Composition for lipase activity inhibition

Abstract

To reveal the health benefits of Ecklonia kurome and to provide compositions that can be used in foods and the like.SOLUTION: Provided is a composition for lipase activity inhibition, lipid absorption inhibition, stool slippage improvement, or intestinal regulation, which contains an effective amount of the water extract of Ecklonia kurome. Provided is a method for producing the water extract or processed product of Ecklonia kurome, including the following step: (1) extracting Ecklonia kurome with hot water at 50-80°C for 1-5 hours.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] Ecklonia kurome (scientific name: Ecklonia kurome) is a species of the genus Ecklonia in the Lessoniaceae family. It is a type of brown algae found in the waters around Japan, from the central and southern Pacific coast of Honshu to Kyushu, the Seto Inland Sea, and the Japan Coast It is distributed throughout Japan, and overseas it has been confirmed to be distributed in places such as Jeju Island in South Korea. Adult size is usually 20-50cm.

[0002] Ecklonia cava and Eisenia bicolor are known as species with similar morphology, but Ecklonia cava has wrinkled leaves. They are distinguished from Kurome in that they cannot be inserted into the roots, and Arame in that their stems are forked at the top.

[0003] Kurome is a type of alga harvested mainly in winter and eaten as food. In addition to chopped raw kurome, the local specialty is also used as seasonings (soy sauce, sauces, etc.) and processed foods. Kurome is rich in amino acids and minerals, and is expected to have health benefits. do.

[0004] However, in Japan as a whole, black seaweed harvested as an incidental product of other seafood harvests is discarded. It is difficult to say that it is a food ingredient that is being fully utilized. Due to the high value of the fish as food, development of aquaculture technology is underway (Non-patent Documents 1 and 2). . [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] Tomokazu Nishigaki et al., "Distribution characteristics and seasonal fluctuations of the brown alga Kurome in the western waters of Wakasa Bay," Research Report of the Marine Research Center, Kyoto Prefectural Agriculture, Forestry and Fisheries Technology Center, No. 37, 2015, pp. 1-6 [Non-patent document 2] Tomokazu Nishigaki et al., "Growth and Morphology of Cultivated Kurome in the Kamai Coast of Kyoto Prefecture," Kyoto Prefectural Oceanic and Fisheries Research Center Research Report, No. 28, 2006, pp. 16-20 [Non-patent document 3] Hiroaki Ueno et al., "Trends and Prospects of New Anti-obesity Drugs," Journal of the Japanese Society of Internal Medicine, Vol. 103, No. 3, 2014, pp. 753-759 Summary of the Invention [Problem to be solved by the invention]

[0006] As mentioned above, Kurome is a seaweed that is often discarded and has not yet been fully utilized in food products. There are few detailed studies or research reports on the health benefits of Kurome.

[0007] Therefore, the present invention clarifies the health benefits of Kurome and provides a composition that can be used in foods, etc. The purpose is to provide. [Means for solving the problem]

[0008] In order to solve the above problems, the present inventors have conducted extensive research and found that the water extract of Kurome has the following properties: The inventors have found that the compound has a significant inhibitory effect on lipase activity, and have thus completed the present invention.

[0009] That is, the present invention provides the compositions listed below. [1] Contains an effective amount of water extract of Ecklonia kurome (scientific name: Ecklonia kurome) A composition for inhibiting lipase activity.

[0010] [2] Contains an effective amount of water extract of Ecklonia kurome (scientific name: Ecklonia kurome) A composition for inhibiting lipid absorption, improving stool lubrication, or regulating the intestines.

[0011] [3] The composition according to [1] or [2], wherein the Ecklonia kuroume is a dried product of Ecklonia kuroume.

[0012] [4] Any of [1] to [3], wherein the extracted part of the Ecklonia kurome is a leaf and / or a stem. The composition according to any one of claims 1 to 10.

[0013] [5] [1] to [4], wherein the growth period of the Ecklonia kurome is 1 to 6 years. The composition described.

[0014] [6] Any of [1] to [5], wherein the extraction temperature for obtaining the extract is 50 to 80°C. 1. The composition described in 1.

[0015] [7] Any one of [1] to [6], wherein the extraction time for obtaining the extract is 1 to 5 hours. The composition described in

[0016] [8] Any one of [1] to [7] that is a drug, quasi-drug, cosmetic, or food or drink The composition described above.

