Hair-consolidating, hair-loss-preventing and hair-blackening composite-effect liposome and preparation method thereof

Abstract

The invention provides a compound-effect lipidosome for fixing hair, preventing hair loss and blackening hair and a preparation method of the compound-effect lipidosome. The compound-effect lipidosome comprises a herbal hair-growing lipidosome 001 and a compound-effect lipidosome 002, cholesterol; a first active extract; the herbaceous hair-growing lipidosome 002 comprises phosphate ester, a hair-growing agent and a hair-growing agent, cholesterol; an active extract; the hair care and blackening liposome comprises phosphate ester; cholesterol; a third active extract; the skin care, hair loss prevention and hair growth liposome-001 is prepared from the following components in parts by mass: phosphate ester; cholesterol; a fourth active extract; the skin care, hair loss prevention and hair growth liposome-002 comprises: phosphate ester; cholesterol; a fifth active extract; the skin care, hair loss prevention and hair growth liposome-003 comprises: phosphate ester; cholesterol; and a sixth active extract. The lipidosome prepared by the invention has the effects of preventing hair loss and fixing hair, also can promote hair blackening, and is suitable for various types of hair loss.

Description

Technical Field

[0001] The invention belongs to the field of hair care products, and more specifically, relates to a hair-fixing, anti-hair-loss and blackening composite functional liposome and a preparation method thereof. Background Art

[0002] At present, there are many products for preventing hair loss and promoting black hair on the market, including plant extracts, synthetic chemicals, etc. However, most of these products have problems such as poor stability, easy degradation of active ingredients, and low absorption rate, resulting in unsatisfactory use effects. As a nanoscale drug carrier, liposomes have gradually become a hot topic of research because they can improve drug stability and bioavailability. There are various methods for preparing liposomes in the prior art, but in the preparation and application of composite efficacy liposomes, there are still problems such as low encapsulation rate, insufficient drug loading, and poor stability. Specifically, the existing liposomes are prone to aggregation and precipitation during storage and use, which affects their stability, causes leakage of effective ingredients, and reduces the effect of the product. In the prior art, the encapsulation rate of liposomes for multiple active ingredients is often not high, especially in composite efficacy products, where the synergistic effect between the ingredients is difficult to give full play. Although liposomes have improved the absorption rate of some drugs, in practical applications, the penetration and targeting capabilities of liposomes still need to be optimized. Summary of the invention

[0003] The purpose of the present invention is to provide a hair-fixing, anti-hair-loss and blackening composite functional liposome and a preparation method thereof, aiming to solve the technical problems of low encapsulation rate, insufficient drug loading and poor stability existing in the prior art.

[0004] To achieve the above object, the technical solution adopted by the present invention is: to provide a hair-fixing, anti-hair loss and black hair composite functional liposome, comprising the following components:

[0005] Herbal hair growth liposome-001, comprising the following components in parts by weight: phosphate; cholesterol; a first active extract; wherein the first active extract comprises: one or more of a colorless salvia miltiorrhiza extract, a ginger extract, an chrysanthemum leaf extract, an chrysanthemum leaf extract, an aconite extract, and a Ligustrum lucidum fruit extract;

[0006] Herbal hair growth liposome-002, comprising the following components in parts by weight: phosphate; cholesterol; a second active extract; wherein the second active extract comprises: one or more of soapberry extract, mint leaf extract, mulberry extract, wolfberry extract, wolfberry extract, angelica extract, and safflower extract;

[0007] The hair care black hair liposome comprises the following components in parts by weight: phosphate; cholesterol; a third active extract; wherein the third active extract comprises: one or more of sh-pentapeptide-1, palmitoyl tetrapeptide-10, and acetyl hexapeptide-1;

[0008] Skin care, anti-hair loss and hair growth liposome-001 comprises the following components in parts by mass: phosphate; cholesterol; fourth active extract; wherein the fourth active extract comprises: one or more of sh-pentapeptide-1, oligopeptide-54, tripeptide-1, AHK-copper peptide, acetyl tetrapeptide-3, and decapeptide-10;

[0009] Skin care, anti-hair loss and hair growth liposome-002 comprises the following components in parts by weight: phosphate; cholesterol; the fifth active extract; wherein the fifth active extract comprises: one or more of sh-pentapeptide-1, oligopeptide-54, tripeptide-1, AHK-copper peptide, acetyl tetrapeptide-3, and decapeptide-10;

[0010] Skin care, anti-hair loss and hair growth liposome-003 includes the following components in parts by mass: phosphate; cholesterol; the sixth active extract; wherein the sixth active extract includes: one or more of sh-pentapeptide-1, octapeptide-2, myristoyl hexapeptide-16, myristoyl pentapeptide-17 and decapeptide-18.

[0011] Preferably, the first active extract comprises the following components in parts by weight:

[0012] 5-15 parts of colorless Salvia miltiorrhiza extract;

[0013] Ginger extract 5-15 parts

[0014] 5-15 parts of Channa ulmoides leaf extract

[0015] 5-15 parts of Thuja sutchuenensis leaf extract

[0016] 5-15 parts of Aconitum extract

[0017] 5-15 parts of Ligustrum lucidum extract.

[0018] Preferably, the third active extract comprises the following components in parts by weight:

[0019] sh-pentapeptide-1 0.1-0.5 parts;

[0020] Palmitoyl Tetrapeptide-10 0.1-0.5 parts;

[0021] Acetyl Hexapeptide-1 0.01-0.1 parts.

[0022] Preferably, the second active extract comprises the following components in parts by weight:

[0023] 5-15 parts of soapberry extract;

[0024] 5-15 parts of peppermint leaf extract;

[0025] 5-15 parts of mulberry extract;

[0026] 5-15 parts of wolfberry extract;

[0027] 5-15 parts of wolfberry extract;

[0028] 5-15 parts of angelica extract;

[0029] 5-15 parts of safflower extract.

