Lipase activity inhibitors and their uses

Abstract

To provide a lipase activity inhibitor of natural origin that strongly inhibits lipase activity and can be effectively used to prevent and treat obesity and diseases that could be caused by the obesity as a risk factor; and a composition that allows industrially effective utilization of the lipase activity inhibitor.SOLUTION: A lipase activity inhibitor contains a plant belonging to Kunzea or a processed product thereof, or suitably a product extracted or purified from leaves and / or bark of Kunzea ericoides, or more suitably, an extracted product obtained by using water, ethanol or a hydrous solvent thereof. Desirably, the inhibitor also comprises polyphenols. There is also provided an edible composition or a pharmaceutical composition containing the inhibitor.SELECTED DRAWING: None

Description

[Technical Field]

[0001] The present invention relates to a lipase inhibitor containing a plant belonging to the genus Kunzea, such as Kanuka, as a raw material, and uses thereof. More specifically, the present invention relates to a lipase inhibitor containing the plant, and oral compositions containing the same, such as foods and beverages, pharmaceuticals, feeds, and pet foods. [Background technology]

[0002] The rapid increase in obesity, not only in developed countries but also in developing countries, has been raised as a major public health problem. In recent years, Western-style eating patterns have become more prevalent in Japan, leading to overeating and increased energy intake. In particular, the number of patients suffering from lifestyle-related diseases such as obesity and hyperlipidemia, which are caused by the long-term intake of high-calorie foods that are high in fat, has been increasing significantly as eating habits have become more Westernized, causing major social problems such as increased national medical expenses.

[0003] Obese people in Japan (BMI ≥ 25 kg / m 2 In 1980, the proportion of obese men was 18.4% and that of women was 20.6%, but by 2017, it was 30.7% and 21.9%. The proportion of obese men has increased significantly, especially since the Heisei era, and has recently reached around 30%. Meanwhile, the proportion of obese women has remained at around 20%. Although the proportion of obese people in Japan is lower than the global proportion, preventive measures, particularly for adult men, are necessary to reduce the risk of obesity-related diseases and functional disorders such as those mentioned above.

[0004] Furthermore, factors that cause obesity and lifestyle-related diseases in modern times include not only poor dietary habits but also chronic lack of exercise. In recent years, advances in science and technology have made various aspects of daily life more convenient. For example, it is clear that the amount of physical activity has decreased due to the automation of housework and work, and the development of transportation, and this lack of exercise, along with changes in dietary habits, is one of the causes of the recent increase in obesity and lifestyle-related diseases.

[0005] According to the National Health and Nutrition Survey (Ministry of Health, Labour and Welfare), the percentage of people who were considering and / or were currently managing their weight was 82.2% for men and 86% for women in 2006, but by 2013 it had risen to 90.2% for men and 93.5% for women, indicating that public health awareness is increasing year by year. However, the percentage of people who have an exercise habit was low at 33.8% for men and 27.2% for women. Thus, while awareness of the health benefits of physical activity and exercise is becoming more widespread among the public, the percentage of people who actually have developed an exercise habit is low.

[0006] Dietary therapy and exercise therapy are mainly used to prevent and treat obesity, but because it takes a long time to see results and requires strong willpower, it is difficult to implement and continue these in today's busy world, and they often result in only temporary weight loss.

[0007] Drug treatments are mainly applied to patients with severe obesity, and there are concerns about side effects of the drugs, so neither method can be considered a general-purpose method. Therefore, there is a need to provide a safer method for preventing and improving obesity.

[0008] To prevent obesity, substances that inhibit the activity of digestive enzymes such as amylase and lipase have been developed to reduce excessive energy intake. Lipids, in particular, are the most calorie-dense food component, and reducing lipid accumulation is thought to be an effective way to prevent obesity. Pancreatic lipase, a lipid-digesting enzyme secreted by the pancreas, acts on triglycerides (neutral lipids), the main component of dietary fat, hydrolyzing ester bonds to produce diglycerides, monoglycerides, fatty acids, and glycerin. These pancreatic lipase degradation products are absorbed by the small intestine and typically serve as an energy source. However, in cases of low calorie consumption due to lack of exercise or excessive fat intake, lipids resynthesized in the body become fat and accumulate in the body, eventually leading to symptoms of obesity. Therefore, substances that inhibit lipase activity are effective in preventing and treating obesity and the associated diseases.

