JP2024064707A - Fat decomposition promoter - Google Patents

Abstract

[Problem] To provide a lipolysis promoter that can effectively promote the breakdown of fats such as neutral fats and body fat after oral ingestion. [Solution] The lipolysis promoter of the present invention contains Chakasaponin as an active ingredient. Here, the saponin is at least one compound selected from the group consisting of Chakasaponin I, Chakasaponin II, and Chakasaponin III. The lipolysis promoter of the present invention is useful as an ingredient for oral compositions such as general foods, supplements, health functional foods, and pharmaceuticals. [Selected Figures] None

Description

The present invention relates to a fat decomposition promoter, and more specifically to a fat decomposition promoter that can effectively decompose neutral fats and body fat in the human body.

With the rise in health consciousness, attention is being focused on the development of oral materials (e.g., food materials and oral pharmaceutical materials) that can act effectively on neutral fats and body fat (hereinafter sometimes referred to as neutral fats, etc.) in the human body. These oral materials can be classified from the perspective of whether they do not take in neutral fats or body fat, whether they do not accumulate in the body, or whether they break down neutral fats or body fat in the body.

For example, various oral materials that can prevent or reduce the intake of neutral fats and other substances into the body have been proposed, such as those that inhibit the activity of fat-decomposing enzymes in the body (pancreatic lipase), those that stabilize fats using micelles, and those that act on the intestinal lymphatic vessels to suppress fat absorption. Specific examples of materials that can prevent or reduce the intake of neutral fats and body fat into the body include resistant dextrin, isomaltodextrin, and Terminalia bellirica extract.

On the other hand, oral materials that can prevent or reduce the accumulation of neutral fats and other substances in the body have been proposed, such as those that act on the liver to suppress fat synthesis in the body. Specific examples include omega-3 fatty acids such as eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA).

As oral materials that can initiate or enhance the breakdown of neutral fats in the body, for example, materials that act directly on fat to break it down themselves have been proposed. Specific examples include polymethoxyflavones, isoflavones derived from kudzu flowers, tiliroside derived from rose hips, and tea catechins.

However, the reality is that none of the oral materials mentioned above provide sufficient effects for those who take them. There is a particular need for the development of oral materials that can initiate or enhance the breakdown of neutral fats and other substances in the body.

The present invention aims to solve the above problems, and its purpose is to provide a lipolysis promoter that can effectively promote the decomposition of fats such as neutral fats and body fat after oral ingestion.

The present invention relates to a fat decomposition promoter containing Chakasaponin as an active ingredient,
This is a lipolysis promoter, wherein the chakasaponin is at least one compound selected from the group consisting of chakasaponin I, chakasaponin II, and chakasaponin III.

In one embodiment, the chakasaponin contains all of the chakasaponin I, the chakasaponin II, and the chakasaponin III.

In a further embodiment, the Chakasaponin I is contained in an amount of 0.04 to 0.71 times the amount of Chakasaponin II, and the Chakasaponin III is contained in an amount of 0.01 to 0.39 times the amount of Chakasaponin II.

In a further embodiment, based on the total mass, the content of Chakasaponin I is 0.11% to 0.57% by mass, the content of Chakasaponin II is 0.51% to 1.47% by mass, and the content of Chakasaponin III is 0.03% to 0.31% by mass.

In a further embodiment, the total content of Chakasaponin I, Chakasaponin II, and Chakasaponin III is 1.00% by mass or more and 6.0% by mass or less based on the total mass.

In one embodiment, the chakasaponin is contained in the form of tea flower extract.

In a further embodiment, the tea flower extract is a hot water extract of tea (Camellia sinensis L.).

The present invention is also a fat decomposition promoter that contains tea flower extract as an active ingredient.

According to the present invention, neutral fats and the like in the body can be effectively broken down after oral ingestion. This makes it possible to inhibit the accumulation of neutral fats and the like in the body. Furthermore, the lipolysis promoter of the present invention can be easily produced from materials such as tea flower extract, is safe for humans, and can be taken by a wide range of ages.

1 is a graph showing the relative values of the amount of inhibition of fat accumulation in each test group obtained in Example 5. 1 is a graph showing the relative values of the free glycerol amount in each test group obtained in Example 6.