[0030] Preferably, the fourth active extract comprises the following components in parts by weight:

[0031] sh-pentapeptide-1 0.1-0.5 parts;

[0032] Oligopeptide-54 0.02-0.2 parts;

[0033] Tripeptide-1 0.1-0.5 parts;

[0034] AHK-Copper Peptide 0.01-0.5 parts

[0035] Acetyl tetrapeptide-3 0.03-0.1 parts;

[0036] Decapeptide-10 0.1-0.5 parts.

[0037] Preferably, the fifth active extract comprises the following components in parts by weight:

[0038] sh-pentapeptide-1 0.1-1 parts;

[0039] Oligopeptide-54 0.1-0.5 parts;

[0040] Tripeptide-1 0.01-0.05 parts;

[0041] AHK-copper peptide 0.1-0.5 parts;

[0042] Acetyl tetrapeptide-3 0.02-0.03 parts;

[0043] Decapeptide-10 0.01-0.05 parts.

[0044] Preferably, the sixth active extract comprises the following components in parts by weight:

[0045] sh-pentapeptide-1 0.1-1 parts;

[0046] Octapeptide-2 0.05-0.1 parts;

[0047] Myristoyl hexapeptide-16 0.1-0.5 parts;

[0048] Myristoyl pentapeptide-17 0.01-0.1 parts;

[0049] Decapeptide-18 0.02-0.2 parts.

[0050] A method for preparing a hair-fixing and hair-loss-preventing composite functional liposome, based on the hair-fixing and hair-loss-preventing composite functional liposome as described in any one of the above, comprising the following steps:

[0051] Complete the preparation of liposomes; the liposomes include: one or more of herbal anti-hair loss and hair growth liposome-001, herbal anti-hair loss and hair growth liposome-002, hair care black hair liposomes, anti-hair loss and hair growth polypeptide liposome-001, anti-hair loss and hair growth polypeptide liposome-002, and anti-hair loss and hair growth liposome-003;

[0052] The prepared liposomes of different types are mixed according to a preset ratio to obtain a final product.

[0053] Preferably, the liposomes include: herbal anti-hair loss and hair growth liposome-001, herbal anti-hair loss and hair growth liposome-002, hair care black hair liposome, anti-hair loss and hair growth polypeptide liposome-001, anti-hair loss and hair growth polypeptide liposome-002 and anti-hair loss and hair growth liposome-003.

[0054] The beneficial effects of the composite functional liposome for hair solidification, hair loss prevention and blackening and the preparation method thereof provided by the present invention are: compared with the prior art, the liposome is prepared by a thin film hydration method, and the formula and preparation conditions are optimized to improve the encapsulation rate, stability and bioavailability of the liposome, thereby solving the problems existing in the prior art. The liposome composition prepared by the present invention not only has the effects of preventing hair loss and hair solidification, but also promotes black hair, is suitable for various types of hair loss, and has significant market application prospects. Specifically, 1. High stability: The liposome of the present invention has higher stability during storage and can maintain the effectiveness of its active ingredients for a long time. 2. High encapsulation rate: By optimizing the preparation conditions, the encapsulation rate of the liposome of the present invention is significantly improved, ensuring the effective synergistic effect of multiple active ingredients. 3. Improved bioavailability: The nanostructure of the liposome helps to improve the skin permeability and bioavailability of the active ingredients, thereby enhancing the actual effect of the product. DETAILED DESCRIPTION

[0055] In order to make the technical problems, technical solutions and beneficial effects to be solved by the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0056] The hair-fixing and hair-loss-preventing composite functional liposome provided by the present invention is now described. The hair-fixing and hair-loss-preventing composite functional liposome comprises the following components in parts by weight:

[0057] Phosphate 0.1-2 parts;

[0058] Cholesterol 0.5-5 parts;

[0059] 1-35 parts of active extract;

[0060] In the above-mentioned hair-fixing, anti-hair-loss and blackening composite functional liposome, as the main component of the liposome, phosphate ester can form a stable bilayer structure; cholesterol is used to regulate the fluidity of the phospholipid bilayer and increase the stability of the liposome; the active extract is used to play at least one of the effects of anti-hair loss, hair-fixing and hair-blackening.

[0061] As a specific implementation of an embodiment of the present invention, the active extracts include: salvia miltiorrhiza extract, colorless salvia miltiorrhiza extract, ginger extract, chrysanthemum intestinal leaf extract, truncatum leaf extract, aconitum extract, glossy privet extract, soapberry extract, mint leaf extract, mulberry extract, wolfberry extract, angelica extract, safflower extract, hair care peptide (sh-pentapeptide-1), black hair peptide (palmitoyl tetrapeptide-10), acetyl hexapeptide-1, oligopeptide-54, tripeptide-1, tripeptide-1, AHK-copper peptide, acetyl tetrapeptide-3, decapeptide-10, octapeptide-2, myristoyl hexapeptide-16, myristoyl pentapeptide-17, decapeptide-18, one or more.

[0062] Specifically, in any specific example, the active extract may be preferably a colorless salvia extract, ginger extract, chrysanthemum leaf extract, chrysanthemum leaf extract, aconite extract and glossy privet extract. In any specific example, the active extract may be preferably a soapberry extract, a mint leaf extract, a mulberry extract, a wolfberry extract, a wolfberry extract, an angelica extract, a safflower extract. In any specific example, the active extract may be a hair care peptide (sh-pentapeptide-1), a black hair peptide (palmitoyl tetrapeptide-10), and an acetyl hexapeptide-1. In any specific example, the active plant extract may be preferably a hair care peptide (sh-pentapeptide-1), an oligopeptide-54, a tripeptide-1AHK-copper peptide, an acetyl tetrapeptide-3, and a decapeptide-10. In any specific example, the active plant extract may be preferably a hair care peptide (sh-pentapeptide-1), an oligopeptide-54, a tripeptide-1, an AHK-copper peptide, an acetyl tetrapeptide-3, and a decapeptide-10. In any specific example, the active plant extract may preferably be hair care peptide (sh-pentapeptide-1), octapeptide-2, myristoyl hexapeptide-16, myristoyl pentapeptide-17, decapeptide-18.