[0009] From these viewpoints, natural components that inhibit lipase activity, as well as various sources and processed products containing such components, have been proposed. For example, epigallocatechin gallate, a component of green tea (Patent Document 1), water extracts of bell peppers, pumpkins, shimeji mushrooms, maitake mushrooms, hijiki seaweed, green tea, black tea, and oolong tea (Patent Document 2), tannins derived from plant bark (Patent Document 3), an ethanol extract of Cassia capillaris containing tannin as an active ingredient (Patent Document 4), a tamarind seed coat extract containing proanthocyanidin as an active ingredient (Patent Document 5), and compounds derived from avocados, onions, and other fruits. Examples of such extracts include hot water extracts of Japanese knotweed, Chinese cabbage, jamboran, myrtle and wax tree (Patent Document 6), 50% ethanol extracts of Japanese knotweed, kidney bean, butterfly staghorn, giant peach, banyan tree, Alpinia speciosa, Hermissenda japonica, Kalanchoe kaempferi, Takasagogi, Nandina, safflower, Senna chinensis, Momordica charantia and Ryukyu pine (Patent Document 7), quinic acid diester derivatives of green coffee beans (Patent Document 8), and extracts from fennel, melon, fig and Centipede tree (Patent Document 9).

[0010] The kanuka plant used in this invention (scientific name: Kunzea ericoides) is a plant belonging to the genus Kunzea in the Myrtaceae family, native to New Zealand and growing wild only there. It is commonly known as kanuka, white manuka, or white tea tree. The Maori, the indigenous people of New Zealand, have used kanuka in traditional medical treatments and rituals. A decoction of the leaves has been used to treat urinary disorders and as an antipyretic. The steam produced when the leaves are boiled has been inhaled to treat coughs and colds. A decoction of the seeds has been used externally to suppress inflammation and as a drink for diarrhea. The leaves and seeds have also been chewed to treat dysentery. The resinous exudate has been used to treat burns; decoctions of the seeds, leaves, and bark have been used to treat inflammation; and a decoction of the bark has been used externally as a sedative, for the treatment of oral and eye diseases, and as a disinfectant (Non-Patent Documents 1 and 2).

[0011] Extracts of kanuka leaves and branches are known to contain trace components such as terpenes, such as globulol, spatulenol, viridiflorol, betulinic acid, oleanolic acid, and ursolic acid, and flavonoids, such as 5,7-dihydroxy-6-methylflavanone, and it is said that these components are responsible for the aforementioned pharmacological effects (Non-Patent Document 1).

[0012] It has also been reported that gallic acid, quercetin, catechin, etc. were detected as major components in an extract of kanuka leaves extracted using subcritical water (Non-Patent Document 3).

[0013] Furthermore, in an attempt to industrially utilize the oil obtained by processing kanuka, a fungicidal emulsion containing kanuka oil has been proposed (Patent Document 10). However, there is no reference to the relationship between the kanuka leaves and bark themselves or extracts and the inhibition of lipase activity. [Prior art documents] [Patent documents]

[0014] [Patent Document 1] Japanese Patent Application Publication No. 3-228664 [Patent Document 2] Japanese Patent Application Publication No. 3-219872 [Patent Document 3] Special Publication No. 60-11912 [Patent Document 4] Japanese Patent Application Laid-Open No. 2012-240928 [Patent Document 5] Japanese Patent Application Publication No. 9-291039 [Patent Document 6] Re-tabled publication No. 2005 / 056031 [Patent Document 7] Japanese Patent Application Laid-Open No. 2010-280734 [Patent Document 8] Japanese Patent Application Laid-Open No. 2011-207799 [Patent Document 9] Japanese Patent Application Publication No. 2017-203032 [Patent Document 10] Special Publication No. 2005-538157 [Non-patent literature]

[0015] [Non-Patent Document 1] Rebecca M. Wyatt et al., Phytother.Res., Vol. 19, pp. 963-970, 2005. [Non-patent document 2] Maria Lis-Balchin, Aromatherapy Science: A Guide for Healthcare Professionals, pp. 217-218, 2006 [Non-patent document 3] Sinemobong Essien et al., Journal of Supercritical Fluids, Volume 158, Page 104721, 2020 Summary of the Invention [Problem to be solved by the invention]

[0016] Lipase activity inhibitors proposed to date, while used as pharmaceuticals, have significant side effects and limitations, such as the need to adhere to dosage instructions under the guidance of a physician. Therefore, there is a need for lipase activity inhibitors that contain active ingredients with proven efficacy and efficacy, have mild effects, and are highly safe even when taken over a long period of time. Extracts and components isolated from food raw materials have advantages, such as fewer side effects than pharmaceuticals, but they are often less effective in practice, are based on experimental results under impractical intake conditions, or require large amounts to be taken in the form of a normal diet. None of these have demonstrated fully satisfactory efficacy. Therefore, the present invention aims to provide a lipase activity inhibitor that potently inhibits lipase and can be effectively used to prevent and treat obesity and various diseases for which obesity is a risk factor, and that is highly safe and has little risk of side effects, as well as oral compositions containing the same, such as foods, beverages, pharmaceuticals, feed, and pet food. [Means for solving the problem]

[0017] In order to solve the above problems, the present inventors have conducted extensive research in order to provide a composition comprising a lipase activity inhibitor and the agent, which is highly safe, exhibits a strong lipase activity inhibitory effect, and can be effectively used in the prevention and treatment of obesity symptoms and various diseases related thereto. As a result, they have found that extremely high lipase activity inhibition is achieved when a specific plant is used, and that this can be effectively used in industrial fields such as food and beverages, pharmaceuticals, quasi-drugs, and animal feed, and have completed the present invention.

