Alternative autophagy activator

Abstract

An alternative autophagy activator comprising at least one selected from the group consisting of: a coffee extract and / or a coffee residue extract; a black pepper extract; a red pine extract; a juniper berry extract; a turmeric extract; and an apple polyphenol extract.

Description

Alternative autophagy activators

[0001] The present invention relates to an alternative autophagy activator. The present invention also relates to a food or beverage composition for activating alternative autophagy.

[0002] Autophagy is known as an intracellular purification and recycling system present in all eukaryotic organisms, including yeast and plants, and research into this subject has been progressing in recent years. Progress in research into the molecular mechanisms of autophagy has led to the identification of over 30 autophagy-related molecules. Among these, Atg5, Atg7, LC3, and other molecules are considered essential for autophagy. However, a novel form of autophagy has recently been discovered that does not require these molecules, originates from the Golgi apparatus or endosomes, and is regulated by molecules such as Rab9 (Non-Patent Document 1). This type of autophagy is called alternative autophagy, in distinction from conventional autophagy dependent on Atg5, Atg7, LC3, and other factors (sometimes simply referred to as Atg5-dependent autophagy).

[0003] In recent years, it has become clear that a decrease in alternative autophagy activity reduces the function of cells and tissues, and Non-Patent Document 2 discloses that enteritis worsens in mice lacking Trim31, an alternative autophagy-related protein. Patent Document 1 also discloses an alternative autophagy inducer and an anticancer agent containing a benzothiophene compound as an active ingredient. Patent Document 2 discloses an inhibitor of ultraviolet light-induced inflammation containing an alternative autophagy inducer as an active ingredient.

[0004] International Publication No. WO 2013 / 118842 International Publication No. WO 2020 / 091070

[0005] Nishida et al., Nature, 2009, 641, 654-658 Yoichi Nibe et al., Grant-in-Aid for Scientific Research Report, 2021, Research Project / Area Number 19K17426

[0006] The problem to be solved by the present invention is to provide a novel component capable of activating alternative autophagy.

[0007] As a result of intensive research to solve the above-mentioned problems, the present inventors discovered that coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, or apple polyphenol extract activates alternative autophagy, and thus completed the present invention.

[0008] The present invention is as follows: [1] An alternative autophagy activator containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [2] The alternative autophagy activator according to [1], wherein at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract activates alternative autophagy by promoting fusion of lysosomal proteins with autophagosomes in a manner independent of Atg5, Atg7, and / or LC3. [3] The alternative autophagy activator according to [1] or [2], which is used for at least one purpose selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. [4] A food or beverage composition for activating alternative autophagy, comprising at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [5] A food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, for at least one effect selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.[6] The food or beverage composition according to [5], wherein the food or beverage composition exhibits at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality, by activating at least one alternative autophagy selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [7] The food or beverage composition according to [5] or [6], wherein at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract promotes the fusion of lysosomal proteins with autophagosomes, independently of Atg5, Atg7, and / or LC3, and thereby exhibits at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.

[0009] The present invention may be as follows: [1A] An alternative autophagy activator containing at least one selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK). [2A] The alternative autophagy activator according to [1A], wherein the at least one selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK) activates alternative autophagy by promoting fusion of lysosomal proteins with autophagosomes, independent of Atg5, Atg7, and / or LC3. [3A] The alternative autophagy activator according to [1A] or [2A], which is used for at least one purpose selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. [4A] A food or beverage composition for activating alternative autophagy, comprising at least one selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK). [5A] A food or beverage composition containing at least one compound selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK), the food or beverage composition being useful for at least one effect selected from the group consisting of regulating gastric tone, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.[6A] The food or beverage composition according to [5A], which exhibits at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing blemishes, wrinkles, and / or dullness of the skin, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells, by activating at least one alternative autophagy pathway selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK). [7A] The food or beverage composition according to [5A] or [6A], wherein at least one selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK) promotes fusion of lysosomal proteins with autophagosomes independently of Atg5, Atg7, and / or LC3, thereby exhibiting at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality. Furthermore, in each of the above aspects [1] to [7], the coffee ground extract may contain DC and / or DK, and the DC and DK may be derived from the coffee ground extract.

[0010] [A1] A method for activating alternative autophagy using at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [A2] The method according to [A1], in which alternative autophagy is activated using at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, by promoting fusion of lysosomal proteins with autophagosomes in a manner independent of Atg5, Atg7, and / or LC3. [A3] The method according to [A1] or [A2], in which alternative autophagy is activated for at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, improving memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. [A4] A method for activating alternative autophagy using a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [A5] A method for achieving at least one effect selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality, using a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.[A6] The method according to [A5], wherein alternative autophagy is activated using a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, thereby resulting in at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality. [A7] The method according to [A5] or [A6], wherein the method uses a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, and promotes fusion of lysosomal proteins with autophagosomes without relying on Atg5, Atg7, and / or LC3, thereby achieving at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.

[0011] [B1] Use of at least one extract selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract for activating alternative autophagy. [B2] Use according to [B1] for activating alternative autophagy by promoting the fusion of lysosomal proteins with autophagosomes without dependence on Atg5, Atg7, and / or LC3. [B3] Use according to [B1] or [B2] for activating alternative autophagy for at least one purpose selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality. [B4] Use of a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract for activating alternative autophagy. [B5] Use of a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract for at least one selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.[B6] The use according to [B5] for achieving at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality, by activating alternative autophagy with at least one extract selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [B7] The use according to [B5] or [B6], wherein at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract promotes the fusion of lysosomal proteins with autophagosomes, independently of Atg5, Atg7, and / or LC3, thereby achieving at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.