[0063] Specifically, a hair-fixing, anti-hair-loss and black-hair-blackening composite liposome comprises:

[0064] The components of the active extracts required for Herbal Hair Growth Liposome-001, where the content units are parts by mass;

[0065] project content Colorless Salvia miltiorrhiza extract 5-15 Ginger Extract 5-15 Channa ulmoides leaf extract 5-15 Pleurotus truncatula leaf extract 5-15 Aconitum Extract 5-15 Ligustrum lucidum extract 5-15

[0066] That is, the herbal hair growth liposome-001 is composed of the following components in parts by mass:

[0067] Phosphate 0.1-2 parts;

[0068] Cholesterol 0.5-5 parts;

[0069] 1-35 parts of the first active extract; wherein the first active extract is composed of the following substances in parts by weight: 5-15 parts of colorless Salvia miltiorrhiza extract; 5-15 parts of ginger extract; 5-15 parts of Channa chuatsi leaf extract; 5-15 parts of Pleurotus ostreatus leaf extract; 5-15 parts of Aconitum foetida extract; 5-15 parts of Ligustrum lucidum fruit extract;

[0070] The components of the active extracts required for Herbal Hair Growth Liposome-002, where the content units are parts by mass;

[0071] project content Soapberry Extract 5-15 Peppermint Leaf Extract 5-15 Mulberry Extract 5-15 Lycium barbarum extract 5-15 Angelica Extract 5-15 Safflower Extract 5-15

[0072] That is, Herbal Hair Growth Liposome-002 is composed of the following components by mass:

[0073] Phosphate 0.1-2 parts;

[0074] Cholesterol 0.5-5 parts;

[0075] 1-35 parts of the second active extract; the second active extract is composed of the following substances in parts by mass: 5-15 parts of soapberry extract; 5-15 parts of peppermint leaf extract; 5-15 parts of mulberry extract; 5-15 parts of wolfberry extract; 5-15 parts of angelica extract; and 5-15 parts of safflower extract.

[0076] The components of the active extracts required for the black hair liposomes, where the content units are parts by mass;

[0077] project content Hair care peptide (sh-pentapeptide-1) 0.1-0.5 Black hair peptide (palmitoyl tetrapeptide-10) 0.1-0.5 Acetyl Hexapeptide-1 0.01-0.1

[0078] That is, the hair care black hair liposome is composed of the following components in parts by mass:

[0079] Phosphate 0.1-2 parts;

[0080] Cholesterol 0.5-5 parts;

[0081] 1-35 parts of the third active extract; the third active extract is composed of the following substances in parts by mass:

[0082] Hair care peptide (sh-pentapeptide-1) 0.1-0.5 parts;

[0083] Black hair peptide (palmitoyl tetrapeptide-10) 0.1-0.5 parts;

[0084] Acetyl hexapeptide-1 0.01-0.1 parts.

[0085] The components of the active extracts required for the skin care, anti-hair loss and hair growth liposome-001, where the content unit is mass parts;

[0086] project content Hair care peptide (sh-pentapeptide-1) 0.1-0.5 Oligopeptide-54 0.02-0.2 Tripeptide-1 0.1-0.5 AHK-Copper Peptide 0.01-0.5 Acetyl Tetrapeptide-3 0.03-0.1 Decapeptide-10 0.1-0.5

[0087] That is, the skin care, anti-hair loss and hair growth liposome-001 is composed of the following components in parts by mass:

[0088] Phosphate 0.1-2 parts;

[0089] Cholesterol 0.5-5 parts;

[0090] 1-35 parts of the fourth active extract; wherein the fourth active extract is composed of the following components in parts by mass: 0.1-0.5 parts of hair care peptide (sh-pentapeptide-1);

[0091] Oligopeptide-54 0.02-0.2 parts;

[0092] Tripeptide-1 0.1-0.5 parts;

[0093] AHK-copper peptide 0.01-0.5 parts;

[0094] Acetyl tetrapeptide-3 0.03-0.1 parts;

[0095] Decapeptide-10 0.1-0.5 parts.

[0096] The components of the active extracts required for the skin care, anti-hair loss and hair growth liposome-002, where the content unit is mass parts;

[0097] project content Hair care peptide (sh-pentapeptide-1) 0.1-1 Oligopeptide-54 0.1-0.5 Tripeptide-1 0.01-0.05 AHK-Copper Peptide 0.1-0.5 Acetyl Tetrapeptide-3 0.02-0.03 Decapeptide-10 0.01-0.05

[0098] That is, the skin care, anti-hair loss and hair growth liposome-002 is composed of the following components in parts by mass:

[0099] Phosphate 0.1-2 parts;

[0100] Cholesterol 0.5-5 parts;

[0101] 1-35 parts of the fifth active extract; wherein the fifth active extract is composed of the following components in parts by mass: 0.1-1 parts of hair care peptide (sh-pentapeptide-1);

[0102] Oligopeptide-54 0.1-0.5 parts;

[0103] Tripeptide-1 0.01-0.05 parts;

[0104] AHK-copper peptide 0.1-0.5 parts;

[0105] Acetyl tetrapeptide-3 0.02-0.03 parts;

[0106] Decapeptide-10 0.01-0.05 parts.

