Autophagy activator and composition for autophagy activation

Abstract

This autophagy activator contains at least one of compounds represented by any one of structural formulas (1)-(6).

Description

Autophagy activator and composition for activating autophagy

[0001] The present invention relates to an autophagy activator and a composition for activating autophagy.

[0002] Autophagy is a mechanism by which unnecessary substances within cells are broken down. Autophagy is known to enhance cellular metabolism and contribute to cellular health. Maintaining autophagy activity is also thought to be involved in the suppression of individual aging and the extension of lifespan, and research is underway to elucidate its detailed mechanisms.

[0003] In Japan, the aging population (proportion of the population aged 65 or over) is increasing year by year, and in an aging society, improving quality of life and extending healthy lifespan are important. For this reason, activation of autophagy has attracted attention.

[0004] To date, autophagy activators have been proposed that contain, as an active ingredient, at least one selected from the group consisting of heparinoids, glycyrrhizinic acid and its salts, tocopherol and its derivatives, ε-aminocaproic acid and its salts, nicotinamide, urea, pantothenyl alcohol, and vitamin A (see, for example, Patent Document 1).

[0005] However, there remains a strong demand for new materials that have excellent autophagy activation properties, are highly safe, and can therefore be widely used as ingredients in foods and beverages, pharmaceuticals, quasi-drugs, research reagents, and more, and there is currently a need for their rapid development.

[0006] Japanese Patent Application Laid-Open No. 2021-172641

[0007] The present invention aims to solve the above-mentioned conventional problems and achieve the following objectives: Namely, the present invention aims to provide an autophagy activator and a composition for activating autophagy that have excellent autophagy activating activity and are highly safe.

[0008] As a result of extensive research conducted by the present inventors to solve the above problems, they discovered that compounds represented by any of the following structural formulas (1) to (6) have excellent autophagy activation effects, are highly safe, and are useful materials for activating autophagy.

[0009] The present invention is based on the above-mentioned findings of the present inventors, and provides the following means for solving the above problems: <1> An autophagy activator characterized by containing at least one compound represented by any one of the following structural formulas (1) to (6). <2> A composition for activating autophagy, comprising the autophagy activator according to <1>.

[0010] The autophagy activator and autophagy activating composition of the present invention can solve the above-mentioned conventional problems and achieve the above-mentioned object, and can provide an autophagy activator and autophagy activating composition that have excellent autophagy activating activity and are highly safe.

[0011] (Autophagy Activator) The autophagy activator of the present invention contains at least one compound represented by any one of the following structural formulas (1) to (6) as an active ingredient, and further contains other ingredients as necessary.

[0012] As used herein, "activating autophagy" refers to enhancing the function of autophagy compared to when the autophagy activator is not used. The degree of activation of autophagy is not particularly limited and can be selected appropriately depending on the purpose.

[0013] <Compound Represented by Structural Formula (1)> The English name of the compound represented by the following structural formula (1) is dihydroferulic acid 4-sulfate.

[0014] <Compound Represented by Structural Formula (2)> The English name of the compound represented by the following structural formula (2) is dihydroferulic acid 4-O-glucronide.

[0015] <Compound Represented by Structural Formula (3)> The English name of the compound represented by the following structural formula (3) is dihydrocaffeic acid.

[0016] <Compound Represented by Structural Formula (4)> The English name of the compound represented by the following structural formula (4) is 3-(4-hydroxyphenyl)propionic acid.

[0017] <Compound Represented by Structural Formula (5)> The English name of the compound represented by the following structural formula (5) is cinnamic acid.

[0018] <Compound Represented by Structural Formula (6)> The English name of the compound represented by the following structural formula (6) is 3-phenylpropionic acid.

[0019] The fact that compounds represented by any of the structural formulas (1) to (6) have excellent autophagy activating activity and are useful as autophagy activators was completely unknown in the past, and is a new finding made by the present inventors.

[0020] The compounds represented by any one of the structural formulas (1) to (6) are known compounds, and commercially available products can be used.

[0021] The compounds represented by any of the structural formulas (1) to (6) have excellent autophagy activating properties and can therefore be used as active ingredients of autophagy activators.

