KR102777094B1 - Composition for improving blood circulation comprising extracts of Camilla japonica nectar - Google Patents

Abstract

The present invention relates to a composition for improving blood circulation including camellia japonica, wherein the camellia japonica has the effect of reducing vascular permeability induced by cytotoxic cytokines, the effect of producing collagen by fibroblasts, the effect of producing nitric oxide, and the effect of increasing the expression of vascular endothelial growth factor by hair papilla cells, thereby effectively improving blood circulation, and thereby not only improving or treating hair loss and dark circles, but also being non-irritating to the skin and having excellent safety for the human body, it can be usefully used in compositions such as cosmetics.

Description

Composition for improving blood circulation comprising extracts of Camilla japonica nectar

The present invention relates to a composition for improving blood circulation, including camellia japonica. More specifically, the present invention relates to a composition for improving blood circulation, including camellia japonica as an effective ingredient, which improves blood flow through the effects of reducing vascular permeability, increasing nitric oxide, increasing collagen, and increasing the expression of vascular endothelial growth factor in hair papilla cells. In addition, the present invention relates to a composition for preventing, improving, or treating hair loss or dark circles by the above-mentioned improvement of blood flow, etc.

Blood circulation is the circulation of blood, which means that blood moves to each part of the body to supply oxygen and nutrients to each organ and remove waste products. Blood cells that make up the blood and blood clotting factors in the plasma interact with each other to maintain blood circulation at a constant level, and this is achieved through the treatment of damaged areas and hemostasis by factors in the blood. In addition to the components of the blood, another part that affects blood circulation is the vascular endothelial cells that make up the inner wall of the blood vessels. The endothelium is a barrier between tissues and an endocrine organ that regulates relaxation, contraction, and proliferation of blood cells, and it is known to be able to locally regulate blood flow by regulating vascular tone. It is also involved in the regulation of inflammatory responses and adhesion, transfer, and activation of blood cells (Krㆌger-Genge et al., Int J Mol Sci. 2019 Sep, 10.3390/ijms20184411).

Poor blood circulation occurs when blood flow is obstructed by the accumulation of LDL cholesterol in the blood vessels, high blood sugar, high blood pressure, smoking, drugs, and stress, or when blood vessel elasticity naturally decreases due to aging. If poor blood circulation persists, symptoms such as memory loss, chronic fatigue, angina, cold hands and feet, and peripheral blood circulation disorders may appear, and in particular, a decrease in blood flow around the hair follicles in the scalp can cause hair loss (Deng et al., J Invest Dermatol. 2022 Aug, 10.1016/j.jid.2022.01.003). To improve this, blood circulation improving agents are sold on the market, and they have the effect of lowering blood circulation disorders by inhibiting the formation of blood clots or lipids, strengthening blood vessel walls to facilitate circulation, or strengthening the elasticity of blood vessel walls.

The barrier function of microvessels affects the physiological interactions between local metabolites. Cytotoxic cytokines such as TNF-α induce microvascular endothelial dysfunction, such as increasing permeability (vascular permeability) through irregular vasodilation or reducing vascular integrity. Therefore, the effect of alleviating microvascular hyperpermeability can improve blood flow circulation and contribute to the alleviation of vascular diseases such as edema.

Collagen is the most abundant protein in mammals and is a family of extracellular matrix proteins that play an important role in maintaining the structure of various tissues. Collagen is involved in the elasticity and motility of skin and blood vessels. In particular, when nutrients are transported in the blood, blood vessels are repeatedly damaged and restored. It is known that collagen binds to platelet receptors to activate platelet aggregation and adhesion and participate in the recovery of damaged blood vessel walls. The reason blood vessels become weaker as we age is also related to the decrease in collagen. Therefore, collagen production is widely discussed in research on strengthening blood vessels and improving blood flow.

Nitric oxide (NO) is a diatomic gas produced by vascular endothelial cells, and plays an important role in many physiological processes such as vascular tone, skeletal muscle contraction, and neurotransmission. Among them, it is known that blood vessels produce cGMP to relax the vascular wall, helping to improve blood flow, and inhibit platelet activity to lower blood clotting. As blood vessels age, the nitric oxide secretion ability of vascular endothelial cells decreases, which can cause vascular dysfunction along with a decrease in vascular elasticity. (Jones et al., Med Sci Sports Exerc. Feb, 10.1249/MSS.0000000000002470) Therefore, research to increase vascular nitric oxide is becoming a target for improving blood flow.

In terms of skin beauty, improved blood circulation may be related to alleviating alopecia and dark circles.

Hair loss was thought to occur more frequently with aging, but the actual onset age was 50% in their 20s and 30s, 29.8 years old and 33.6 years old for men and women, respectively (Health Insurance Review and Assessment Service). Causes of hair loss include the use of chemical products such as dyeing and perms, side effects of medication, hormonal changes, imbalanced eating habits, and stress. Hair repeats growth and regression through a certain cycle (hair cycle), which is the growth phase (anagen) in which hair grows, the regression phase (catagen) in which the metabolic process slows down and the hair bulb shrinks, and the resting phase (telogen) in which old hair falls out. It is known that if the growth phase is relatively shortened or the regression or resting phase is long, hair loss (alopecia) occurs (Marlon R Schneider et al., Curr Biol. 2009 Feb, 10.1016/j.cub.2008.12.005).

A representative hair loss prevention agent is Minoxidil, an FDA-approved topical skin agent. Minoxidil is a potent vasodilator that opens potassium channels in the smooth muscles of peripheral arteries and causes hyperpolarization of cell membranes. It is known to promote hair growth by increasing blood circulation in the scalp, increasing blood flow and nutrient supply. In addition, there are results showing that Minoxidil activates the signal of beta-catenin to promote hair follicle proliferation, thereby extending the growth phase of dermal papilla cells, and increasing the expression of vascular endothelial growth factor to increase the proliferation of hair follicle cells (Suchonwanit et al., Drug Des Devel Ther., 2019 Aug, 10.2147/DDDT.S214907. Mi Hee Kwack et al., J Dermatol Sci., 2011 Jun, 10.1016/j.jdermsci.2011.01.013). However, minoxidil can cause skin problems such as rashes and itchiness when used, and may have side effects such as hair growth in areas other than the scalp.

