KR20230051105A - Composition for growth promotion and proliferation of hair comprising a hydrolyzed DNA fragment mixture as an active ingredient - Google Patents

Abstract

The present invention relates to a composition for promoting hair growth and growth comprising a mixture of hydrolyzed DNA fragments and a method for preparing the same.

Description

Composition for growth promotion and proliferation of hair comprising a hydrolyzed DNA fragment mixture as an active ingredient

The present invention relates to a composition for promoting hair growth and growth comprising a mixture of hydrolyzed DNA fragments and a method for preparing the same.

The scalp is composed of epidermis, dermis, and subcutaneous tissue, and the outermost tissue is the epidermis. The epidermis serves to protect the body's sensitive tissues from damage. However, various physical and chemical external stimuli, such as changes in external temperature and humidity, pollutants, stress, lack of influence, etc. may cause loss of function of the scalp, and may further affect hair loss.

In addition, in modern society where beauty is emphasized, many people, regardless of age or gender, use hair dyes, permanent agents, etc. to satisfy the desire for individual expression. However, in the case of some sensitive scalps, chemical procedures such as dyeing and perming cause irritation and inflammation such as itching or stinging of the scalp, which leads to environmental hair loss.

In this way, research on ingredients that can protect the scalp from environmental stimulation, stress, chemical procedures, etc. caused by various causes, promote hair growth while preventing hair loss, and are ecologically friendly and harmless to the environment and humans continues. It is becoming.

Nucleic acids are DNA and RNA present in the nucleus of cells, and contain purine bases (adenine, A; guanine, G) or pyrimidine bases (cytosine, C; thymine, T; uracil, It is a polymer of nucleotides composed of U), pentose sugar and phosphoric acid. These nucleic acids have been recognized as a material for storing and transmitting genetic information, but recently, nucleic acids are broken down into small pieces outside the cell and selectively combine with receptors on the cell surface to activate signals into the cell and small pieces of nucleic acid As they are known to move into cells and play a role in rapidly inducing DNA synthesis with little energy, new functions and technologies using them are being developed.

Polydeoxyribonucleotide, a kind of mixture of DNA fragments with specific specifications extracted from animals or plants, stimulates adenosine A2 receptors to promote anti-inflammation and cell proliferation , It is known to have a tissue regeneration effect (Kim, Y.H., et al., The wound healing effect of PDRN (polydeoxyribonucleotide) material on full thickness skin defect in the mouse, J. Korean Soc. Plast. Reconstr. Surg., 37(3), 220-226, 2010.).

Accordingly, there is a need for a composition that can be used to promote hair proliferation and growth using such a mixture of hydrolyzed DNA fragments for cell regeneration or cell growth.

An object of the present invention is to provide a composition for promoting hair growth and growth comprising a hydrolyzed DNA fragment mixture as an active ingredient.

Another object of the present invention is to provide a quasi-drug, cosmetic composition, health functional food composition or pharmaceutical composition comprising the composition as an active ingredient.

A detailed description of this is as follows. Meanwhile, each description and embodiment disclosed in the present invention may also be applied to each other description and embodiment. That is, all combinations of the various elements disclosed herein fall within the scope of the present invention. In addition, it cannot be seen that the scope of the present invention is limited by the specific descriptions described below.

In addition, those skilled in the art will recognize, or be able to ascertain using no more than routine experimentation, many equivalents to the specific embodiments of the invention described herein. Also, such equivalents are intended to be included in this invention.

One aspect of the present invention provides a composition for promoting hair growth and growth comprising a hydrolyzed DNA fragment mixture as an active ingredient.

In the present invention, the term "hair growth" or "hair growth promotion" includes all activities that increase hair growth from hair follicles. For example, it may mean any action that increases the total amount of hair, such as the growth of hair follicle tissue, the proliferation of hair follicle cells, the activity of hair follicle cells, or the effect of promoting the growth of hair follicle cells. It also means that individual hairs are formed by rapidly differentiating cells and pushing other cells out of the hair follicle.

The term "hair" is an element constituting the skin, an organ with the largest area of the human body, and is structurally composed of cuticle, cortex, and medulla, and has a physical impact reduction as well as an aesthetic function. It plays a variety of roles, such as regulating body temperature, blocking ultraviolet rays, discharging waste products, and strengthening sensory abilities. The hair to which the composition of the present invention is applied includes hair roots and hair follicles of the head, hair, eyelashes, eyelashes, beard, armpits, pubic hair, and all parts of the body with hair roots and hair follicles.