[0012] [C1] Use of at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract for the manufacture of an alternative autophagy activator. [C2] Use according to [C1] for the manufacture of an alternative autophagy activator that activates alternative autophagy by promoting the fusion of lysosomal proteins with autophagosomes, independent of Atg5, Atg7, and / or LC3. [C3] The use according to [C1] or [C2] for the manufacture of an alternative autophagy activator that activates alternative autophagy for at least one application selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. [C4] The use of at least one extract selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract for the manufacture of a food or beverage composition for activating alternative autophagy. [C5] Use of at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract for the production of a food or beverage composition for at least one selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, improving memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells.[C6] Use according to [C5] for the production of a food or beverage composition, which exhibits at least one effect selected from the group consisting of regulating gastrointestinal condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality, by activating at least one alternative autophagy pathway selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [C7] The use according to [C5] or [C6] for the production of a food or beverage composition, wherein at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract promotes the fusion of lysosomal proteins with autophagosomes, independently of Atg5, Atg7, and / or LC3, and thereby exhibits at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.

[0013] [D1] Coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, which are used to activate alternative autophagy. [D2] Coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract according to [D1], which are used to activate alternative autophagy by promoting the fusion of lysosomal proteins with autophagosomes in a manner independent of Atg5, Atg7, and / or LC3. [D3] The coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract according to [D1] or [D2], which is used to activate alternative autophagy for at least one application selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. [D4] A food or beverage composition containing at least one extract selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, which is used to activate alternative autophagy.[D5] A food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, which is used for at least one selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, enhancing motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or shine, extending healthy lifespan, and improving the quality of germ cells. [D6] The food or beverage composition according to [D5], which is used to bring about at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality, by activating at least one alternative autophagy selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. [D7] The food or beverage composition according to [D5] or [D6], wherein at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract is used to achieve at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality by promoting the fusion of lysosomal proteins with autophagosomes, independent of Atg5, Atg7, and / or LC3.

[0014] In the above [A1] to [D7], instead of the specified extract, a food or beverage composition containing, as its active ingredient, at least one selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK), or at least one selected from the group consisting of dehydrocafestol (DC) and dehydrokahweol (DK), may be used, and the coffee residue extract may contain DC and / or DK.

[0015] In the present invention, particularly in the above [A1] to [D7], the inventions may be therapeutic or non-therapeutic. That is, they may be inventions of therapeutic or non-therapeutic methods, inventions of therapeutic or non-therapeutic uses, or inventions of therapeutic or non-therapeutic products. Furthermore, in the present invention, a "non-therapeutic invention" means an invention that does not involve medical practice when administered to a subject or ingested by a subject, i.e., an invention that does not involve methods of surgery, treatment, or diagnosis of a subject, particularly a human. In some cases, the subject may also include non-human animals. More specifically, a "non-therapeutic invention" means an invention that does not involve methods of surgery, treatment, or diagnosis performed on a human by a physician or a person under the physician's supervision. Examples of "non-therapeutic inventions" include inventions aimed at health promotion or cosmetic purposes. Furthermore, a "non-therapeutic invention" may also be a "prophylactic invention" as long as it does not fall under the category of a "therapeutic invention."

[0016] The present invention makes it possible to provide a novel component capable of activating alternative autophagy.

[0017] Figure 1 shows the results of evaluating alternative autophagy activity. Figure 2 shows the results of evaluating alternative autophagy activity.

[0018] Hereinafter, an embodiment of the present invention (hereinafter referred to as "the present embodiment") will be described, but the scope of the present invention should not be interpreted as being limited to the following embodiment.

[0019] One embodiment of the present invention relates to an alternative autophagy activator containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

[0020] As used herein, "extract" refers to a composition containing specific components separated from a raw material, and a food or beverage composition containing an extract is distinguished from the raw material itself. In this embodiment, the extract (including fractions) may be prepared using any method or apparatus known to those skilled in the art. Examples of preparation methods include solvent extraction, supercritical extraction, steam distillation, and pressing. Further extracts (including fractions) may be prepared from extracts (which may be fractions (including pressed products)) obtained by pressing. When preparing extracts (including fractions), particularly solvent extraction, an appropriate solvent may be used depending on the raw material and the specific components to be extracted. Examples of solvents include water, acetic acid, ethanol, methanol, acetone, hexane, benzene, and diethyl ether. A solvent containing an organic solvent is preferred for extraction from coffee grounds. The resulting extract (including fractions) may be in any form, such as liquid, solid, or powder. When in a liquid form, the extract may be in the same solvent as used for extraction, or may be a liquid concentrate obtained by concentration. Alternatively, the extract (including fractions) may be in the form of a suspension in which a portion of the extract (including fractions) has precipitated during the concentration process.The liquid extract (including fractions) may be subjected to a drying process such as freeze-drying or spray-drying, either directly or together with an excipient, to form a solid extract (including fractions).

[0021] In this specification, "coffee" refers to the seeds (coffee beans) of plants belonging to the genus Coffea. In this embodiment, only one species of Coffea may be used, or two or more species may be used in combination.

[0022] As used herein, "black pepper" refers to a climbing plant belonging to the genus Piper, whose scientific name is Piper nigrum.

[0023] In this specification, "Japanese red pine" refers to a coniferous tree whose scientific name is Pinus densiflora, among evergreen coniferous trees belonging to the genus Pinus.

[0024] As used herein, "juniper berry" refers to the fruit of Juniperus communis, a coniferous tree belonging to the Juniperus genus.

[0025] In this specification, "turmeric" refers to a perennial plant belonging to the genus Curcuma, and is sometimes called turmeric. Examples include turmeric (C. longa), C. aromatica, C. xanthorrhiza, and C. zedoaria. "Turmeric" is preferably turmeric (C. longa). In this embodiment, only one species of the genus Curcuma may be used, or two or more species may be used in combination.

[0026] In this specification, "apple" refers to the fruit of a deciduous tree belonging to the genus Malus. In this embodiment, only one species of Malus may be used, or two or more species may be used in combination.