[0107] The components of the active extracts required for the skin care, anti-hair loss and hair growth liposome-003, where the content unit is mass parts;

[0108] project content Hair care peptide (sh-pentapeptide-1) 0.1-1 Octapeptide-2 0.05-0.1 Myristoyl Hexapeptide-16 0.1-0.5 Myristoyl Pentapeptide-17 0.01-0.1 Decapeptide-18 0.02-0.2

[0109] That is, the skin care, anti-hair loss and hair growth liposome-003 is composed of the following components in parts by mass:

[0110] Phosphate 0.1-2 parts;

[0111] Cholesterol 0.5-5 parts;

[0112] 1-35 parts of the sixth active extract; wherein the sixth active extract is composed of the following components in parts by mass:

[0113] Hair care peptide (sh-pentapeptide-1) 0.1-1 part;

[0114] Octapeptide-2 0.05-0.1 parts;

[0115] Myristoyl hexapeptide-16 0.1-0.5 parts;

[0116] Myristoyl pentapeptide-17 0.01-0.1 parts;

[0117] Decapeptide-18 0.02-0.2 parts.

[0118] It should be noted that the preparation of the hair-fixing, anti-hair-loss and black-hair-blackening composite functional liposome can be achieved by any one of the following methods: thin film hydration method, double emulsification method and reverse phase evaporation method. The method is selected according to the requirements of the target product. At the same time, the type and proportion of the plant extract can be adjusted according to market demand and efficacy requirements.

[0119] Instruments and Equipment

[0120] N-1100 rotary evaporator, Shanghai Ailang Instrument Co., Ltd.;

[0121] SHZ-D (Ⅲ) type circulating water multi-purpose vacuum pump, Shanghai Jinfu Experimental Instrument Equipment Co., Ltd.;

[0122] VC505 cell disruptor, Suzhou Renode Biotechnology Co., Ltd.;

[0123] CF16RN high-speed refrigerated centrifuge, Hitachi (China) Co., Ltd.;

[0124] SpectraMax microplate reader, Bio-Rad, USA;

[0125] ZS90 laser nanoparticle size analyzer, Malvern Instruments Ltd., UK;

[0126] JEM2100F transmission electron microscope, JEOL Co., Ltd.

[0127] TenaorⅡ infrared spectrometer, Bruker, Germany;

[0128] The present invention also provides a method for preparing a hair-fixing, hair-loss-preventing and black-hair-darkening composite functional liposome, which is used to prepare the hair-fixing, hair-loss-preventing and black-hair-darkening composite functional liposome described in any one of the above items, comprising the following steps:

[0129] S1. Complete the preparation of liposomes; the liposomes include: one or more of herbal anti-hair loss and hair growth liposome-001, herbal anti-hair loss and hair growth liposome-002, hair care black hair liposomes, anti-hair loss and hair growth polypeptide liposome-001, anti-hair loss and hair growth polypeptide liposome-002, and anti-hair loss and hair growth liposome-003.

[0130] The implementation process of step S1 is:

[0131] S1.1, dissolving components. Dissolve lecithin and cholesterol in an organic solvent in proportion. Dissolve the active extract in the aqueous phase; it should be noted that when preparing a specific liposome, it corresponds to the active extract dissolved in the aqueous phase in this step. Specifically, taking the preparation of herbal anti-hair loss and hair growth liposome-001 as an example, the active extracts in this step are: colorless salvia miltiorrhiza extract, ginger extract, chrysanthemum intestinale leaf extract, chrysanthemum intestinale leaf extract, Aconitum scutellariae extract and Ligustrum lucidum fruit extract.

[0132] S1.2, film formation: the dissolved organic solution was placed on a rotary evaporator at a speed of 150 rpm, a temperature of 45°C, and a time of 3 hours to remove the solvent and form a uniform lipid film on the bottle wall;

[0133] S1.3, hydration: add aqueous solution to the lipid film, and hydrate it on a rotary evaporator at a speed of 150 rpm and a temperature of 37°C to form a liposome suspension;

[0134] S1.4, ultrasonic treatment: The liposome suspension is subjected to ultrasonic treatment at a frequency of 2.4 KHz for 30 minutes. The ultrasonic treatment can reduce the particle size and improve the uniformity.

[0135] Exemplarily, 50 mg of lecithin (preferably egg yolk lecithin), 8 mg of cholesterol, 10 μL of Tween-80 are added, a certain amount of fat-soluble raw materials are ultrasonically dissolved in 3 mL of anhydrous ethanol as an organic phase. 10 mL of the dissolved water-soluble raw materials are weighed and dissolved in 20 mL of phosphate buffer (PBS buffer) with a concentration of 0.01 mol / L, preheated at 50 ° C, and formed into a uniform vortex by magnetic stirring as the aqueous phase. The ethanol in the organic phase is removed by rotary evaporation, so that the lipid forms a uniform film at the bottom of the flask. The aqueous phase is added to the flask, stirred and hydrated for 1 hour, and the probe is ultrasonicated for 12 minutes to obtain the liposome product.

[0136] As a specific implementation of the embodiment of the present invention, the preparation method further includes: S1.4, encapsulation efficiency detection. The encapsulation efficiency of the test liposome is detected to ensure that the components in the composite functional liposome are evenly distributed and well encapsulated.

[0137] S2. Mix the prepared different types of liposomes according to a preset ratio. The specific steps are as follows: first, accurately weigh herbal hair growth liposome-1 and herbal hair growth liposome-2 of different volume fractions, stir and mix in a magnetic stirrer, then add hair care black hair liposomes of corresponding volume fractions, stir and mix in a magnetic stirrer, then add skin care anti-hair loss and hair growth liposome-1, skin care anti-hair loss and hair growth liposome-2, and skin care anti-hair loss and hair growth liposome-3 of corresponding volume fractions at a time, finally stir in a magnetic stirrer at 25°C for 1h to completely mix them to obtain the final product.

[0138] Table 7 Different liposome ratios (%, v / v)

[0139]

[0140]

[0141] Different liposomes were prepared by the above method, and the average particle size, PDI, Zeta potential and encapsulation efficiency of the liposomes prepared above were tested. The experimental results are shown in Table 1.