[0022] The autophagy activator may consist solely of at least one of the compounds represented by any one of structural formulas (1) to (6), or may be a formulation of at least one of the compounds represented by any one of structural formulas (1) to (6). The compounds represented by any one of structural formulas (1) to (6) may be used singly or in combination of two or more. Furthermore, when two or more compounds are used in combination, the mass ratio of the compounds represented by any one of structural formulas (1) to (6) is not particularly limited and can be selected appropriately depending on the purpose.

[0023] At least one of the compounds represented by any of the structural formulae (1) to (6) can be formulated into any dosage form, such as powder, granules, tablets, or liquid, using pharmaceutically acceptable carriers such as dextrin and cyclodextrin, and any other auxiliary agents, according to standard methods. Examples of auxiliary agents that can be used in this formulation include excipients, binders, disintegrants, lubricants, stabilizers, and flavorings / odor masking agents. The autophagy activator can be incorporated into other compositions (e.g., the autophagy activating composition described below) and used as oral preparations such as tablets, powders, capsules, granules, extracts, and syrups; parenteral preparations such as injections, infusions, and suppositories; ointments, eye drops, topical liquids, and patches.

[0024] The content of at least one of the compounds represented by any of the structural formulae (1) to (6) in the formulated autophagy activator is not particularly limited and can be selected appropriately depending on the purpose.

[0025] <Other Components> The other components are not particularly limited as long as they do not impair the effects of the present invention and can be appropriately selected depending on the form of use of the autophagy activator, and examples include the above-mentioned components that can be used when formulating the drug. The other components may be used alone or in combination of two or more.

[0026] The content of the other components in the autophagy activator is not particularly limited and can be appropriately selected depending on the purpose.

[0027] The method of use of the autophagy activator is not particularly limited and can be appropriately selected depending on the purpose. Examples of methods include oral administration, parenteral administration, and external application, with oral administration being preferred.

[0028] The dosage form of the autophagy activator is not particularly limited, and known dosage forms can be appropriately selected depending on the purpose. The method for producing the autophagy activator in a desired dosage form is not particularly limited, and known methods can be appropriately selected.

[0029] The dose, administration site, administration period, administration interval, etc. of the autophagy activator are not particularly limited and can be appropriately selected depending on the purpose.

[0030] The autophagy activator can activate autophagy through the autophagy-activating activity of at least one of the compounds represented by any of structural formulas (1) to (6). This can enhance cellular metabolism, improve cell health, enhance autophagy activity that has declined with age, or prevent the decline in autophagy activity due to age, thereby suppressing aging and extending lifespan. However, in addition to these uses, the autophagy activator can also be used for any other uses in which exerting autophagy activation is meaningful.

[0031] Because the autophagy activator has excellent autophagy activation activity and is highly safe, it can be used in a wide range of applications, such as pharmaceuticals, quasi-drugs, oral compositions, cosmetics, and reagents for research into the mechanism of autophagy activation. For example, it can be suitably used as an active ingredient in the autophagy activating composition described below. In this case, at least one of the compounds represented by any of the structural formulas (1) to (6) may be blended directly, or at least one of the compounds represented by any of the structural formulas (1) to (6) may be blended in a formulation.

[0032] If necessary, the autophagy activator can be used as an active ingredient by blending other components having autophagy activating activity with at least one of the compounds represented by any of structural formulas (1) to (6).

[0033] The autophagy activator of the present invention is preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as its functional effects are maintained.

[0034] (Composition for Activating Autophagy) The composition for activating autophagy of the present invention contains the autophagy activator of the present invention, and further contains other components as necessary.

[0035] <Autophagy Activator> The autophagy activator is the above-described autophagy activator of the present invention.

[0036] The content of the autophagy activator in the autophagy activating composition is not particularly limited and can be adjusted appropriately depending on the form of the autophagy activating composition, but is preferably 0.0001% to 20% by mass, and more preferably 0.0001% to 10% by mass, calculated as the amount of at least one of the compounds represented by any of structural formulas (1) to (6). The autophagy activating composition may consist solely of the autophagy activator.

[0037] <Other Components> The other components in the autophagy activating composition are not particularly limited and can be selected appropriately depending on the form of use of the autophagy activating composition, and examples include the same other components as those described in the section on autophagy activators above. These may be used alone or in combination of two or more.

[0038] The content of the other components in the composition for activating autophagy is not particularly limited and can be selected appropriately depending on the purpose.