[0017] [9] Any of [1] to [8] whose daily intake is 500 mg or more in terms of dry solids. 1. The composition according to claim 1.

[0018] The present invention also provides the following production methods.

[10] The following steps: (1) Extracting Ecklonia kurome with hot water at 50 to 80 ° C for 1 to 5 hours; A method for producing a water extract or processed product of Ecklonia kuroume.

[0019]

[11] The method according to

[0010] , which is a water extract or processed product of Ecklonia kuroume, which has an enhanced lipid absorption inhibitory effect.

[0020] The present invention also provides the following processed products of Ecklonia kurome.

[0021]

[12] Contains an effective amount of water extract of Ecklonia kurome (scientific name: Ecklonia kurome) , The extraction temperature for obtaining the extract is 50 to 80°C; The extraction time for obtaining the extract is 1 to 5 hours. A processed product from the Ecklonia genus. [Effects of the Invention]

[0022] According to the present invention, the water extract of Kurome, which can be used in foods, etc., inhibits lipase activity. It is possible to provide a composition for preventing bowel movements, inhibiting lipid absorption, improving stool lubrication, or regulating the intestines. do. [Brief explanation of the drawings]

[0023] [Figure 1] FIG. 1 is a graph showing the results of a lipase activity inhibition test of water extracts (test samples) of Kurome obtained at various temperatures and extraction times in Test Example 2. DETAILED DESCRIPTION OF THE INVENTION

[0024] [Lipase activity inhibiting composition] In the present invention, the composition for inhibiting lipase activity is Ecklonia kurome (scientific name: Ecklonia kurome) of the genus Ecklonia kurome (scientific name: Ecklonia kurome) It contains an effective amount of a water extract of (Ilia kurome).

[0025] Kurome is a type of brown algae belonging to the genus Ecklonia in the family Lessoniaceae. The part of the lome is not limited as long as it can produce the effects of the present invention. For example, the lome may be a leaf or a stem. (core), and bud, and at least one selected from the group consisting of thallus, and / or stems are preferred.

[0026] Kurome is widely distributed in the waters around Japan, from the central and southern Pacific coast of Honshu to Kyushu, the Seto Inland Sea, and the Japan Coast. It is widely distributed, and overseas it has been confirmed to be distributed in places such as Jeju Island in Korea. The growth of seaweed and the amount of nutrients vary depending on the strength of the ocean current, water temperature, and the height of the seabed. The origin of Kurome is not limited as long as the effects of the present invention are achieved. For example, Tottori Examples include those produced in Tottori Prefecture, Oita Prefecture, Kumamoto Prefecture, Ehime Prefecture, and Aichi Prefecture, and those produced in Tottori Prefecture, Oita Prefecture, and It is preferable that the product is from Kumamoto Prefecture, more preferably from Tottori Prefecture or Oita Prefecture, and particularly preferably from Tottori Prefecture. Here, prefecture-specific products refer to the sea areas managed by the prefecture (port areas, fishing port areas, etc.). This refers to items that can be harvested either in the general sea area or on the coast (coastal conservation area, general public coastal area). Although not limited thereto, from the viewpoint of significantly exhibiting the effects of the present invention, the harvesting time of Kurome is In the case of Kurome grown in Tottori Prefecture, for example, February to August is preferred, and April to June is more preferred.

[0027] Although Kurome is a perennial plant, its growing period is not limited as long as the effects of the present invention are achieved. The growing period of Kurome is, for example, preferably 1 to 6 years, more preferably 1 to 5 years. Preferably, the period is 1 to 4 years, more preferably 1 to 3 years, and most preferably 1 to 2 years. In the first year of growth, the Kurome plant has a bamboo leaf-like shape, and from the second year onwards, the upper part of the stem The pinnate leaf parts on the stem indicate that the Kurome is in its first or second year of growth. It is possible.

[0028] The water extract of Kurome can be extracted with water by a conventional method as long as the effects of the present invention are achieved. The extract may be prepared by the above method, and may be a liquid extract or a dried product thereof.