[0027] As used herein, coffee residue extract refers to components separated by extraction from coffee residue after extraction with water or the like, and coffee extract refers to components separated by extraction from coffee beans (e.g., ground coffee beans; the same applies hereinafter). The coffee residue extract may be, for example, an ethanol or acetone extract of the residue (also referred to as coffee extract residue) obtained by extracting coffee beans with water or the like. The concentration of the solvent used for extraction is not particularly limited, but may be 50-100%, 70-100%, 70-99.5%, or 80-99.5%. The coffee residue extract may also be, for example, a pressed product of the residue obtained by extracting coffee beans with water or the like (pressed coffee residue product), or coffee oil derived from the coffee extract residue. The alcohol-soluble component of the pressed coffee residue product may also be used as the coffee residue extract. Examples of alcohols include methanol and ethanol. The coffee extract and / or coffee residue extract may be a mixture of a coffee extract and a coffee residue extract. The coffee beans may be raw beans (including those that have been fermented; the same applies hereinafter), roasted beans, or beans containing both raw and roasted beans. The coffee beans may also be raw coffee beans or roasted and ground into powder. The coffee beans may consist of one type of coffee beans, or two or more types of beans.

[0028] The coffee extract or coffee residue extract may contain dehydrocafestol (DC) and / or dehydrokahweol (DK). Furthermore, in this specification, an invention relating to a coffee extract or coffee residue extract may be understood as an invention in which "coffee extract" or "coffee residue extract" is read as "dehydrocafestol (DC) and / or dehydrokahweol (DK)."

[0029] The structure of DC is shown below.

[0030]

[0031] The structure of DK is shown below.

[0032]

[0033] The method for obtaining a coffee residue extract containing DC and / or DK is not particularly limited. The method described below for obtaining a coffee residue extract containing DC and / or DK can also be used as a method for obtaining a coffee residue extract. The extraction solvent for obtaining the coffee residue is preferably water, and the extraction temperature may be, for example, 70°C or higher, or a low temperature of approximately 5 to 40°C. The coffee beans used are preferably roasted coffee beans. Roasting conditions can be determined appropriately depending on the type of coffee beans, but may be, for example, roasted for 8 minutes or longer at a temperature of 200°C or higher. Alternatively, commercially available roasted (preferably dark-roasted) coffee beans (or their grounds) may be used. The obtained coffee residue may be used directly to prepare a coffee residue extract. To obtain a coffee residue extract containing DC and / or DK at a higher concentration, the coffee residue may be further dried. Drying methods such as air drying, dry heat drying, vacuum drying, and freeze drying may be used. After obtaining the coffee residue, a coffee residue extract may be prepared using methods and equipment known to those skilled in the art, for example, by solvent extraction, which may use a solvent containing an organic solvent. The obtained coffee residue extract preferably contains DC and / or DK.

[0034] Specifically, a coffee residue extract containing DC and / or DK may be produced as follows: Coffee beans, including Arabica beans, are roasted at 220 to 260°C for 8 minutes or longer. The roasted coffee beans are ground in a coffee mill, and then the ground coffee beans are filtered using a filter to drip-feed with water at 75 to 95°C to obtain a coffee residue. The resulting coffee residue is dried at less than 100°C for 5 to 20 hours until the moisture content is 10% or less. 70 to 95% (v / v) ethanol is added to the dried coffee residue, and the mixture is stirred at 40 to 60°C for 1 to 6 hours, filtered using filter paper, and the filtrate is recovered. The coffee residue remaining on the filter paper is subjected to the same procedure as above, and the filtrate is recovered, repeated 1 to 3 times. The recovered filtrates are combined, and the combined filtrates are concentrated using an evaporator until the ethanol is removed, to obtain a coffee residue extract containing DC and / or DK.

[0035] DC and DK may be isolated from coffee residue or the like as a raw material, or may be commercially available. The method for isolating DC and DK from coffee residue is not particularly limited, and they can be isolated, for example, from an extract or compressed product of coffee residue using chromatography or the like. When isolating by chromatography, a solvent such as water, acetonitrile, or alcohol can be used as the mobile phase. A single solvent or a mixed solvent of two or more solvents can be used, and elution can be performed with a mobile phase under gradient conditions.

[0036] DC and DK can be isolated using coffee residue as a raw material, for example, as follows. Coffee residue with a moisture content of 10% or less is obtained by the method described above. The obtained coffee residue is heated to 80 to 100°C in a roasting pot, and the heated coffee residue is pressed one to three times to recover the oil. The recovered oil is filtered to obtain compressed coffee residue oil. 2 to 6 times the amount of hexane is added to the compressed coffee residue oil and mixed, and then 1 to 3 times the amount of methanol as the compressed coffee residue oil is added and mixed. After allowing to stand for a while, separation into two layers is confirmed, and the methanol layer is recovered. An equal amount of methanol to the recovered methanol layer is added again and mixed. The above procedure is repeated one to three times to obtain a methanol layer. Distilled water in an amount 1 / 5 to 1 / 15 of the amount of the compressed coffee residue oil is added to this methanol layer, and mixed. After allowing to stand to separate, the methanol layer is recovered. The recovered methanol layer is concentrated to dryness to obtain a dried product, which is then fractionated by reverse phase chromatography. Ultrapure water, acetonitrile, and isopropanol are used as the mobile phase, and gradient elution is performed from ultrapure water-acetonitrile (95:5) to 100% acetonitrile. Isopropanol is then added, and the eluted fraction is concentrated to dryness to obtain a dried product. The dried product is then subjected to HPLC to isolate DC and DK. Ultrapure water and acetonitrile are used as the mobile phase, and ultrapure water-acetonitrile (60:40) is run, and the eluate from each peak is collected. The components in each peak are identified, and the eluates identified as DC and DK are used as DC and DK isolation solutions, respectively.

[0037] As used herein, black pepper extract, red pine extract, juniper berry extract, and turmeric extract refer to components isolated by extraction from the raw materials black pepper, red pine, juniper berry, and turmeric, respectively. The raw plant material for each extract may be the plant itself, a dried plant, the plant itself or the dried plant cut or shredded, or powder.