[0142] Table 1 Detection of physical properties of different liposomes

[0143] Liposomes Particle size / nm PDI Zeta point / mV Encapsulation rate% Herbal anti-hair loss liposome-001 100.2 0.214 -28.4 84.3 Herbal anti-hair loss liposome-002 105.4 0.226 -24.1 85.6 Hair Care Black Hair Liposome 100.6 0.216 -23.2 89.4 Anti-hair loss and hair growth polypeptide liposome-001 104.3 0.223 -22.6 81.3 Anti-hair loss and hair growth polypeptide liposome-002 106.2 0.229 -20.17 88.5 Anti-hair loss liposome-003 100.3 0.227 -27.8 86.7

[0144] The results showed that the average particle size of these liposomes was about 100 nm, and the PDI was less than 0.3, indicating that the particle size distribution was uniform. The Zeta potential was negative, indicating that these liposomes were negatively charged and had stable dispersibility. The encapsulation efficiency was more than 80%, among which the encapsulation efficiency of the hair care black hair liposome was the highest, reaching 89.4%. In general, the liposomes prepared by the thin film hydration method provided by the present invention showed good stability and high encapsulation efficiency.

[0145] The egg yolk lecithin content (20, 40, 60, 80, 100 mg), the cholesterol to egg yolk lecithin mass ratio (1:2, 1:3, 1:4, 1:5, 1:6), and the effective ingredient (the effective ingredient refers to the extract and polypeptide component in the table above) to egg yolk lecithin mass ratio (1:2, 1:3, 1:4, 1:5, 1:6) were selected as single factor indicators to analyze the effects of different factors on different liposomes.

[0146] Table 1 Effect of preparation conditions on average particle size and encapsulation efficiency of herbal hair growth liposome-001 product

[0147]

[0148]

[0149] Different lecithin mass, cholesterol to lecithin mass ratio, and active ingredient to lecithin mass ratio had a significant effect on the encapsulation efficiency of herbal hair growth liposome-1. When the lecithin mass was 60 mg and the cholesterol to lecithin mass ratio was 1:4, the liposome encapsulation efficiency was the highest, at 86.22% and 82.22%, respectively. When the mass ratio of active ingredient to lecithin was 1:4, the encapsulation efficiency reached 80.22%. These results show that under optimized preparation conditions, the encapsulation efficiency of herbal hair growth liposomes can be significantly improved.

[0150] Table 2 Effect of preparation conditions on average particle size and encapsulation efficiency of herbal hair growth liposome-002 product

[0151]

[0152] Similar to Table 2, different preparation conditions also had a significant effect on the encapsulation efficiency of herbal hair growth liposome-2. When the mass of lecithin was 60 mg and the mass ratio of cholesterol to lecithin was 1:4, the liposome encapsulation efficiency was the highest, which was 85.42% and 81.26%, respectively. When the mass ratio of active ingredient to lecithin was 1:4, the encapsulation efficiency reached 82.56%. These results further verified the importance of appropriate preparation conditions in improving the liposome encapsulation efficiency.

[0153] Table 3 Effect of preparation conditions on average particle size and encapsulation efficiency of hair care black hair liposome products

[0154]

[0155] The encapsulation efficiency of the hair care black hair liposomes varied significantly under different preparation conditions. When the mass of lecithin was 40 mg, the encapsulation efficiency reached 84.31%, which was the best performance. When the mass ratio of cholesterol to lecithin was 1:3, the encapsulation efficiency was 76.45%. When the mass ratio of the active ingredient to lecithin was 1:4, the encapsulation efficiency was the highest, reaching 83.21%. These results show the significant effect of preparation conditions on the encapsulation efficiency of the hair care black hair liposomes.

[0156] Table 4 Effect of preparation conditions on the average particle size and encapsulation efficiency of skin care, anti-hair loss and hair growth liposome-001 product

[0157]

[0158] When the mass of lecithin was 40 mg, the mass ratio of cholesterol to lecithin was 1:3, and the mass ratio of active ingredient to lecithin was 1:4, the encapsulation rates of skin care, anti-hair loss and hair growth liposome-1 were 85.19%, 83.54% and 88.71%, respectively, all showing high encapsulation rates. These data show that the encapsulation efficiency of skin care, anti-hair loss and hair growth liposomes can be significantly improved by properly adjusting the preparation conditions.

[0159] Table 5 Effect of preparation conditions on average particle size and encapsulation efficiency of skin care, anti-hair loss and hair growth liposome-002

[0160]

[0161] The best encapsulation rate of skin care, anti-hair loss and hair growth liposome-2 appeared when the mass of lecithin was 40 mg, the mass ratio of cholesterol to lecithin was 1:3, and the mass ratio of active ingredients to lecithin was 1:4, reaching 86.19%, 79.54% and 86.19% respectively. These results are consistent with the previous table, further proving that the optimization of preparation conditions is crucial to the improvement of encapsulation rate.

[0162] Table 6 Effect of preparation conditions on the average particle size and encapsulation efficiency of skin care, anti-hair loss and hair growth liposome-003 product

[0163]

[0164]

[0165] When the mass of lecithin was 40 mg, the mass ratio of cholesterol to lecithin was 1:3, and the mass ratio of active ingredient to lecithin was 1:4, the encapsulation rates of skin care, anti-hair loss and hair growth liposome-3 were 86.54%, 87.42% and 86.51%, respectively. These results show that the liposomes prepared under these conditions have a high encapsulation rate, indicating that these conditions are the key to optimizing the preparation of skin care, anti-hair loss and hair growth liposomes.