[0018] That is, the present invention is characterized by the following points. (1) The lipase activity inhibitor of the present invention contains, as an active ingredient, a plant belonging to the genus Kunzea of ​​the family Myrtaceae or a processed product thereof. (2) The inhibitor preferably contains, as an active ingredient, a dried powder of kanuka (Kunzea ericoides) leaves and / or bark, an extract thereof, or a purified product thereof. (3) The lipase inhibitor is preferably pancreatic lipase. (4) In the inhibitor, the extract is preferably extracted using water and / or a hydrophilic organic solvent. (5) In the inhibitor, the hydrophilic organic solvent is preferably ethanol or a water-containing product thereof (water content: 30 to 99% by mass). (6) It is more preferable that the inhibitor further comprises a polyphenol in combination. (7) It is more preferable that the polyphenols be one or more selected from the group consisting of proanthocyanidins, tannins, catechins, flavones, flavonols, isoflavones, and glycosides thereof. (8) The oral composition of the present invention contains the lipase activity inhibitor. (9) The oral composition is preferably in the form of a food or drink or a pharmaceutical product. (10) The oral composition is preferably used for obesity prevention or dieting. (11) The present invention also features a method for preventing or ameliorating obesity symptoms (excluding medical procedures) by orally ingesting the lipase activity inhibitor. [Effects of the Invention]

[0019] The lipase activity inhibitory effect of the kanuka leaf extract according to the present invention is extremely strong compared to the inhibitory effect of citrus rhizome extract and guava leaf extract, which are known to have lipase activity inhibitory effect. Therefore, the extract according to the present invention has an excellent lipase activity inhibitory effect and is effective in preventing obesity and diseases such as lifestyle-related diseases caused by obesity by inhibiting the absorption of dietary fat. Furthermore, in the fields of foods and beverages, pharmaceuticals, quasi-drugs, feeds, etc., which contain the agent, it can be effectively used as is or in a form blended with various conventional products in the above fields. DETAILED DESCRIPTION OF THE INVENTION

[0020] First, the lipase activity inhibitor of the present invention is characterized by containing a plant belonging to the genus Kunzea or a processed product thereof. As mentioned above, plants belonging to the genus Kunzea are classified in the Myrtaceae family, and while there are various examples of plants that are relevant to the present invention as described below, a representative example is kanuka (scientific name: Kunzea ericoides). Kanuka is native to New Zealand and grows wild only in that country. The indigenous Maori people of New Zealand used kanuka in traditional treatments and rituals. Later, early European settlers also used it as a medicinal plant for muscle pain, rheumatoid arthritis, and other conditions.

[0021] Examples of plants in the Kunzea genus include Kunzea acicularis, Kunzea acuminata, Kunzea affinis, Kunzea ambigua, Kunzea baxteri, Kunzea bracteolata, Kunzea calida, Kunzea cambagei, Kunzea capitata, Kunzea ericifolia, Kunzea ericoides, Kunzea eriocalyx, Kunzea flavescens, Kunzea glabrescens, Kunzea graniticola, Kunzea jucunda, Kunzea micrantha, and Kunzea. Examples of such species include Kunzea micromera, Kunzea montana, Kunzea muelleri, Kunzea newbyi, Kunzea obovata, Kunzea opposita, Kunzea parvifolia, Kunzea pauciflora, Kunzea pomifera, Kunzea preissiana, Kunzea pulchella, Kunzea recurva, Kunzea rupestris, Kunzea spicata, Kunzea strigosa, and Kunzea villiceps.

[0022] The preferred embodiment of the Kunzea plant, such as Kanuka, used in the present invention is the leaves or bark of the plant. This may be the leaves and / or bark themselves, those processed by drying, shredding, or crushing, extracts obtained by extracting these with a solvent, extracts obtained by removing the solvent from the extracts, or purified products obtained by subjecting the extracts to purification treatments such as column chromatography or solvent fractionation using adsorbents such as silica gel, magnesium silicate, ion exchange resins, activated alumina, cellulose, or activated carbon. For food applications, it is preferable, from the standpoint of convenience and production cost, to dry and appropriately crush the leaves and / or bark of the plant and prepare a powder, or to extract the finely shredded or powdered dried product with water or a hydrophilic organic solvent. For pharmaceutical applications, the extracts, extracts, or highly purified products are preferred.