[0029] Although not limited thereto, for example, the kurome in a raw state, dried matter, or crushed matter is soaked in water. The water extract of Kurome is prepared by immersion extraction under optimal temperature and time conditions. Although not limited thereto, in the example (Test Example 6) described later, It has been confirmed that the use of dried products provides a greater effect of the present invention. The method for obtaining the dried product is not particularly limited, and known methods can be used. For example, drying in the sun can be used. Drying the kurome in the sun is also possible. In this case, the conditions will vary depending on the weather, season, time of day, etc. and may be adjusted accordingly, but at least It is recommended to dry the product in the sun for 6 hours or more, 12 hours or more, or 24 hours (1 day) or more. When a dryer is used, the drying conditions are not particularly limited, but may be freeze-drying, heat drying, etc. The heating temperature when drying by heating is preferably, for example, 30 to 50°C. The temperature is more preferably 5 to 40° C. The heating time when drying by heating is, for example, 1 to 24 hours. It is preferable that the time is 1 to 10 hours, more preferable that the time is 1 to 5 hours.

[0030] In another embodiment, for example, kurome is pressed to obtain juice, and the juice separated by pressing is The pressed lees are soaked in water and extracted under optimal temperature and time conditions. After the extracts are obtained, they may be mixed together and used as a water extract of Kurome.

[0031] The temperature conditions for the above immersion extraction are not limited as long as the effects of the present invention are achieved. However, for example, the temperature may be room temperature or may be heated. The temperature is, for example, preferably 30 to 120°C, more preferably 40 to 110°C, and 90°C is more preferable, 50 to 80°C is even more preferable, and 60 to 80°C is even more preferable. Preferably, 60 to 75°C is even more preferred, 65 to 75°C is even more preferred, and 65 to 70 °C (e.g., about 70°C) is most preferred.

[0032] The time conditions for the above immersion extraction are not limited as long as the effects of the present invention are achieved. However, the extraction time is preferably, for example, 10 minutes to 24 hours, and more preferably 30 minutes to 10 hours. The time is preferably 1 to 5 hours (for example, 1 to 3 hours), more preferably 2 to 4 hours (for example, 2.5 hours to 3.5 hours, 3 hours) is particularly preferred.

[0033] [Application] The composition for inhibiting lipase activity of the present invention inhibits the activity of a lipid-degrading enzyme (lipase) in vivo. The lipase activity inhibitory effect can be measured by the method described in the Examples below. It is possible to evaluate it more precisely.

[0034] The composition for inhibiting lipase activity of the present invention inhibits the activity of a lipid-degrading enzyme (lipase) in vivo. By exerting an effect of inhibiting lipid absorption, inhibiting visceral fat accumulation, improving obesity, etc. It can also be used effectively for intestinal regulation, improving intestinal bacteria, and improving the intestinal environment. can.

[0035] Furthermore, the present invention can suppress lipid decomposition and promote lipid excretion from the body, thereby reducing the amount of lipid in the feces. It also increases the amount of lipids in the body, which is good for promoting bowel movements and improving the smoothness of stool during bowel movements. This means that pharmaceutical preparations with a lipase activity inhibitory mechanism can be used in It is used as an anti-obesity drug by inhibiting the absorption of glycerides, and is effective in increasing bowel movements and reducing oily stools. This is also evident from reports that the condition is accompanied by an increase in the number of cases (Non-Patent Document 3).

[0036] Furthermore, the present invention can suppress lipid breakdown and promote lipid excretion from the body, thereby reducing oily skin. It can also be used to improve skin.

[0037] The composition of the present invention can be used in a variety of applications, including, but not limited to, pharmaceuticals, quasi-drugs, cosmetics, foods and beverages, feeds, Alternatively, it can be prepared and processed as a pet food.

[0038] For example, when prepared as a food or drink, it has the potential to inhibit lipid absorption, inhibit visceral fat accumulation, improve obesity, and Foods and beverages that claim to improve the intestinal flora, intestinal environment, and stool lubrication as functional foods and beverages. In other words, to be used as health foods, functional foods, foods for the sick, and foods for specified health uses, etc. can be done.

[0039] Health foods, functional foods, foods for the sick, and foods for specified health uses are specifically solid preparations. (tablets, granules, fine granules, powders, capsules, chewable tablets, etc.) and liquids (syrups, It can be used in various formulations such as suspensions, liquid foods, etc. It can be manufactured in the same manner as known pharmaceutical preparations, and contains an active ingredient and a carrier acceptable for food, e.g. For example, it can be prepared by mixing with suitable excipients and the like, and then using conventional means.