Dark circles are relatively dark areas under the eyes, and are one of the major aesthetic concerns along with hair loss. Dark circles can be largely divided into three types depending on their causes: vascular dark circles caused by poor blood circulation, pigmentation dark circles caused by excessive pigmentation, and structural dark circles caused by deep sunken areas under the eyes (Huang et al., Int J Dermatol, 2014 Feb, 10.1111/j.1365-4632.2012.05701.x.). In particular, when blood circulation is not smooth and blood stagnates under the eyes, the supply of nutrients becomes insufficient, causing a deterioration of complexion due to oxygen deficiency and the deposition of a pigment called hemosiderin in the blood on the skin, forming dark circles. Or, vascular permeability increases due to decreased adhesion between capillary endothelial cells caused by aging and stimuli such as ultraviolet rays, which delays blood flow and creates dark circles.

Treatment methods for dark circles include surgical methods to reposition fat under the eyes or laser treatments to shrink dilated blood vessels under the eyes and skin. In the case of dark circles caused by excessive melanin pigmentation, vitamin C penetration treatment is also performed (Friedmann et al., Clin Plast Surg, 2015 Jan, 10.1016/j.cps.2014.08.007). However, surgical treatments can affect daily life and are expensive, making them difficult to choose.

Meanwhile, nectar is honey produced in flowers and is known to originate from phloem sap. When sugar compounds are produced by photosynthesis, they accumulate in parenchyma cells located near nectar glands. Parenchyma cells absorb water as osmotic pressure increases, are hydrolyzed, and secreted into nectar glands, which is called nectar. Nectar mainly contains water and sugar components, and also contains some amino acids, organic acids, proteins, fats, vitamins, minerals, and secondary metabolites. Flowering plants provide nectar and pollen as a reward for nutrients to attract insect pollinators as a means of pollination and fertilization. Plants develop various forms of flower structure, size, scent, and color to attract pollinators, and the nutritional components and composition of nectar and pollen also differ depending on the species. Pollen is released only once from flowers, but nectar is a renewable floral resource that is secreted or replenished daily in long-lived flowers. The amount and pattern of secreted nectar, the content and composition of free sugars, are different depending on the harvesting time and tree species, so the selection of tree species is one of the important factors in collecting nectar. According to recent research results, the amount of nectar secreted per flower of Camellia japonica was 357.1㎕, and it was evaluated as a major nectar source in the evaluation of nectar sources.

Honey is nectar produced by flowers, collected by bees, converted by their own enzymes, and stored in the honeycomb. Nectar is mainly composed of sucrose, a disaccharide, which is stored in the honey sac of bees and then hydrolyzed by the enzyme into glucose and fructose, which are monosaccharides, and stored. Since nectar is absorbed and processed by bees, it shows differences not only in sugar content but also in other components, and thus shows many differences in physiological activity.

There have been various studies on the cosmetic use of honey, but there has been little research on nectar. For example, LVMH RECHERCHE in France studied whether nectar from Trifolium repens or its extracts are useful for the proliferation of fibroblasts in the skin (Korean Patent No. 10-1844598), and the Korea Beauty Industry Promotion Agency studied whether a mixed extract of green tangerine, turmeric, and citrus flower honey relieves inflammation and has an antibacterial effect on C. acnes , a type of acne bacteria (Korean Patent No. 10-2228257).

Accordingly, the inventors of the present invention selected camellia flowers, which mainly contain nectar, and conducted research on their physiological activities, and discovered that the camellia nectar developed through this research has high stability and safety, has no skin side effects, and can simultaneously exhibit the efficacy of improving blood flow by decreasing vascular permeability, increasing nitric oxide, increasing collagen, and increasing vascular endothelial growth factor of hair papilla cells, thereby revealing that it is a useful ingredient for improving blood circulation, and completing the present invention.

One object of the present invention is to provide a composition for improving blood circulation containing camellia militaris as an effective ingredient.

Another object of the present invention is to provide a composition for preventing, improving, or treating hair loss or dark circles, comprising camellia japonica extract as an effective ingredient.

Another object of the present invention is to provide a composition for improving blood circulation or preventing, improving, or treating hair loss or dark circles as a cosmetic, pharmaceutical, skin external preparation, or food composition.

Another object of the present invention is to provide a method for improving blood circulation in a subject by administering the composition for improving blood circulation to the subject.

Another object of the present invention is to provide a method for preventing, improving, or treating hair loss or dark circles in a subject by administering to the subject a composition for preventing, improving, or treating hair loss or dark circles.

The present invention relates to a composition for improving blood circulation, which contains camellia japonica as an effective ingredient. More specifically, the present invention relates to a composition for improving blood circulation, which contains camellia japonica as an effective ingredient, which is excellent in human safety as it does not cause skin irritation and cytotoxicity, and which can improve and treat blood circulation by regulating blood circulation by increasing blood flow through inhibition of vascular permeability, increasing collagen, and promoting nitric oxide production.

In addition, the present invention relates to a composition for preventing, improving, or treating hair loss or dark circles, comprising camellia japonica as an effective ingredient. More specifically, the present invention relates to a composition for preventing, improving, or treating hair loss or dark circles, comprising camellia japonica as an effective ingredient, which is excellent in human safety as it is free of skin irritation and cytotoxicity, and can prevent, improve, or treat hair loss or dark circles by improving blood circulation, increasing proliferation of hair papilla cells, and increasing expression of vascular endothelial growth factor of hair papilla cells, as described above.

In the present invention, the term "camellia nectar" refers to nectar derived from camellia. Nectar refers to a liquid substance secreted from the nectary glands to attract insects such as honeybees, bees, moths, and animals such as magpies and hummingbirds to reproduce, and is a different substance from honey (honey) that honeybees collect from various plants, convert with their own enzymes, and store in their hives.

