Peptide-binding liposome composition having excellent scalp penetration effect, and hair growth efficacy and hair loss alleviation targeting technology using same

Abstract

The present invention relates to a composition for alleviating hair loss or promoting hair growth, and more particularly, to a composition including an active ingredient in a liposome to which a PTD peptide and palmitoyl pentapeptide-4 are bound, and thus capable of effectively delivering the active ingredient into the skin, and having an excellent hair loss alleviation or hair growth promotion effect.

Description

Peptide-conjugated liposome composition with excellent scalp penetration and technology targeting hair growth and hair loss relief utilizing the same

[0001] The present invention relates to a composition for alleviating hair loss or promoting hair growth, and more specifically, to a composition capable of effectively delivering an active ingredient into the skin by including the active ingredient in a liposome combined with a PTD peptide and palmitoyl pentapeptide-4, and having an excellent effect of alleviating hair loss or promoting hair growth.

[0002] Human hair primarily serves a defensive function, protecting the scalp from external irritants like UV rays, and also plays an aesthetic role in expressing an individual's external image. However, in modern society, hair loss is becoming more common due to natural factors like environmental pollution and strong UV rays, as well as physiological factors like stress and hormonal imbalances. Unlike in the past, when hair loss was limited to adult men, hair loss is now affecting people of all ages and genders, creating a pressing need for preventative and therapeutic solutions.

[0003] Hair follicles, physiological organs in the body that produce hair, are involved in the growth, maintenance, and loss of hair by repeating the hair growth cycle, which is divided into the anagen phase, in which hair is produced and actively grows, the catagen phase, in which hair degenerates, the telogen phase, in which hair falls out, and the exogen phase, in which hair loss occurs. Many studies have revealed that the activity of these hair follicles is caused by hair dermal papilla cells, and in particular, the proliferation and differentiation of dermal papilla cells are primarily involved in the progression of the hair growth cycle and hair formation. During the anagen phase, when hair actively grows, dermal papilla cells actively proliferate and differentiate, and during the catagen phase, when hair growth stops and hair loss occurs, these cells die. Therefore, since hair growth and hair loss are closely related to the proliferation and death of hair papilla cells, inducing their proliferation to prolong the growth phase or suppressing cell death to shorten the regression phase, resting phase, and hair loss phase will be a way to improve and treat hair loss.

[0004] Alopecia is a condition in which more than 100 hairs fall out per day, the number of hairs in the growth phase decreases, and the number of hairs in the catagen or resting phase increases. As hair loss progresses, the proportion of hairs in the resting phase increases rapidly, and the proportion of hairs in the growing phase decreases correspondingly. Hair loss can generally be categorized into hereditary androgenetic alopecia (baldness), alopecia areata, tinea capitis caused by a fungal infection, telogen effluvium, trichotillomania, and hair growth disorders. The most common types of hair loss include baldness (male pattern baldness), female pattern baldness, alopecia areata, and telogen effluvium.

[0005] Accordingly, many studies are being conducted to alleviate or treat hair loss, and in particular, research is being conducted to increase the efficiency of substance delivery into the scalp.

[0006] Accordingly, the inventors of the present invention attempted to deliver an active ingredient into the scalp by using liposomes, and in order to increase the delivery efficiency thereof, they produced a PTD peptide having a novel sequence containing a large amount of arginine and lysine, and further combined palmitoyl pentapeptide-4 with the liposomes, thereby completing the present invention.

[0007] The present invention aims to provide a liposome composition comprising a PTD (Protein transduction domain) peptide represented by sequence number 1; and palmitoyl pentapeptide-4 (PP).

[0008] The technical problems of the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0009] According to an embodiment of the present invention, a liposome composition is provided, comprising a PTD (Protein transduction domain) peptide represented by SEQ ID NO: 1; and palmitoyl pentapeptide-4 (PP).

[0010] According to another embodiment of the present invention, a cosmetic composition for improving hair loss or inducing hair growth, comprising the liposome composition, is provided.