Hair follicles can go through cyclical cycles of rest (telogen phase), regeneration (anagen phase), and regression (catagen phase). Hair follicle stem cells (HFSCs) exist in hair follicles and are involved in maintaining cell homeostasis of hair follicles. Hair follicle stem cells exhibit the ability to shuttle back and forth between proliferative and resting states in vivo . Various factors act on the hair cycle to cause hair loss and hair loss. When hair is induced into the anagen phase, hair follicle cells are rapidly differentiated and push other cells out of the hair follicle, thereby promoting the formation of individual hair.

The catagen phase of the hair follicle is a time when the hair that has passed the growth phase gradually faces the degeneration phase and the metabolic process slows down. Cell division or proliferation is reduced, and eventually hair growth stops. Therefore, reducing the catagen phase of hair follicles means that hair growth arrest and hair loss can be delayed.

In one embodiment of the present invention, the hair follicle elongation effect was confirmed by administering the composition of the present invention, and the composition of the present invention can be usefully used for promoting hair proliferation and growth.

In one embodiment, the composition for promoting hair proliferation and growth may be used to improve hair loss. In the present invention, the term "improvement" may mean any activity that at least reduces a parameter related to alleviation or treatment of a condition, for example, the severity of a symptom. In the present invention, the term "improvement of hair loss" may comprehensively mean a process or effect of treating, reducing, alleviating a hair loss condition.

In addition, in one embodiment of the present invention, the effect of reducing the catagen phase of hair follicles was confirmed by administering the composition of the present invention, and the composition of the present invention can be used to improve hair loss.

The composition may include the DNA fragment mixture in an amount of 5 parts by weight or more and 50 parts by weight or less based on 1 part by weight of the cationic additive.

The DNA fragment mixture in the composition may be present in the form of monodisperse particles.

The inventors of the present invention confirmed that a composition containing a DNA fragment mixture and a cationic additive in a specific amount contained the DNA fragment mixture in the form of monodisperse particles, and confirmed that such a composition promotes hair growth and hair growth. Provides a novel hair proliferation and/or growth promoting use of the composition of

In the present invention, "DNA fragment mixture" refers to a form in which DNA corresponding to a biopolymer composed of phosphoric acid, four bases, and deoxyribose is extracted and exists in a mixture in the form of DNA fragments having various molecular weights. do.

In one embodiment, the DNA fragment mixture may be in a hydrolyzed form.

In one embodiment, the DNA fragment mixture may be at least one selected from the group consisting of polydeoxyribonucleotide (PDRN) and polynucleotide.

In the present invention, the term "PDRN" refers to polydeoxyribonucleotide, which is a mixture of short deoxyribonucleotides. PDRN is a type of low molecular weight DNA complex made by fractionating a DNA chain into a certain size.

In the present invention, the term "polynucleotide" refers to a DNA or RNA strand of a certain length or longer as a polymer of nucleotides in which nucleotide monomers are connected in a chain shape by covalent bonds. For example, the polynucleotide may have a longer nucleic acid length or a higher molecular weight than polydeoxyribonucleotides, but is not limited thereto.

In any one of the above embodiments, the DNA fragment mixture of the present invention may have a molecular weight of about 1 to 100,000 kDa, specifically about 5 to 50,000 kDa, about 50 to 50,000 kDa, more specifically about It may be 50 to 10,000 kDa, but is not limited thereto.

In any one of the above embodiments, the DNA fragment mixture of the present invention may be a DNA fragment mixture obtained by extracting from fish testis or semen. Specifically, the fish may be salmon family fish. More specifically, it may be salmon or trout, but is not limited thereto.

In one embodiment, the cationic additive of the present invention may be at least one selected from the group consisting of cationic peptides, cationic lipids and cationic polymers.

For example, the cationic peptide may be at least one selected from the group consisting of polylysine, protamine, and KALA peptide (cationic fusogenic peptide).