The present invention is described in detail below.

The lipolysis promoter of the present invention contains chakasaponin as an active ingredient.

Chakasaponin is a glycoside (saponin) composed of sapogenin and sugar. In the present invention, an example of chakasaponin is chakasaponin I represented by the following formula (I):

Chakasaponin II represented by the following formula (II):

Chakasaponin III represented by the following formula (III):

and combinations thereof.

In the present invention, it is preferable that the chakasaponin contains all of the above chakasaponin I, chakasaponin II, and chakasaponin III, because these can be easily obtained from natural materials as described below, and because a synergistic improvement in the lipolytic activity of each component can be expected.

In one embodiment, when Chakasaponin contains all of Chakasaponin I, Chakasaponin II, and Chakasaponin III, the relationship between their contents can be explained, for example, based on the content of Chakasaponin II contained in the lipolysis promoter of the present invention.

That is, the content of Chakasaponin I contained in the lipolysis promoter of the present invention is preferably 0.04 to 0.71 times, more preferably 0.14 to 0.56 times, the content of Chakasaponin II. The content of Chakasaponin III contained in the lipolysis promoter of the present invention is preferably 0.01 to 0.39 times, more preferably 0.04 to 0.32 times, the content of Chakasaponin II. By having the contents of Chakasaponin I and Chakasaponin III independently fall within the above ranges relative to the content of Chakasaponin II, the lipolysis promoter of the present invention can more effectively break down fat in the body and reduce the amount of fat accumulation.

Alternatively, in one embodiment, when Chakasaponin contains all of Chakasaponin I, Chakasaponin II, and Chakasaponin III, the content of Chakasaponin I is preferably 0.11% to 0.57% by mass, more preferably 0.18% to 0.51% by mass, based on the total mass of the lipolysis promoter of the present invention. On the other hand, the content of Chakasaponin II is preferably 0.51% to 1.47% by mass, more preferably 0.68% to 1.29% by mass, based on the total mass of the lipolysis promoter of the present invention. Furthermore, the content of Chakasaponin III is preferably 0.03% to 0.31% by mass, more preferably 0.07% to 0.28% by mass, based on the total mass of the lipolysis promoter of the present invention. By containing Chakasaponin I, Chakasaponin II, and Chakasaponin III within the above ranges as saponins, the lipolysis promoter of the present invention can more effectively break down fat in the body, thereby reducing the amount of fat accumulation.

Alternatively, when the lipolysis promoter of the present invention contains chakasaponin I, chakasaponin II, and chakasaponin III as chakasaponins, the lower limit of the total chakasaponin content is preferably 1.00% by mass or more, more preferably 1.13% by mass or more, based on the total mass of the lipolysis promoter of the present invention, because the chakasaponins can be particularly effective in promoting the decomposition of neutral fats in the body. Also, the upper limit of the total chakasaponin content containing chakasaponin I, chakasaponin II, and chakasaponin III is preferably 6.0% by mass or less, more preferably 3.6% by mass or less, based on the total mass of the lipolysis promoter of the present invention, from the viewpoint of ensuring safety to the ingesting body and/or preventing overdosing.

The lipolysis promoter of the present invention may also contain the above-mentioned chakasaponin in the form of tea flower extract.

Tea flowers are obtained from the flower buds of tea (Camellia sinensis L.). Tea (Camellia sinensis L.) is a perennial evergreen tree belonging to the Theaceae family, native to the areas from Fujian, Yunnan, and Sichuan provinces in the People's Republic of China to northern Myanmar. Tea flowers are known to contain functional components such as Chakasaponins, such as Chakasaponin I, Chakasaponin II, and Chakasaponin III, as well as saponins such as floratheasaponins, and are commercially available in the form of, for example, a dried product (dried flowers).

