Elastase activity inhibitor

Abstract

To provide an elastase activity inhibitor comprising a plant extract of high safety as an active ingredient.SOLUTION: An elastase activity inhibitor comprises, as an active ingredient, an extract derived from one or two or more plants selected from the group consisting of chamomile, mugwort, and adlay.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to an elastase activity inhibitor containing a highly safe plant extract as an active ingredient. [Background technology]

[0002] The dermis and epidermis of the skin are composed of epidermal cells, fibroblasts, and the extracellular matrix, such as elastin and collagen, which is located outside these cells and supports the skin structure. In young skin, these skin tissues maintain homeostasis, ensuring moisture retention, flexibility, elasticity, etc., and the skin is maintained in a firm, glossy, and moist appearance.

[0003] However, due to the influence of certain external factors such as ultraviolet radiation, extremely dry air, and excessive skin washing, as well as the progression of aging, the production of elastin, collagen, and other major components of the dermal extracellular matrix decreases, leading to their degeneration and degradation. As a result, the keratin begins to peel abnormally, the skin loses its firmness and luster, and symptoms of aging such as rough skin and wrinkles appear.

[0004] Thus, changes associated with skin aging, such as wrinkle formation, loss of firmness, and reduced elasticity, are associated with the decrease and degeneration of dermal extracellular matrix components such as elastin and collagen.

[0005] Among these matrix components, elastin is a fiber that provides elasticity to skin tissue. When elastin breaks down with aging, skin loses its firmness and elasticity decreases. Elastase, an enzyme that breaks down elastin, is activated by ultraviolet light, which accelerates the breakdown of elastin, causing skin to lose its firmness and elasticity. Therefore, inhibiting the activity of elastase suppresses the breakdown of elastin, which is thought to prevent and improve skin aging symptoms such as loss of firmness and loss of elasticity.

[0006] It is also known that smoking and other factors increase elastase activity in the body. Because the lung matrix is ​​composed of collagen and elastin, increased elastase activity due to smoking and other factors is thought to destroy the alveolar walls and lead to emphysema and other conditions. Furthermore, increased elastase activity is thought to destroy pulmonary capillaries, potentially leading to acute respiratory distress syndrome (ARDS), including pulmonary edema.

[0007] Therefore, if the activity of elastase in the body can be inhibited, it is thought that respiratory diseases such as emphysema, pulmonary edema, etc. Conventionally, substances known to have an elastase activity inhibitory effect include, for example, star fruit extract (see Patent Document 1) and extracts of the genus Artemisia (see Patent Document 2). [Prior art documents] [Patent documents]

[0008] [Patent Document 1] Japanese Patent Application Laid-Open No. 2003-300893 [Patent Document 2] Japanese Patent Application Laid-Open No. 2003-2820 Summary of the Invention [Problem to be solved by the invention]

[0009] The present invention aims to find a highly safe plant extract having an elastase activity inhibitory effect, and to provide an elastase activity inhibitor containing the same as an active ingredient. [Means for solving the problem]

[0010] In order to achieve the above object, the elastase activity inhibitor of the present invention is characterized by containing as an active ingredient an extract from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears. [Effects of the Invention]

[0011] According to the present invention, it is possible to provide an elastase activity inhibitor containing a highly safe plant extract as an active ingredient. DETAILED DESCRIPTION OF THE INVENTION

[0012] Hereinafter, an embodiment of the present invention will be described. The elastase activity inhibitor of the present invention contains, as an active ingredient, an extract from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears.

[0013] In the present invention, "an extract from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears" includes an extract obtained using one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears as an extraction raw material, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, or a crude or purified product thereof.

[0014] The extraction raw materials used in this embodiment are chamomile (scientific name: Matricaria chamomilla), mugwort (scientific name: Artemisia princeps Pampanini), and Job's tears (scientific name: Coix lachryma-jobi L.). [Plant-derived extract] (chamomile extract) Chamomile (Matricaria chamomilla, also known as chamomile) is a hardy annual plant of the Asteraceae family. Parts of the plant that can be used as raw materials for extraction include leaves, stems, flowers, roots, and seeds, with flowers being preferred.

[0015] (Mugwort extract) Mugwort (Artemisia princeps Pampanini) is a perennial herb belonging to the genus Artemisia in the family Asteraceae, which is distributed throughout Japan. Parts that can be used as raw materials for extraction include leaves, stems, flowers, roots, seeds, etc., with leaves being preferred.