[0038] In this specification, each extract may be, for example, as follows. The red pine extract may be an ethanol extract of red pine or an ethanol extract of red pine powder. The ethanol concentration used for the extraction is not particularly limited, but may be 50-100%, 70-100%, 70-99.5%, or 80-99.5%. The juniper berry extract may be an ethanol extract of juniper berries or an ethanol extract of juniper berry powder. The ethanol concentration used for the extraction is not particularly limited, but may be 50-100%, 70-100%, 70-99.5%, or 80-99.5%. The apple polyphenol extract may be a fraction containing apple polyphenols (apple polyphenol fraction derived from apples). The black pepper extract may be an ethanol extract of black pepper or an ethanol extract of black pepper powder. The concentration of ethanol used for extraction is not particularly limited, but may be 50 to 100%, 70 to 100%, 70 to 99.5%, or 80 to 99.5%. The black pepper extract may contain piperine as a main component.

[0039] In this embodiment, the coffee residue extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract may each be commercially available. For example, commercially available products containing piperine or curcumin as the main component may be used as the black pepper extract or turmeric extract, respectively. Alternatively, a prepared extract and a commercially available product may be mixed and used as an extract (including fractions). Specific examples of turmeric extract include products containing three compounds (curcuminoids): curcumin, demethoxycurcumin (DMC), and bisdemethoxycurcumin (BDMC), and standardized to 95% or more curcuminoids. Products derived from dried Curcuma longa rhizomes may also be used. For the apple polyphenol extract, commercially available products containing apple polyphenols as the main component may also be used.

[0040] The alternative autophagy activator in this embodiment may contain any amount of at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, as long as it can exert an alternative autophagy activating effect. It may also contain other active ingredients as long as they do not impair the effect. The other active ingredients may be other components known as alternative autophagy activators, or other components not known to have or do not have the ability to activate alternative autophagy. The alternative autophagy activator may contain only at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, or may contain only two or more selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

[0041] The alternative autophagy activator in this embodiment is capable of activating alternative autophagy by including at least one selected from the group consisting of coffee residue extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract. In this embodiment, activation of alternative autophagy may induce or promote the degradation of intracellular molecules such as proteins, organelles, or bacteria or viruses that have invaded from outside the cell, for example, without relying on Atg5, Atg7, and / or LC3. Activation of alternative autophagy may also be induced by the fusion of lysosomal proteins with autophagosomes, without relying on Atg5, Atg7, and / or LC3. That is, activation of alternative autophagy may be induced by fusion of a lysosomal protein with an autophagosome independent of at least one molecule selected from the group consisting of Atg5, Atg7, and LC3, or by fusion of a lysosomal protein with an autophagosome independent of all of Atg5, Atg7, and LC3. The alternative autophagy activator in this embodiment may be an alternative autophagy activator that activates alternative autophagy by promoting fusion of a lysosomal protein with an autophagosome with at least one selected from the group consisting of coffee residue extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, independent of Atg5, Atg7, and / or LC3, or independent of at least one molecule selected from the group consisting of Atg5, Atg7, and LC3. Furthermore, in this embodiment, activation of alternative autophagy may induce or promote the fusion of lysosomal proteins with autophagosomes, without relying on autophagy-related molecules including Atg5, Atg7, and LC3, and may induce or promote the degradation of intracellular molecules such as proteins, organelles, and bacteria and viruses that have invaded from outside the cell.That is, activation of alternative autophagy in this embodiment may not involve the molecular mechanism of autophagy.The alternative autophagy activator in this embodiment may be an alternative autophagy activator that does not activate autophagy.

[0042] Activation of alternative autophagy results in the breakdown of intracellular substances, and the reuse of these degraded substances promotes intracellular metabolism. It also prevents the accumulation of abnormal proteins and maintains immune function by eliminating foreign invaders. Therefore, activation of alternative autophagy improves functions such as maintaining internal homeostasis and eliminating abnormal molecules, thereby improving the condition of all cells and organs in the body. This makes it possible to prevent various diseases and infections and alleviate their symptoms. Specifically, activation of alternative autophagy results in the following effects: regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, improving memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing blemishes, wrinkles, and / or dullness of the skin, improving hair color, moisture, and / or luster, extending healthy lifespan, and / or improving the quality of germ cells.

[0043] As one embodiment of the present invention, there is provided an alternative autophagy activator which contains at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, and which is used for at least one purpose selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. Alternatively, alternative autophagy may be induced by the fusion of lysosomal proteins with autophagosomes, independent of Atg5, Atg7, and / or LC3. Therefore, one embodiment of the present invention provides a method for producing an autophagosome-derived extract containing at least one extract selected from the group consisting of coffee residue extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, which is capable of fusion with lysosomal proteins without relying on Atg5, Atg7, and / or LC3. The present invention provides an alternative autophagy activator that is used for at least one application selected from the group consisting of activating alternative autophagy by promoting autophagosome fusion, regulating gastrointestinal condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, improving memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, reducing skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving the quality of germ cells. Specifically, the alternative autophagy activator of this embodiment or the food or beverage composition of this embodiment contains at least one of the extracts, thereby enabling it to activate alternative autophagy, and can be used for the applications listed in Table 1.

[0044]

[0045] In this embodiment, the alternative autophagy activator may be an agent that promotes the fusion of lysosomal proteins and autophagosomes, an agent that enhances the degradative action of alternative autophagy, or an agent that promotes intracellular metabolism.