[0011] The liposome composition according to the present invention comprises a novel sequence of PTD peptides containing large amounts of arginine and lysine, and palmitoyl pentapeptide-4, positioned in the outer layer of the liposome, thereby effectively penetrating the stratum corneum of the scalp, thereby delivering active ingredients deep into the scalp. Therefore, it can be utilized as a cosmetic composition for promoting the growth or proliferation of hair papilla cells, strengthening hair roots, and suppressing hair loss.

[0012] In order to more fully understand the drawings cited in the detailed description of the present invention, a brief description of each drawing is provided.

[0013] Figure 1 is a diagram showing the results of confirming the cytotoxicity of the PTD-PP-liposome composition according to the present invention.

[0014] Figure 2 is a diagram showing the results comparing the amount of adenosine permeated per area after 10 hours.

[0015] Figure 3 is a diagram showing the results of comparing the degree of skin penetration of adenosine administered for 24 hours.

[0016] According to an embodiment of the present invention, a liposome composition is provided, comprising a PTD (Protein transduction domain) peptide represented by SEQ ID NO: 1; and palmitoyl pentapeptide-4 (PP).

[0017] Additionally, the PTD peptide may be located in the outer layer of the liposome and may have a positive charge.

[0018] According to another embodiment of the present invention, a cosmetic composition for improving hair loss or inducing hair growth, comprising the liposome composition, is provided.

[0019] In addition, the liposome composition contains an active ingredient therein, and the active ingredient may be at least one selected from the group consisting of biotin, adenosine, kaempferol, Streptococcus thermophilus ferment, sabal palm fruit extract, and apple extract.

[0020] Additionally, the composition may promote growth or proliferation of mammary gland cells.

[0021] Additionally, the composition may strengthen hair roots.

[0022] Additionally, the liposome composition may be included in an amount of 0.01 to 10 wt% based on the total weight of the entire composition.

[0023] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains. In general, the nomenclature used herein is well known and commonly used in the art. In addition, when describing embodiments of the present invention, if a detailed description of a related known structure or function is judged to hinder the understanding of the embodiments of the present invention, a detailed description thereof will be omitted. In addition, although embodiments of the present invention will be described below, the technical idea of ​​the present invention is not limited or restricted thereto, and can be modified and implemented in various ways by those skilled in the art.

[0024] When a part in this specification is said to include a certain component, this does not exclude other components, unless otherwise specifically stated, but rather means that other components may be included. In this specification, the term "and / or" includes a combination of multiple related items or any one of multiple related items.

[0025]

[0026] According to an embodiment of the present invention, a liposome composition is provided, comprising a PTD (Protein transduction domain) peptide represented by SEQ ID NO: 1; and palmitoyl pentapeptide-4 (PP).

[0027] The liposome composition according to the present invention is characterized in that it has a positive charge by binding the PTD peptide and palmitoyl pentapeptide-4 to the outer layer of the liposome.

[0028] In the present invention, the PTD (Protein transduction domain) peptide is composed of sequence number 1 (RKKRRQRRR), and is a material transfer peptide that transfers extracellular substances into cells. It contains a large amount of arginine and lysine, so it has a positive charge and has the characteristic of being well attached through interaction with the phosphate group of the head portion of the phospholipid of the cell membrane of skin keratinocytes, which has a negative charge.

[0029] In the present invention, the palmitoyl pentapeptide-4 (PP) is a collagen peptide that promotes collagen production and helps cell proliferation, and when bound to liposomes, helps to improve the delivery of substances into cells.

[0030] Throughout this specification, the conventional one-letter and three-letter codes for naturally occurring amino acids are used, as well as generally accepted three-letter codes for other amino acids, such as Aib (α-aminoisobutyric acid) and Sar (N-methylglycine). Furthermore, amino acids referred to herein by abbreviations are described according to the IUPAC-IUB nomenclature as follows:

[0031] Alanine: A, arginine: R, asparagine: N, aspartic acid: D, cysteine: C, glutamic acid: E, glutamine: Q, glycine: G, histidine: H, isoleucine: I, leucine: L, lysine: K, methionine: M, phenylalanine: F, proline: P, serine: S, threonine: T, tryptophan: W, tyrosine: Y, and valine: V.