[0016] (Job's tears extract) Job's tears (Coix lachryma-jobi L.) is an annual plant belonging to the genus Job's tears of the family Poaceae. Parts that can be used as raw materials for extraction include leaves, stems, flowers, roots, and seeds, with seeds being preferred.

[0017] The one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears used as the extraction raw material are suitably dried and pulverized immediately after harvesting. Drying may be performed in the sun or using a commonly used dryer. The plants may also be used as the extraction raw material after pretreatment such as degreasing with a nonpolar solvent such as hexane. Pretreatment such as degreasing allows efficient extraction of one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears with a polar solvent.

[0018] Although the details of the elastase activity inhibitor contained in the extract from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears are unknown, an extract having elastase activity inhibitory activity can be obtained from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears by an extraction method commonly used for plant extraction.

[0019] The extraction solvent may be either a polar solvent or a non-polar solvent.

[0020] Water that can be used as an extraction solvent includes pure water, tap water, well water, mineral water, and hot spring water. This includes water that has been subjected to various treatments, such as hot water, spring water, and fresh water. Treatments for water include, for example, purification, heating, sterilization, filtration, ion exchange, adjustment of osmotic pressure, and buffering. Therefore, water that can be used as an extraction solvent in the present invention also includes purified water, hot water, ion-exchanged water, physiological saline, phosphate buffer, phosphate-buffered physiological saline, and the like.

[0021] Examples of hydrophilic organic solvents that can be used as extraction solvents include lower aliphatic alcohols having 1 to 5 carbon atoms, such as methanol, ethanol, propyl alcohol, and isopropyl alcohol; lower aliphatic ketones, such as acetone and methyl ethyl ketone; and polyhydric alcohols having 2 to 5 carbon atoms, such as 1,3-butylene glycol, propylene glycol, and glycerin.

[0022] When a mixture of two or more polar solvents is used as the extraction solvent, the mixing ratio can be adjusted appropriately. For example, when a mixture of water and a lower aliphatic alcohol is used, it is preferable to mix 10 parts by volume of water with 1 to 90 parts by volume of the lower aliphatic alcohol, when a mixture of water and a lower aliphatic ketone is used, it is preferable to mix 10 parts by volume of water with 1 to 40 parts by volume of the lower aliphatic ketone, and when a mixture of water and a polyhydric alcohol is used, it is preferable to mix 10 parts by volume of water with 10 to 90 parts by volume of the polyhydric alcohol.

[0023] On the other hand, when a non-polar solvent is used as the extraction solvent, examples of such a solvent include aliphatic hydrocarbons such as petroleum ether, n-hexane, n-pentane, n-butane, n-octane, cyclohexane, squalane, and squalene; aromatic hydrocarbons such as benzene and toluene; halogenated hydrocarbons such as dichloromethane, chloroform, dichloroethane, and carbon tetrachloride; chain ethers such as diethyl ether; edible oils such as rapeseed oil and soybean oil, and fats and oils such as diacylglycerol (DAG) and medium-chain fatty acid oils; and the like. These can be used alone or in combination of two or more.

[0024] Among these, aliphatic hydrocarbons and oils and fats are preferred, and as aliphatic hydrocarbons, squalane and squalene are preferred. Examples of squalane and squalene include those derived from sharks and those derived from plants (e.g., olives), and either can be used.

[0025] In this embodiment, the extraction solvent used for extracting mugwort and Job's tears is preferably a polar solvent, more preferably a mixed solvent of water and a hydrophilic organic solvent, and particularly preferably a mixed solution of water and a lower aliphatic alcohol or a polyhydric alcohol, while the extraction solvent used for extracting chamomile is preferably a non-polar solvent, more preferably a saturated or unsaturated hydrocarbon, edible oil, or fat or oil, and particularly preferably squalane.

[0026] The extraction process is not particularly limited as long as it allows the soluble components contained in one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears to be dissolved in the extraction solvent, and can be carried out according to conventional methods. For example, an extract can be obtained by immersing one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears in an extraction solvent in an amount (mass ratio) 1 to 20 times the amount of the extraction material, extracting the soluble components at room temperature or under reflux, and then filtering to remove the extraction residue. The obtained extract can be subjected to conventional treatments such as dilution, concentration, drying, and purification to obtain a diluted or concentrated solution of the extract, a dried product of the extract, or a crude or purified product thereof.