In the present invention, for example, a tea flower extract containing the above-mentioned chakasaponins (e.g., chakasaponin I, chakasaponin II, and chakasaponin III) can be obtained by extracting the dried tea flowers in a solvent consisting of water or ethanol, or a combination of these. The temperature used for extracting the tea flowers is preferably 90°C to 98°C, more preferably 93°C to 96°C. In the present invention, the tea flower extract is preferably a hot water extract of tea (Camellia sinensis L.), because it can be extracted so as to contain chakasaponin I, chakasaponin II, and chakasaponin III in a rich and effective ratio.

In the present invention, the tea flower extract can be separated and purified into Chakasaponin I, Chakasaponin II, and Chakasaponin III by methods known to those skilled in the art and used as Chakasaponin, but from the perspective of increasing production efficiency, the tea flower extract (for example, an extract that may contain other components in addition to Chakasaponin I, Chakasaponin II, and Chakasaponin III) may be used as it is as a lipolysis promoter.

The lipolysis promoter of the present invention may also contain other active ingredients having a lipolysis promoting effect in addition to the above-mentioned Chakasaponin. Examples of other active ingredients having a lipolysis promoting effect include tea catechin, rosehip-derived tiliroside, acetic acid, kudzu flower-derived isoflavone, enokitake mushroom-derived fatty acid (e.g., linoleic acid, α-linolenic acid), forskolin, red paprika-derived xanthophyll, polymethoxyflavone, chlorogenic acid, apple-derived procyanidin, hydroxycitric acid, licorice-derived glabridin, alanine, arginine, phenylalanine, psicose, medium-chain fatty acid (e.g., octanoic acid, decanoic acid), and HMB calcium, as well as combinations thereof. The content of such other active ingredients having a lipolysis promoting effect is not particularly limited, and an appropriate amount can be selected by a person skilled in the art within a range that does not inhibit the effect of the above-mentioned saponin.

The lipolysis promoter of the present invention may also contain excipients, lubricants, binders, disintegrants, and surfactants, as well as combinations thereof, in addition to the chakasaponin.

Excipients include, for example, maltodextrin, lactose, trehalose, L-cysteine, trehalose, maltitol, sorbitol, and corn starch, and combinations thereof.

Lubricants include, for example, magnesium stearate, calcium stearate, sodium stearyl fumarate, sucrose fatty acid esters, light anhydrous silicic acid, and talc, and combinations thereof.

Examples of binders include crystalline cellulose, dextrin, cyclodextrin, starch, sucrose, pregelatinized starch, hydroxypropyl cellulose, gelatin, gum arabic, polyvinylpyrrolidone, pullulan, methylcellulose, ethylcellulose, hydroxymethylcellulose, hydroxypropyl cellulose, hydroxypropylmethylcellulose, polyvinyl alcohol, and polyethylene glycol, as well as combinations thereof.

Disintegrants include, for example, low-substituted hydroxypropylcellulose, partially pregelatinized starch, carboxymethylcellulose, sodium carboxymethylcellulose, and potassium carboxymethylcellulose, and combinations thereof.

Examples of surfactants include lecithin.

The amount of these components contained in the lipolysis promoter of the present invention is not particularly limited, and an appropriate amount can be selected by a person skilled in the art within a range that does not inhibit the effects of the above-mentioned chakasaponin.

The lipolysis promoter of the present invention may further contain other additives known in the art. Such other additives include pH adjusters, antioxidants, UV absorbers, colorants, flavorings, preservatives, and fragrances, as well as combinations thereof. The content of other additives in the present invention is not particularly limited, and an appropriate amount can be selected by a person skilled in the art within a range that does not inhibit the effects of the above-mentioned chakasaponin.

The lipolysis promoter of the present invention can be processed into a dosage form such as a powder, granules, powder, pill, or plain tablet by combining Chakasaponin with the above-mentioned various components. Alternatively, it may be processed into a capsule dosage form housed in a hard capsule or soft capsule.

The dosage of the lipolysis promoter of the present invention is, for example, 1.00 mg to 6.00 mg, preferably 1.13 mg to 3.60 mg, per day as the total amount of the active ingredient chakasaponin (for example, the total amount of chakasaponin I, chakasaponin II, and chakasaponin III). When a living body (for example, a human) ingests the lipolysis promoter of the present invention at a dose within the above range per day, lipolysis in the living body can be carried out safely and efficiently.