For example, the cationic lipid is N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (N-[1-(2,3-dioleoyloxy)propyl ] -N,N,N-trimethylammonium chloride), 1,2-dioleoyloxy-3-(trimethylammonio)propane (1,2-dioleoyloxy-3-(trimethylammonio)propane), 1,2-dioleo Il-3- (4'-trimethylammonio) butanoyl-sn-glycerol (1,2-dioleoyl-3- (4'-trimethylammonio) butanoyl-sn-glycerol), 1,2-diacyl-3-dimethyl Ammonium-propane (1,2-diacyl-3-dimethylammonium-propane), 1,2-diacyl-3-trimethylammonium-propane (1,2-diacyl-3-trimethylammonium-propane), 1,2- Diacyl-sn-glycerol-3-ethylphosphocholine (1,2-diacyl-sn-glycerol-3-ethylphosphocholine), 3β-[N-(N',N'-dimethylaminoethane)-carbamoyl] It may be one or more selected from the group consisting of cholesterol (3β-[N-(N',N'-dimethylaminoethane)-carbamoyl]cholesterol, dimethyldioctadexylammonium bromide, and copolymers thereof.

For example, the cationic polymer may be at least one selected from the group consisting of chitosan, spermine, putrescine, cadaverine, and spermidine. .

In any one of the above embodiments, the cationic additive of the present invention is at least one selected from spermine, spermidine, and dimethyldioctadexylammonium bromide. it could be

In any one of the above embodiments, the cationic additive of the present invention may include spermine, but is not limited thereto.

The composition of the present invention may include a cationic additive and a DNA fragment mixture in a specific content ratio, so that the DNA fragment mixture exists in the form of monodisperse particles.

In one embodiment, the composition of the present invention may include 2.5 parts by weight or more of the DNA fragment mixture based on 1 part by weight of the cationic additive. Specifically, it may include about 3 parts by weight or more, about 4 parts by weight or more, about 4.5 parts by weight or more, or about 5 parts by weight or more of the DNA fragment mixture based on 1 part by weight of the cationic additive, but is not limited thereto. don't

The term “about” includes all ranges of ±0.5, ±0.4, ±0.3, ±0.2, ±0.1, etc., and includes all ranges equivalent to or similar to the ranges following the term “about”. Not limited.

Based on the content of the cationic additive, the upper limit of the content of the DNA fragment mixture is not particularly limited as long as the DNA fragment mixture is included in the form of monodisperse particles. For example, the content of the DNA fragment mixture based on 1 part by weight of the cationic additive may be about 200 parts by weight or less. In another example, the content of the DNA fragment mixture based on 1 part by weight of the cationic additive is about 100 parts by weight or less, about 50 parts by weight or less, about 40 parts by weight or less, about 35 parts by weight or less, about 30 parts by weight or less, about 25 parts by weight or less. or less, or about 20 parts by weight or less, but is not limited thereto.

In any one of the above embodiments, the composition of the present invention may include 0.01% to 5.0% by weight of the DNA fragment mixture based on the total composition.

As any one of the above embodiments, the composition of the present invention may include 0.0005% to 2.0% by weight of the cationic additive based on the total composition.

In the present invention, the term "monodisperse" refers to uniformity of size and shape. As an example, particles having a narrow size distribution in the manner of size distribution, such as the mean of the size distribution, may be referred to as "monodisperse". As an example, a series of polymer molecules having similar molecular weights may be referred to as “monodisperse”.

In any one of the above embodiments, the shape of the monodisperse particles may be a condensed form rather than an amorphous dispersion form, for example, a sphere.

In the present invention, the shape of a particle is "spherical" means that it is observed in a circular or spherical shape at a specific magnification on an electron microscope, and particles such as oval, toroid, irregularly shaped, etc. It is not meant to exclude other forms. That is, the term "spherical" to express the shape of a specific particle in the present invention does not necessarily mean that the particle has a shape in which sphericity is 1. In one embodiment, in the present invention, "spherical shape" " can be used as a term referring to a particle shape with a sphericity (equivalent circle diameter/maximum diameter) of 0.7 to 1.0. In one embodiment, the spherical particles of the present invention may be solid or hollow particles. However, it is not limited thereto.

In any one of the foregoing embodiments, the surface potential of the monodisperse particles may be about 0 mV to -60 mV. Specifically, it may be about 0 mV to -30 mV, but is not limited thereto.

In any one of the above embodiments, the monodisperse particle may have a structure in which a mixture of DNA fragments is condensed around a cationic additive forming a core. However, it is not limited thereto. In the present invention, the form in which the DNA fragment mixture is condensed may be used interchangeably with the terms 'condensed hydrolyzed DNA', 'condensed DNA fragment mixture', or 'n-PDRN'.