[0039] <Aspects> The aspect of the autophagy-activating composition is not particularly limited and can be appropriately selected depending on the purpose, and examples include pharmaceuticals, quasi-drugs, foods and beverages, etc. The autophagy-activating composition of the present invention can be used daily, and can highly effectively exert various physiological activities, including autophagy activation, due to the action of at least one of the compounds represented by any of the structural formulas (1) to (6) as active ingredients.

[0040] The autophagy-activating composition of the present invention is preferably applied to humans, but can also be applied to animals other than humans (e.g., mice, rats, hamsters, dogs, cats, cows, pigs, monkeys, etc.) as long as the respective functional effects are achieved.

[0041] The method of use of the autophagy-activating composition of the present invention is not particularly limited and can be selected appropriately depending on the purpose. Examples of methods include oral administration, parenteral administration, and topical administration, with oral administration being preferred.

[0042] The oral composition is not particularly limited and can be appropriately selected depending on the purpose, and examples thereof include orally administered agents, foods and beverages, etc. Here, foods and beverages refer to those that are unlikely to be harmful to human health and are taken orally or by administration through the digestive tract in normal social life, and are not limited to administrative classifications such as foods, medicines, and quasi-drugs. Therefore, the foods and beverages refer to a wide range of orally ingested general foods, health foods (functional foods and beverages), health functional foods (foods for specified health uses, foods with nutrient functions, foods with functional claims), quasi-drugs, medicines, etc.

[0043] The type of the oral composition is not particularly limited and can be appropriately selected depending on the purpose. Examples of the oral composition include beverages such as tea drinks, soft drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, alcoholic drinks, coffee drinks, and coffee-containing soft drinks (including concentrated liquids and powders for adjusting these beverages); frozen desserts such as ice cream, ice sherbet, and shaved ice; noodles such as soba noodles, udon noodles, vermicelli, gyoza wrappers, shumai wrappers, Chinese noodles, and instant noodles; confectioneries such as candy, candy, gum, chocolate, candy tablets, snacks, biscuits, jelly, jam, cream, baked goods, and bread; seafood such as crab, salmon, clams, tuna, sardines, shrimp, bonito, mackerel, whale, oysters, saury, squid, ark shells, scallops, abalone, sea urchin, salmon roe, and tokobushi sea bream; and fish products such as kamaboko (fish paste), hamachi (fish paste), and the like. processed seafood and livestock foods such as sausages and other seafood; dairy products such as processed milk and fermented milk; oils and fats and oil-processed foods such as salad oil, tempura oil, margarine, mayonnaise, shortening, whipped cream, and dressings; condiments such as sauces and dressings; retort pouch foods such as curry, stew, oyakodon (chicken and egg rice bowl), rice porridge, rice porridge, Chinese rice bowl, pork cutlet rice bowl, tempura rice bowl, eel rice bowl, hayashi rice, oden, mapo dolphin, beef bowl, meat sauce, egg soup, omelet rice, gyoza (dumplings), shumai (steamed dumplings), hamburger steak, and meatballs; side dishes such as salads and pickles; health, beauty, and nutritional supplements in various forms; pharmaceuticals and quasi-drugs such as tablets, powders, capsules, granules, extracts, syrups, drinks, lozenges, and mouthwash; oral fresheners used in the oral cavity, such as mouth fresheners and breath fresheners, and toothpaste.

[0044] The method for producing the autophagy-activating composition is not particularly limited and can be appropriately selected depending on the form of the autophagy-activating composition, etc.

[0045] The amount, duration, and interval of use of the autophagy-activating composition are not particularly limited and can be appropriately selected depending on the purpose.

[0046] As described above, the autophagy activator and composition for activating autophagy of the present invention have excellent autophagy activating properties. Therefore, the present invention also relates to a method for activating autophagy, which comprises administering at least one of the autophagy activator and the composition for activating autophagy to an individual.

[0047] Test examples and formulation examples of the present invention will be explained below, but the present invention is not limited to these test examples and formulation examples.

[0048] (Test Example 1: Evaluation of autophagy activity by tfLC3 test) Compounds represented by any of the structural formulas (1) to (6) were used as test samples, and the autophagy activation effect was tested by the tfLC3 test. - Compound represented by structural formula (1) (manufactured by Toronto Research Chemicals) - Compound represented by structural formula (2) (manufactured by Toronto Research Chemicals) - Compound represented by structural formula (3) (manufactured by Tokyo Chemical Industry Co., Ltd.) - Compound represented by structural formula (4) (manufactured by Sigma-Aldrich) - Compound represented by structural formula (5) (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) - Compound represented by structural formula (6) (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.)