The above term "camellia" refers to the flower of the camellia tree ( Camellia japonica ) belonging to the Theaceae family.

The present inventors have discovered that camellia honey has excellent effects in reducing excessive vascular permeability, increasing the production of nitric oxide and collagen, which contribute to vascular tone and elasticity, increasing the expression of vascular endothelial growth factor in hair papilla cells, and increasing the proliferation of hair papilla cells.

Therefore, camellia honey is very useful as an effective ingredient in a composition for improving blood circulation. In addition, camellia honey is very useful as an effective ingredient in a composition for preventing and improving hair loss and preventing and improving dark circles due to the effect of improving blood circulation.

The camellia nectar of the present invention may be commercially produced or collected from camellia. The collection may be performed, for example, on the second or third day after the camellia has fully bloomed.

In the present invention, the term "improving blood circulation" means improving blood circulation disorders that occur due to blood flow obstruction caused by the accumulation of LDL cholesterol in blood vessels, hyperglycemia, high blood pressure, smoking, drugs, stress, or decreased blood vessel elasticity due to aging, by inhibiting the accumulation of lipids in blood vessels or increasing the barrier function and elasticity of blood vessels. Therefore, "improving blood circulation" of the present invention can be considered to mean inhibiting the production and accumulation of lipids such as blood clots or cholesterol in blood vessels and reinforcing the structure or elasticity of blood vessel walls to facilitate or improve circulation. In addition, the "improving blood circulation" can be used herein as blood flow regulation or improvement.

The above improvement in blood circulation can be achieved by the effective ingredient of the present invention, camellia japonica, causing an inhibitory effect on vascular permeability induced by cytotoxic cytokines, an increase in nitric oxide, and an increase in collagen production, thereby improving blood flow.

In the present invention, the term "dark circle" refers to a perobital dark circle around the eye, in which the skin color around the eye becomes relatively darker than that of other parts of the face.

The above dark circles can be prevented and improved by alleviating vascular permeability around the eyes and facilitating blood circulation using camellia militaris, an effective ingredient of the present invention, thereby preventing and improving excessive pigmentation.

In the present invention, hair loss refers to a state in which there is no hair in an area where hair should normally exist, and specifically refers to the loss of terminal hair (thick, black hair) on the scalp.

The above hair loss can be prevented and improved by the effective ingredient of the present invention, camellia japonica, by alleviating the vascular permeability of the scalp and improving blood flow, as well as by promoting the proliferation of hair papilla cells and the expression of vascular endothelial growth factors in hair papilla cells.

In the present invention, the term "prevention" refers to reducing the possibility of dark circles and/or hair loss occurring.

In the present invention, the term "improvement" refers to regulating blood flow in blood vessels to be smooth by inhibiting vascular permeability and increasing vascular elasticity, angiogenesis, etc., and restoring blood circulation to a healthy state, reducing the area or degree of hair loss, and/or facilitating blood circulation around the eyes and reducing the degree of pigmentation. The above "improvement" may also have the same meaning as "treatment" in the present specification.

In the present invention, the cells include all cells existing within an individual ( in vivo ) or cultured outside an individual ( in vitro ). The cells can be classified in various ways based on cell shape and location, and examples thereof include skin cells, hair cells, fat cells, stem cells, vascular cells, immune cells, etc.

In one specific example, the degree of decrease in vascular permeability according to the concentration of camellia japonica honey of the present invention was examined by increasing vascular permeability with cytotoxic cytokines in human capillary endothelial cells. As a result, it was found that vascular permeability was significantly reduced by camellia japonica honey, confirming that camellia japonica honey has the effect of inhibiting vascular permeability increased by cytotoxic cytokines.

In another specific example, the degree of promotion of collagen synthesis according to the concentration of camellia japonica in human fibroblasts was examined, and collagen synthesis was found to increase, confirming that camellia japonica has an effect of promoting collagen synthesis in relation to blood vessel elasticity or strengthening.

In another specific example, the effect of nitric oxide production from immune macrophages was examined, and it was found that nitric oxide production was increased by camellia honey, confirming that camellia honey has the effect of promoting nitric oxide production.

In another specific example, the degree of cell proliferation and vascular endothelial growth factor expression was confirmed according to the concentration of camellia japonica in human breast papilla cells, and it was confirmed that the proliferation of breast papilla cells and the expression of vascular endothelial growth factor were increased.

Camellia japonica honey contained in the above composition is not particularly limited thereto, but may be included in an amount of 0.0001 to 20.0 wt%, preferably 0.001 to 10.0 wt%, based on the total composition weight. If the content of camellia japonica honey is less than 0.0001 wt%, the blood circulation improvement effect, hair loss prevention and improvement effect, and dark circle prevention and improvement effect cannot be expected, and if it exceeds 20.0 wt%, the increase in the blood circulation improvement effect due to the increase in the content is minimal, and difficulty occurs in manufacturing the formulation.

As one specific embodiment, the blood circulation improving composition of the present invention can be provided as a cosmetic composition for improving blood circulation, preventing and improving hair loss, or preventing and improving dark circles, containing camellia japonica as an effective ingredient.

The above camellia honey, its blood circulation improving effect, its hair loss prevention and improvement effect, and its dark circle prevention and improvement effect are as described above.

The cosmetic composition of the present invention can be used for the purpose of improving blood circulation in the skin, preventing and improving hair loss, and/or preventing and improving dark circles. Specifically, the composition can be applied to a subject suffering from edema, numbness in the extremities, cold extremities, stiff shoulders and neck, hair loss, dark circles, etc. due to decreased blood circulation in the skin capillaries.

The above cosmetic composition may additionally contain ingredients commonly used in cosmetic compositions in addition to the effective ingredient, camellia japonica. For example, it may contain conventional auxiliary agents and carriers such as antioxidants, stabilizers, solubilizers, vitamins, pigments, and fragrances.