The composition of the present invention can be used as a cosmetic composition and can be formulated in various forms. In one embodiment, the cosmetic composition of the present invention is a product having professional functionality with an emphasis on physiological activity and effect, a product that helps improve the scalp, a product that helps hair growth, and promotes hair growth. Among the products that help to do so, it may be included in cosmetics determined by the Ordinance of the Ministry of Health and Welfare. However, it is not limited thereto.

In one embodiment, the cosmetic composition of the present invention is a solution, external ointment, cream, foam, nutrient lotion, softening lotion, pack, softening water, emulsion, makeup base, essence, soap, liquid cleanser, bath additive, sunscreen cream, sunscreen It can be prepared in a formulation selected from the group consisting of oil, suspension, emulsion, paste, gel, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation, patch and spray; , but not limited thereto.

More specifically, 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, hair oil, hair drying agent, hair preservative, hair dye, hair waving agent, hair bleach, hair gel, hair glaze, hair dressing, hair lacquer, hair moisturizer, It may be included in formulations such as hair mousse, hair spray, eyelash extensions, or mascara, but is not limited thereto.

In addition, the composition may be used as an external skin preparation. Specifically, creams, gels, patches, sprays, ointments, warning agents, lotions, liniment agents, pasta agents, or cataplasma agents may be manufactured and used as pharmaceuticals or quasi-drugs for external use on the skin, but are not limited thereto.

In any one of the foregoing embodiments, the composition of the present invention may further include a physiologically active substance.

The physiologically active substance is, for example, an antibody, protein, peptide, miRNA (microRNA), siRNA (small interfering RNA), aptamer, synthetic drug (chemical drug), etc. It may be, but is not limited thereto, and may further include an appropriate material depending on the intended use.

In any one of the foregoing embodiments, the composition of the present invention may further include one or more of a carrier, an excipient, and a diluent.

Examples of the carrier, excipient, and diluent include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, undecided quality cellulose, polyvinyl pyrrolidone, purified water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil; and the like. For example, when the composition of the present invention is formulated and used, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, and surfactants may be included in the composition of the present invention, but are not limited thereto.

Another aspect of the present invention provides a quasi-drug composition comprising a composition for promoting hair growth and hair growth, including the DNA fragment mixture provided in the present invention and a cationic additive.

In the present invention, the term "quasi-drugs" refers to items that are less active than pharmaceuticals among items used for the purpose of diagnosing, treating, improving, mitigating, treating or preventing human or animal diseases. For example, according to the Pharmaceutical Affairs Act of the Republic of Korea Quasi-drugs are items excluding items used for pharmaceutical purposes, and include products used for the treatment or prevention of diseases in humans and animals, and products with minor or no direct action on the human body.

In one embodiment, the quasi-drug composition of the present invention may be prepared in a form selected from the group consisting of body cleanser, shampoo, conditioner, foam, soap, mask, ointment, cream, lotion, essence and spray, but is not limited thereto no.

The composition for promoting hair growth and growth of the present invention may be used as a transdermal administration method such as direct application or spraying to the skin, but is not limited thereto.

As used herein, the term "administration" means introducing the composition of the present invention by any suitable method, and the administration route of the composition of the present invention can be administered through any general route as long as it can reach the target tissue. there is. Specifically, transdermal administration is possible, and among them, topical application is most preferred. The number of applications of the composition of the present invention may be determined according to prescription, necessity or desire. The amount of the composition of the present invention can be appropriately adjusted according to individual differences such as age, severity of lesions, etc., or dosage form, and it is preferable to apply an appropriate amount to the scalp once or several times a day and use it for one week to several months, but it is limited thereto. It doesn't work.

In addition, the composition may be used as a composition for raw materials for medical devices. For the purpose of the present invention, the medical device may include, but is not limited to, a wound dressing, a patch, an implantable medical device, and the like. The composition can be loaded into various medical devices.

In addition, the composition can be used as a health functional food composition. Since the food composition of the present invention can be consumed on a daily basis, it is very useful because it can be expected to promote hair proliferation and growth or prevent hair loss.

The food composition of the present invention includes forms such as pills, powders, granules, precipitates, tablets, capsules or liquids, and as foods to which the composition of the present invention can be added, for example, various foods, for example, There are beverages, chewing gum, tea, vitamin complexes, and health supplements.

There are no particular restrictions on other ingredients except for containing the Hwansam vine extract or a fraction thereof as an essential ingredient that can be included in the food composition of the present invention, and various herbal extracts, food additives, or natural carbohydrates are added as in conventional foods. It can be contained as a component. In addition, the food auxiliary additives include food auxiliary additives common in the art, such as flavoring agents, flavoring agents, coloring agents, fillers, stabilizers, and the like.