[0049] The tfLC3 assay uses tfLC3, an autophagosome marker protein fused in tandem with the fluorescent proteins mRFP (RFP; Red Fluorescent Protein) and EGFP (GFP; Green Fluorescent Protein) (Autophagy 2007; 3:452-460. (PubMed: 17534139) DOI: 10.4161 / auto.4451). In cells undergoing autophagy, LC3 dots increase, but even inhibition of autophagic degradation leads to LC3 accumulation. Therefore, an increase in LC3 dots alone does not necessarily indicate enhanced autophagy. Therefore, by examining the maturation process of autophagosomes (the fusion process between autophagosomes and lysosomes, or degradation within the autolysosome after fusion), it is possible to distinguish between enhanced and inhibited autophagy. The GFP signal (green) of tfLC3 becomes less fluorescent in the lysosomal environment, whereas the RFP (red) remains stable in the same environment. Taking advantage of the property of tfLC3 that the autophagosome turns red due to quenching of GFP as it matures into an autolysosome, the tfLC3 test determines whether a test sample added to culture medium has the ability to activate autophagy by analyzing the intracellular GFP / RFP ratio. The specific experimental method is as follows.

[0050] HeLa cells stably expressing tfLC3 were seeded in a 96-well plate (Perkin Elmer #6055302 Cell Carrier 96 Ultra Black 96-well, Clear Bottom, with Lid) and incubated at 37°C for 1 hour at 5% CO ２ After culturing in growth medium (DMEM (Sigma D6546) supplemented with 10% FBS and 4 mM L-glutamine) in an incubator for 24 hours, the medium was replaced with the medium containing the test sample (see Table 1 for the final concentration of the test sample) and cultured for another 24 hours. Cells cultured in medium containing 0.1% dimethyl sulfoxide (DMSO) served as a negative control.

[0051] The GFP / RFP value measured in the negative control was set to 1, and the GFP / RFP value measured in the cells cultured in the medium containing the test sample (hereinafter sometimes referred to as the "ratio relative to DMSO") was calculated. The results are shown in Table 1. Test samples that had a lower GFP / RFP ratio than the negative control could be determined to have an autophagy-activating effect.

[0052]

[0053] From the above results, it was confirmed that the compounds represented by any of the structural formulas (1) to (6) have an autophagy activating effect.

[0054] Formulation Example 1 Tablets having the following composition were produced by a conventional method: Compound represented by the structural formula (1) 50.0 mg Grape extract 100.0 mg β-nicotinamide mononucleotide 5.0 mg Dolomite 23.4 mg (containing 20% ​​calcium and 10% magnesium) Vitamin C 13.4 mg Maltitol 96.2 mg Sucrose fatty acid ester 12.0 mg

[0055] Formulation Example 2 An oral liquid formulation having the following composition was produced by a conventional method. <Composition in 1 ampoule (100 mL per ampoule)> - 0.2% by mass of compound represented by the structural formula (2) - 0.1% by mass of compound represented by the structural formula (3) - 12.0% by mass of sorbitol - 0.1% by mass of sodium benzoate - 1.0% by mass of flavoring - 0.5% by mass of calcium sulfate - purified water, balance

[0056] (Formulation Example 3) A coffee drink having the following composition was produced by a conventional method: 0.1% by mass of the compound represented by the structural formula (4) 40% by mass of coffee extract (L (lightness) = 20, Brix = 3) 2% by mass of maltitol an appropriate amount of flavoring water, balance

[0057] Formulation Example 4 Capsules having the following composition were produced by a conventional method. No. 1 hard gelatin capsules were used. <Composition per capsule (200 mg per tablet)> Compound represented by the structural formula (5) 15.0 mg Compound represented by the structural formula (6) 15.0 mg Corn starch 70.0 mg Lactose 80.0 mg Calcium lactate 10.0 mg Hydroxypropyl cellulose (HPC-L) 10.0 mg

[0058] This application claims priority based on Japanese Patent Application No. 2023-105885, filed on June 28, 2023, the entire contents of which are incorporated herein by reference.

Claims

1. An autophagy activator comprising at least one compound represented by any one of the following structural formulas (1) to (6):

2. A composition for activating autophagy, comprising the autophagy activator described in claim 1.

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