The above cosmetic composition can be manufactured into any formulation commonly manufactured in the art. For example, it can be formulated into a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant-containing cleansing, oil, powder foundation, emulsion foundation, wax foundation or spray. More specifically, it can be manufactured into a formulation such as a nourishing cream, astringent toner, flexible toner, lotion, essence, nourishing gel or massage cream. However, it is not particularly limited thereto.

When the formulation of the above cosmetic composition is a paste, cream or gel, animal oil, vegetable oil, wax, paraffin, starch, tragacanth, cellulose derivatives, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

When the formulation of the above cosmetic composition is a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane/butane or dimethyl ether may be additionally included.

When the formulation of the above cosmetic composition is a solution or emulsion, a solvent, a solubilizer or an emulsifier may be used as a carrier component. For example, water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan may be used.

When the formulation of the above cosmetic composition is a suspension, liquid diluents such as water, ethanol or propylene glycol, suspending agents such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as carrier components.

When the formulation of the above cosmetic composition is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkyl amidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, lanolin derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

Camellia japonica honey contained in the above cosmetic composition is not particularly limited thereto, but may be included in an amount of 0.0001 to 20.0 wt%, preferably 0.001 to 10.0 wt%, based on the total composition weight. If the content of camellia japonica honey is less than 0.0001 wt%, the blood circulation improvement effect, hair loss prevention and improvement effect, and dark circle prevention and improvement effect cannot be expected, and if it exceeds 20.0 wt%, the increase in the above effects due to the increase in the content is minimal, and it becomes difficult to manufacture a cosmetic formulation.

As another specific embodiment, the present invention can be provided as a pharmaceutical composition for improving or treating blood circulation, preventing and treating hair loss, or preventing and treating dark circles, containing camellia militaris as an effective ingredient.

The above camellia honey, its blood circulation improving effect, its hair loss prevention and treatment effect, and its dark circle prevention and treatment effect are as described above.

The term "pharmaceutical composition" of the present invention means a composition manufactured for the purpose of preventing or treating a disease, and may be formulated and used in various forms according to conventional methods. For example, it may be formulated as an oral formulation such as a powder, granule, tablet, capsule, suspension, emulsion, syrup, aerosol, etc., and may be formulated and used as a parenteral formulation using a diluent or excipient such as a lubricant, a wetting agent, a flavoring agent, an emulsifier, a suspending agent, a preservative, a surfactant, etc. In addition, it may be formulated and used in the form of an external preparation, a suppository, an external skin preparation, and a sterile injection solution.

As a specific example, the composition of the present invention can be used in an oral dosage form for oral administration. As another specific example, the composition of the present invention can be used by methods such as direct application or spraying on skin areas where edema, numbness of the extremities, cold extremities, stiff shoulders and neck, etc. occur, hair or scalp where hair loss occurs, and around the eyes where dark circles occur.

In addition, depending on each formulation, it can be manufactured by additionally including pharmaceutically acceptable carriers, such as buffers, analgesics, solubilizers, isotonic agents, stabilizers, and carriers known in the art.

The term "pharmaceutically acceptable carrier" used in the present invention may mean a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity and properties of the active ingredient to be injected. The type of the carrier usable in the present invention is not particularly limited, and any carrier that is commonly used in the relevant technical field and is pharmaceutically acceptable may be used. Non-limiting examples of the carrier include saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol, and the like. These may be used alone or in combination of two or more. The carrier may be a non-naturally occurring carrier.

The composition of the present invention can be administered in a pharmaceutically effective amount. The pharmaceutically effective amount means an amount sufficient to treat a disease at a reasonable benefit/risk ratio applicable to medical treatment and not causing side effects, and the effective dosage level can be determined according to factors including the patient's health condition, the type and severity of the disease, the activity of the drug, the sensitivity to the drug, the administration method, the administration time, the administration route and the excretion rate, the treatment period, the drugs used in combination or simultaneously, and other factors well known in the medical field. Specifically, the dosage is generally 0.01 mg to 5000 mg per 1 kg of the body weight of the administered subject per day, and may be administered once or several times a day at regular intervals depending on the judgment of a doctor or pharmacist, but is not limited thereto.

Camellia japonica honey contained in the above composition is not particularly limited thereto, but may be included in an amount of 0.0001 to 20.0 wt%, preferably 0.001 to 10.0 wt%, based on the total composition weight. If the content of camellia japonica honey is less than 0.0001 wt%, the blood circulation improvement effect, hair loss prevention and treatment effect, and dark circle prevention and treatment effect cannot be expected, and if it exceeds 20.0 wt%, the increase in the above effects due to the increase in the content is minimal, and difficulty occurs in manufacturing the formulation.

In addition, the pharmaceutical composition of the present invention can be used alone or in combination with other pharmaceutically active compounds that exhibit the effects of improving blood circulation, preventing or treating hair loss, or forming an appropriate group.

The pharmaceutical composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. And may be administered singly or in multiple doses. It is important to administer an amount that can achieve the maximum effect with the minimum amount without causing side effects by considering all of the above factors, and this can be easily determined by those skilled in the art.

In the present invention, the term "administration" means introducing the pharmaceutical composition of the present invention into a subject by any appropriate method, and the route of administration of the composition of the present invention may be through various routes, such as oral or parenteral, as long as it can reach the target tissue.

The method of administration of the pharmaceutical composition according to the present invention is not particularly limited, and may follow a method commonly used in the relevant technical field. As a non-limiting example of the above-mentioned method of administration, the composition may be administered orally or parenterally.

The pharmaceutical composition according to the present invention can be prepared in various dosage forms depending on the intended administration method.

The frequency of administration of the composition of the present invention is not particularly limited, but may be administered once a day or administered several times in divided doses.

In another specific embodiment, the present invention provides a skin external composition for improving or treating blood circulation, preventing, improving, or treating hair loss, or preventing, improving, or treating dark circles, comprising camellia japonica extract as an effective ingredient.

The above camellia honey, its blood circulation improving effect, its hair loss prevention and improvement effect, and its dark circle prevention and improvement effect are as described above.