[0166] Storage stability analysis of liposomes obtained in different embodiments

[0167] Liposome preparations are easily affected by the environment during daily processing, transportation and storage, resulting in aggregation, precipitation and other phenomena, which may lead to leakage of embedded substances and changes in particle size distribution. The composite functional liposomes prepared under optimized conditions were divided into two parts, and stored at 4 and 25 ° C in the dark for a certain period of time. The changes in particle size distribution and encapsulation efficiency of the composite functional liposomes under different storage conditions were regularly tested to determine the most suitable storage conditions for the samples. The storage stability experiment is shown in the table below. The particle size of SCSP-LIP stored at 4 ° C did not change significantly. In contrast, the particle size of the composite functional liposomes stored at 25 ° C increased significantly; with the extension of storage time, the encapsulation efficiency of the composite functional liposomes stored at 25 ° C was significantly lower than that of the composite functional liposomes stored at 4 ° C. The relatively high stability of the composite functional liposomes at low temperature (compared with other storage conditions in the same study) can be attributed to the maintenance of Zeta potential, the reduction of bilayer fluidity and the delay of unsaturated fatty acid oxidation in the membrane material under low temperature. Higher ambient temperature will change the lipid crystal structure, soften the lipid membrane and increase its fluidity, causing the embedded substances to leak out of the liposomes faster.

[0168] Table 9 Storage stability analysis of herbal hair growth liposome-1

[0169]

[0170] Table 10 Storage stability analysis of herbal hair growth liposome-2

[0171]

[0172] Table 11 Storage stability analysis of hair care black hair liposome

[0173]

[0174] Table 12 Storage stability analysis of skin care, anti-hair loss and hair growth liposome-1

[0175]

[0176] Table 13 Storage stability analysis of skin care, anti-hair loss and hair growth liposome-2

[0177]

[0178]

[0179] Table 14 Storage stability analysis of skin care, anti-hair loss and hair growth liposome-3

[0180]

[0181] The above stability results show that the particle size and encapsulation efficiency of liposomes change to varying degrees at both 4°C and 25°C. Overall, liposomes stored at 4°C show higher stability in terms of particle size and encapsulation efficiency. After 7 days of storage, the particle size increases slightly and the encapsulation efficiency decreases slightly. However, the particle size of liposomes stored at 25°C increases significantly during storage, and the encapsulation efficiency decreases significantly. The results show that storage conditions at 4°C are more conducive to maintaining the stability of liposomes and extending their shelf life.

[0182] Comparison of the efficacy of liposome compositions obtained from different liposome ratios

[0183] Hair follicle cell proliferation: The effect of liposomes on hair follicle cell proliferation was determined using the CCK-8 method. Detailed steps: Preheat the culture medium to 37°C. Inoculate hair follicle cells into 96-well plates at a density of 5000-10000 cells / well, and add 100 μL of cell suspension to each well. Incubate the cells in an incubator at 37°C and 5% CO2 for 24 hours until the cells adhere to the wall. Use a pipette to add liposome samples of predetermined concentrations to different wells, and set at least 3 replicate wells for each group of treatments. Continue to culture in the incubator for a specified time (e.g., 12 hours, 24 hours, 48 ​​hours, 72 hours). At each time point, remove the culture medium and add 10 μL of CCK-8 reagent to each well. Place the 96-well plate in the incubator again for 2-4 hours to ensure sufficient reaction. Use a microplate reader to read the absorbance value of each well at a wavelength of 450nm to calculate cell viability (relative to the control group).

[0184] Effects of four kinds of compound liposomes on the activity of hair follicle cells

[0185]

[0186]

[0187] The effect of liposomes on hair follicle cell proliferation was determined by using the CCK-8 method. The results showed that different liposome samples significantly promoted the proliferation of hair follicle cells. After 12 hours of treatment, the cell activity increased significantly, especially the effect of liposome C was the most significant, and the cell activity reached 354.1±0.33% at 72 hours. The effects of liposomes A, B, and D were also good, with cell activities of 289.5±0.74%, 205.4±0.91%, and 316.4±0.51%, respectively. These results show that liposomes can effectively promote the proliferation of hair follicle cells, and thus have the potential to promote hair growth.

[0188] Hair loss inhibition test. Method: Annexin V / PI staining was used to detect the effect of liposomes on the apoptosis rate of hair follicle cells. Detailed steps: Hair follicle cells were seeded into 6-well plates and cultured to 70-80% confluence. The culture medium was removed and the cells were washed twice with PBS. A predetermined concentration of liposome samples was added to the cells and cultured for a specified time (e.g., 12 hours, 24 hours, 48 ​​hours, 72 hours). Suspended cells and adherent cells were collected and transferred to a centrifuge tube. Centrifuge at 300xg for 5 minutes and the supernatant was discarded. Resuspend the cells with cold PBS and centrifuge again. Resuspend the cell suspension in 500 μL of binding buffer, add 5 μL of Annexin V-FITC and 5 μL of PI dye, and incubate in the dark for 15 minutes. The proportion of apoptotic cells was analyzed using a flow cytometer.

[0189] Effects of four kinds of compound liposomes on apoptosis rate of hair follicle cells

[0190]

[0191] The effect of liposomes on the apoptosis rate of hair follicle cells was detected by using Annexin V / PI staining. The results showed that different liposome samples had a significant inhibitory effect on the apoptosis of hair follicle cells. After 12 hours of treatment, liposome C showed the lowest apoptosis rate (2.19±0.26%), while the apoptosis rates of liposomes A, B and D were also significantly lower than those of the control group. At 72 hours, the apoptosis rate of liposome C was 10.17±0.66%, which was significantly lower than that of other samples. This shows that liposome C has the best effect on inhibiting apoptosis of hair follicle cells and may have the effect of inhibiting hair loss.