[0040] For example, in the case of tablets, the active ingredient in powder form and pharmaceutically acceptable carrier ingredients (excipients, etc.) It can be prepared by mixing and compressing, and candy and other confectionery tablets are molded. Tablets may be prepared by injection into the granules. Furthermore, the tablet may be a single-layer tablet or a layered tablet such as a double-layer tablet. stomach.

[0041] Granules and other powders are produced using various granulation methods (extrusion granulation, grinding granulation, dry compaction granulation, flow granulation, etc.). The tablets may be prepared by the above-mentioned methods. It can be prepared by appropriately combining granulation and tableting methods (wet tableting, direct tableting), etc.

[0042] The capsules are prepared by placing powdered granules (powder) in capsules (soft or hard capsules) using a conventional method. It can be prepared by filling the tablet with a pharmaceutical preparation (such as a tablet, granules, etc.) or a liquid preparation.

[0043] Liquid formulations are prepared by dissolving each ingredient in an aqueous medium (purified water, purified water containing ethanol, etc.) which is a carrier component. The solution is dissolved or dispersed, filtered or sterilized as necessary, filled into a specified container, and then sterilized. It can be prepared by:

[0044] Soft capsules have a smooth surface and are easy to swallow, making them popular with users. Examples of methods for producing the coating agent include a flat plate method, a rotary method, and a seamless method.

[0045] The rotary method (punching method) is a manufacturing method in which the sheet-shaped capsule shell is pressed against the flowing filling material. The object is sandwiched between the mold and formed into a capsule shape along the hole of a rotating cylindrical mold. The capsule shell composition and contents are dispensed from multiple concentric nozzles in the drip-less method. are simultaneously ejected and formed into a seamless capsule shape.

[0046] The base material for the soft capsule shell is not particularly limited, but may be starch, pullulan, cellulose, etc. Starch, polyvinyl alcohol, gelatin, succinated gelatin, etc. can be used. Preferred are cellulose, gelatin, and succinated gelatin, and more preferred are gelatin and succinated gelatin. These may be used alone or in combination of two or more.

[0047] The preferred dosage form of the solid preparation of the present invention is a capsule or a tablet, and a soft capsule (soft capsule) It is more preferable that the formulation is a capsule or soft capsule.

[0048] Also, soups, juices, fruit juice drinks, milk, dairy drinks, whey drinks, lactic acid bacteria drinks, tea drinks Food, alcoholic beverages, coffee drinks, carbonated drinks, soft drinks, water drinks, cocoa drinks, jelly Liquid drinks such as liqueur drinks, sports drinks, diet drinks, etc., and semi-solid drinks such as pudding and yogurt. The composition of the present invention can be produced as a food product, noodles, confectionery, spreads, etc. .

[0049] Examples of soups include miso soup, Kurome soup, Kurome chazuke, etc. However, the Kurome algae (or dried algae) can be soaked in water for 1 to 5 hours. By heating the product at 0-80℃, the useful components in the kurome are extracted, and this extract is It is possible to produce ingredients for soups containing the algae. The crushed material can be used as soup stock in various Japanese dishes, including clear soup. Dried or crushed Kurome algae can be added to soy sauce and other foods to be used as a seasoning. By performing heat treatment within the above range of conditions, lipase activity can be inhibited. It is expected that the effective ingredients will be dissolved and the ingredients will be efficiently absorbed.

[0050] The composition of the present invention can also be prepared as a stew. As one of the raw materials, it is boiled with water, dashi, etc., and seasoned with salt, sake, sugar, mirin, etc. Examples of simmered dishes include Kurome tsukudani, Kurome simmered, and Kurome simmered. Examples of simmered dishes include simmered romes, simmered kurome, simmered kurome, and simmered kurome. However, from the viewpoint of long-term heat treatment, black tuna is boiled in soy sauce, stewed, simmered in soy sauce, or Simmered dishes are preferred, and tsukudani of Kurome or simmered dishes are more preferred.