[0023] The extraction solvent used to obtain the extract from the above-mentioned plants is preferably water and / or a hydrophilic organic solvent, including lower alcohols such as ethanol, water, etc., with water being particularly preferred. When the hydrophilic organic solvent is a water-containing substance, the water content is 30 to 99% by mass, more preferably 50 to 99% by mass, for example, when the hydrophilic organic solvent is water-containing ethanol. If the water content is outside this range, the desired effects of the present invention will be reduced.

[0024] The heating temperature when obtaining an extract from the above plants is preferably 1 to 90°C under normal pressure or 1 to 121°C under high pressure when water is used as the extraction solvent, and preferably 1 to 70°C under normal pressure when a hydrophilic organic solvent such as ethanol is used as the extraction solvent.

[0025] To obtain the extract of the present invention simply and efficiently, extraction with water or aqueous ethanol is preferred. The extraction process involves adding the extraction solvent in an amount approximately 1 to 20 times the mass of the raw material, and extracting for 10 minutes to 1 hour under normal or increased pressure, at room temperature or under heat, with appropriate stirring. Insoluble matter is removed by filtration or centrifugation, and if necessary, the extraction procedure is repeated using the above method to obtain an extract. The extract of the present invention can be obtained by further removing the solvent from the extract using known means such as vacuum distillation, freeze-drying, or spray-drying.

[0026] Next, the lipase activity inhibitor of the present invention is preferably a combination of the aforementioned kanuka plant or a processed product thereof with polyphenols. While known polyphenols can be used, plant polyphenols are readily available and convenient. These plant polyphenols include hydrolyzed tannins, catechins, condensed catechins (procyanidins, etc.), flavones, flavonols, isoflavones, etc. Examples of hydrolyzed tannins include gallotannins (gallnut tannin, gall tannin, etc.), ellagitannins (pedenculagin, tellimagladin, cajuarinin, etc.), ellagin oligomers (agrimoniin, oenothein B, etc.), and caffe-tannins (rosmarinic acid, lithospermic acid, etc.). Examples of catechins include epicatechin, epigallocatechin, epigallocatechin gallate, etc., and condensed tannins are catechin condensates, such as proanthocyanidins such as propelargonidin, procyanidin, and prodelphinidin, and their acid decomposition products (anthocyanidins).Further examples of flavones include apigenin and luteolin, flavonols include quercetin, quercitrin, isoquercitrin, rutin, and kaempferol, and isoflavones include daidzein, genistein, and isogenin, as well as glycosides thereof.

[0027] In the present invention, hydrolyzed tannins, catechins, proanthocyanidins, flavonols, isoflavones, proanthocyanidins, flavonols, isoflavones, and glycosides thereof, more preferably hydrolyzed tannins, catechins, and flavonols, are used alone or in combination with two or more of the above-mentioned plants of the genus Kunzea to form lipase activity inhibitors.

[0028] The polyphenols used in the present invention may be chemically synthesized pure products, or natural products containing these, more preferably plants, as raw materials, and are preferably dried powders, extracts, and purified products obtained by subjecting the raw materials to known processes such as drying, crushing, extraction, concentration, and distillation. That is, while there is no particular limitation on the plant as long as it contains the polyphenols, it is preferable to use one or more raw materials selected from grape seeds, guava leaves, unripe apples or skins, Eucommia leaves, Pu-erh tea leaves, Mallotus japonicus bark, green tea leaves, mulberry leaves, whole buckwheat plants, soybeans, French maritime pine, and cocoa beans, and the following forms can be used:

[0029] The form may be any of the following: the plant body has been processed by drying, shredding, pulverization, or the like; an extract obtained by adding a solvent to the plant body and subjecting it to extraction; an extract obtained by removing the solvent from the extract; or a purified product obtained by subjecting the extract to purification treatment such as column chromatography filled with an adsorbent such as silica gel, magnesium silicate, ion exchange resin, activated alumina, cellulose, activated carbon, or solvent fractionation. In order to achieve the desired effects of the present invention, when used in food applications, a dried powder or an extract extracted with water and / or a hydrophilic organic solvent is simple and convenient, while when used in pharmaceutical applications, the extract or purified product is preferable.

[0030] The extraction process uses hydrophilic organic solvents or aqueous solvents such as lower monohydric alcohols (e.g., methanol, ethanol, propanol, isopropanol), polyhydric alcohols (e.g., propylene glycol, 1,3-butanediol, glycerin), acetone, methyl ethyl ketone, and mixtures thereof. The extraction solvent is added in an amount equal to or approximately 100 times the mass of the raw material, and the extraction process is carried out under normal or increased pressure, at room temperature, or under heat, with appropriate stirring for 10 minutes to several days. Insoluble matter is removed by centrifugation or filtration to obtain an extract containing polyphenols. The solvent can then be removed from the extract by vacuum distillation, freeze-drying, spray-drying, or other means to prepare a polyphenol-containing extract. These solvents may also be commercially available.