[0032] The term “peptide” used in the present invention refers to a linear amino acid polymer formed by amino acid residues being linked to each other by peptide bonds, and may include not only natural amino acids but also non-protein amino acids as components.

[0033] In the present invention, the peptide preferably has the amino acid sequence described above, but is not limited thereto. According to a preferred embodiment of the present invention, the peptide preferably has a high ratio of the amino acid sequence of 50% or more, preferably 60% or more, more preferably 70% or more, more preferably 80% or more, more preferably 90% or more, and most preferably 100%.

[0034] Even if the present invention describes a 'peptide composed of a specific sequence number', if it has the same or corresponding activity as a peptide composed of an amino acid sequence of the sequence number, it does not exclude meaningless sequence additions before and after the amino acid sequence of the sequence number, mutations that may occur naturally, or silent mutations thereof, and it is clear that even if it has such sequence additions or mutations, it falls within the scope of the present invention.

[0035] In the present invention, modification of the peptide sequence may be one in which some amino acids are modified through any one of substitution, addition, deletion, and modification, or a combination of these methods. Such modifications include modifications using L- or D-amino acids, and / or non-natural amino acids; and / or modifications of the native sequence, for example, modification of side chain functional groups, intramolecular covalent bonds such as ring formation between side chains, methylation, acylation, ubiquitination, phosphorylation, aminohexanoic acid oxidation, biotinylation, etc. The substituted or added amino acids may be the 20 amino acids commonly observed in human proteins, as well as atypical or non-naturally occurring amino acids.

[0036] In the present invention, the peptide may also include a targeting sequence, a tag, a labeled residue, or an additional amino acid sequence designed for a specific purpose, such as increasing the half-life or stability of the peptide. Furthermore, the peptide of the present invention may be linked to coupling partners, such as effectors, drugs, prodrugs, toxins, peptides, or delivery molecules.

[0037] In the present invention, the peptide can be prepared in the form of a pharmaceutically acceptable salt. Specifically, a salt can be formed by adding an acid, for example, an inorganic acid (e.g., hydrochloric acid, hydrobromic acid, phosphoric acid, nitric acid, sulfuric acid, etc.), an organic carboxylic acid (e.g., haloacetic acids such as acetic acid, trifluoroacetic acid, propionic acid, maleic acid, succinic acid, malic acid, citric acid, tartaric acid, salicylic acid), and an organic sulfonic acid including an acidic sugar (e.g., glucuronic acid, galacturonic acid, gluconic acid, ascorbic acid), an acidic polysaccharide (e.g., hyaluronic acid, chondroitin sulfate, arginic acid), a sulfonic acid sugar ester such as chondroitin sulfate (e.g., methanesulfonic acid, p-toluenesulfonic acid), etc.

[0038] Peptides according to the present invention can be prepared by chemical synthesis methods known in the art, standard synthetic methods, recombinant expression systems, or any other method in the art.

[0039] The liposome composition according to the present invention exhibits excellent skin barrier permeability, enabling the active ingredients within the liposomes to be effectively delivered to target cells located deep within the scalp's stratum corneum via two pathways: intercellular and transcellular. Furthermore, the liposomes according to the present invention have a size of 100 to 200 nm and exhibit excellent storage stability, making them suitable for use in cosmetic compositions.

[0040]

[0041] According to another embodiment of the present invention, a cosmetic composition for improving hair loss or inducing hair growth is provided, comprising the liposome composition.