[0051] Although not limited to this, the algae body of Kurome (or dried algae body) is soaked in water and used for soup stock or By heating it with seasonings for 1 to 5 hours at 50 to 80°C, the It is possible to extract the useful components from lome and produce stews containing this extract. By carrying out the heat treatment within the above conditions, the components effective in inhibiting lipase activity are obtained. It is expected that the ingredients will be dissolved and efficiently ingested. It may be boiled down until the liquid is gone, but it should be heated within the above conditions. By processing, the ingredients that are effective in inhibiting lipase activity are extracted and then boiled down. When lowering the temperature after boiling, the active ingredients will soak into the blackcurrant and other ingredients. This leads to efficient intake of the ingredients.

[0052] The above soups and simmered dishes can also be sold as prepared foods or ready-to-eat meals. In the case of processed foods using conventional kurome, high heat (90- In the present invention, cooking is carried out under conditions of a temperature of 100°C or less and a short time (for example, within 10 minutes). The longer the cooking time and lower the heat, which are not possible under conventional cooking conditions, the more pronounced the lipase activity. It is now possible to prepare processed products from Ecklonia kurome, which has enhanced activity inhibitory effects and health functions.

[0053] When the composition of the present invention is prepared as a food or drink, various food additives may be added thereto. Food additives include, for example, antioxidants, colorings, flavorings, seasonings, sweeteners, acidulants, pH Examples include regulators, quality stabilizers, preservatives, etc.

[0054] The composition of the present invention can be prepared as a health food, a functional food, a food for the sick, or a food for specified health uses. When preparing the product, other fat absorption inhibitors may be added. For example, indigestible dextrin , isomaltodextrin, EPA, DHA, gallic acid, gall derived from Terminalia bellirica gallic acid from the fruit of Scutellaria Baicalensis, valine-valine-tyrosine Examples include lolin, ellagic acid derived from African mango, inulin, and gymnemic acid. Not limited to.

[0055] When the composition of the present invention is prepared as a drug or quasi-drug, the active ingredient It can be prepared as a formulation containing a water extract of lome and preferably a pharmaceutically acceptable carrier. Pharmaceutically acceptable carriers are generally inert substances that do not react with the active ingredient. Non-toxic, solid or liquid fillers, diluents or encapsulating materials, such as , water, ethanol, polyols, suitable mixtures thereof, vegetable oils, and other solvents or dispersion media. Examples include the body.

[0056] Medicines and quasi-drugs are administered orally or parenterally, for example, into the oral cavity or the digestive tract. Oral administration preparations include solid preparations (tablets, granules, fine granules, powders, etc.). These include tablets (tablets, capsules, chewable tablets, etc.) and liquids (syrups, suspensions, inhalants, etc.). Examples of parenteral administration preparations include infusions, nasal drops, and injections.

[0057] Drugs and quasi-drugs may contain additives commonly used in the pharmaceutical field. Such additives include, for example, excipients, binders, disintegrants, lubricants, antioxidants, and colorants. Coloring agents, flavoring agents, etc. can be used as needed. Therefore, they can be coated with known retardants, etc. If necessary, other additives or drugs, such as antacids and gastric mucosa protecting agents, may be added.

[0058] Pharmaceuticals and quasi-drugs can be applied in the form of oral compositions, oral compositions, etc. Furthermore, pharmaceuticals and quasi-drugs may be used therapeutically or non-therapeutically.

[0059] In addition, when used in the form of medicines, quasi-drugs, cosmetics, etc., the production of microorganisms on the skin Lipases and other enzymes in the skin produce fatty acids from sebum, which then activate neutrophils and help keep them active. Activated neutrophils release active oxygen, which can be a cause of rough skin. By incorporating it into topical formulations, it has lipase activity that is effective against rough skin such as acne. Inhibitory compositions may be provided.

[0060] The daily oral intake or dosage of Kurome water extract for adults varies depending on the individual's condition and body condition. Weight, sex, age, activity of ingredients, route of ingestion or administration, schedule of ingestion or administration, formulation The daily intake of the water extract of Kurome for an adult is The oral intake or dosage of, for example, 500 mg / day or more in dry solids equivalent. Preferably, 800 mg / day or more is more preferred, and 1000 mg / day or more is even more preferred. A dose of 1500 mg / day or more is particularly preferred, and 2000 mg / day or more is most preferred.