The lipolysis promoter of the present invention is delivered to the body of each living organism, for example, a human, a pet animal, livestock, poultry, farmed fish, etc., by oral ingestion.

Next, the present invention will be further explained by giving examples, but the present invention is not limited to these examples.

(Example 1: Preparation of tea flower extract)
Dried tea flowers (1500 kg) from the Fujian Province of the People's Republic of China (Camellia sinensis L.) were crushed and extracted with hot water at 95° C. for 2 hours to obtain an extract. The extract was then dried with a spray dryer to obtain 375 kg of pure extract. Maltodextrin was added to the pure extract as an excipient for stabilization to obtain 750 kg of tea flower extract.

The obtained tea flower extract was analyzed using high performance liquid chromatography (SPD-20A, manufactured by Shimadzu Corporation) under the following measurement conditions.
Measurement condition:
(Column) COSMCIL Code No. 38020-41, size 4.6 x 250 mm, Type 5C18-MS-II Waters (manufactured by Nacalai Tesque, Inc.)
(Detector) Ultraviolet absorption photometer (measurement wavelength: 230 nm)
(Column temperature) Constant temperature around 40°C (Mobile phase) 65:35 mixture of methanol/0.01 mol/L phosphate buffer (pH 2.3) (Flow rate) 1.0 mL/min (Analysis time) 60 min

As a result of the analysis, it was confirmed that the tea flower extract obtained above contained 0.34% by mass of chakasaponin I, 0.99% by mass of chakasaponin II, and 0.17% by mass of chakasaponin III (total saponins: 1.50% by mass) based on the total mass of the extract. In other words, the content of chakasaponin I contained in the obtained tea flower extract was 0.35 times the content of chakasaponin II (0.99% by mass), and the content of chakasaponin III was 0.18 times the content of chakasaponin II.

(Examples 2 to 4: Purification of Chasaponins I, II, and III)
3.0 g of the tea flower extract obtained in Example 1 was added to ion-exchanged water to prepare a concentration of 20 mg/mL. Then, 150 mL of a mixed solution of water, methanol, and acetic acid mixed in a volume ratio of 30/70/1 was added, and ultrasonic treatment was performed for 10 minutes. Then, unnecessary substances were filtered using a PVDF filter (0.2 μm) to obtain a raw material solution.

Next, 120 mL of the raw material solution was subjected to separation and purification by reverse phase chromatography under the following conditions.
Column: Triart PreP C18-S (10 μm, 12 nm)
250 x 50 mm I.D. (manufactured by YMC Co., Ltd.)
Load: 40mL
Runs: 3

As a result, 97.83 mg of Chakasaponin I (Example 2), 215.66 mg of Chakasaponin II (Example 3), and 32.21 mg of Chakasaponin III (Example 4) were obtained as purified products.

(Example 5: Confirmation test of inhibition of fat accumulation in 3T3-L1 cells)
Culture and differentiation induction of mouse-derived 3T3-L1 preadipocytes (American Type Culture Collection) were performed according to existing methods. 3T3-L1 cells were grown in 24-well plates in DMEM medium (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (FBS) and antibiotics at 37°C under 5% _CO2 conditions, and when the cells reached confluence, the medium was replaced with a differentiation induction medium (10% FBS, 0.5 μM dexamethane, 10 μg/ml insulin, 0.5 mM 3-isobutyl-1-methylxanthine), and differentiation induction was performed for 2 days. The adipocyte differentiation induction medium was replaced with adipocyte differentiation medium, and the tea flower extract of Example 1 and Chakasaponins I to III of Examples 2 to 4 dissolved in phosphate buffered saline (PBS) were added at the prescribed concentrations (5 ppm, 50 ppm, and 250 ppm for the tea flower extract of Example 1; 5 ppm and 20 ppm for Chakasaponins I to III of Examples 2 to 4), and the cells were further cultured for 2 days.