The composition for external application of the skin according to the present invention can be formulated containing a cosmetically or dermatologically acceptable medium or base. It can be provided in any formulation suitable for topical application, for example, in the form of a solution, a gel, a solid, a pasty anhydrous product, an emulsion obtained by dispersing an oil phase in an aqueous phase, a suspension, a microemulsion, a microcapsule, a microgranule, or a vesicular dispersion of ionic (liposomes) and nonionic types, or in the form of a cream, a skin, a lotion, a powder, an ointment, a spray, a pack or a concealer stick. It can also be used in the form of a foam or in the form of an aerosol composition further containing a compressed propellant. These compositions can be prepared according to conventional methods in the art.

In addition, the skin external composition according to the present invention may contain adjuvants commonly used in the fields of cosmetology or dermatology, such as fatty substances, organic solvents, solubilizers, thickeners, gelling agents, emollients, antioxidants, suspending agents, stabilizers, foaming agents, fragrances, surfactants, water, ionic or nonionic emulsifiers, fillers, metal ion sequestering agents, chelating agents, preservatives, vitamins, blocking agents, humectants, essential oils, dyes, pigments, hydrophilic or lipophilic active agents, lipid vesicles or any other ingredients commonly used in cosmetics. The adjuvants are introduced in an amount commonly used in the fields of cosmetology or dermatology.

Camellia japonica honey contained in the above composition is not particularly limited thereto, but may be included in an amount of 0.0001 to 20.0 wt%, preferably 0.001 to 10.0 wt%, based on the total composition weight. If the content of camellia japonica honey is less than 0.0001 wt%, the blood circulation improvement effect, hair loss prevention and improvement effect, and dark circle prevention and improvement effect cannot be expected, and if it exceeds 20.0 wt%, the increase in the above effects due to the increase in the content is minimal, and difficulty occurs in manufacturing the formulation.

In another specific embodiment, the composition for improving blood circulation of the present invention provides a food composition for improving blood circulation, preventing and improving hair loss, or preventing and improving dark circles, containing camellia japonica as an effective ingredient.

The above camellia honey, its blood circulation improving effect, its hair loss prevention and improvement effect, and its dark circle prevention and improvement effect are as described above.

When the composition of the present invention is used as a food additive, the camellia militaris can be added as is or used together with other foods or food ingredients, and can be used appropriately according to a conventional method. The mixing amount of the effective ingredient can be appropriately determined according to the purpose of use (prevention, health, or improvement treatment), and a food-related acceptable food auxiliary additive can be additionally included. Since the composition of the present invention uses a substance derived from a natural product as an effective ingredient, there is no problem in terms of stability, so there is no significant limitation on the mixing amount.

The food composition of the present invention can include all foods in the conventional sense, and can be used interchangeably with terms known in the art, such as functional food and health functional food.

The term "functional food" of the present invention means a food manufactured and processed using raw materials or ingredients having functionality useful to the human body, and "functionality" means consumption for the purpose of obtaining a useful effect for health purposes such as regulating nutrients for the structure and function of the human body or physiological effects.

In addition, the term "health functional food" of the present invention refers to a food manufactured or processed by using a specific ingredient as a raw material or by extracting, concentrating, refining, mixing, etc. a specific ingredient contained in a food raw material for the purpose of health supplementation, and refers to a food designed and processed so that the above-mentioned ingredient can sufficiently exert a bioregulatory function, such as biological defense, biological rhythm regulation, disease prevention and recovery, on the body, and the above-mentioned health food composition can perform functions related to disease prevention and disease recovery.

There is no limitation on the types of foods in which the composition of the present invention can be used for improving blood circulation, preventing and improving hair loss, or preventing and improving dark circles. In addition, the composition containing the camellia militaris of the present invention as an active ingredient can be prepared by mixing other appropriate auxiliary ingredients that can be contained in foods and known additives according to the selection of a person skilled in the art. Examples of foods to which the composition can be added include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes, and can be prepared by adding the extract according to the present invention and the fraction thereof as a main ingredient to juice, tea, jelly, and juice.

In addition, foods applicable to the present invention may include all foods, such as special nutritional foods (e.g., formulated milk, infant/toddler food, etc.), processed meat products, fish products, tofu, starch jelly, noodles (e.g., ramen, noodles, etc.), health supplements, seasoned foods (e.g., soy sauce, soybean paste, red pepper paste, mixed sauce, etc.), sauces, confectionery (e.g., snacks), processed dairy products (e.g., fermented milk, cheese, etc.), other processed foods, kimchi, pickled foods (various kimchi types, pickled vegetables, etc.), beverages (e.g., fruit and vegetable beverages, soy milk, fermented beverages, etc.), natural seasonings (e.g., ramen soup, etc.).

The content of camellia militaris included in the above food composition is not particularly limited thereto, but may be included in an amount of 0.00001 to 100 wt%, preferably 0.0001 to 20 wt%, more preferably 0.001 to 15 wt%, and even more preferably 0.001 to 10 wt% based on the total weight of the composition.

In another aspect, the present invention provides a method for improving blood circulation in a subject by administering to the subject a composition for improving blood circulation containing camellia militaris as an effective ingredient.

In addition, the present invention provides a method for preventing, improving, or treating hair loss or dark circles by administering to a subject a solution containing camellia japonica extract as an effective ingredient for preventing, improving, or treating hair loss or dark circles.

As described above, the composition for improving blood circulation contains camellia japonica as an active ingredient, and is a composition that improves blood circulation in the skin and scalp by causing the effect of reducing vascular permeability, promoting collagen synthesis, producing nitric oxide, and increasing the expression of vascular endothelial growth factor in hair papilla cells through the effective ingredient camellia japonica.

In addition, the composition for preventing, improving, or treating hair loss or dark circles is a composition that can prevent, improve, and treat hair loss and dark circles by improving blood circulation in the skin and scalp and promoting proliferation of hair papilla cells and expression of vascular endothelial growth factors.