[0061] The daily oral intake or dosage of the water extract of Kurome for adults is, for example, 45 g. / day or less is preferred, 40g / day or less is more preferred, and 35g / day or less is even more preferred, 30 g / day or less is particularly preferred, and 25 g / day or less is most preferred.

[0062] The daily oral intake or dosage of the water extract of Kurome for adults is, for example, 500 mg to 45g / day is preferable, 800mg to 40g / day is more preferable, and 1000mg to 35 g / day is more preferable, 1500 mg to 30 g / day is particularly preferable, and 2000 mg to 25g / day is most preferred.

[0063] The content of the water extract of Kurome can be set to the amount that corresponds to the intake or administration amount described above. The oral intake or dosage per day for adults is determined according to the dosage form, e.g., capsules. If it is a tablet, 1 to 6 capsules, 1 to 4 capsules, 1 to 3 capsules, or 1 to 2 capsules It may be taken in divided doses.

[0064] The composition of the present invention can be administered once to several times a day, typically 1 to 6 times a day, 1 to 3 times a day, or 1 to 2 times a day. It may be taken or administered once or twice or at any time and interval.

[0065] [Method of manufacturing water extract or processed product of Ecklonia kuroume] In the present invention, the method for producing a water extract or a processed product of Ecklonia kurome comprises the steps of: (1) extracting Ecklonia kurome from Ecklonia kurome; The method includes a step of extracting Kurome with hot water at 50 to 80°C for 1 to 5 hours.

[0066] The above steps are carried out in accordance with the temperature conditions, time conditions, etc. described in the description of the composition for inhibiting lipase activity. It is possible to set it accordingly as appropriate. [Example]

[0067] Next, the present invention will be described in detail with reference to examples, but the present invention is not limited to the following examples. In addition, the amount of extract shown in the examples is the amount of dry solids unless otherwise specified. This is the amount converted into minutes.

[0068] [Test Example 1: Examination of extraction conditions for water extract of Kurome] Ecklonia kurome (scientific name: Ecklonia kurome) algae (produced in Tottori Prefecture, at the time of harvest) (season: April, part: leaf and stem, growth period: 2-5 years) and cut into small pieces of 5g each. The water extracts used as test samples were prepared by combining the appropriate temperature with an extraction time of 1 hour or 3 hours. The extraction conditions were examined using the amount of extract and viscosity as indicators. The results regarding the amount of extract are shown in Table 1. Shown below.

[0069] [Table 1]

[0070] As shown in Table 1, the water extract of Kurome exhibited temperature- and extraction time-dependent results. The amount of extract increases as the temperature increases, and the longer the extraction time at each extraction temperature, the greater the amount of extract. It was confirmed that the amount of extract obtained was increased.

[0071] The results regarding viscosity are shown in Table 2. The viscosity was evaluated by filling 30 mL of each test sample. When using a filter (mesh size: 3 μm, manufactured by ADVANTEC), the time required for filtration to be completed It takes more than 15 minutes, and you should replace the filter every time it gets clogged, and use more than three filters. The samples that were found to be viscous were evaluated as "sticky: x".

[0072] [Table 2]

[0073] As shown in Table 2, the temperature was 50°C (1 hour treatment and 3 hours treatment), 60°C (1 hour treatment and The water extract of Kurome at 70°C (treated for 3 hours) and 70°C (treated for 1 hour) was viscous and could be filtered. On the other hand, the filtration process took time. The water extract of lome does not show any obvious viscosity when filtered, and is relatively easy to prepare. From the viewpoint of extract preparation and convenience as a raw material, we were able to ) was found to be more desirable.

[0074] [Test Example 2: Examination of the lipase activity inhibitory effect depending on extraction conditions] Using the water extracts (test samples) of Kurome obtained at each extraction temperature and extraction time, lipase The lipase activity inhibition test was carried out using 4-methylumbelliferone. 4-MUO was used as the substrate. 0.05 ml of 4-MUO solution (1 0 -4 0.1 ml of the test sample was added to the 100 mM PBS, mixed, and then 3 U / mL of pancreatic lipase solution was added. After adding 0.05 ml of the solution and reacting at 37°C for 20 minutes, 0.1 ml of hydrochloric acid was added. The reaction was stopped by the reaction in the solution obtained by mixing the reaction solution and the citric acid solution. Fluorescence of the resulting 4-methylumbelliferone (excitation wavelength 320 nm, emission wavelength 450 nm) The activity was measured using a fluorescence microplate reader. The activity value without the addition of the test sample was set at 100%. The activity value (IC50) of each test sample was calculated as follows. The lower the activity value, the greater the lipase inhibitory effect. The results are shown in Figure 1. In Figure 1, the dashed line indicates the results for an extraction time of 1 hour. The solid line shows the results for an extraction time of 3 hours.