[0027] Purification can be carried out by, for example, treatment with activated carbon, treatment with adsorption resin, treatment with ion exchange resin, etc. The obtained extract can be used as it is as an active ingredient of an elastase activity inhibitor, but it is easier to use in the form of a concentrated liquid or a dried product.

[0028] Extracts from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears have unique colors, odors, etc., and can be purified for the purpose of decolorization, deodorization, etc., to the extent that this does not result in a decrease in their physiological activity. However, since they are not used in large quantities when blended into pharmaceuticals, quasi-drugs, cosmetics, foods and beverages, etc., there is no practical problem with leaving them unpurified.

[0029] The extracts obtained in the manner described above from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears have an elastase activity inhibitory effect, and can be used as active ingredients of elastase activity inhibitors by utilizing this effect.

[0030] Extracts from one or more plants selected from the group consisting of chamomile, artemisia princeps, and Job's tears can be formulated into any dosage form, such as powder, granules, tablets, or liquid, using pharmaceutically acceptable carriers such as dextrin and cyclodextrin, and any other auxiliary agents, according to conventional methods. This allows for the production of elastase activity inhibitors containing extracts from one or more plants selected from the group consisting of chamomile, artemisia princeps, and Job's tears as active ingredients. Examples of auxiliary agents that can be used in this process include excipients, binders, disintegrants, lubricants, stabilizers, and flavoring agents. Extracts from one or more plants selected from the group consisting of chamomile, artemisia princeps, and Job's tears can be incorporated into other compositions (e.g., pharmaceuticals, quasi-drugs, cosmetics, foods, beverages, etc.) and can also be used as ointments, topical liquids, patches, etc.

[0031] The elastase activity inhibitor of the present invention can be used as an active ingredient in combination with other plant extracts having an elastase activity inhibitory effect, if necessary.

[0032] The elastase activity inhibitor of the present invention can generally be administered by transdermal administration, oral administration, intravascular (intravenous) administration, etc., and a method suitable for the prevention, treatment, etc. of the disease can be appropriately selected depending on the type of disease. The dosage of the elastase activity inhibitor of the present invention can also be increased or decreased as appropriate depending on the type and severity of the disease, individual differences in patients, administration method, administration period, etc.

[0033] The elastase activity inhibitor of the present invention can prevent and improve skin aging symptoms such as loss of firmness and elasticity by inhibiting the degradation of elastin through the elastase activity inhibitory effect of the active ingredient, an extract from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears. However, in addition to these uses, the elastase activity inhibitor of the present invention can also be used for any other uses where exerting an elastase activity inhibitory effect is significant.

[0034] For example, the elastase activity inhibitor of the present invention can be used as an active ingredient in pharmaceuticals or quasi-drugs for the prevention and treatment of diseases caused by increased elastase activity, such as respiratory diseases such as emphysema and pulmonary edema.

[0035] The elastase activity inhibitor of the present invention can also be incorporated into cosmetics, whereby the elastase activity inhibitor can be imparted to the cosmetics with an elastase activity inhibitory effect.

[0036] Cosmetics that can be formulated with the elastase activity inhibitor of the present invention are not particularly limited, and examples include skin cosmetics such as ointments, creams, emulsions, lotions, packs, jellies, lip balms, lipsticks, eyeliners, mascaras, nail polishes, sunscreens, foundations, primers, and bath additives; and hair cosmetics such as shampoos, rinses, and hair styling products.

[0037] The amount of elastase activity inhibitor in a cosmetic can be adjusted as appropriate depending on the type of cosmetic, the physiological activity of the extract, etc. The blending rate of the elastase activity inhibitor in a cosmetic is preferably 0.0001 to 10 mass%, and particularly preferably 0.001 to 1 mass%, calculated as the dry matter of the extract from one or more plants selected from the group consisting of chamomile, artemisia princeps, and Job's tears. By blending within the above range, the elastase activity inhibitory effect of the extract from one or more plants selected from the group consisting of chamomile, artemisia princeps, and Job's tears can be effectively imparted to the cosmetic.