In the present invention, the term "subject" means a mammal including a human, such as a monkey, cow, horse, pig, sheep, dog, cat, rat, mouse, chimpanzee, etc., which can achieve the purpose of improving blood circulation, preventing, improving, and treating hair loss and dark circles by administering the composition for improving blood circulation of the present invention, or the composition for preventing, improving, or treating hair loss or dark circles.

In the present invention, the term "administration" means introducing a given substance into a subject by any suitable method, and the route of administration of the composition of the present invention may be oral or parenteral administration through any common route as long as it can reach the target tissue. In addition, the composition of the present invention may be administered by any device through which the active substance can travel to a target object, for example, a cell.

The appropriate dosage of the composition of the present invention varies depending on the condition and weight of the subject, the degree of the disease, the formulation, and the time, but can be appropriately selected by a person skilled in the art. The daily dosage of the composition for improving blood circulation is 1.0 to 3.0 ml for adults, and can be administered by application 1 to 5 times a day as needed, but is not necessarily limited thereto.

Meanwhile, camellia honey according to the present invention is a natural substance, harmless to the human body, and has almost no toxicity or side effects, so it can be used safely even for long-term use, and thus can be safely applied to cosmetics, pharmaceuticals, external skin preparations, and food compositions containing it.

Camellia japonica, an effective ingredient of the present invention, does not irritate the skin and has excellent safety for the human body, while simultaneously having the effects of inhibiting vascular permeability increased by cytotoxic cytokines, increasing collagen synthesis, promoting nitric oxide production, promoting proliferation of hair papilla cells, and increasing expression of vascular endothelial growth factor in hair papilla cells, and thus can be usefully used as a cosmetic, pharmaceutical, external skin preparation, or food composition for improving blood flow that effectively improves blood flow. In addition, the composition for improving blood flow can prevent and improve hair loss and also prevent and improve dark circles due to the effects of improving blood flow, proliferation of hair papilla cells, and increasing expression of vascular endothelial growth factor.

Figure 1 shows the results of confirming the cytotoxicity of camellia japonica in human capillary endothelial cells according to one embodiment of the present invention.
Figure 2 is a result showing the effect of camellia japonica extract on inhibiting high vascular permeability induced by TNF-α in human capillary endothelial cells according to one embodiment of the present invention.
Figure 3 is a result showing the effect of camellia japonica extract on promoting collagen synthesis in human fibroblasts according to one embodiment of the present invention.
Figure 4 is a result showing the effect of camellia japonica honey on increasing nitric oxide production in immune macrophages according to one embodiment of the present invention.
FIG. 5 is a result showing the effect of camellia japonica extract on increasing the expression of vascular endothelial growth factor in human breast papilla cells according to one embodiment of the present invention.
Figure 6 shows the results of confirming the cell proliferation effect of camellia japonica in human breast papilla cells according to one embodiment of the present invention.

Hereinafter, in order to help understand the present invention, examples and the like will be described in detail. However, the examples according to the present invention may be modified in various different forms, and the scope of the present invention should not be construed as being limited to the following examples. The examples of the present invention are provided to more completely explain the present invention to a person having average knowledge in the art.

Example 1: Preparation of camellia honey

Camellia flowers were harvested in the morning on the 2nd or 3rd day, when they began to bloom and were fully bloomed. Immediately after harvest, the flowers were cut in half, and nectar was collected from the nectar glands inside the flowers using a pipette. The nectar was heat-treated at 80°C for 30 minutes and then filtered through filter paper (Advantec, No. 2) to produce camellia nectar.

Example 2: Measurement of cytotoxicity of Camellia sinensis on capillary endothelial cells

To investigate the effect of Camellia sinensis on the proliferation of capillary endothelial cells, the following experiment was conducted. Human dermal microvascular endothelial cells (HMEC-1) were purchased from ATCC (American Tissue Culture Collection, USA) and prepared, and cultured in MCDB131 medium supplemented with 10% FBS (Fetal Bovine Serum), glutamine, EGF, and hydrocortisone. Specifically, 2 × 10 ⁴ cells/well of capillary endothelial cells HMEC-1 were seeded in a 24-well plate and cultured at 37℃ in a CO ₂ incubator with a 5% concentration for 24 hours. Afterwards, the same medium was replaced with the medium treated with the camellia japonica extract of Example 1 at different concentrations (0.01 v/v%, 0.1 v/v%, and 1 v/v%), and cultured again for the next 24, 48, and 72 hours. After that, the medium was replaced with MCDB131 medium containing MTT (1 μg/ml) and reacted for 3 hours. After the reaction, the medium was removed, 0.5 ml of DMSO was added to each well to dissolve the formazan formed within the cells, and the absorbance was measured at 570 nm, and the cell proliferation rate was calculated using the following empirical formula 1.

[Experimental Formula 1]

Cell proliferation rate (Proliferation Pattern, %) = [(Absorbance value of treated sample) / (Absorbance value of control group) × 100]

As shown in Fig. 1, it was confirmed that camellia honey according to the present invention is not toxic to capillary endothelial cells.

Example 3: Evaluation of the efficacy of Camellia sinensis extract on vascular permeability reduction

In order to investigate the effect of Camellia sinensis on vascular permeability inhibition, the following experiment was conducted. To evaluate vascular permeability, HMEC-1 capillary endothelial cells (2 × 10 ⁵ cells/well) were placed in the cell insert of a 24-well plate and attached. After 72 hours, the medium was replaced with 1% FBS, and 200 ng/ml TNF-α, which induces an increase in vascular permeability, and HRP to check the degree of reaction were added. After treating with Camellia sinensis at a concentration that had no cytotoxic effect in Example 2, the cells were cultured for 24 hours. After that, the culture medium was collected from the plate well outside the insert. TMB was added to the harvested medium, and the absorbance was measured at 450 nm, and the results are shown in Fig. 2. Here, the positive control group is a group treated with TNF-α alone.

