KR20100090531A - Extracts of morinda officinalis and oral composition for preventing and improving periodontal disease and cosmetic composition for preventing and improving skin aging - Google Patents

Abstract

PURPOSE: A composition containing Morinda officinalis extract is provided to suppress matrix metalloproteinase activity and to prevent and treat various diseases. CONSTITUTION: A Morinda officinalis extract suppresses matrix metalloproteinase(MMP) activity. The matrix metalloproteinase(MMP) is collaganse. An oral composition for preventing and treating periodontal diseases contains 0.001-20.0 weight parts of Morinda officinalis extract. A cosmetic composition for preventing and treating skin aging contains 0.001-20.0 weight parts of Morinda officinalis extract.

Description

Extract of pagokcheon, oral composition for improving and preventing periodontal disease and cosmetic composition for preventing and improving skin aging including the same {Extracts of Morinda officinalis and oral composition for preventing and improving periodontal disease and cosmetic composition for preventing and improving skin aging}

The present invention relates to pageokcheon extract, oral composition for improving and preventing periodontal disease comprising the same and a cosmetic composition for preventing and improving skin aging. More specifically, the present invention inhibits the activity of matrix metalloproteinase (MMP) to prevent and alleviate the inflammatory response of tissues, prevent the destruction of collagen, and high human safety even after long-term use. Pageokcheon extract, it relates to a composition for oral cavity for preventing and improving periodontal disease comprising the same and a cosmetic composition for preventing and improving skin aging.

Periodontal disease can not be explained only one cause of it, it can be seen as a chronic disease caused by a combination of causes. An important enzyme in periodontal disease is collagen hydrolase secreted by islet infantry cells, polymorphonuclear leukocytes, epithelial cells and macrophages. When the periodontal tissue is infected with Gram-negative bacteria, endotoxins such as lipopolysaccharide are secreted and the tissues are destroyed or cytokines such as prostaglandins and interleukins are inflamed by the body's immunity. Collagen, which is a substrate of periodontal tissue, is degraded by collagen hydrolase and bacterial collagen hydrolase secreted by the stimulation of these inflammatory mediators, so that the gums are torn down and left unattended, leading to periodontal disease.

Skin aging is divided into physiological aging process and external environmental aging process over time. External factors affecting skin aging include wind, temperature, humidity, tobacco smoke, pollution, ultraviolet light, etc. In particular, skin aging caused by ultraviolet light is called photoaging. Biochemical phenomena seen in chronic photo-damaged skin include abnormal elastotic material deposition in the upper collagen of the dermis, an increase in proteoglycans and a decrease in the collagen, a major protein in the dermis. In general, the dermal layer is composed of a large amount of type 1 collagen and a small amount of type 3 collagen, elastin, proteoglycan, fibronectin, and the like. Collagen accounts for more than 90% of these, plays an important role in the strength and tension of the skin and protects the skin from external irritation or force. Thus, the reduction of collagen is closely related to skin aging.

Matrix metalloproteinases (MMPs) are a type of zinc-containing endopeptidase that can degrade large protein-based components such as collagen, gelatin, and proteoglycans. Collagen hydrolase, gelatin hydrolase, stromellisin and cell membranes Can be classified as type MMP. They are all expressed in the form of pro-enzymes and then become active by partly spontaneously cutting off.

Collagen hydrolase acts on triple helix collagen and gelatin, and three types of fibroblast collagenase (MMP-1), neutrophil collagenase (MMP-8), and collagenase-3 (MMP-13) are known. . They have at least 50% gene sequence identity with each other and X-ray crystallographic studies of the sites of action of MMP-1 and MMP-8 have revealed that they have two zinc ion-synthesizing sites and one to two calcium ion-binding sites.

Gelatin hydrolase is an enzyme that degrades denatured collagen and 4, 5, 7, 10 collagen. There are two types of gelatin hydrolase released from fibroblasts (MMP-2) and mononuclear phagocytes (MMP-9). In particular, these enzymes are the most important enzymes for cancer cell infiltration and metastasis.

Stromlycine originally had the characteristics of proteoglycanase, but actually has a broader protein degradation spectrum, stromelysin-1 (MMP-3), stromelysin-2 (MMP-10), and stromelysin-3 (MMP-11) and act on fibronectin.