[0046] The activation of alternative autophagy may be evaluated by methods known to those skilled in the art. Examples include evaluation of alternative autophagy activity when autophagosomes fuse with lysosomes, or evaluation using fluorescence intensity or electron microscopy. Alternatively, alternative autophagy activity may be measured by measuring the gene expression level or protein expression level of a group of alternative autophagy-related proteins. Specifically, with reference to Patent Document 1 and the like, evaluation can be performed using cells deficient in Atg5, Atg7, and / or LC3 that express lysosomal proteins tagged with fluorescent proteins. Specifically, evaluation can be performed using the fluorescence intensity generated when alternative autophagy is induced and lysosomal proteins aggregate as an indicator. The generated fluorescence intensity may be quantified and used for evaluation. Furthermore, with reference to Patent Document 1 and the like, alternative autophagy can be evaluated as being activated if the fluorescence intensity due to lysosomal protein aggregation is greater when the alternative autophagy activator of the present embodiment is administered compared to when the alternative autophagy activator of the present embodiment is not administered. Furthermore, alternative autophagy can be evaluated as being activated, for example, when the fluorescence intensity is increased by 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, or 500% or more. Preferably, alternative autophagy can be evaluated as being activated when the fluorescence intensity is increased by 30% or more.

[0047] The alternative autophagy activator can exert its function, for example, by being administered directly or indirectly to an organ or cell in the body.

[0048] The dosage form of the alternative autophagy activator may be any dosage form as long as it can exert its function, and may be administered in dosage forms such as oral preparations such as tablets, capsules, granules, fine granules, powders, liquids, syrups, chewable tablets, and troches, as well as ointments, gels, creams, injections, sublingual preparations, inhalants, and suppositories. To obtain these dosage forms, conventionally known additives may be used, and, without particular limitation, industry-known excipients, binders, diluents, disintegrants, buffers, thickeners, stabilizers, emulsifiers, dispersants, suspending agents, and / or preservatives may be used as needed.

[0049] The dosage and frequency of administration of the alternative autophagy activator may be appropriately determined depending on various conditions, such as the route of administration and the age, weight, and symptoms of the subject.

[0050] The subjects to which the alternative autophagy activator is administered are preferably mammals, more preferably humans, monkeys, cats, pigs, horses, cows, mice, rats, guinea pigs, dogs, and rabbits, and even more preferably humans.

[0051] One embodiment of the present invention provides a food, beverage, feed, cosmetic, pharmaceutical, quasi-drug, or the like containing the alternative autophagy activator of this embodiment. The food, beverage, feed, cosmetic, pharmaceutical, quasi-drug, or the like may have the same dosage form as the dosage form described above for the alternative autophagy activator.

[0052] In another embodiment of the present invention, there is provided a food or beverage composition for activating alternative autophagy, which contains at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

[0053] Food or beverage compositions can be in a form suitable for the intended use, including, but not limited to, oral preparations such as tablets, capsules, granules, fine granules, powders, liquids, syrups, chewable tablets, and lozenges, as well as ointments, gels, creams, injections, sublingual preparations, inhalants, and suppositories. Food or beverage compositions can be in an edible form, including, but not limited to, solids, liquids, granules, powders, capsules, creams, pastes, and jellies. For example, capsules may be coated with one or more layers as needed. These dosage forms and shapes can be achieved using conventionally known additives, including, but not limited to, excipients, binders, diluents, disintegrants, buffers, thickeners, stabilizers, emulsifiers, dispersants, suspending agents, and / or preservatives known in the food industry. The food or beverage composition is not particularly limited, and may be, for example, a health food, a functional food, a food for specified health uses, a nutrient-functional food, a health supplement, or a supplement. Examples of the food or beverage composition include food, beverages, seasonings, and food additives. Specific examples of foods include confectioneries, dairy products, breads, noodles, rice, soups, and prepared dishes. Specific examples of beverages include milk drinks, soft drinks, fruit juice drinks, carbonated drinks, sports drinks, energy drinks, and alcoholic beverages.

[0054] If necessary, coloring agents, flavoring agents, sweeteners, bittering agents, acidulants, preservatives, antioxidants, pH adjusters, and / or thickening stabilizers may be added to the food or beverage composition.

[0055] The food or beverage composition may be provided after heat sterilization, or may be provided in a sealed form in a bag or container. When provided, it may be labeled with at least one claim selected from the group consisting of "regulating stomach condition," "maintaining intestinal function," "rejuvenating the intestines," "improving cognitive function," "improving memory," "removing waste products from the brain," "maintaining healthy liver function," "improving motor function," "maintaining skin moisture and elasticity," "improving skin blemishes, wrinkles, and / or dullness," "improving hair color, moisture, and / or luster," "extending healthy lifespan," and "improving germ cell quality."

[0056] In another embodiment of the present invention, the food or beverage composition of this embodiment may be a food or beverage composition that activates alternative autophagy by promoting the fusion of lysosomal proteins with autophagosomes, independent of Atg5, Atg7, and / or LC3.

[0057] The food or beverage composition in this embodiment contains at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, thereby enabling activation of alternative autophagy and providing at least one effect selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, improving memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality. Therefore, this embodiment also provides a food or beverage composition containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, for at least one selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestines, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality.The food or beverage composition of this embodiment may exhibit at least one effect selected from the group consisting of regulating gastric condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing waste products from the brain, maintaining healthy liver function, improving motor function, maintaining skin moisture and elasticity, improving skin blemishes, wrinkles, and / or dullness, improving hair color, moisture, and / or luster, extending healthy lifespan, and improving germ cell quality, by activating alternative autophagy by at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract, or by promoting the fusion of lysosomal proteins with autophagosomes independent of Atg5, Atg7, and / or LC3, or by activating alternative autophagy by promoting the fusion of lysosomal proteins with autophagosomes independent of Atg5, Atg7, and / or LC3. Furthermore, by including at least one of the extracts in the food or beverage composition of this embodiment, it becomes possible to activate alternative autophagy, and each food or beverage composition containing at least one of the extracts provides the effects described in Table 1. Therefore, this embodiment also provides a food or beverage composition that provides the effects described in Table 1 above, containing at least one selected from the group consisting of coffee ground extract, coffee extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

[0058] The food or beverage composition can exert its alternative autophagy activating effect when orally ingested.