[0042] The cosmetic composition for improving hair loss or inducing hair growth according to the present invention relates to root vital technology, which refers to a targeting technology that applies root vital code, which adds an antioxidant effect to a peptide-bonded liposome component so that it can be delivered deep into the scalp. The root vital code refers to a hair loss-only care composition proposed by the inventors of the present invention, which is characterized by including the active ingredients described below (e.g., biotin, adenosine, kaempferol, Streptococcus thermophilus ferment, sabal palm fruit extract, apple extract, etc.). More specifically, the root vital code is composed of three steps, and the first step is a step for organizing the scalp composed of ingredients that provide nutrition to the scalp. Specifically, the ingredients may include antioxidant-based substances such as apple water and apple extract, and scalp care ingredients that promote hair follicle cell growth, such as kaempferol, sabal palm fruit extract, and Streptococcus thermophilus ferment. Next, the second step is a step for maintaining and caring for weak hair to be strong, and may specifically include adenosine as an ingredient, and lastly, the third step is a step for strengthening and caring for hair to be strong, and may specifically include biotin as an ingredient. That is, the cosmetic composition for improving hair loss or inducing hair growth according to the present invention can effectively deliver active ingredients deep into the scalp by including PTD peptide and palmitoyl pentapeptide-4 in liposomes containing various active ingredients.

[0043] The above liposome composition contains an active ingredient therein, and the active ingredient may be at least one selected from the group consisting of biotin, adenosine, kaempferol, Streptococcus thermophilus ferment, sabalpam fruit extract, and apple extract, but is not limited thereto. For example, it can be effectively applied to synthetic raw materials such as minoxidil, finasteride, and aminexil, and natural extracts such as nettle, walnut, and natural grass, and synthetic pharmaceuticals. Any method known in the art can be used for producing liposomes or including an active ingredient inside liposomes, but is not limited thereto. The active ingredient may be included in an amount of 0.1 to 5 wt%, specifically, 0.5 to 5 wt%, based on the total weight of the entire composition, but is not limited thereto.

[0044] The liposome composition may be included in an amount of 0.01 to 10 wt% relative to the total weight of the entire composition, but is not limited thereto. If the liposome concentration is less than 0.01 wt%, it is difficult to expect a significant effect, and if it is 10 wt% or more, cytotoxicity may be exhibited.

[0045] The above composition exhibits an effect of improving hair loss or inducing hair growth by promoting the growth or proliferation of hair papilla cells and strengthening hair roots.

[0046] The above hair loss may be at least one selected from the group consisting of androgenetic alopecia, telogen effluvium, drug-induced alopecia, mechanical alopecia, traumatic alopecia, compression alopecia, anagen alopecia, pityriasis versicolor, syphilitic alopecia, seborrheic alopecia, symptomatic alopecia, cicatricial alopecia, congenital alopecia, alopecia areata, tinea capitis, alopecia totalis, hypotrichosis, hereditary hypotrichosis simplex, and alopecia universalis.

[0047] The above cosmetic composition can be manufactured into any formulation commonly manufactured in the technical field to which the present invention pertains. For example, the cosmetic composition according to the present invention can be manufactured into the formulation of hair tonic, hair conditioner, hair essence, hair lotion, hair nutrition lotion, hair shampoo, hair rinse, hair treatment, hair cream, hair nutrition cream, hair moisture cream, hair massage cream, hair wax, hair aerosol, hair pack, hair nutrition pack, hair soap, hair cleansing foam, scalp shampoo, scalp hair pack, scalp ampoule, hair oil, hair drying agent, hair preservation treatment agent, hair dye, hair waving agent, hair bleaching agent, hair gel, hair glaze, hair dressing agent, hair lacquer, hair moisturizer, hair mousse, or hair spray, but is not limited thereto.

[0048] The cosmetic composition of the present invention may include other additives such as excipients and carriers, and it is possible to apply and mix as much as necessary the usual ingredients mixed in general skin cosmetics.

[0049]

[0050] Hereinafter, examples and experimental examples are presented to explain the present invention more specifically, but the present invention is not limited thereto. 1

[0051]

[0052] [Manufacturing example]