These various forms of matrix metalloproteinases are known to have substrate specificities and are expressed in metabolism when there is a need to degrade extracellular matrix or other collagen structures in normal cells. In particular, the activity of collagen hydrolase and stromelysin in fibroblasts isolated from inflamed gums was confirmed, and the amount of enzyme was correlated with the degree of gingivitis observed (J. Periodontal Res. 22, 81-88). 1987). In addition, when the skin is exposed to ultraviolet rays, the activity of the matrix metalloproteinase increases markedly and collagen is degraded in the skin, thereby promoting skin aging. Therefore, the matrix metalloproteinase is closely related to skin aging. .

In order to solve the above problems, an object of the present invention is to inhibit the activity of matrix metalloproteinase (MMP) to prevent and alleviate the inflammatory response of tissues, to prevent the destruction of collagen, even in the long-term use It is to provide a high safety paepukcheon extract, oral composition for improving and preventing periodontal disease comprising the same and a cosmetic composition for preventing and improving skin aging.

The present invention to achieve the above object,

Provided are paepukcheon extracts that inhibit the activity of matrix metalloproteinases.

The substrate metalloproteinase may be a collagen hydrolase.

The cheonpokcheon extract is a group consisting of purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, chloroform, diethyl ether, dichloromethane and hexane It can be extracted alone or a mixture thereof selected from the extraction solvent.

The present invention to achieve another object of the present invention,

It provides a composition for the prevention and improvement of periodontal disease, including the paepukcheon extract.

The cheonpokcheon extract may be included in 0.001 to 20 parts by weight based on 100 parts by weight of the composition for oral cavity.

The present invention to achieve another object of the present invention,

It provides a skin composition for preventing and improving skin aging comprising the paepukcheon extract.

The paepukcheon extract may be included in 0.001 to 20 parts by weight based on 100 parts by weight of the cosmetic composition.

Papokcheon extract of the present invention is effective in the improvement and prevention of periodontal disease by inhibiting the activity of the matrix metalloproteinase to alleviate the inflammatory response of periodontal tissue.

Papokcheon extract of the present invention is effective in preventing and improving skin aging by preventing the reduction of collagen in the skin.

Papocheon extract of the present invention can be used in oral compositions for the improvement and prevention of periodontal disease and compositions for the prevention and improvement of skin aging, and can be formulated in various forms.

Papokcheon extract of the present invention has a low toxicity and excellent safety to the human body, so it is harmless to human oral mucosa even after long-term use, and bacteria in the oral cavity is not resistant.

The cosmetic composition comprising the extract of Pakcheoncheon of the present invention does not cause skin irritation and does not cause problems such as skin troubles even for a long time use. Therefore, it is possible to prepare a cosmetic composition that can be applied to various skin types.

Hereinafter, the present invention will be described in detail so that those skilled in the art can easily practice.

Papocheon extract of the present invention inhibits the activity of the substrate metalloproteinase. In particular, paepukcheon extract of the present invention inhibits the activity of collagen hydrolase which is one of the substrate metalloproteinases. The paepukcheon extract can be effectively used for the prevention and improvement of various diseases by inhibiting the activity of the substrate metalloproteinase involved in a number of physiological processes. In addition, it is safe for the human body as a plant extract, so there are almost no side effects even when used for a long time.

The Morinda officinalis is said to be an Indian mulberry in India, a Pantsian in China, a painkiller tree in the Caribbean, a cheese fruit in Australia, and a Nono in Tahiti. In Dongbogam, it is called Haepukgeok (海巴 戟) or Paegukcheon (巴戟 天). Its height is 3-12m, and its size is various. The leaves are egg-shaped, 30cm long, 15cm butterfly, the flowers are small and white. They come out from the leaf axes in a bunch and bloom mainly from summer to autumn. The fruit is about 10cm in size, looks like a potato, and has a bumpy surface and a dent. The fruit contains a lot of fiber and juice, especially when the fruit is ripe, the yellowish-white peel appears thin and transparent. At this time, the fruit, which could not be tasted, has an unpleasant taste and smells like a rotten cheese. The fruit can be seen almost year round and contains a lot of brown seeds. Leaves, stems, flowers, fruits, and seeds have been used in folk medicine, and ancient literature in the South Pacific is the best natural remedy. In fact, it contains ingredients such as anthraquinone and serotonin, which help digestion, reduce pain, and are effective in hypertension and cancer.