[0059] The present invention will be described in more detail below with reference to specific examples, although the present invention is not limited thereto.

[0060] (1) Preparation of samples for measuring alternative autophagy activity

[0061] (Black Pepper Extract) We used pepper extract (piperine, HJK-3458) from Inabata Koryo, BioPerine from Sabinsa Japan Corporation, black pepper extract powder (95%) from Vidya Japan, black pepper extract powder (GIH-95) from GOA International, and an in-house extracted black pepper ethanol extract.

[0062] (Pinus densiflora extract) Whole dried Pinus densiflora was crushed into powder, and 3.0 g of this powder was added with 30 mL of 99.5% ethanol and stirred at room temperature for 30 minutes to perform extraction. After that, the residue was removed by a conventional method, and the mixture was dried to obtain 0.116 g of a dried product.

[0063] (Juniper berry extract) Whole dried juniper berries were crushed into powder, and 3.0 g of this powder was added with 30 mL of 99.5% ethanol and stirred at room temperature for 30 minutes to perform extraction. After removing the residue by a conventional method, the extract was dried to obtain 0.948 g of a dried product.

[0064] (Turmeric Extract) Curcumin C3 Complex (registered trademark) from Sabinsa Japan Corporation was used.

[0065] (Pressed Coffee Residue) A blend of Arabica and Robusta beans, roasted and milled, was extracted with hot water to produce a residue, which was then dried in a dryer at 80°C for 30 hours to obtain a coffee extract residue with a moisture content of 3.8%. 10 kg of the coffee extract residue was heated to 100°C in a roasting kettle, and after heating, it was pressed using an oil press (H-54, Hander Oil Press) to obtain oil. Three presses were performed to recover a total of 700 g of oil. The recovered oil was filtered using No. 2 filter paper, and 425 g of coffee oil (pressed coffee residue) was finally obtained.

[0066] (Fraction of pressed coffee residue) 5.0 g of coffee oil was weighed into a 100 mL separatory funnel, and 20 mL of hexane was added and mixed thoroughly. 10 mL of methanol was then added and mixed. After visually confirming that the mixture had separated into two layers, the methanol layer was recovered. This process of adding another 5 mL of methanol, mixing, and recovering the methanol layer was repeated twice. The recovered methanol layer was placed in a 100 mL separatory funnel, and 0.5 mL of distilled water was added and mixed. The hexane layer separated, and the methanol layer was recovered in a weighed 50 mL eggplant flask. The methanol layer was evaporated to dryness, yielding 0.42 g of dried material.

[0067] (80 v / v % ethanol extract of coffee grounds) 900 mL of 80 v / v % ethanol was added to 100 g of coffee extract grounds. Extraction was carried out for 1 hour using a stirrer in a thermostatic bath at 30°C, and then filtration was carried out using a Buchner funnel lined with No. 2 filter paper. The filtered permeate was collected and added to an eggplant flask, and the ethanol was removed by vacuum concentration to obtain a concentrated solution. The concentrated solution was frozen at -80°C, and the frozen concentrated solution was dried using a freeze dryer.

[0068] (99.5 v / v % ethanol extraction of coffee residue) 900 mL of 99.5 v / v % ethanol was added to 100 g of coffee extract residue. Extraction was carried out for 1 hour using a stirrer in a thermostatic bath at 30°C, and then filtered using a Buchner funnel lined with No. 2 filter paper. The filtered permeate was collected and added to a recovery flask, and the ethanol was removed by vacuum concentration to obtain a concentrated solution.

[0069] (Acetone Extraction of Coffee Residue) 800 mL of acetone was added to 200 g of coffee extract residue. Extraction was carried out for 1 hour using a stirrer in a thermostatic bath at 30°C, and then filtered using a Buchner funnel equipped with No. 2 filter paper. The filtered permeate was collected and added to a recovery flask, and the acetone was removed by vacuum concentration to obtain a concentrated solution.

[0070] (Dehydrocafestol (DC) and dehydrokahweol (DK)) A coffee extract residue with a moisture content of 10% or less was obtained by drying the residue produced when roasted coffee beans (after milling) blended with Arabica and Robusta beans were extracted with hot water at 80°C for 16 hours. The dried coffee extract residue was heated to 100°C in a roasting kettle and, after heating, pressed using an oil press (H-54, Hander Oil Press). Three pressings were performed, and a total of 700 g of oil was recovered. The recovered oil was filtered using No. 2 filter paper, and 425 g of coffee residue compressed oil was finally obtained. Five grams of coffee residue compressed oil was added to a separatory funnel, and four times the amount of hexane was added and mixed. Next, two times the amount of methanol as the coffee residue compressed oil was added and mixed. After allowing to stand for a while to confirm separation into two layers, the methanol layer was recovered. To the recovered methanol layer, an equal amount of methanol to the coffee grounds compressed oil was added again and mixed. The above procedure was repeated twice to obtain a methanol layer. To this methanol layer, distilled water in an amount 1 / 10 the amount of the coffee grounds compressed oil was added and mixed, and after leaving to stand for separation, the methanol layer was recovered. The recovered methanol layer was concentrated to dryness, yielding 0.42 g of a dried product. This dried product was fractionated using a medium-pressure preparative liquid chromatograph (Yamazen Corporation, EPCLC-W-Prep 2XY) under the following conditions: The column used was a Universal Premium ODS-SM 30 μm 120A, 2L size (3 x 20 cm, 51 g) (Yamazen Corporation). The mobile phase was ultrapure water, acetonitrile, and isopropanol. Elution was performed at a flow rate of 20 mL / min with a linear gradient from ultrapure water-acetonitrile (95:5) to 100% acetonitrile over 40 minutes, followed by a 12-minute run of isopropanol, with fractions collected every 10 mL. The fractions eluted between 37 and 45 minutes were concentrated to dryness. DC and DK were isolated from the dry product using a preparative HPLC (Shimadzu Corporation, Prominence) under the following conditions:A SunFire 5 μm (19 × 150 mm) (Waters) column was used, and ultrapure water and acetonitrile were used as the mobile phase. Ultrapure water-acetonitrile (60:40) was run over 160 minutes at a column temperature of 30°C and a flow rate of 17 mL / min, and the peak eluted between 116 and 126 minutes (component A) and the peak eluted between 128 and 141 minutes (component B) were collected, yielding 5 mg and 14 mg of concentrated dried products of components A and B, respectively. NMR and mass spectrometry were performed on components A and B isolated as described above. NMR measurements were performed using an AVANCE NEO 400 MHz instrument (Bruker), and mass spectrometry was performed using a Vanquish Flex liquid chromatograph (Thermo Fisher Scientific) and a ThermoFisher Scientific OrbiTrap Explorers 240 mass spectrometer (Thermo Fisher Scientific). The following data were obtained, and component A was identified as dehydrokahweol (DK) and component B as dehydrocafestol (DC). The concentrated dried products of components A and B were designated as in-house purified DK and DC, respectively. Commercially available DC was also used as a standard sample to evaluate alternative autophagy activity.