[0053] Example 1. Preparation of a liposome composition containing a peptide

[0054] The present inventors attempted to use liposomes because the absorption rate of active ingredients into the skin is low when directly applied, and in order to increase the substance delivery efficiency compared to existing liposomes, the present inventors newly designed a substance delivery peptide (protein transduction domain, PTD) containing a large amount of arginine (R) and lysine (K): RKKRRQRRR (SEQ ID NO: 1). The peptide exhibits a positive charge, and the arginine of the peptide increases the distance between the guanidine group and the peptide backbone, enabling penetration into cells, and in particular, assisting the delivery of substances through electrical interaction with the proteoglycans of the outer cell membrane. Next, by conjugating palmitoyl pentapeptide-4 (PP), a collagen peptide, to the liposome, it was made to play a role in promoting collagen production and assisting cell proliferation. The PTD of the present invention is located in the outer layer of the liposome and adheres well to the cell through interaction with the negative charge of the phospholipids of the skin keratinocytes. Specifically, oleic acid, polyglyceryl-10 stearate, hydrogenated lecithin, and hydroxypropyl cyclodextrin were included to form the liposome, and finally, as shown in Table 1, a liposome composition containing PTD and PP together with an active ingredient for preventing hair loss or promoting hair growth was prepared.

[0055] [Table 1]

[0056]

[0057]

[0058] Preparation of Comparative Examples 1 to 4

[0059] Liposome compositions were prepared in the same manner as in Example 1, but without including PP or PTD, to prepare compositions of Comparative Examples 1 to 3. In addition, a sample consisting solely of palmitoyl pentapeptide-4 was designated Comparative Example 4. Specific information is shown in Table 2.

[0060] [Table 2]

[0061]

[0062]

[0063] Example 2. Preparation of a cosmetic composition comprising a liposome composition

[0064] In order to prepare a cosmetic composition for preventing hair loss or promoting hair growth, including the liposome composition prepared in Example 1, a cosmetic composition was prepared by mixing root vital codes as shown in Table 3. Specifically, root vital codes include apple water, apple extract, and Streptococcus thermophilus fermentation product obtained by distilling apples known to have antioxidant effects, to which the liposome composition prepared in Example 1 was added.

[0065] [Table 3]

[0066]

[0067]

[0068] [Experimental Example]

[0069] Experimental Example 1. Cell Proliferation Evaluation

[0070] Human dermal papilla (HDP) cells were treated with the samples of Example 1 and Comparative Examples 1 to 4 at a concentration of 1% and then cultured in a cell culture medium. EPG was used as a positive control. Afterwards, WST-1 solution was added and the absorbance was measured. The experimental results are shown in Table 4.

[0071] [Table 4]

[0072]

[0073] As shown in Table 4, the PP4 powder according to Comparative Example 4 did not pass through the cell membrane well and thus did not show any effect on cell proliferation, and Comparative Example 1 (PTPPX) containing only the active ingredient and Comparative Example 2 (PPO) not containing PTD also did not show any direct effect on cell proliferation, whereas the PTD-PP-liposome composition (PTPPO) according to the present invention was confirmed to effectively increase the proliferation of hair papilla cells by delivering the active ingredient well into the cells.

[0074]

[0075] Experimental Example 2. Evaluation of hair root strengthening

[0076] Human dermal papilla (HDP) cells were treated with the sample (PTPPO) of Example 1 at a concentration of 1% and then cultured in a cell culture medium. Adenosine was used as a positive control. RNA was then extracted from the cells, cDNA was synthesized, and VEGF expression was confirmed using a real-time PCR device. VEGF is a gene that promotes hair thickness and strengthens hair follicles by generating microvessels around the hair follicle and increasing their distribution, thereby supplying nutrients to the hair follicle. The results are shown in Table 5.

[0077] [Table 5]

[0078]

[0079] As shown in Table 5, it was confirmed that the PTD-PP-liposome composition (PTPPO) according to the present invention has an excellent hair root strengthening effect by increasing VEGF expression.

[0080]

[0081] Experimental Example 3. Evaluation of hair loss relief

[0082] Human dermal papilla (HDP) cells were treated with Substance P and the sample of Example 1 (PTPPO) at a concentration of 0.5% and then cultured in a cell culture medium. Caffeine was used as a positive control. RNA was then extracted from the cells, cDNA was synthesized, and TGF-β2 expression was confirmed using a real-time PCR device. TGF-β2 is a gene that induces hair loss by shortening the hair follicle growth phase and promoting the transition to the catagen phase. The results are shown in Table 6.