[0031] A preferred embodiment of the lipase activity inhibitor of the present invention comprises, as an essential ingredient, the leaves and / or bark of a plant belonging to the genus Kunzea, such as kanuka, obtained as described above, or a dried product, extract, or purified product thereof. A more preferred embodiment comprises a plant derived from the genus Kunzea as the essential main ingredient, in combination with polyphenols, and most preferably, an extract or purified product of kanuka leaves and / or bark is used as the main ingredient, in combination with one or more dried powders, extracts, or purified products selected from the group consisting of grape seeds, guava leaves, unripe apples or skins, duzhong leaves, Pu-erh tea leaves, Mallotus japonicus bark, green tea leaves, and cocoa beans.

[0032] The lipase activity inhibitory effect is more pronounced when a material derived from a plant of the genus Kunzea is used in combination with a material containing polyphenols. The blending ratio of the two materials in this lipase activity inhibitor can be varied appropriately depending on the intended use and application, production costs, etc. to achieve the desired effect. The ratio of the material derived from a plant of the genus Kunzea to the material containing polyphenols is 100 / 0 to 0 / 100 (mass ratio; the same applies hereinafter), more preferably 100 / 0 to 50 / 50, and most preferably 90 / 10 to 60 / 40.

[0033] The lipase activity inhibitor of the present invention can be formulated in combination with various raw materials and ingredients as long as it does not violate the spirit of the present invention. Examples include excipients, moisture-proofing agents, preservatives, strengthening agents, thickeners, emulsifiers, antioxidants, sweeteners, acidulants, seasonings, coloring agents, and fragrances commonly used in foods and pharmaceuticals. The use of other known materials that have lipase activity inhibitory effects in combination with the lipase activity inhibitors of the present invention is also a desirable embodiment of the present invention.

[0034] As described above, the present invention provides a lipase activity inhibitor containing a dried product, extract, or purified product of a plant belonging to the genus Kunzea, such as Kanuka, or containing the same together with a dried product, extract, or product of a polyphenol-containing substance. This lipase activity inhibitor has excellent lipase activity inhibitory properties and suppresses the breakdown of dietary fat, thereby reducing the absorption of lipids in the body and promoting improved blood lipid metabolism, enabling the prevention of obesity and various related diseases. Therefore, while the lipase activity inhibitor of the present invention may be used directly for the desired purpose of the present invention, the present invention also provides compositions containing the same. These compositions can be used in a variety of forms, including foods and beverages, food additives, livestock feed, and pet food, with edible or pharmaceutical compositions being preferred.

[0035] The edible composition of the present invention is characterized by comprising the aforementioned lipase inhibitor, which contains as an essential ingredient an extract or purified product of a plant belonging to the genus Kunzea, such as Kanuka, preferably in the form of leaves and / or bark, or the essential ingredient and a dried powder, extract, or purified product of a plant containing polyphenols.

[0036] The edible composition may take the form of the above-mentioned dried powder, extract or purified product as is, or the lipase activity inhibitor may be added to liquid, gel, powder or solid foods, such as fruit drinks, soft drinks, tea, soup, jelly, yogurt, pudding, cake mix, seasonings such as furikake, miso, soy sauce, dressing, mayonnaise and yakiniku sauce, noodles, processed meat and fish foods such as ham and sausage, powdered, solid or liquid dairy products such as jam, milk, cream, butter and cheese, margarine, bread, cake, cookies and the like.

[0037] Furthermore, if necessary, the composition may be processed into powders, granules, pellets, tablets, etc., together with excipients such as dextrin, lactose, starch or processed materials thereof, cellulose powder, vitamins, minerals, animal, plant, or seafood oils and fats, proteins, carbohydrates, colorants, flavorings, or other food additives, or may be coated with gelatin or formed into capsules, or made into drinks, and used as a nutritional supplement or health food. In this case, a composition containing a known edible material having lipase activity inhibitory activity is preferred. The edible composition of the present invention can take a wide variety of forms, and is not limited to the above examples. However, from the perspective of suppressing the absorption of dietary fat in the body and improving lipid metabolism, it is desirable to add the composition to foods containing large amounts of fats and oils or carbohydrates, or to process it into the form of the nutritional supplement or health food.

[0038] The amount of the lipase activity inhibitor of the present invention in the foods or edible compositions described above is difficult to uniformly define, depending on the type, form, and intended use of the edible composition, as well as the type and form of the lipase activity inhibitor. However, when added to general processed foods, for example, if the lipase activity inhibitor of the present invention is a 50:50 (mass ratio) extract of kanuka leaves extracted with water and an extract of guava leaves extracted with aqueous ethanol having a water content of 50% by mass, the amount is generally 0.01 to 50% by mass, and more preferably 0.1 to 30% by mass. If the amount is outside this range, the desired effect of the present invention upon oral ingestion is diminished. Conversely, if the amount is too high, depending on the type of food, the flavor may be impaired or the food may become unprepared. The lipase activity inhibitor of the present invention may be consumed as is.