[0071] The results of NMR and mass spectrometry of DK are shown below. １ H-NMR (DMSO-d6, 400MHz, corrected with solvent): δ (ppm) 7.50 (1H, d, J = 1.5Hz, H19), 6.45 (1H, brs, H18), 6.29 (1H, d, J = 10.0Hz, H2), 5.93 (1 H, d, J=10.1Hz, H1), 5.34 (1H, s, H15), 4.65 (1H, brt, -OH, H17), 3.99 (2H, s, H17), 2. 54-2.50 (2H, overlapped, H5, 13), 2.01 (1H, d, J = 10.2Hz, H14a), 1.86 (1H, m, H6a), 1 .78-1.33 (9H, overlapped, H6b, 7a, 7b, 9, 11a, 11b, 12a, 12b, 14b), 0.94 (3H, s, H20) １３C-NMR (DMSO-d6, 100 MHz, solvent correction): δ (ppm) = 149.37 (C3), 147.22 (C16), 141.72 (C19), 138.60 (C1), 133.60 (C15), 121.43 (C4), 115.03 (C2), 109.05 (C18), 59.05 (C17), 48.62 (C8), 44.43 (C14), 43.72 (C5), 41.23 (C10), 40.36 (C13), 40.00 (C9), 37.26 (C7), 24.86 (C12), 20.62 (C6), 19.02 (C11), 15.18 (C20) MS (m / z) C ２０ H ２３ O([M+H-H2O] ＋ ), theoretical value 279.1743, experimental value 279.1742; C ２０ H ２５ O ２ ([M+H] ＋ ) Theoretical value 297.1849, experimental value 297.1847

[0072] The results of DC NMR and mass analysis are shown below. １ H-NMR (DMSO-d6, 400MHz, solvent correction): δ (ppm) = 7.39 (1H, d, J = 1.4Hz, H19), 6.29 (1H, d, J = 1.7Hz, H18), 5.32 (1H, s, H15), 4.62 (1H, brt, -OH, H17), 3.99 (2H, brs, H17), 2.57-2.55 (2H, m, H2), 2.21 (1H, dd, J = 12.5, 1 . 9Hz, H5), 2.11 (1H, d, J=10.2Hz, H14a), 2.03 (1H, dt , J=12.8, 4.2Hz, H1a), 1.79-1.33 (10H, overlapped , H6a, 6b, 7a, 7b, 11a, 11b, 12, 14b), 1.24-1.17 (1H, m, H1b), 1.14 (1H, d, J=8.4Hz, H9), 0.79 (3H, s, H20) １３C-NMR (DMSO-d6, 100MHz, corrected with solvent): δ (ppm) = 148.00 (C3), 147.04 (C16), 140. 84 (C19), 133.97 (C15), 119.88 (C4), 108.50 (C18), 59.08 (C17), 48.24 (C8) , 44.48 (C14), 43.62 (C9), 43.47 (C5), 40.48 (C13), 38.02 (C10), 37.81 (C7) , 35.17 (C1), 25.01 (C12), 21.56 (C6), 20.20 (C2), 18.95 (C11), 12.93 (C20) ESI MS (m / z) C ２０ H ２５ O([M+H-H2O] ＋ ), theoretical value 281.1900, actual value 281.1899; C ２０ H ２７ O ２ ([M+H] ＋ ) Theoretical value 299.2006, measured value 299.2005

[0073] (DC and DK-Rich Residue Extract) 200 g (dry weight) of the coffee extract residue obtained above, with a moisture content adjusted to 2.5%, was added with 70 v / v% ethanol to a total volume of 1 L. The mixture was stirred at 250 rpm for 2 hours at 50°C using a Three-One Motor (BL-600, manufactured by Shinto Scientific Co., Ltd.). After 2 hours of extraction, the mixture was filtered using No. 2 filter paper, and the filtrate and the extraction residue on the filter paper were collected. The extracted residue on the filter paper was added with 70 v / v% EtOH to a total volume of 1 L, and extracted again for 1 hour at 50°C with stirring. After 1 hour of extraction, the mixture was filtered using No. 2 filter paper, and the filtrate was collected. The first and second filtrates were combined and concentrated using an evaporator until the EtOH was removed, and the concentrate was freeze-dried. 7.9 g of the freeze-dried extract was obtained. This extract was designated the DC and DK-rich residue extract.

[0074] (Apple Polyphenol Extract) Apple Phenon (registered trademark) (manufactured in China) sold by BGG Japan was used.

[0075] (2) Evaluation of alternative autophagy activity

[0076] DMEM (containing 4.5 g / L glucose) medium (Sigma Aldrich) containing 10% FBS (Sigma Aldrich), 1 mM sodium pyruvate solution (Nacalai Tesque), 1 mM HEPES buffer (Nacalai Tesque), 0.1 mM non-essential amino acid solution (Fujifilm Wako Pure Chemical Industries), 0.1 mM MEM Vitamin solution (Invitrogen), 100 U / mL penicillin-100 μg / mL streptomycin (Nacalai Tesque), and 0.05 mM 2-mercaptoethanol (Bio-Rad) was used.