[0192] Black hair effect test: Method: Use tyrosinase activity to determine the promoting effect of liposomes on melanin synthesis. Detailed steps: Melanocytes are inoculated into 6-well plates and cultured to 80-90% fusion. Remove the culture medium and wash the cells twice with PBS. Add liposome samples of different concentrations to the cells and continue to culture for a specified time (e.g. 12 hours, 24 hours, 48 ​​hours, 72 hours). At the specified time point, collect the cells, lyse the cells with lysis buffer, and incubate on ice for 30 minutes. Further lyse the cells by ultrasonic cell disruptor. Centrifuge at 12000xg for 10 minutes and collect the supernatant. Use a tyrosinase activity assay kit and perform the test according to the instructions of the kit to record the tyrosinase activity.

[0193] Effects of four kinds of compound liposomes on tyrosinase activity of hair follicle cells

[0194]

[0195] The liposomes were evaluated for their promoting effect on melanin synthesis by measuring tyrosinase activity. The results showed that different liposome samples could significantly increase the tyrosinase activity in melanocytes, especially liposome C, which had the most significant effect. After 48 hours of treatment, the tyrosinase activity of liposome C reached 65.32±3.16U / L, and the activity of other liposomes also increased significantly. After 72 hours of treatment, the tyrosinase activity of liposome C was further increased to 75.44±2.61U / L. These results indicate that liposomes can promote the synthesis of melanin and may have a positive effect on hair darkening.

[0196] In vivo animal experiment. Hair growth experiment. Methods: The effect of liposomes on promoting hair growth was tested in a shaved mouse model.

[0197] Detailed steps: Select healthy adult mice and shave the hair on their backs. The shaved area is about 2x2 cm. 2 . The mice were randomly divided into groups, and different liposome samples were applied to the shaved area on the back of each group of mice to ensure uniform coverage of each group of samples. The liposomes were applied once a day for 5 weeks. The length of hair growth was measured with a ruler or caliper every week, and photos were taken to record the hair growth. At the end of the experiment, the new hair was shaved, and the hair length and weight were measured to evaluate the hair growth effect of the liposomes.

[0198] Effects of four kinds of compound liposomes on hair growth rate in mice

[0199]

[0200] In the shaved mouse model, after applying different liposomes, it was observed that the liposomes had a significant promoting effect on hair growth. Liposome C had the highest hair growth rate at week 5, reaching 168.4±3.15%, which was significantly higher than other liposomes. The hair growth rates of liposomes A, B, and D at the same time point were 87.4±1.89%, 80.1±2.54%, and 98.2±2.55%, respectively. These results show that liposome C has the most significant effect on promoting hair growth and has good hair growth potential.

[0201] Hair growth experiment. Method: The effect of liposomes on hair growth was tested in a shaved mouse model. Detailed steps: Select healthy adult mice and shave the hair on the back. The shaved area is about 2x2 cm 2 The mice were randomly divided into groups, and different liposome samples were applied to the shaved area on the back of each group of mice to ensure uniform coverage of each group of samples. The liposomes were applied once a day for 5 weeks. The length of hair growth was measured with a ruler or caliper every week, and photos were taken to record the hair growth. At the end of the experiment, the new hair was shaved off, the hair length was measured, and the hair growth effect of the liposomes was evaluated.

[0202] Effects of four kinds of compound liposomes on the hair removal area in mice

[0203]

[0204] Effects of four kinds of compound liposomes on hair growth rate in mice

[0205]

[0206] In the glucocorticoid-induced mouse hair loss model, the liposome samples showed different degrees of hair loss inhibition effects. Liposome C showed the most significant inhibitory effect at week 5, with the hair loss area of ​​the mice reduced to 0.89±0.39cm 2 The hair removal areas of liposomes A, B and D at the same time point were 3.76±0.26cm 2 、3.54±0.22cm 2 and 2.55±0.55cm 2 At the same time, the number of hair follicles in the C liposome group increased significantly to 82.4±1.85n / cm 2 , indicating that its effect on hair regeneration and hair loss inhibition is better than other samples.

[0207] Black hair experiment. Method: Test the effect of liposomes on hair color in mice. Detailed steps: Select healthy adult mice and shave the hair on the back. The shaved area is about 2x2 cm 2The mice were randomly divided into groups, and different liposome samples were applied to the shaved area on the back of each group of mice for 5 weeks. At the end of the experiment, the hair from the shaved area was taken, washed with PBS, dried and weighed. The melanin content in the hair was measured using a spectrophotometer, and the changes in melanin concentration were recorded.

[0208] Effects of four kinds of compound liposomes on the content of melanin in mouse hair

[0209]

[0210] In the mouse black hair experiment, the liposome samples had a significant effect on the melanin content of mouse hair. After 5 weeks of treatment, the melanin content of mouse hair treated with liposome C was the highest, reaching 33.4±3.19pg / g, which was significantly better than other liposomes. The melanin content of liposomes A, B and D at the same time point was 18.2±0.42pg / g, 20.1±0.51pg / g and 29.5±1.84pg / g, respectively. This shows that liposome C has the most significant effect on hair color and may have potential in promoting black hair.

[0211] The present invention provides a hair-fixing, anti-hair-loss and black-hair composite functional liposome and a preparation method thereof. Compared with the prior art, the liposome is prepared by a thin film hydration method, and the formula and preparation conditions are optimized to improve the encapsulation rate, stability and bioavailability of the liposome, thereby solving the problems existing in the prior art. The liposome composition prepared by the present invention not only has the efficacy of preventing hair loss and fixing hair, but also can promote black hair, is suitable for various types of hair loss, and has significant market application prospects. Specifically, 1. High stability: The liposome of the present invention has higher stability during storage and can maintain the effectiveness of its active ingredients for a long time. 2. High encapsulation rate: By optimizing the preparation conditions, the encapsulation rate of the liposome of the present invention is significantly improved, ensuring the effective synergistic effect of multiple active ingredients. 3. Improved bioavailability: The nanostructure of the liposome helps to improve the skin permeability and bioavailability of the active ingredients, thereby enhancing the actual effect of the product.