The health functional food (functional food) is the same term as food for special health use (FoSHU), and is a medicine processed to efficiently display bioregulatory functions in addition to nutritional supply, and has high medical effect. means food. Here, "function (sex)" means to obtain useful effects for health purposes such as regulating nutrients for the structure and function of the human body or physiological functions. The food of the present invention can be prepared by a method commonly used in the art, and can be prepared by adding raw materials and ingredients commonly added in the art during the preparation. In addition, the formulation of the food may also be prepared without limitation as long as the formulation is recognized as a food. The food composition of the present invention can be prepared in various types of formulations, and unlike general drugs, it has the advantage of not having side effects that may occur when taking drugs for a long time using food as a raw material, and has excellent portability, so that the composition of the present invention The food can be consumed as an adjuvant to enhance the effect of promoting hair proliferation and growth or preventing hair loss.

In addition, the composition may be used as a pharmaceutical composition for preventing or treating hair loss. In the present invention, the term “prevention” refers to all actions of suppressing or delaying hair loss by weakening the degenerative phase of the hair follicle and inducing the anagen phase of the hair follicle by the composition of the present invention. The term suppresses hair loss caused by a decrease in the specific gravity of anagen hair, and may have the same meaning as “relieving hair loss”, “improving hair loss”, and “treating hair loss”.

In the present invention, the term "treatment" refers to all activities in which the symptoms of hair loss are improved or beneficially changed by administration of the composition of the present invention.

In the present invention, the pharmaceutical composition can be prepared as a pharmaceutical.

In the present invention, the pharmaceutical composition may further include a pharmaceutically acceptable carrier.

In the present invention, the term "pharmaceutically acceptable carrier" refers to a carrier or diluent that does not stimulate living organisms and does not inhibit the activity and properties of preventing or treating hair loss of the composition of the present invention. In the composition formulated as a liquid solution, acceptable pharmaceutical carriers are sterile and biocompatible, and include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, and ethanol. And one or more of these components may be mixed and used, and other conventional additives such as antioxidants, buffers, and bacteriostatic agents may be added if necessary.

In addition, pharmaceutically acceptable carriers of the present invention may include non-natural carriers.

The pharmaceutical composition of the present invention can be administered in a pharmaceutically effective amount, the term "pharmaceutically effective amount" is sufficient to treat or prevent disease with a reasonable benefit / risk ratio applicable to medical treatment or prevention. It means the amount, and the effective dose level is the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the administration time of the composition of the present invention used, the route of administration and the excretion rate, the treatment period , It may be determined according to factors including drugs used in combination or simultaneous use with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a known therapeutic agent for pterygium. It is important to administer the amount that can obtain the maximum effect with the minimum amount without side effects in consideration of all the above factors.

The composition of the present invention has increased hair proliferation and hair growth promotion, and can be usefully used in various fields such as quasi-drugs, cosmetics, health functional foods, pharmaceuticals, and raw materials for medical devices.

1a and 1b show the results of analyzing the size distribution of a DNA fragment mixture using DLS.
Figure 2 shows the result of confirming the shape of the DNA fragment mixture using TEM.
3 shows human hair follicles isolated for hair transplantation.
Fig. 4(a) shows the anagen phase, early catagen phase, terminal catagen phase, damaged hair follicles and hair follicle organs including sebaceous glands, and Fig. 4(b) shows the morphological changes of the anagen phase and catagen phase hair follicles.
FIG. 5 shows the results of determining whether growth and catagen progresses according to concentration-specific n-PDRN treatment through hair follicle photography.
Figure 6 shows the proliferation effect of DP cells according to n-PDRN treatment by concentration.
7 shows the molecular weight distribution of the DNA fragment mixture used in the present invention.

Hereinafter, the present invention will be described in more detail through Examples and Experimental Examples. However, these Examples and Experimental Examples are intended to illustrate the present invention, and the scope of the present invention is not limited to these Examples and Experimental Examples.