[0039] The pharmaceutical composition of the present invention is prepared by adding known excipients and additives, as necessary, to the lipase activity inhibitor, provided that they do not deviate from the spirit of the present invention, and processing them by conventional methods to form tablets, capsules, granules, powders, and other preparations. It is primarily administered orally to inhibit the absorption of ingested fat into the body, improve blood lipid balance, and treat obesity and various associated diseases such as hyperlipidemia, and the amount of the lipase activity inhibitor of the present invention is difficult to uniformly determine due to factors such as its form and the type, form, dosage, and administration of the pharmaceutical preparation, but is generally 0.01 to 50% by mass. The dosage for oral administration is not particularly limited, but for example, based on a lipase activity inhibitor obtained by extracting Kanuka leaves with aqueous acetone having a water content of 20% by mass, and further separating the extract with ethyl acetate, and extracting grape seeds with ethanol, in a ratio of 75:25 (by mass), the dosage is 0.01 to 20 g, more preferably 0.1 to 10 g, per day for an adult (body weight 50 kg). If the dosage is too low outside this range, the desired effect will decrease, and if the dosage is too high, further effect cannot be expected. [Example]

[0040] The present invention will now be described in more detail with reference to examples, but the present invention is not limited or restricted by these examples. In each example, percentages, parts and ratios are all by weight.

[0041] Manufacturing Example 1 Kanuka leaves and kanuka leaves or bark were semi-dried in the sun and then chopped or crushed to produce dried kanuka leaves or bark.

[0042] Manufacturing Example 2 330 mL of water was added to 30 g of the dried kanuka leaves described in Preparation Example 1, and extraction was carried out under pressure at 121°C for 1 hour. The mixture was then cooled to room temperature and filtered to separate the filtrate. 240 mL of water was added to the residue, and the mixture was heated in the same manner. After cooling, the mixture was filtered and the filtrate was collected. The two filtrates were combined and concentrated under reduced pressure, freeze-dried, and pulverized to obtain 8.4 g of an extract according to the present invention (Sample A-1).

[0043] Manufacturing Example 3 220 mL of water was added to 20 g of the dried kanuka leaves described in Preparation Example 1, and extraction was carried out at 85 to 90°C for 1 hour. The mixture was then cooled to room temperature and filtered to separate the filtrate. 160 mL of water was added to the residue, and the mixture was heated in the same manner. After cooling, the mixture was filtered to separate the filtrate. The two filtrates were combined and concentrated under reduced pressure, freeze-dried, and pulverized to obtain 3.5 g of an extract according to the present invention (Sample A-2).

[0044] Production Example 4 160 mL of 50% ethanol (water content: 50%) was added to 20 g of the dried kanuka leaves described in Preparation Example 1, and extraction was carried out at 70°C for 1 hour. The mixture was then cooled to room temperature and filtered to separate the filtrate. Another 120 mL of 50% aqueous ethanol was added to the residue, and the mixture was heated in the same manner. After cooling, the mixture was filtered to collect the filtrate. The two filtrates were combined and concentrated under reduced pressure, freeze-dried, and pulverized to obtain 5.5 g of an extract according to the present invention (Sample A-3).

[0045] Production Example 5 160 mL of 70% ethanol (water content: 30%) was added to 20 g of the dried kanuka leaves described in Preparation Example 1, and extraction was carried out at 70°C for 1 hour. The mixture was then cooled to room temperature and filtered to separate the filtrate. Another 120 mL of 70% aqueous ethanol was added to the residue, and the mixture was heated in the same manner. After cooling, the mixture was filtered to collect the filtrate. The two filtrates were combined and concentrated under reduced pressure, freeze-dried, and pulverized to obtain 5.9 g of an extract according to the present invention (Sample A-4).

[0046] Manufacturing Example 6 600 mL of water was added to 50 g of the dried kanuka bark described in Preparation Example 1, and extraction was carried out under pressure at 121°C for 1 hour. The mixture was then cooled to room temperature and filtered to separate the filtrate. 400 mL of water was added to the residue, and the mixture was heated in the same manner. After cooling, the mixture was filtered and the filtrate was collected. The two filtrates were combined and concentrated under reduced pressure, freeze-dried, and pulverized to obtain 7.5 g of an extract according to the present invention (Sample A-5).

[0047] Manufacturing Example 7 Sample A-1 and (-)-epigallocatechin gallate (Fujifilm Wako Pure Chemical Industries, Ltd.: HPLC purity 99.0%) were mixed in a ratio of 90 / 10 and processed in the usual manner to prepare a homogeneously mixed powder composition (Sample A-6).