[0075] As shown in Figure 1, the water extract of Kurome prepared at 70°C for 3 hours had the most significant lipase activity. Interestingly, when the temperature was shifted from 70°C to lower or higher temperatures, It was found that the degree of inhibition of lipase activity weakened as the temperature increased.

[0076] It is expected that the components present in the viscous solution at low temperature and the solution at high temperature will be different. However, similar results were obtained. From this, it can be seen that the extraction conditions at 70°C are It was suggested that the ratio of the components was well balanced. Based on the results of enzyme activity inhibition evaluation, it was determined that extraction at 70°C for 3 hours was optimal. A higher inhibitory effect was observed even at temperatures up to 80°C for 1 to 3 hours compared to other extraction temperatures. Therefore, the extraction conditions for large-scale preparation of water extract of Kurome should be controlled within this range. It was confirmed that this was appropriate.

[0077] Test Example 3: Comparison of lipase activity inhibitory effects with orlistat and water extract of Kurome Consideration of daily dosage of substances] Using orlistat and a water extract of Kurome, lipase activity was measured in the same manner as in Test Example 2. Orlistat inhibits lipid metabolism by inhibiting lipase. It is a component known to have an anti-absorption effect.

[0078] The water extract of Kurome was prepared at 70°C for 3 hours. The comparative results of the enzyme activity inhibition test (IC50) for both components are shown in Table 3.

[0079] [Table 3]

[0080] As shown in Table 3, when comparing IC50 values, orlistat was found to be 26% of the water extract of Kurome. It was 8.3 times.

[0081] Regarding the effective dose of orlistat, "J. Zhi et al., Retrospective population-b ased analysis of the dose-response (fecal fat excretion) relationship of orlis tat in normal and obese volunteers Clin Pharmacol Ther. 1994 Jul;56(1):82-5. In Table II, the 95% confidence interval for ED50 is 30.2 to 166.0 (mg / day).

[0082] Based on the effective dose of orlistat described in the above paper and the results in Table 3, the water extract of Kurome The effective dose (ED50) of the substance is 8g / day to 45g / day, as shown in Table 4. This range of values ​​is equivalent to the efficacy of the drug orlistat. Therefore, when used as a food, the daily intake should be 500 mg or more. Above this level, appropriate health benefits are expected.

[0083] [Table 4]

[0084] [Test Example 4: Examination of lipase activity depending on collection time] The Kurome algae collected from the coastal area of ​​Tottori Prefecture at the end of April 2020 were lightly washed with water and stored at 37°C. Drying was carried out in a dryer (manufacturer: ETTAS (AS ONE) model number: OFW-450B) under the following conditions: After drying, 0.5 g was weighed out and finely crushed in a food processor. 35 mL of distilled water was added to the kurome, and the mixture was left to stand in a thermostatic bath at 70°C for 3 hours for extraction. Then, the extract is filtered by suction using a glass filter and a water aspirator. The extract was then suction filtered using 3 μm and 0.45 μm filters. The filtered extract was frozen in liquid nitrogen and then freeze-dried for 3 days. In the same manner as in Example 2, the activity value (IC50) of each test sample was calculated.

[0085] [Table 5]

[0086] As shown in Table 5, when Kurome harvested at the end of April was used as the raw material, significant lipase activity was observed. Although the results are not shown, the Kurome harvested up until March was used as the raw material. When used in the same way as in the previous study, the lipase activity inhibitory effect was confirmed. It was found that when used in

[0087] [Test Example 5: Examination of lipase activity depending on growth period] Among the Kurome plants collected from the coastal areas of Tottori Prefecture, the algae with a growth period of about 1 to 2 years were After lightly washing with water, the water was drained and the mixture was finely ground in a food processor. 35 mL of distilled water was added to 5 g of kromme, and the mixture was left to stand in a thermostatic bath at 70°C for 3 hours to perform extraction. After extraction, the extract is filtered using a glass filter and a water aspirator. The residue was removed, and the extract was then suction filtered using 3 μm and 0.45 μm filters. The filtered extract was frozen in liquid nitrogen and then freeze-dried for 3 days. The activity value (IC50) of each test sample was calculated in the same manner.