The paepukcheon extract means that it is obtained by extracting various organs or parts of paepukcheon, that is, leaves, flowers, fruits, stems, roots, branches and shells, preferably means obtained by extracting the roots of pagokcheon.

The matrix metalloproteinase (MMP) is a family of metalloproteinases having zinc at the active site, and thus, membrane collagen, aggrecan, and fibro, which form structural proteins in the extracellular matrix. It is an enzyme that breaks down and reconstructs proteins such as nectin (fibronectin) and laminin. Overexpression and activation of the matrix metalloproteinase causes tissue degradation and plays an important role in the development of chronic diseases such as malignant tumor growth and metastasis, multiple sclerosis, arthritis, fibrosis and other inflammations. Therefore, it is involved in various diseases of pathological processes such as cancer metastasis, periodontal disease, rheumatoid arthritis, inflammation, hyperparathyroidism, diabetes, corneal ulcer, osteoporosis, gastric ulcer, trauma, wrinkle, acne, AIDS, burns, arteriosclerosis, fracture . Therefore, the paepukcheon extract of the present invention that inhibits the activity of the substrate metalloproteinase can be effectively used for the prevention and improvement of the various diseases.

The pulpocheon extract inhibits the activity of collagen hydrolase, which is one of the matrix metalloproteinases. Collagen hydrolase acts on triple helix collagen and gelatin, and three types of fibroblast collagenase (MMP-1), neutrophil collagenase (MMP-8), and collagenase-3 (MMP-13) are known. . Since the destruction of collagen causes periodontal disease and skin aging, it is effective in preventing and improving periodontal disease and skin aging by inhibiting the activity of collagen hydrolase.

According to one embodiment of the invention, the method for producing the cheonpokcheon extract is a crushing step (S1); Extraction step (S2); Filtration step (S3); Concentration step S4; And a powdering step (S5). By the above method, it is possible to obtain an extract of Papocheon which inhibits the activity of matrix metalloproteinase, in particular collagen hydrolase, which is known to be mainly involved in the destruction of collagen in the inflammatory reaction process.

The crushing step (S1) is a crushing step of breaking cloth. The rupture cloth may be used as it is for extraction or crushed into small pieces or powdered by methods known in the art. In the case of crushing the rupture cloth finely, the extraction speed can be increased and the extraction efficiency can be increased when immersed in the extraction solvent. The rupture cloth may be used by drying it or without drying it, and preferably, it may be dried. The drying method of the ruptured cloth can be used without limitation methods known in the art. After washing and grooming the pagoda cloth, it can be dried by hot air drying, high temperature drying, natural drying, and the like, and preferably by natural drying in sunlight.

The extraction step (S2) is a step of obtaining an extract by extracting the dried pagoda cloth prepared in step S1 with a solvent. The extraction method may be a method known in the art such as cold needle extraction, hot water extraction, ultrasonic extraction, reflux cooling extraction, heating extraction. Specifically, it is possible to carry out 1 to 4 days by immersing the dried catapult fabric prepared in step S1 in the extraction solvent at 5 to 80 ℃, it can be extracted repeatedly 2 to 3 times. Preferably it can be extracted by immersion in ethanol for 3 to 4 days at room temperature.

The extraction solvent consists of purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, chloroform, diethyl ether, dichloromethane and hexane Single or mixtures thereof selected from the group can be used. Preferably, single or a mixture thereof may be used selected from the group consisting of polar solvents such as purified water, methanol, ethanol, acetone, glycerol, propylene glycol, 1,3-butylene glycol, and the like. The polar solvent extract is more excellent in inhibiting substrate metal protein activity than the non-polar solvent extract, and is easy to formulate due to its high solubility and emulsification.

The filtration step (S3) is a step of filtering the extract. The filtration can be carried out by methods known in the art, and usually the extract is filtered through filter paper. The filtration step can be omitted if necessary.

The concentration step (S4) is a step of obtaining a concentrated liquid by concentrating the paekcheoncheon extract or filtrate. It is preferable to perform the said concentration by vacuum distillation.