[0048] Manufacturing Example 8 Sample A-1 and Sample B-1 described in Comparative Production Example 1 below were mixed in a mixing ratio of 75 / 25 and treated in a conventional manner to prepare a homogeneous mixed powder composition (Sample A-7).

[0049] Manufacturing Example 9 Sample A-1 and Sample C-1 described in Comparative Production Example 2 below were mixed in a ratio of 60 / 40 and treated in a conventional manner to prepare a homogeneous mixed powder composition (Sample A-8).

[0050] Comparative Manufacturing Example 1 Dried rhizomes of Astilbe thunbergii (SIEB. et ZUCC) MIQ. were crushed and further processed in a grinder to produce a fine powder that passed a 200 Tyler mesh. 1 kg of this was placed in a stainless steel extraction kettle, and 10 L of aqueous ethanol (45% water content) was added. Extraction was carried out at 70°C for 6 hours with periodic stirring. The residue was then filtered to obtain an extract. The solvent was then distilled off under reduced pressure to obtain 85 g of a reddish-brown extract (Sample B-1). The content of Astilbe thunbergii polyphenols in this extract (colorimetric method) was 21%.

[0051] Comparative Manufacturing Example 2 Fresh sun-dried guava (Psidium guajava L.) leaves were crushed and further pulverized to a fine powder passing a 200-mesh Tyler mesh. 1 kg of this was placed in a stainless steel extraction kettle, and 20 L of a 1:1 water / ethanol mixed solvent was added. Extraction was carried out under reflux for 5 hours. The residue was filtered to obtain an extract. The solvent was then distilled off under reduced pressure to obtain 380 g of a tannin-containing extract (Sample C-1) derived from brown guava leaves. The tannin content of this extract was 19%.

[0052] Test Example 1 The effects of various processed products of kanuka and polyphenol-containing substances according to the present invention on lipase activity were investigated using the method described below. Lipase activity was measured using Lipase Kit S (manufactured by DS Pharma Biomedical Co., Ltd.). The measurement method was according to the instructions attached to the kit, and the outline thereof is as follows.

[0053] 25 μL of each diluted sample was mixed with 10 μL of 0.1 mg / mL pancreatic lipase solution (5,5'-dithiobis[2-nitrobenzoic acid]) to prepare 500 μL of color-developing solution, and the mixture was preheated at 30°C for 5 minutes. 50 μL of substrate solution (dimethylmercaprol tributyrate, sodium dodecyl sulfate) was then added and mixed. The mixture was incubated at 30°C for 30 minutes in the dark, followed by the addition of 600 μL of stop solution. 200 μL of this reaction mixture was transferred to a 96-well plate, and the absorbance at 420 nm was measured using a plate reader. As a control, the same reaction was performed using extraction solvent (2% DMSO) instead of the test sample, and the absorbance at 420 nm (control absorbance) was measured. Test sample and control blanks were prepared by the same procedure as above, but without the addition of substrate solution, followed by the addition of stop solution. The absorbance of the test sample blank and control blank was measured. From the absorbance thus obtained, the lipase inhibition rate was calculated according to the following formula (1).

[0054]

number

[0055] The symbols in the formula indicate the following: S: absorbance of test sample reaction solution SB: absorbance of test sample blank C: Control absorbance CB: Absorbance of control blank

[0056] The lipase activity inhibition test was carried out according to the above-mentioned method for each of the preparations obtained in Production Examples 1 to 9 and Comparative Production Examples 1 and 2. The results are shown in Table 1.

[0057] [Table 1]

[0058] The data in Table 1 reveal that pancreatic lipase activity is significantly inhibited by the kanuka-derived test substance (Sample A-1), depending on the concentration of the substance. In contrast, the inhibitory effect of processed products of other materials (Samples B-1 and C-1) is weaker. Among the kanuka-derived test substances (Samples A-1 to A-5), the lipase activity inhibitory effect of test substances extracted using water (Samples A-1, A-2, and A-5) was greater, while that of test substances extracted using aqueous ethanol (Samples A-3 and A-4) was weaker. Furthermore, the lipase activity inhibitory effect tended to decrease depending on the ethanol concentration of the extraction solvent, suggesting that water-soluble components of kanuka leaves and bark are involved in the lipase activity inhibitory effect. Furthermore, the combined use of the water extract and polyphenols (Samples A-6 to A-8) further enhanced the inhibition of lipase activity.

[0059] It is generally known that almost all plant tissues, extracts, and purified products thereof contain tannins, proanthocyanidins, catechins, etc., and it is natural to expect that the kanuka plant and processed products thereof of the present invention will also contain such known components. However, they also contain a wide variety of other components and unknown components, and it is not realistic to elucidate these components and individually identify the active ingredients of the present invention, as this would require complicated work, long-term effort, and significant costs.

[0060] The IC is the concentration of the test sample in the reaction mixture when lipase activity is inhibited by 50% (lipase activity inhibition rate 50%). 50 The results are shown in Table 2.