[0083] [Table 6]

[0084]

[0085] As shown in Table 6, it was confirmed that the PTD-PP-liposome composition (PTPPO) according to the present invention has an excellent effect of suppressing and alleviating hair loss by suppressing TGF-β2 expression.

[0086]

[0087] Experimental Example 4. Cytotoxicity Evaluation

[0088] HaCaT cells were treated with the sample (PTPPO) of Example 1 at various concentrations (0.5 to 10%) and cultured in a cell culture incubator. After 24 hours, an MTT assay was performed according to a known method. The results are shown in Figure 1.

[0089] As shown in Fig. 1, it was confirmed that the PTD-PP-liposome composition (PTPPO) according to the present invention did not exhibit cytotoxicity up to 5% and exhibited slight cytotoxicity at 10%.

[0090]

[0091] Experimental Example 5. Percutaneous Absorption Evaluation

[0092] To evaluate percutaneous absorption, material permeability was assessed for 24 hours using Neoderm-ED artificial skin. Neoderm-ED is an experimental skin model that recreates the dermis and epidermis using human skin cells. It is composed of living keratinocytes and is physiologically and morphologically very similar to actual human skin. All samples contained adenosine within liposomes. Sample A was the PTD-PP-liposome composition according to Example 1, Sample B was a liposome composition that did not contain any peptides, and Sample C was a PTD-liposome composition. The results are shown in Figures 2 and 3 and Table 7.

[0093] [Table 7]

[0094]

[0095] As shown in Fig. 2, after 10 hours, the material permeability was confirmed, and it was confirmed that the permeability per unit area was significantly higher when using the PTD-PP-liposome composition according to the present invention compared to other compositions. In particular, the PTD-liposome composition (sample C) showed a superior permeability per unit area compared to the liposome composition (sample B) that did not contain any peptides, but there was a significant difference from the PTD-PP-liposome composition (sample A) according to the present invention.

[0096] In addition, as shown in FIG. 3, the PTD-PP-liposome composition according to the present invention (sample A) was compared with the amount of adenosine administered for 24 hours to determine that adenosine was detected after 1 hour, and that it exhibited significantly superior permeability compared to the liposome composition (sample B) containing no peptides in which adenosine was detected after 4 hours and the PTD-liposome composition (sample C) in which adenosine was detected after 2 hours.

[0097] In addition, as shown in Table 7, the permeation amount and permeation rate of adenosine were confirmed after 24 hours, and the effect of the PTD-PP-liposome composition according to the present invention was confirmed to be significantly superior to the liposome composition containing no peptide and the PTD-liposome composition. Consequently, it was confirmed that PTD and PP exhibited a remarkable effect on substance permeation, i.e., substance transfer, by working together with liposomes.

[0098]

[0099] Through the above experimental results, it was confirmed that the PTD-PP-liposome composition according to the present invention has an excellent effect of stably delivering an active ingredient into the skin, and can be used as a cosmetic composition for alleviating hair loss or promoting hair growth.

[0100]

[0101] While specific aspects of the present invention have been described in detail above, it will be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. Therefore, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. A liposome composition comprising a PTD (Protein transduction domain) peptide represented by sequence number 1; and palmitoyl pentapeptide-4 (PP).

2. In paragraph 1, A liposome composition wherein the above PTD peptide is located in the outer layer of the liposome and has a positive charge.

3. A cosmetic composition for improving hair loss or inducing hair growth, comprising a liposome composition according to claim 1 or 2.

4. In paragraph 3, The above liposome composition contains an active ingredient therein, A cosmetic composition wherein the above active ingredient is at least one selected from the group consisting of biotin, adenosine, kaempferol, Streptococcus thermophilus ferment, sabal palm fruit extract, and apple extract.

5. In paragraph 3, A cosmetic composition wherein the composition promotes the growth or proliferation of breast papilla cells.

6. In paragraph 3, A cosmetic composition, wherein the composition strengthens hair roots.

7. In paragraph 3, A cosmetic composition, wherein the liposome composition is included in an amount of 0.01 to 10 wt% based on the total weight of the entire composition.

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