[0038] The above-mentioned cosmetic preparations may be used in combination with various main ingredients and auxiliary agents commonly used in the manufacture of cosmetics, such as astringents, bactericides / antibacterial agents, whitening agents, ultraviolet absorbers, moisturizers, cell activators, anti-inflammatory / antiallergic agents, antioxidants / active oxygen scavengers, etc., as long as they do not interfere with the elastase activity inhibitory effect of the extract from one or more plants selected from the group consisting of chamomile, mugwort, and Job's tears, which is the active ingredient of the elastase activity inhibitor. Using such combinations will result in a more general-purpose product, and the synergistic effect with the other active ingredients used in combination may bring about better effects than would normally be expected.

[0039] Although the elastase activity inhibitor of the present invention is preferably applied to humans, it can also be applied to animals other than humans as long as the desired action and effect is achieved. For example, the elastase activity inhibitor of the present invention can be added to food for pets and livestock, such as dog food and cat food, to prevent and treat various diseases in pets and livestock. [Example]

[0040] The present invention will be specifically explained below by showing production examples, test examples and formulation examples, but the present invention is not limited to the following examples in any way.

[0041] [Production Example 1] Production of chamomile extract Squalane was added to 100 g of dried chamomile flowers, extracted by heating, and filtered. The resulting extract was dried to obtain chamomile extract (2 g, Sample 1). [Production Example 2] Production of mugwort extract 1500 mL of 80% by volume ethanol was added to 100 g of mugwort leaves, and extraction was carried out under reflux at 80-90°C for 2 hours, followed by hot filtration. The resulting extract was dried to obtain mugwort extract (7.5 g, sample 2). [Production Example 3] Production of Job's Tears extract 1500 mL of 50% by volume 1,3-butylene glycol was added to 100 g of Job's tears seeds (Coix seed kernel) with the seed coat removed, followed by extraction with heating and filtration. The resulting extract was dried to obtain Job's tears extract (6 g, sample 3).

[0042] [Test Example 1] Elastase activity inhibitory effect test The extracts (samples 1 to 3) obtained in Production Examples 1 to 3 were tested for their elastase activity inhibitory effect as follows.

[0043] Fifty microliters of sample solution (Samples 1-3) prepared in 0.1 mol / L HEPES buffer (pH 7.5) and 25 μL of 6 μg / mL human neutrophil-derived elastase (Elastase, Human Leukocyte, manufactured by Elastin Products) solution were mixed in a 96-well plate. Then, 25 μL of a 2 mmol / L solution of N-METHOXYSUCCINYL-ALA-ALA-PRO-VAL-p-NITROANILIDE (manufactured by SIGMA) substrate prepared in the above buffer was added, and the mixture was allowed to react for 15 minutes at 25°C. After the reaction, the absorbance at 415 nm was measured.

[0044] Similar measurements were also performed on a reaction solution without the addition of sample solution as a sample-free control. Furthermore, similar measurements were performed on a sample solution and a sample-free solution without the addition of enzyme to serve as blanks for the sample-added and sample-free controls. The elastase activity inhibition rate (%) was calculated from the obtained measurement results using the following formula. The results are shown in Table 1.

[0045] Elastase activity inhibition rate (%) = {1-(AB) / (CD)} x 100 In the formula, "A" represents the absorbance at a wavelength of 415 nm when the sample and enzyme are added, "B" represents the absorbance at a wavelength of 415 nm when the sample and enzyme are not added, "C" represents the absorbance at a wavelength of 415 nm when the sample is not added but the enzyme is added, and "D" represents the absorbance at a wavelength of 415 nm when the sample and enzyme are not added.

[0046] [Table 1] Sample number Concentration (μg / mL) Elastase activity inhibition rate (%) 1 1.56 15.8 6.25 44.8 25 71.9 100 86.8 400 89.6 2 25 5.7 100 12.2 400 56.4 3 100 17.1 400 73.6

[0047] Referring to Table 1, it was confirmed that each of the extracts (samples 1 to 3) above had excellent elastase activity inhibitory activity. [Industrial Applicability]

[0048] The elastase activity inhibitor of the present invention can make a significant contribution to the prevention and improvement of skin aging.

Claims

[Claim 1] An elastase activity inhibitor characterized in that it contains as an active ingredient an extract from one or more plants selected from the group consisting of chamomile, mugwort and Job's tears.