The powdering step (S5) is a step of powdering the concentrate. The powdering is preferably a method of removing moisture by lyophilization. The concentrate may be used in liquid form without being powdered.

The composition for oral cavity for preventing and improving periodontal disease of the present invention comprises the paepukcheon extract. The pagokcheon extract inhibits the activity of matrix metalloproteinase, prevents the destruction of collagen and is effective for the prevention and improvement of periodontal disease, and it is not toxic to the human body, so it may be harmless to the human oral mucosa even after long-term use. In addition, the bacteria in the oral cavity are not resistant to the composition of the present invention, and thus its effect is not reduced even in the long term use.

The cheonpokcheon extract may be included in an amount of 0.001 to 20.0 parts by weight based on 100 parts by weight of the total composition. Preferably it may be included in 0.5 to 10.0 parts by weight. If it is included less than 0.001 parts by weight of the activity inhibitory effect of the substrate metalloproteinase is insignificant, if it contains more than 20.0 parts by weight it may be difficult to ensure the stability of the formulation and formulation for a long time to increase the viscosity of the composition.

The oral composition for preventing and improving periodontal disease may be prepared in any shape such as paste, solid, liquid, or gum. Specific formulations include toothpaste, tooth powder, jelly, caramel, candy, oral rinse, mouthwash, gum, and the like.

The composition for oral cavity for preventing and improving periodontal disease may include other components conventionally used within the range of not impairing the object of the present invention, in addition to the paepukcheon extract. For example, an aqueous component, an oily component, surfactant, a humectant, a thickener, a preservative fragrance, a pigment | dye, etc. can be included. Specifically, when the composition for oral cavity is a toothpaste, abrasives such as alumina, silica gel, precipitated silica, calcium carbonate, calcium dihydrogen phosphate, sodium bicarbonate, etc. which are typically contained in toothpaste; Wetting agents, such as sorbitol, glycerin, polyethylene glycols; Foaming agents such as sodium lauryl sulfate, sodium lauryl sarcosinate, dodecylbenzenesulfonate and the like; Binders such as carboxymethyl cellulose, carrageenan, polyacrylate, and the like; Sweetening agents such as saccharin, aspartame, stevioside and the like; Preservatives such as methyl paraoxybenzoate and propyl paraoxybenzoate; Medicinal agents such as sodium fluoride, sodium fluoride phosphate, vitamins, salts and the like; Spices; Pigments; And pH adjusters.

Skin aging prevention and improvement of the cosmetic composition of the present invention comprises the paepukcheon extract. The pagokcheon extract is effective in preventing and improving skin aging because it inhibits the activity of matrix metalloproteinases that destroy collagen in the skin, which protects the skin from external stimuli, especially when the skin is exposed to ultraviolet light. Since proteolytic enzymes are more activated, they are effective in preventing and improving photoaging due to ultraviolet rays. In addition, there is no irritation on the skin, so long-term use does not have side effects.

The cheonpokcheon extract may be included in an amount of 0.001 to 20.0 parts by weight based on 100 parts by weight of the total composition. Preferably it may be included in 0.5 to 10.0 parts by weight. If it is contained less than 0.001 parts by weight of the activity of inhibiting the activity of the substrate metalloproteinase is insufficient to prevent and improve the skin aging, if contained in more than 20 parts by weight may be difficult to secure long-term stability of the formulation, The fragrance may cause a feeling of use of the cosmetic composition.

The cosmetic composition for preventing and improving skin aging may be prepared in any formulation commonly prepared in the art, for example, suspensions, emulsions, pastes, gels, creams, lotions, powders, soaps, oils, powders -Can be formulated as a foundation, spray, or the like. More specifically, it may be variously formulated into a flexible lotion, nutrition lotion, nutrition cream, massage cream, essence, pack, cleansing cream, cleansing foam, spray, powder and the like.

The cosmetic composition for preventing and improving skin aging may be combined with components conventionally used in cosmetics within the range of not impairing the object of the present invention in addition to the paepukcheon extract. For example, it may include conventional auxiliaries such as antioxidants, stabilizers, solubilizers, humectants, vitamins, pigments, fragrances, preservatives, and carriers.