[0061] Thus, it was revealed that sample A-1 according to the present invention had 2.64 times and 4.18 times higher inhibitory effects than samples B-1 and C-1.

[0062] [Table 2]

[0063] Test Example 2 150 mg of Sample A-1, 50 mg of beeswax, and 50 mg of rapeseed oil were heated to 60-65°C and mixed homogeneously. The mixture was then loaded into a capsule filling machine and prepared into gelatin-coated capsules containing 250 mg of content per capsule by standard methods. Volunteers (5 men and 5 women, aged 20-60 years, with a BMI of 25 or greater) wishing to lose weight (diet) orally ingested this capsule formulation, one capsule at a time, two capsules per day, for three weeks, and weight change was assessed. Seven subjects experienced weight loss, while three subjects experienced no weight change. Capsules were prepared in the same manner, except for replacing Sample A-1 with Sample A-7. These capsules were then orally administered to other volunteers (5 men and 5 women, aged 20-55 years, with a BMI of 25 or greater) under the same conditions. Nine subjects experienced weight loss, and one subject experienced no weight change. From the above results, it was confirmed that these preparations can be used as orally ingestible food compositions or pharmaceutical compositions for anti-obesity or dietary purposes.

[0064] Prototype example 1 (tablet) 5.0 kg of the lipase activity inhibitor of the present invention, consisting of Sample A-1: ​​oolong tea leaf powder = 3:1, was placed in a blender together with 3.5 kg of modified starch (manufactured by Matsutani Chemical Industry Co., Ltd., trade name: Pine Flow, registered trademark), 0.3 kg of tribasic calcium phosphate, 0.3 kg of vitamin B1, 0.2 kg of vitamin B2, 0.3 kg of vitamin B6, and 0.4 kg of vitamin C, and mixed with stirring for 10 minutes. The mixture was fed into a direct compression tableting machine to produce tablets measuring 7 mm in diameter, 4 mm in height, and 150 mg in weight, which were then coated with a thin film of shellac in a coating machine to produce tablet-shaped food samples.

[0065] Prototype 2 (cookies) 120 g of butter, 100 g of shortening, 90 g of caster sugar, and 100 mL of milk were placed in a household whisk, and one egg was added while stirring and thoroughly mixed. Then, 190 g of soft flour, 1 g of baking powder, and 10 g of the lipase activity inhibitor of the present invention (Sample A-1) were added and thoroughly kneaded. After leaving to rest for 30 minutes, the mixture was divided into 50 pieces using a mold and baked in an oven to produce butter cookies.

[0066] Prototype 3 (juice) 5 g of the lipase activity inhibitor of the present invention, consisting of Sample A-1 and grape seed extract (manufactured by Interhealth, product name: Activin, registered trademark) in a ratio of 2:1, was added to 1 L of commercially available apple juice and mixed to produce a prototype of anti-obesity apple juice. This was in no way inferior to the original apple juice.

[0067] Prototype 4 (Pet food) 6.2 kg of wheat flour, 1 kg of defatted soybeans, 500 g of beef tallow, 360 g of meat meal, 200 g of vitamins and minerals, and 150 g of the lipase activity inhibitor of the present invention (Sample A-1) were mixed thoroughly, and then 2 kg of water was added and kneaded thoroughly. After shaping and baking, a biscuit-type pet food sample was produced. [Industrial Applicability]

[0068] The lipase activity inhibitor of the present invention, which is characterized by containing a plant belonging to the genus Kunzea, such as kanuka, or a processed product thereof, or a combination of these with polyphenols, when taken orally, potently inhibits the hydrolysis of dietary lipids by pancreatic lipase, thereby suppressing lipid absorption and preventing the accumulation of body fat, thereby helping to improve obesity symptoms, making it useful in the fields of anti-obesity and dieting. Furthermore, foods and beverages containing the lipase activity inhibitor have milder effects than existing obesity treatments such as orlistat, and are highly safe because they are extracts from biological resources that have long been used for medicinal purposes in New Zealand, making them suitable for daily consumption.

Claims

1. A lipase activity inhibitor comprising, as an active ingredient, a water extract of Kunzea spp., which belongs to the genus Kunzea of ​​the Myrtaceae family.

2. 2. The method according to claim 1, wherein the extract is a water extract of kanuka leaves and / or bark. Inhibitor of lipase activity.

3. The lipase activity inhibitor according to claim 1 or 2, wherein the lipase is pancreatic lipase.

4. An oral composition comprising the lipase activity inhibitor according to any one of claims 1 to 3.

5. The oral composition according to claim 4, wherein the composition is a food, drink or pharmaceutical product.

6. A lipase inhibitor described in any one of claims 1 to 3, used for preventing or improving obesity symptoms.

7. An oral composition described in any one of claims 4 or 5, used for preventing or improving obesity symptoms.

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