Figure 2 is a graph showing the vascular permeability of capillary endothelial cells by Camellia japonica extract. As seen here, compared to the group treated with TNF-α, which is a positive control group, it was confirmed that the high vascular permeability induced by TNF-α was significantly alleviated in capillary endothelial cells treated with 0.1% and 1% Camellia japonica extract.

Example 4: Collagen synthesis promotion effect of camellia honey

In order to investigate the effect of Camellia sinensis on promoting collagen synthesis, the following experiment was conducted. Human fibroblasts were seeded at 2 × 10 ⁴ per well in a 24-well plate containing DMEM medium supplemented with 10% FBS, and cultured in a 5% CO ₂ incubator at 37℃ for 24 hours. Thereafter, the test substance was dissolved in DMEM medium without FBS and treated. TGF-β (Transforming growth factor-β) was used as a positive control for promoting collagen synthesis. After 72 hours, the cell culture supernatant was collected to prepare a sample. The amount of collagen synthesis of the sample prepared above was measured for the amount of procollagen type I C-peptide (PICP, Type I C-peptide) using a collagen measurement kit (MK101, Procollagen Type I C-peptide EIA kit) from TAKARA. The results are shown in Fig. 3.

As shown in Figure 3, it was confirmed that camellia honey promoted collagen synthesis in the human fibroblast culture supernatant.

Example 5: Effect of Camellia japonica honey on increasing nitric oxide production

To measure the effect of Camellia japonica on increasing nitric oxide production, the following experiment was conducted. Mouse macrophages (Macrophages, Raw 264.7 cells, Korean Cell Line Bank) were seeded at a density of approximately 2 × 10 ⁵ cells per well in a 24-well microplate containing DMEM medium and cultured in a CO ₂ incubator with a 5% concentration at 37°C for 24 hours. After treatment with Camellia japonica at various concentrations and cultured for 48 hours, the culture solution was collected and mixed with the same amount of Griess reagent (5% (w/v) phosphoric acid, 1% (w/v) sulfanilamide, 0.1% (w/c) N-[1-naphthyl] ethylene diamine dihydrochloride) for 10 minutes, and measured at 540 nm. Macrophages not treated with Camellia japonica were used as a control. The results are shown in Fig. 4.

As shown in Figure 4, it was confirmed that camellia honey increased the production of nitric oxide at 0.1% and 1%.

Example 6: Effect of Camellia sinensis on increasing vascular endothelial growth factor expression

To investigate the effect of Camellia japonica on increasing the expression of vascular endothelial growth factor in hair follicle dermal papilla cells, the following experiment was conducted. Human hair follicle dermal papilla cells (HFDPC, PromoCell) were cultured in hair follicle dermal papilla cell growth medium containing supplementMix, seeded in 24-well plates (2 × 10 ⁴ cells/well), and cultured in a 5% CO ₂ incubator at 37°C for 24 h. Camellia japonica was dissolved in DMEM medium containing 1% FBS, treated to each cultured well, and cultured again for 72 h. After 72 h, the cell culture supernatant was removed, treated with Trizol reagent, and collected into a tube. After adding chloroform (Sigma-Aldrich), the separated supernatant was mixed with 100% isopropyl alcohol (DAEJUNG, Siheung, Korea). The supernatant was removed, and the RNA pellet was washed with 70% ethanol, and the purified total RNA was used for cDNA synthesis. At this stage, TaKaRa Thermal Cycler Dice Touch (TP350, TAKARA BIO INC) and amfiRivert cDNA Synthesis Platinum Master Mix (GenDEPOT) were used, and finally, gene expression was confirmed with 7500 Real-Time PCR System (Applied Biosystems), and is shown in Fig. 5.

As shown in Figure 5, it was confirmed that camellia honey increased the expression of vascular endothelial growth factor in mammary papilla cells by about 2 times compared to the control group.

Example 7: Effect of Camellia japonica on proliferation of human mammary papilla cells

To determine whether Camellia sinensis affects the proliferation of hair papilla cells, which are the main cells for hair growth, the proliferation capacity of hair papilla cells was measured. Specifically, human hair papilla cells, HFDPC, were cultured in a hair papilla cell growth medium containing SupplementMix, seeded in a 24-well plate (1.5 × 10 ⁴ cells/well), and cultured in a 5% CO ₂ incubator at 37°C for 24 hours. Camellia sinensis was dissolved in DMEM medium containing 1% FBS, treated to each cultured well, and cultured again for 72 hours. After 72 hours, the cell culture supernatant was removed and washed once with PBS, and then MTT reagent dissolved at 1 mg/mL was diluted 10-fold and mixed with the medium, treated, and cultured for 1 hour. After 1 hour, the MTT reagent was removed, 200 μL of DMSO was added per well, formazan was dissolved, and the absorbance was measured at 540 nm, and the values are shown in Figure 6.

As shown in Figure 6, camellia honey exhibited a mammary gland papilla cell proliferation ability of about 20% compared to the control group.

Example 8: Safety test of camellia honey on human skin

In order to verify whether camellia honey, which has been proven to have an excellent anti-aging effect according to the present invention, is safe for human skin, a skin topical preparation containing camellia honey was prepared and a skin safety verification experiment was conducted using the product.

Example 1 A skin external preparation containing 1% camellia militaris was prepared with the ingredient contents shown in Table 1 below. To prepare the skin external preparation, purified water, glycerin, and butylene glycol were mixed and dissolved at 70°C (water phase part), and the remaining ingredients except for the above three ingredients and trimethanolamine were dissolved at 70°C (oil phase part). The oil part was added to the water phase and stirred with a homomixer (Tokushu Kika, Japan) to perform primary emulsification, and trimethanolamine was finally added thereto. After removing the air bubbles formed in the mixture, the mixture was cooled to room temperature to prepare a skin external preparation.

ingredient Content (%) Control group Test group Purified water 72 71 glycerin 8.0 8.0 Butylene glycol 4.0 4.0 Camellia honey 0 1.0 Caprylic Capri Triglyceride 8.0 8.0 Squalane 5.0 5.0 Cetearyl Glucuronide 1.5 1.5 Sorbitan stearate 0.4 0.4 Cetearyl alcohol 1.0 1.0 Trimethanolamine 0.1 0.1 Total weight 100 100

Each skin topical preparation manufactured as described above was applied as a 24-hour occlusive patch to the back of 30 healthy adults to determine whether camellia honey irritates the skin.