Example 1: Preparation of composition comprising condensed DNA fragment mixture and confirmation of particle morphology of DNA fragment mixture

Hydrolyzed DNA (manufactured by Pharma Research Co., Ltd.) was used as the DNA fragment mixture, and spermine (Spermine-4HCl) was used as the cationic additive. The DNA fragment mixture used in the present invention shows a molecular weight distribution as shown in FIG. 7 . Specifically, as a result of profiling the size of the DNA fragment mixture using DNA size markers, it was confirmed that DNA fragments ranging in size from 15 bp to 20,000 bp or more were present in the mixture. Convert the molecular weight to kDa ( https://www.thermofisher.com/kr/ko/home/references/ambion-tech-support/rnatools-and-calculators/dna-and-rna-molecular-weights-and-conversions .html ), it can be seen that, for example, 20,000 bp is converted to approximately 12,150 kDa.

Purified water was added to the components having the contents shown in Table 1 below to prepare a 1ml composition.

Content (% by weight) Experimental example control group DNA Fragment Mixture 0.5 0.5 0.5 0.5 0.5 NaCl 0.9 0.9 0.9 0.9 0.9 Spermine-4HCl 0.01 0.05 0.1 0 0.2

The particle size distribution was confirmed by a dynamic light scattering method using a particle size analyzer, DLS. Specifically, the size of the Brownian motion sample having irregular motion in the colloidal state was measured by the diffusion coefficient, and the particle size was calculated by the following equation.

D=kT/6πηR

(D = diffusion coefficient, k = Boltzmann constant, T = temperature, η = solution viscosity, R = particle diameter)

Here, the particle distribution was confirmed by applying a temperature of 25°C and a scattering angle of 90 degrees. In order to compare the relative size of the particles, a DNA fragment mixture prepared without cationic additives (hereinafter referred to as Hydrolyzed DNA) and a DNA fragment mixture prepared with a relatively high content (0.2%) of cationic additives were used as a control group to compare the size of the particles. did The results of analyzing the size distribution of the DNA fragment mixture using DLS are shown in FIGS. 1a and 1b.

Since most of the DNA fragment mixtures generally have non-uniform base lengths, it was confirmed that the hydrolyzed DNA was polydisperse (FIG. 1a). However, when a cationic additive was used, it was confirmed that the content of the experimental example was monodisperse (Fig. 1b).

In addition, as a result of confirming the shape of DNA through TEM (FIG. 2), when the cationic additive was not used, the DNA had an irregular shape, whereas when the cationic additive was used as the content of the experimental example, the DNA was condensed. I was able to confirm.

The composition containing the condensed DNA fragment prepared in this example (DNA fragment mixture 0.5%, spermine salt 0.05%, NaCl 0.9% content; hereinafter referred to as condensed Hydrolyzed DNA, or n-PDRN) was tested in the following example. was used in

Example 2: Confirmation of hair follicle lengthening effect of a composition containing a mixture of condensed hydrolyzed DNA fragments

It was confirmed whether the composition containing the condensed DNA fragment mixture (n-PDRN) of the present invention was effective in hair follicle length growth.

As shown in Table 2 below, the hair follicle organ test group was a non-treated group (G1), and a positive control group with minoxidil 20μM (G2), n-PDRN 0.01% (G3), n-PDRN 0.1% (G4), and n-PDRN Experiments were conducted with 6 strands of hair follicles per group at 0.2% (G5). Minoxidil (cat. no. M4145, sigma), a positive control, was dissolved in DMSO, and the hair follicle organ was treated at a final concentration of 20 μM.

military name military explanation Treatment substance and concentration test
number of hair follicles G1 untreated group - 6 G2 positive control minoxidil 6 G3 Low concentration of test substance n-PDRN 0.01% 6 G4 Median concentration of test substance n-PDRN 0.1% 6 G5 High concentration of test substance n-PDRN 0.2% 6

The hair follicle tissue of the occipital biopsy was placed in saline and trimmed to the follicular unit, and additionally, the fat tissue around the hair follicle was removed as much as possible. As shown in FIG. 3, the hair follicle organ was cultured using only the lower part from which the upper part including the sebaceous gland was removed. The hair follicles shown in FIG. 3 represent human hair follicles isolated for hair transplantation.

Hair follicles were cultured by adding L-glutamin (2 mM), insulin (10 μg/ml), hydrocortisone (10 ng/ml), penicillin/streptomycin, and Fungizone (2.5 μg/ml) to Williams E medium. The hair follicle organ culture medium was cultured for up to 8 days by putting 1ml in a 24 well plate. The hair follicles were photographed using a stereomicroscope (OLYMPUS, SZ61) on days 0, 4, 6, and 8 of culture, and the hair follicle length was measured through the photos.