The present invention will be described through the following examples. The following examples are only for the description of the present invention, but the scope of the present invention is not limited to the following examples.

Example 1 Preparation of Pageokcheon Extract

2 g of ethanol (95%) was added to 100 g of powdered and dried pulverized dried product, dried at room temperature for 3 to 4 days, extracted twice, filtered, and the filtrate was distilled under reduced pressure to obtain 17.2 g of concentrated solution. Got it. This concentrate was dissolved in 10% DMSO solution and used for the evaluation.

<Examples 2 to 5> Preparation of the oral cleanser containing paepukcheon extract

Oral cleansing agent containing the paepukcheon extract prepared in Example 1 was prepared in the composition shown in Table 1. The unit in Table 1 is parts by weight.

Example 6 Preparation of Toothpaste Containing Pagokcheon Extract

Toothpaste containing the paepukcheon extract prepared in Example 1 was prepared in the composition shown in Table 2. The unit in Table 2 is parts by weight.

Example 7 Preparation of Oral Rinse Containing Pagokcheon Extract

Oral rinse containing the paepukcheon extract of Example 1 was prepared in the composition of Table 3 below. The units in Table 3 below are parts by weight.

Example 8 Preparation of Gum

The gum containing the extract of Pajucheoncheon of Example 1 was prepared in the composition of Table 4 below. The unit in Table 4 is parts by weight.

Example 9 Preparation of a Cream Containing Pajukcheon Extract

A cream containing the extract of Pajucheoncheon of Example 1 was prepared in the composition of Table 5 below. The units in Table 5 below are parts by weight.

<Example 10> Preparation of a lotion containing paepukcheon extract

The lotion containing the cheonpokcheon extract of Example 1 was prepared in the composition of Table 6 below. The unit in Table 6 is parts by weight.

<Examples 11 to 14> Preparation of the essence containing paepukcheon extract

Essence comprising the paepukcheon extract of Example 1 was prepared in the composition of Table 7. The unit in Table 7 is parts by weight.

Example 15 Preparation of Cleansing Foam Containing Pagokcheon Extract

The cleansing foam containing the paepukcheon extract of Example 1 was prepared in the composition of Table 8. The units in Table 8 below are parts by weight.

<Comparative Examples 1 and 2>

Oral cleaning agents were prepared in the composition shown in Table 9. The unit in Table 9 is parts by weight.

<Comparative Examples 3 and 4>

To the essence was prepared in the composition of Table 10. The unit in Table 10 is parts by weight.

Experimental Example 1 Evaluation of Inhibition of Substrate Metalloproteinase Activity

Substrate metalloproteinase activity inhibition inhibition evaluation was performed using a commercially available Assay Kit (Biomol AK-405, USA). Substrate metalloproteinase (MMP-1) diluted with buffer solution in each well of a 96-well microplate, substrate metalloproteinase (MMP-1) diluted with 10% DMSO solution, evaluation sample (papocheon extract), conventional 20 μl each of a substrate metalloproteinase inhibitor (NNGH, N-Isobutyl-N- (4-methoxyphenylsulfonyl) -glycyl hydroxamic acid), which was known to have been injected, was pre-incubated at 37 ° C. for 30-60 minutes. As a final step, each well was injected with a fluorogenic substrate, and then the fluorescence size was measured at intervals of 1 minute for 10 minutes at 37 ° C. The effect of the evaluation sample was measured based on the degree of inhibition of the activity of NNGH, an existing substrate metalloproteinase inhibitor used as a comparative sample. In the same manner, the activity inhibition evaluation of the substrate metalloproteinase MMP-8 was performed. The results of evaluation of inhibition of activity of MMP-1 are shown in Table 11 below, and the results of evaluation of inhibition of activity of MMP-8 are shown in Table 12 below.

The optical values of the fluorescence labeled on the substrate in Tables 11 and 12 are detected in response to light of a specific wavelength when the substrate is decomposed by the enzyme activity, so that the smaller the fluorescence value, the more the enzyme activity is inhibited. According to Tables 11 and 12, in the case of paepcheoncheon extract of Example 1 showed an activity inhibitory effect similar to NNGH, a substrate metalloproteinase inhibitor used in the past.