The patch application method used a chamber (IQ Ultimate™) from Chemotechnique MB Diagnostics AB. 20 μL of each of the above skin external agents was placed in the chamber and occlusively applied for 24 hours. Skin reactions were observed 30 minutes and 24 hours after removal of the patch, and the presence and degree of skin irritation were visually evaluated according to the evaluation criteria. The skin irritation evaluation index was calculated according to the judgment criteria in Table 2 based on Frosch & Kligman (1979) and CTFA guideline (2007). Then, the average skin irritation index of each skin external agent was calculated using the following Empirical Formula 2, and the skin irritation level was evaluated based on the skin irritation index in Table 3 classified by applying the Draize method.

[Experimental Formula 2]

Average Skin Irritation Index Calculation Formula

∑ [(Grade × Number of subjects who responded) / (4; highest grade × total number of subjects)] ÷ Number of evaluations

sign Grade Stimulus evaluation - 0 No response + 1 A mild but noticeable erythema ++ 2 Erythematous reaction with marked erythema and papules or edema +++ 3 Severe erythematous reaction with edema and papules ++++ 4 Severe erythematous reaction with swelling and blistering

Skin irritation index Skin irritation assessment 0.00-0.25 Non-irritating 0.26-1.00 Mild irritation 1.01-2.50 Moderately irritating 2.51-4.00 Strong stimulation

test
substance Number of subjects who responded average
Skin irritation
jisoo 30 minutes later 24 hours later 1 2 3 4 1 2 3 4 Control group - - - - - - - - 0.00 Test group - - - - - - - - 0.00 Number of subjects 30 30 30 30 30 30 30 30

As can be seen in Table 4 above, no irritation reactions were observed in either the control or test groups. According to the above-described formula, the average reaction rate was 0.00, so the skin topical preparation containing camellia honey (test group) was judged to be a non-irritating substance and safe for human skin, just like the control skin topical preparation.

Manufacturing Example 1: Cosmetic preparation

Manufacturing Example 1-1: Flexible Toner

As shown in Table 5 below, a flexible toner containing camellia militaris as an effective ingredient was manufactured according to a conventional method.

ingredient weight% Camellia honey 0.01 glycerin 3.0 Butylene glycol 2.0 Propylene glycol 2.0 Carboxyvinyl polymer 0.1 Ethanol 10.0 Triethanolamine 0.1 Preservatives, trace colors, trace flavors and trace purified water 82.79 aggregate 100.0

Manufacturing Example 1-2: Nutritious Toner

As shown in Table 6 below, a nutritious toner containing camellia honey as an effective ingredient was manufactured according to a conventional method.

ingredient weight% Camellia honey 0.01 beeswax 4.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 5.0 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 3.0 Butylene glycol 3.0 Propylene glycol 3.0 Carboxyvinyl polymer 0.1 Triethanolamine 0.2 Preservatives, trace colors, trace flavors and trace purified water 69.69 aggregate 100.0

Manufacturing Example 1-3: Nourishing Cream

A nutritional cream containing camellia honey as an active ingredient was manufactured according to a conventional method, as shown in Table 7 below.

ingredient weight% Camellia honey 0.01 beeswax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.5 Liquid paraffin 10.0 Squalane 5.0 Caprylic/Capric Triglyceride 5.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, trace colors, trace flavors and trace purified water 56.79 aggregate 100.0

Manufacturing Example 1-4: Massage Cream

A nutritional cream containing camellia honey as an effective ingredient was manufactured according to a conventional method, as shown in Table 8 below.

ingredient weight% Camellia honey 0.01 beeswax 10.0 Polysorbate 60 1.5 Sorbitan sesquioleate 0.8 Liquid paraffin 40.0 Squalane 5.0 Caprylic/Capric Triglyceride 4.0 glycerin 5.0 Butylene glycol 3.0 Propylene glycol 3.0 Triethanolamine 0.2 Preservatives, trace colors, trace flavors and trace purified water 27.49 aggregate 100.0

Manufacturing Example 1-5: Pack

A pack containing camellia honey as an effective ingredient was manufactured according to a conventional method, as shown in Table 9 below.

ingredient weight% Camellia honey 0.01 polyvinyl alcohol 13.0 Sodium carboxymethyl cellulose 0.2 Allantoin 0.1 Ethanol 5.0 Nonylphenyl ether 0.3 Preservatives, trace colors, trace flavors and trace purified water 81.39 aggregate 100.0

Claims (7)

A cosmetic composition for improving blood circulation, preventing or improving hair loss, or preventing or improving dark circles, containing camellia japonica extract as an effective ingredient. A cosmetic composition according to claim 1, characterized in that the composition inhibits vascular permeability of vascular endothelial cells, increases collagen production, increases nitric oxide production, promotes proliferation of hair papilla cells, and promotes expression of vascular endothelial growth factors in hair papilla cells. A cosmetic composition, characterized in that in claim 1, the camellia jasmine is contained in an amount of 0.0001 to 20 wt% based on the total weight of the entire cosmetic composition. A cosmetic composition according to claim 1, characterized in that the cosmetic composition is any one of the group consisting of a solution, a suspension, an emulsion, a paste, a gel, a cream, a lotion, a powder, a soap, a surfactant-containing cleansing, an oil, a powder foundation, an emulsion foundation, a wax foundation, and a spray. An external preparation composition for improving blood circulation, preventing or improving hair loss, or preventing or improving dark circles, containing camellia japonica extract as an effective ingredient. A pharmaceutical composition for improving blood circulation or preventing or treating hair loss, containing camellia japonica extract as an effective ingredient. A food composition for improving blood circulation, preventing or improving hair loss, or preventing or improving dark circles, containing camellia militaris as an effective ingredient.