Statistical analysis of hair follicle length growth was performed using the Prism version 5.0 (GraphPad Software Inc., La Jola, CA, USA) program. In each experiment, the significance according to the sample application time was verified through the Wilcoxon signed rank test with the number of samples less than 10. Statistical significance before/after use between each sample was verified using the Mann-whitney U test, a non-parametric verification method.

As a result, as shown in Table 3 below, it was confirmed that the group treated with n-PDRN 0.1% showed the most similar hair follicle length growth as the positive control group.

G1 (untreated) G2 (Minoxidil 20μM) G3 (n-PDRN 0.01%) G4 (n-PDRN 0.1%) G5 (n-PDRN 0.2%) Day0 Total length(mm) 4.65 ± 0.20 4.75 ± 0.19 4.88 ± 0.10 4.91 ± 0.16 4.85 ± 0.11 Day4 Total length(mm) 5.14 ± 0.36 5.26 ± 0.38 5.31 ± 0.26 5.40 ± 0.26 5.22 ± 0.26 Elongation
(mm) 0.44 ± 0.21 0.44 ± 0.24 0.44 ± 0.17 0.46 ± 0.16 0.32 ± 0.21 Elongation (%) 9.01 ± 4.21
(-) 8.59 ± 4.76
(0.971) 8.32 ± 3.32
(0.193) 8.79 ± 3.07
(0.971) 6.28 ± 4.08
(0.28) Day6 Total length(mm) 5.33 ± 0.46 5.54 ± 0.45 5.47 ± 0.41 5.56 ± 0.32 5.36 ± 0.37 Elongation
(mm) 0.67 ± 0.36 0.84 ± 0.42 0.73 ± 0.45 0.87 ± 0.32 0.59 ± 0.45 Elongation (%) 13.66 ± 7.39
(-) 16.56 ± 8.23
(0.39) 13.98 ± 8.66
(0.57) 16.43 ± 6.07
(0.68) 11.57 ± 8.84
(0.31) Day 8 Total length(mm) 5.32 ± 0.50 5.52 ± 0.46 5.41 ± 0.48 5.65 ± 0.36 5.33 ± 0.34 Elongation
(mm) 0.67 ± 0.49 0.70 ± 0.42 0.59 ± 0.53 0.77 ± 0.34 0.55 ± 0.42 Elongation
(%) 13.70 ± 9.93
(-) 13.90 ± 8.30
(0.708) 11.15 ± 10.11
(0.436) 14.60 ± 6.44
(0.977) 10.63 ± 8.14
(0.286)

Total length (mm): total hair follicle length

Elongation (mm): Dx-D0 ± S.D.

Elongation (%): {(Dx-D0)/DO}*100 ± S.D.

Standard deviation (SD)

p-value: Mann-Whitney U test. **p ≤ 0.0001, comparison to G1 vs G2 or G3 or G4 or G5

In particular, as a result of observing the hair follicle growth rate on the 6th day of hair follicle culture, G1 was 13.91 ± 8.88%, G2 was 16.56 ± 8.23%, G3 was 13.98 ± 8.66%, G4 was 16.43 ± 6.07%, and G5 was 11.54 ± 8.84%. As a result, hair follicle length growth was observed in G2 (minoxidil) and G4 (n-PDRN 0.1%) compared to G1 (negative control).

When hair is induced into the anagen phase, hair follicle cells are rapidly differentiated and push other cells out of the hair follicle, thereby showing the effect of promoting the formation of individual hair. It can be seen that it is stimulating.

Example 3: Evaluation of hair follicle organ degeneration of a composition containing a mixture of condensed hydrolyzed DNA fragments

An experiment was conducted to evaluate the degree of catagen progression of the hair follicle organ of the composition containing the condensed DNA fragment mixture of the present invention.

Length growth was measured using the hair follicle organ used in Example 2, and hair follicle tissue was observed.

Hair follicles in the follicular organ regression phase were photographed using a stereomicroscope (OLYMPUS, SZ61) on days 0, 4, 6, and 8 of culture, and the hair follicle length was measured and the hair follicle tissue was observed in the photographed photos. Specifically, as shown in FIG. 4, (a): growth phase, early catagen phase, terminal catagen phase, damaged hair follicles and hair follicle organs including sebaceous glands, and (b): morphological changes of anagen phase hair follicles and catagen phase hair follicles were observed.

Growth phase and catagen progression were determined through hair follicle photography, and follicle period culture photos were shown in FIG. 5.