[0077] ATG5-KO / GFP-Lamp1 was prepared as follows, with reference to Patent Document 1 (WO 2013 / 118842). Mizushima, N. et al. Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells. J. Cell Biol. 152, 657-668 (2001). Mouse embryonic fibroblasts (Mouse Embryonic Fibroblasts, MEFs) established from embryonic day 14.5 ATG5-deficient C57 / B6J mice prepared by the method described in were immortalized by electroporation using the Nucleofector system (Amaxa) to express the SV40 T antigen. Furthermore, a retrovirus created by introducing an MSCV-Lamp1-GFP (zeocin-resistant) vector into Plat E cells, which are retrovirus packaging cells, was used to introduce a gene expressing a fusion protein of Lamp1 and GFP (also referred to as "GFP-Lamp1"). Then, cells with high expression of GFP-Lamp1 were cloned and prepared as ATG5-KO / GFP-Lamp1.

[0078] ATG5-KO / GFP-Lamp1 at 4.0 × 10 ４ The cells were seeded at a concentration of 1000 cells / mL in a 384-well plate (Greiner), 30 μL per well, and incubated at 37°C, 5% CO ２The cultures were cultured for 24 hours in a 1000-milliliter incubator. Furthermore, the prepared coffee ground extract, coffee ground pressed product, dehydrocafestol (DC), dehydrokahweol (DK), DC and DK-rich residue extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract were added. After 24 hours, 12 μL of 4% paraformaldehyde-phosphate buffer (Fujifilm Wako Pure Chemical Industries) was added to each well and the cultures were left to stand at room temperature for 15 minutes. After washing the cultures with HBSS (Fujifilm Wako Pure Chemical Industries), 30 μL of HBSS mixed with 30 μL of Hoechst (Invitrogen) diluted 2000-fold was added to each well and the cultures were left to stand at room temperature for 15 minutes. Microscopic images were then obtained using a confocal quantitative image cytometer CQ1 (Yokogawa Electric Corporation). With reference to Patent Document 1, the integrated value of the fluorescence intensity of GFP-Lamp1 fluorescent granules per cell was output from microscopic images, and this value was divided by the vehicle group to evaluate alternative autophagy activity as an index. The results are shown in Table 2 and Figure 1. The results for DC, DK, and DC and DK-rich residue extracts are shown in Figure 2. Similar experiments were performed using available extracts for the extract disclosed in Patent Document 2, but the results showed that they were not sufficiently effective in activating alternative autophagy (results not disclosed).

[0079]

[0080] The results in Table 2 and Figures 1 and 2 confirm that coffee ground extract, pressed coffee grounds, dehydrocafestol (DC), dehydrokahweol (DK), DC and DK-rich residue extract, black pepper extract, red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract have the function of activating alternative autophagy.

[0081] <Examples> The changes in DC and DK contents in the coffee extract residue obtained above under each drying condition were investigated, and the results are shown in Table 3. The DC + DK contents (%) are shown, with the DC and DK contents per extract when the freeze-dried residue was extracted with 75 v / v % EtOH being set at 100%.

[0082]

Claims

1. An alternative autophagy activator containing at least one selected from the group consisting of coffee extract and / or coffee residue extract, black pepper extract, Japanese red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

2. The alternative autophagy activator according to claim 1, wherein at least one selected from the group consisting of coffee extract and / or coffee residue extract, black pepper extract, Japanese red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract activates alternative autophagy by promoting the fusion of lysosomal proteins and autophagosomes without depending on Atg5, Atg7, and / or LC3.

3. The alternative autophagy activator according to claim 1 or 2, which is used for at least one application selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing brain waste, maintaining the function of a healthy liver, improving motor function, maintaining skin moisture and elasticity, improving skin spots, wrinkles and / or dullness, improving hair color, moisture and / or gloss, extending healthy lifespan, and improving the quality of germ cells.

4. A composition for food or beverage for activating alternative autophagy, containing at least one selected from the group consisting of coffee extract and / or coffee residue extract, black pepper extract, Japanese red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

5. A composition for food or beverage for at least one selected from the group consisting of regulating stomach condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing brain waste, maintaining the function of a healthy liver, improving motor function, maintaining skin moisture and elasticity, improving skin spots, wrinkles and / or dullness, improving hair color, moisture and / or gloss, extending healthy lifespan, and improving the quality of germ cells, containing at least one selected from the group consisting of coffee extract and / or coffee residue extract, black pepper extract, Japanese red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

6. The food or beverage composition according to claim 5, which exhibits at least one effect selected from the group consisting of regulating the stomach condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing brain waste, maintaining the function of a healthy liver, improving motor function, maintaining skin moisture and elasticity, improving skin spots, wrinkles and / or dullness, improving hair color, moisture and / or gloss, extending health span, and improving the quality of germ cells, by activating autophagy with at least one selected from the group consisting of coffee extract and / or coffee residue extract, black pepper extract, Japanese red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract.

7. The food or beverage composition according to claim 5 or 6, wherein at least one selected from the group consisting of coffee extract and / or coffee residue extract, black pepper extract, Japanese red pine extract, juniper berry extract, turmeric extract, and apple polyphenol extract promotes the fusion of lysosomal proteins and autophagosomes without depending on Atg5, Atg7, and / or LC3, and exhibits at least one effect selected from the group consisting of regulating the stomach condition, maintaining intestinal function, rejuvenating the intestine, improving cognitive function, enhancing memory, removing brain waste, maintaining the function of a healthy liver, improving motor function, maintaining skin moisture and elasticity, improving skin spots, wrinkles and / or dullness, improving hair color, moisture and / or gloss, extending health span, and improving the quality of germ cells.

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