Experimental Example 2 Evaluation of Inhibition of Substrate Metal Protein Activity of Oral Composition and Cosmetic Composition Containing Pajukcheon Extract

In the same manner as in Experimental Example 1, the effects of inhibiting the activity of the substrate metalloproteinase (MMP-8) of the compositions of Examples 2 to 5, Examples 11 to 14 and Comparative Examples 1 to 4 were evaluated. Table 13 shows.

According to the above Table 13, Examples 2 to 5 of the composition for oral cavity containing the pakchekcheon extract and Examples 11 to 14 of the cosmetic composition showed a smaller fluorescence value than the composition of the comparative example. That is, it can be seen that the effect of inhibiting the activity of the substrate metalloproteinase is excellent. On the other hand, Comparative Example 1, which does not contain the pakcheoncheon extract, showed no similar value to the buffer solution containing no inhibitor and only MMP-8, and did not inhibit the activity of the substrate metalloproteinase. Comparative Examples 2 and 4 were used in the evaluation of inhibition of substrate metal protein activity due to the high viscosity of the pakcheoncheon extract contained in excess.

Experimental Example 3 Human Safety Evaluation

The MTT assay, which is generally used for toxicity evaluation, was used to determine whether human extracts were safe. After dissolving the Papocheon extract of Example 1 dissolved in a 10% DMSO solution at an appropriate concentration again with a PBS solution, a sample solution of various concentrations was prepared through step dilution. Each well of the 96-well microplate was inoculated with 1-4 × 10 ⁴ cells / ml of oral mucosal cells (KB cell. Incubated for 2 to 3 days at% CO ₂ conditions. 0.1 mg of MTT was added to all wells and incubated at 37 ° C. for 4 hours. The plate was centrifuged lightly, the medium in each well was aspirated, DMSO solution was added and gently shaken for 10 minutes until the formazan crystals dissolved. Absorbance was measured in a microplate reader (540 nm) and the results are shown in FIG. 1.

In Figure 1, the axis of ordinates indicates absorbance, which indicates the amount of MTT degraded by cells to formazan (blue) and is proportional to the number of living cells present in each well. According to FIG. 1, the pagokcheon extract showed a level of cytotoxicity almost close to the PBS solution used as the buffer solution at a concentration of about 1%. In addition, when the safety level of the DMSO solution used to dissolve the paekcheoncheon extract is significantly lowered at 1% concentration, it is expected that the human safety level of the paepcheoncheon extract itself is better than the above results. From the above results, it can be seen that paepukcheon extract is excellent in the safety of the oral mucosa as well as the inhibitory effect on the substrate metalloproteinase activity.

As described above, paepukcheon extract of the present invention can be used to prevent and improve the disease associated with the enzyme by inhibiting the activity of the matrix metalloproteinase, in particular it is effective in the prevention and improvement of periodontal disease and skin aging have. In addition, the safety of the human body is excellent, there is no irritation to the oral mucosa and skin even after long-term use.

1 is a result of evaluating the human stability of pagokcheon extract of the present invention.

Claims (7)

Papokcheon extract, characterized in that to inhibit the activity of the matrix metalloproteinase. The method of claim 1, Papokcheon extract, characterized in that the substrate metalloproteinase is collagen hydrolase. The method of claim 1, The cheonpokcheon extract is a group consisting of purified water, methanol, ethanol, propanol, isopropanol, butanol, acetone, glycerol, propylene glycol, 1,3-butylene glycol, methyl acetate, ethyl acetate, chloroform, diethyl ether, dichloromethane and hexane Paekukcheon extract, characterized in that the extraction is carried out using a single or a mixture thereof selected as an extraction solvent. Oral composition for the prevention and improvement of periodontal disease, characterized in that it comprises a paepukcheon extract of any one of claims 1 to 3. The method of claim 4, wherein Oral composition for the prevention and improvement of periodontal disease, characterized in that the paepukcheon extract is contained in 0.001 to 20.0 parts by weight based on 100 parts by weight of the total composition. A cosmetic composition for preventing and improving skin aging, characterized in that it comprises a paepukcheon extract of any one of claims 1 to 3. The method of claim 6, Skin aging prevention and improvement cosmetic composition, characterized in that the paepukcheon extract is contained in an amount of 0.001 to 20.0 parts by weight based on 100 parts by weight of the total composition.

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