As a result of confirming the rate of catagen progression in the hair follicle organ culture evaluation, on the 6th day of hair follicle organ culture, G1 (untreated group) 33.33%, G2 (minoxidil) 50.00%, G3 (n-PDRN 0.01%) 66.67%, G4 (n-PDRN 0.01%) 66.67%, respectively. PDRN 0.1 %) 33.33%, G5 (n-PDRN 0.2 %) 16.67%, respectively, on the 8th day, G1 (untreated group) 66.67%, G2 (minoxidil) 66.70%, G3 (n-PDRN 0.01%) 66.70%, G4 (n-PDRN 0.1%) 50.00%, and G5 (n-PDRN 0.2%) 16.67%, respectively, compared to the non-treated group, the highest catagen inhibition was observed in n-PDRN 0.1% and n-PDRN 0.2%. .

Reducing the catagen phase of hair follicles indicates that hair growth arrest and hair loss can be delayed, and from the results of suppressing the catagen phase of the present invention, it can be seen that there is an effect of preventing hair loss.

Example 4: Evaluation of hair follicle cell proliferation of a composition containing a mixture of condensed hydrolyzed DNA fragments

Experiments were conducted to evaluate the degree of hair follicle cell proliferation of the composition containing the condensed DNA fragment mixture of the present invention.

Specifically, as a result of treating DP cells (Dermal Papila Cell; DPC), known as core cells responsible for hair growth, with PNRN concentrations of 0.01%, 0.02%, 0.03%, 0.05%, and 0.1%, respectively, at each concentration At a concentration of 108.25 ± 11.24, 110.15 ± 8.84, 111.37 ± 3.29, 134.55 ± 0.59, cell growth was observed in a concentration-dependent manner by n-PDRN treatment (FIG. 6).

The above results suggest that hair follicle growth is activated when hair follicle length growth is remarkably increased and at the same time the catagen inhibition of hair follicle organs acts in combination. Therefore, if the hair follicle length growth effect can be increased and the time of the degenerative phase of the hair follicle organ can be delayed, it can be helpful in treating and suppressing hair loss.

From the above description, those skilled in the art to which the present invention pertains will be able to understand that the present invention may be embodied in other specific forms without changing its technical spirit or essential features. In this regard, it should be understood that the embodiments described above are illustrative in all respects and not limiting. The scope of the present invention should be construed as including all changes or modifications derived from the meaning and scope of the claims to be described later and equivalent concepts rather than the detailed description above are included in the scope of the present invention.

Claims (13)

A composition for promoting hair growth and growth comprising a DNA fragment mixture having a molecular weight of 50 to 10,000 kDa and a cationic additive, wherein the composition includes 5 parts by weight or more and 50 parts by weight or less of the DNA fragment mixture based on 1 part by weight of the cationic additive. And, the DNA fragment mixture in the composition is present in the form of monodisperse particles.
The composition according to claim 1, wherein the particles are in a condensed form.
The composition of claim 1, wherein the content of the DNA fragment mixture of the composition is 0.01% to 5.0% by weight based on the total composition.
According to claim 1, wherein the content of the cationic additive of the composition is 0.0005% by weight to 2.0% by weight based on the total composition, the composition.
The composition according to claim 1, wherein the DNA fragment mixture is at least one selected from the group consisting of polydeoxyribonucleotides and polynucleotides.
The composition according to claim 1, wherein the DNA fragment mixture is isolated from fish testis or semen.
The composition according to claim 6, wherein the fish is salmonid.
According to claim 1, wherein the cationic additive is spermine (spermine), spermidine (spermidine) or dimethyldioctadecylammonium bromide (dimethyloctadexylammonium bromide) of one or more species selected from, the composition.
A cosmetic composition comprising the composition of claim 1 as an active ingredient.
A quasi-drug composition comprising the composition of claim 1 as an active ingredient.
A health functional food composition comprising the composition of claim 1 as an active ingredient.
A pharmaceutical composition for preventing or treating hair loss, comprising the composition of claim 1 as an active ingredient.
Condensing the DNA fragment mixture into monodisperse particles by mixing a DNA fragment mixture having a molecular weight of 50 to 10,000 kDa based on 1 part by weight of cationic additive in an amount of 5 parts by weight or more and 50 parts by weight or less, A method for preparing a composition for promoting hair growth and growth comprising a DNA fragment mixture and a cationic additive.

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