The adipocyte differentiation medium was then replaced with a passage medium (DMEM, 10% FBS, 10 μg/ml insulin), and the cells were then cultured for 7 days. The effect on the amount of fat accumulation inhibition was examined by staining fat with Oil Red O (Fujifilm Wako Pure Chemical Industries, Ltd.), recovering the cells with a cell scraper, disrupting the cells by sonication, and measuring the absorbance at 520 nm. As a control, a group (PBS) without tea flower extract or Chakasaponin I-III was used, and the absorbance was measured in the same manner as above. The results are shown in Figure 1 and Table 1.

In Figure 1, the amount of fat accumulation suppression in each test group was compared relative to the control (no addition) which was set at 100%. The test results in Table 1 represent the results of a test (Student's t-test) compared to the control.

As is clear from Figure 1 and Table 1, the tea flower extract obtained in Example 1, Chakasaponin I obtained in Example 2, Chakasaponin II obtained in Example 2, and Chakasaponin III obtained in Example 3 all inhibit fat accumulation in a concentration-dependent manner, indicating that all of them can effectively function as fat decomposition promoters.

(Example 6: Confirmation test of promotion of lipolysis by 3T3-L1 cells)
Mouse-derived 3T3-L1 preadipocytes (American Type Culture Collection) were cultured and induced to differentiate according to existing methods. The cells were grown in DMEM medium (Fujifilm Wako Pure Chemical Industries, Ltd.) containing 10% fetal bovine serum (FBS) and antibiotics at 37°C under 5% _CO2 conditions, and when the cells reached confluence, the medium was replaced with a differentiation induction medium (10% FBS, 0.5 μM dexamethane, 10 μg/ml insulin, 0.5 mM 3-isobutyl-1-methylxanthine), and differentiation induction was carried out for 2 days. The adipocyte differentiation induction medium was replaced with adipocyte differentiation medium, and the tea flower extract of Example 1 and Chakasaponin I-III of Examples 2-4 dissolved in phosphate buffered saline (PBS) were added at the respective prescribed concentrations (5 ppm, 50 ppm, and 250 ppm for the tea flower extract of Example 1; 5 ppm and 20 ppm for Chakasaponin I-III of Examples 2-4), and the cells were further cultured for 2 days (48 hours). The culture medium was then collected after culture, and the amount of free glycerol in the medium was measured using an F-kit (R-Biopharm). The results are shown in Figure 2 and Table 2.

In Figure 2, the amount of free glycerol in each test group was compared relative to the control (no addition) which was set at 100%. The test results in Table 2 show the results of a test (Student's t-test) compared to the control.

As is clear from Figure 1 and Table 1, the tea flower extract obtained in Example 1, Chakasaponin I obtained in Example 2, Chakasaponin II obtained in Example 2, and Chakasaponin III obtained in Example 3 all released glycerol in a concentration-dependent manner, indicating that all of them can effectively function as lipolysis promoters.

The present invention is useful in technical fields such as food and pharmaceuticals.

Claims (8)

A fat decomposition promoter containing Chakasaponin as an active ingredient,
A lipolysis promoter, wherein the chakasaponin is at least one compound selected from the group consisting of chakasaponin I, chakasaponin II, and chakasaponin III. The lipolysis promoter according to claim 1, wherein the chakasaponin contains all of the chakasaponin I, the chakasaponin II, and the chakasaponin III. The lipolysis promoter according to claim 2, wherein the Chakasaponin I is contained in an amount of 0.04 to 0.71 times the amount of Chakasaponin II, and the Chakasaponin III is contained in an amount of 0.01 to 0.39 times the amount of Chakasaponin II. The lipolysis promoter according to claim 2, wherein the content of Chakasaponin I is 0.11% to 0.57% by mass, the content of Chakasaponin II is 0.51% to 1.47% by mass, and the content of Chakasaponin III is 0.03% to 0.31% by mass, based on the total mass. The lipolysis promoter according to claim 2, wherein the total content of the chakasaponin I, the chakasaponin II, and the chakasaponin III is 1.00% by mass or more and 6.0% by mass or less based on the total mass. The lipolysis promoter according to claim 1, wherein the chakasaponin is contained in the form of tea flower extract. The lipolysis promoter according to claim 6, wherein the tea flower extract is a hot water extract of tea (Camellia sinensis L.). A fat breakdown promoter that contains tea flower extract as an active ingredient.

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