseng saponins R1, R2, ST-4, R4, Fc, Fe, Ft1, K, or Fd.

2. The liposomes according to claim 1, characterized in that, The liposomes satisfy at least one of conditions A1 to A3: A1. The particle size of the liposomes is 150~250 nm; A2. The PDI of the liposomes is 0.25~0.28; A3. The Zeta point of negatively charged liposomes is -33.0 to -36.0 mV.

3. The method for preparing hair follicle-targeting liposomes according to claim 1 or 2, characterized in that, The preparation method includes dissolving lecithin and notoginseng saponins in an organic solvent according to the formula amount, rotary evaporating to form a dispersion film, injecting water, and incubating with ultrasound to obtain hair follicle-targeting liposomes; After injecting water, vitamin A is added, followed by incubation with ultrasound.

4. The preparation method according to claim 3, characterized in that, The organic solvent comprises 1,000 to 3,000 parts by weight of dichloromethane and 600 to 2,000 parts by weight of anhydrous ethanol.

5. The application of the hair follicle-targeting liposome according to claim 1 or 2, or the hair follicle-targeting liposome prepared by the preparation method according to claim 3 or 4, in the preparation of hair follicle-targeting products.

6. A product characterized in that, It includes an active ingredient and a carrier, wherein the carrier is a hair follicle-targeting liposome as described in claim 1 or 2, or a hair follicle-targeting liposome prepared by the preparation method described in claim 3 or 4.

7. The product according to claim 6, characterized in that, The active ingredient loading of the hair follicle-targeting liposomes is 5%~20% w / w; The active ingredients include at least one of the following: hair loss treatment drugs, whitening functional ingredients, antioxidant functional ingredients, anti-aging functional ingredients, or soothing and antipruritic functional ingredients.

8. The method for preparing the product according to claim 6 or 7, characterized in that, The preparation method includes dissolving the active ingredients together with lecithin and notoginsenosides in an organic solvent; The amount of the active ingredient added is 5 to 15 parts by weight.

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