[0088] [Table 6]

[0089] As shown in Table 6, when Kurome, which has a growth period of about 1 to 2 years, is used as the raw material, the yield is higher. A strong inhibitory effect on lipase activity was demonstrated.

[0090] [Test Example 6: Examination of lipase activity by drying after collection] Kurome collected from the coastal areas of Tottori Prefecture is lightly washed with water and then dried in the sun outdoors. After drying, 0.5 g of dried Kurome was weighed out and finely ground in a food processor. After grinding, 35 mL of distilled water was added to the ground kurome, and the mixture was left to stand in a thermostatic chamber at 70°C for 3 hours. After water extraction, the extract was filtered using a glass filter with a water aspirator. The extract was then filtered through 3 μm and 0.45 μm filters to remove seaweed residue. The filtered extract was frozen in liquid nitrogen and then freeze-dried for 3 days. Thereafter, the activity value (IC50) of each test sample was calculated in the same manner as in Test Example 2.

[0091] [Table 7]

[0092] As shown in Table 7, the kurome was dried before the extraction process and the dried kurome was used as the raw material. When the hydroxybenzoate was added, a higher lipase activity inhibitory effect was observed.

[0093] [Test Example 7: Comparison of lipase activity with ethanol extract] To prepare the water extract, the algae of Kurome collected from the coastal area of ​​Tottori Prefecture were lightly washed with water and then left for 3 Drying machine (Manufacturer: ETTAS (AS ONE) Model: OFW-450) at 7°C After drying, 0.5 g of dried Kurome was weighed and put into a food processor. After grinding, 35 mL of distilled water was added to the kurome, and the mixture was left to stand in a thermostatic chamber at 70°C for 3 hours. After extraction, the extract was filtered using a glass filter and a water aspirator. The extract was then filtered through a 3 μm and 0.45 μm filter to remove seaweed residue. The filtered extract was frozen in liquid nitrogen and then freeze-dried for 3 days. The test sample was then subjected to the same procedure as in Test Example 2 to obtain a freeze-dried powder. The activity value (IC50) was calculated.

[0094] To prepare the ethanol extract, the algae of Kurome collected from the coastal area of ​​Tottori Prefecture were lightly soaked in water. After washing and removing the water, the sample was weighed out to approximately 5 g. Then, it was frozen at -30°C and dried in a freeze dryer (manufacturer's - EYELA Model No. FDU-2110 (similar to other freeze-drying methods) for 3 days After drying, 0.5 g of the dried kurome was weighed out and crushed in a food processor. After crushing, 35 mL of ethanol was added to the kurome and the mixture was left to stand in a thermostatic chamber at 70°C for 3 hours to perform extraction. The test sample obtained by freeze-drying after ethanol extraction was the same as that obtained by water extraction. Since the amount was less than 100g, 4 sets of (0.5g / 35mL) were prepared. After extraction, the glass filter The extract was filtered by suction using a water aspirator to remove seaweed residue. When the four sets were combined, the total volume of the extract was 120 mL. The solution was concentrated and the organic solvent was removed using a rotary evaporator. After 6 times concentration, Add 50-60 mL of distilled water in several portions (120 mL → 20 mL) and add ethanol. The extract was concentrated and the organic solvent removed until the odor was no longer detectable. After freezing, the mixture was freeze-dried for 3 days to obtain 37.8 mg of freeze-dried powder. The activity value (IC50) of the test sample was calculated in the same manner as above. In addition, since the extraction yield with ethanol is significantly lower than that with water, the extraction yield with water or ethanol was The IC50 was calculated based on the weight of dried kurome before extraction.

[0095] [Table 8]

[0096] As shown in Table 8, when Kurome is used as a raw material, the extraction yield and lipase activity inhibitory effect are Therefore, it was found that the water extract was preferable.

Claims

[Claim 1] The invention described herein.

Images