[0212] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions and improvements made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A hair-fixing and hair-loss-preventing composite liposome, characterized in that: Includes the following components: Herbal hair growth liposome-001, comprising the following components in parts by weight: phosphate; cholesterol; a first active extract; wherein the first active extract comprises: one or more of a colorless salvia miltiorrhiza extract, a ginger extract, an chrysanthemum leaf extract, an chrysanthemum leaf extract, an aconite extract, and a Ligustrum lucidum fruit extract; Herbal hair growth liposome-002, comprising the following components in parts by weight: phosphate; cholesterol; a second active extract; wherein the second active extract comprises: one or more of soapberry extract, mint leaf extract, mulberry extract, wolfberry extract, wolfberry extract, angelica extract, and safflower extract; The hair care black hair liposome comprises the following components in parts by weight: phosphate; cholesterol; a third active extract; wherein the third active extract comprises: one or more of sh-pentapeptide-1, palmitoyl tetrapeptide-10, and acetyl hexapeptide-1; Skin care, anti-hair loss and hair growth liposome-001 comprises the following components in parts by mass: phosphate; cholesterol; fourth active extract; wherein the fourth active extract comprises: one or more of sh-pentapeptide-1, oligopeptide-54, tripeptide-1, AHK-copper peptide, acetyl tetrapeptide-3, and decapeptide-10; Skin care, anti-hair loss and hair growth liposome-002 comprises the following components in parts by weight: phosphate; cholesterol; the fifth active extract; wherein the fifth active extract comprises: one or more of sh-pentapeptide-1, oligopeptide-54, tripeptide-1, AHK-copper peptide, acetyl tetrapeptide-3, and decapeptide-10; Skin care, anti-hair loss and hair growth liposome-003 includes the following components in parts by mass: phosphate; cholesterol; the sixth active extract; wherein the sixth active extract includes: one or more of sh-pentapeptide-1, octapeptide-2, myristoyl hexapeptide-16, myristoyl pentapeptide-17 and decapeptide-18.

2. The hair-fixing, anti-hair loss and blackening composite functional liposome according to claim 1, characterized in that: The first active extract comprises the following components in parts by weight: The first active extract comprises the following components in parts by weight: 5-15 parts of colorless Salvia miltiorrhiza extract; 5-15 parts of ginger extract; 5-15 parts of extract of Channa chuatsi leaf; 5-15 parts of Pleurotus eryngii leaf extract; 5-15 parts of Aconitum extract; 5-15 parts of Ligustrum lucidum extract.

3. The hair-fixing, anti-hair loss and blackening composite functional liposome according to claim 1, characterized in that: The third active extract comprises the following components in parts by weight: sh-pentapeptide-1 0.1-0.5 parts; Palmitoyl Tetrapeptide-10 0.1-0.5 parts; Acetyl Hexapeptide-1 0.01-0.1 parts.

4. The hair-fixing, anti-hair loss and blackening composite functional liposome according to claim 1, characterized in that: The second active extract comprises the following components in parts by weight: 5-15 parts of soapberry extract; 5-15 parts of peppermint leaf extract; 5-15 parts of mulberry extract; 5-15 parts of wolfberry extract; 5-15 parts of angelica extract; 5-15 parts of safflower extract.

5. The hair-fixing, anti-hair loss and blackening composite functional liposome according to claim 1, characterized in that: The fourth active extract comprises the following components in parts by weight: sh-pentapeptide-1 0.1-0.5 parts; Oligopeptide-54 0.02-0.2 parts; Tripeptide-1 0.1-0.5 parts; AHK-Copper Peptide 0.01-0.5 parts Acetyl tetrapeptide-3 0.03-0.1 parts; Decapeptide-10 0.1-0.5 parts.

6. The hair-fixing, anti-hair-loss and black-hair-blackening composite liposome according to claim 1, characterized in that: The fifth active extract comprises the following components in parts by weight: sh-pentapeptide-1 0.1-1 parts; Oligopeptide-54 0.1-0.5 parts; Tripeptide-1 0.01-0.05 parts; AHK-copper peptide 0.1-0.5 parts; Acetyl tetrapeptide-3 0.02-0.03 parts; Decapeptide-10 0.01-0.05 parts.

7. The hair-fixing, anti-hair-loss and black-hair-blackening composite liposome according to claim 1, characterized in that: The sixth active extract comprises the following components in parts by weight: sh-pentapeptide-1 0.1-1 parts; Octapeptide-2 0.05-0.1 parts; Myristoyl hexapeptide-16 0.1-0.5 parts; Myristoyl pentapeptide-17 0.01-0.1 parts; Decapeptide-18 0.02-0.2 parts.

8. A method for preparing a hair-fixing and hair-loss-preventing composite functional liposome, based on a hair-fixing and hair-loss-preventing composite functional liposome as claimed in any one of claims 1 to 7, comprising the following steps: Complete the preparation of liposomes; the liposomes include: One or more of herbal anti-hair loss and hair growth liposome-001, herbal anti-hair loss and hair growth liposome-002, hair care black hair liposome, anti-hair loss and hair growth polypeptide liposome-001, anti-hair loss and hair growth polypeptide liposome-002, and anti-hair loss and hair growth liposome-003; The prepared liposomes of different types are mixed according to a preset ratio to obtain a final product.

9. The method for preparing the hair-fixing, anti-hair-loss and black-hair-blackening composite functional liposome according to claim 8, characterized in that: The liposomes include: herbal anti-hair loss and hair growth liposome-001, herbal anti-hair loss and hair growth liposome-002, hair care black hair liposome, anti-hair loss and hair growth polypeptide liposome-001, anti-hair loss and hair growth polypeptide liposome-002 and anti-hair loss and hair growth liposome-003.

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