JP2023067303A - Glycyrrhiza extract and manufacturing method thereof as well as intestinal bacteria-adjusting composition, intestinal barrier function-enhancing composition, antiobesity composition, antidiabetic composition, and nlrp3 inflammasome activity-controlling composition - Google Patents

Abstract

To provide a composition that provides a novel effect of Glycyrrhiza extract and compounds contained therein and exerts a novel medicinal effect.SOLUTION: A method for producing a Glycyrrhiza extract in one aspect of the present invention has a step of removing glycyrrhizic acid. A Glycyrrhiza extract according to another aspect contains isoliquiritigenin (ILG), and further contains at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). In addition, an intestinal barrier function-enhancing composition according to another aspect contains ILG as an active ingredient. In addition, an intestinal bacteria-adjusting composition according to another aspect contains a Glycyrrhiza extract, or at least one of L and ILG as an active ingredient. In addition, an anti-obesity composition, an anti-diabetic composition, or a NLRP3 inflammasome activity-controlling composition according to another aspect contains ILG, and additionally contains as an active ingredient a Glycyrrhiza extract containing at least one of L, IL and LG.SELECTED DRAWING: None

Description

The present invention provides a licorice extract, a method for producing the same, a composition for regulating intestinal bacteria, a composition for enhancing intestinal barrier function, an anti-obesity composition, an anti-diabetic composition, and a composition for controlling NLRP3 inflammasome activity. Regarding.

Glycyrrhiza (scientific name: G. glabra, G. uralensis, G. echinata, G. inflata, G. yunnanensis, G. eurycarpa, G. aspera, G. pallidiflora, Japanese name: licorice, English name: Licorice) is grown in northeastern and northern China. It is a perennial herb that is widely distributed in , northwestern Japan, Mongolia, Siberia, etc. It is characterized by a large rhizome, underground stems running in all directions, and a long taproot reaching 1 to 2 m. Glycyrrhiza is mainly used in the roots and stolons, and is listed in pharmacological encyclopedias around the world, both in the West and the East, and in Japan, for example, in the Japanese Pharmacopoeia (see, for example, Non-Patent Document 1).

In addition, licorice is known to exhibit physiological activities such as anti-ulcer action and antitussive action, and is currently widely used in cold medicines, chronic hepatitis medicines, health drinks, and the like. In other words, licorice is widely used in daily life as it is often used as a sweetener in addition to cosmetics and quasi-drugs.

https://jpdb. nihs. go. jp/jp/DetailList_ja. aspx? submit = %e8%a9%b3%e7%b4%b0%e6%a4%9c%e7%b4%a2 (%e6%97%a5%e6%9c%ac%e8%aa%9e%e5%90% 8d) &keyword=%e3%82%ab%e3%83%b3%e3%82%be%e3%82%a6

As described above, various uses of licorice have been investigated, but the efficacy of licorice extract is still largely unknown. In addition to further increasing usefulness, it will lead to the promotion of human health.

Therefore, in view of the above problems, it is an object of the present invention to provide a composition that provides new effects of licorice extract and compounds contained therein and exhibits new medicinal effects.

The present inventors have made intensive studies on the above problems, and found that the licorice extract obtained by a predetermined production method and the compounds contained therein have an intestinal bacterium-regulating action, an intestinal barrier function-enhancing action, an anti-obesity action, and an anti-obesity action. The inventors have discovered that it exhibits diabetic activity and NLRP3 inflammasome activity regulating activity, leading to the completion of the present invention.

That is, the method for producing a licorice extract according to one aspect of the present invention includes a step of removing glycyrrhizic acid.

A method for producing a licorice extract according to another aspect of the present invention includes a step of removing glycyrrhizic acid so that the concentration of glycyrrhizic acid is 1% by weight or less.

A method for producing a licorice extract according to another aspect of the present invention includes a step of removing glycyrrhizic acid so that the concentration of glycyrrhizic acid is 0.5% by weight or less.

In addition, the licorice extract according to another aspect of the present invention contains isoliquiritigenin (ILG), and further contains at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). It is something to do.

From this point of view, although not limited, it preferably contains at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG).

In this aspect, although not limited, isoliquiritigenin (ILG) is 0.1% by weight or more and 30% by weight or less, liquiritin is 10% by weight or less, and isoliquiritin (IL) is 15% by weight. and Liquiritigenin (LG) in an amount of 30% by weight or less.

In addition, a licorice extract according to another aspect of the present invention is a licorice extract produced through a step of removing glycyrrhizic acid, containing isoliquiritigenin (ILG) and liquiritin (L) , isoliquiritin (IL) and liquiritigenin (LG).

In this aspect, although not limited, isoliquiritigenin (ILG) is 0.1% by weight or more and 30% by weight or less, liquiritin (L) is 10% by weight or less, and isoliquiritin (IL) is It preferably contains 15% by weight or less and 30% by weight or less of Liquiritigenin (LG).

A composition for enhancing intestinal barrier function according to another aspect of the present invention contains isoliquiritigenin (ILG) as an active ingredient.

A composition for regulating intestinal bacteria according to another aspect of the present invention contains at least one of liquitin (L) and isoliquiritigenin (ILG) as an active ingredient.

From this point of view, although not limited, it is preferable to further contain at least one of isoliquiritin (IL) and liquiritigenin (LG) as an active ingredient.

A composition for adjusting intestinal bacteria according to another aspect of the present invention contains a licorice extract as an active ingredient.

In addition, although the composition for adjusting intestinal bacteria according to the above aspect is not limited, it is preferably a pharmaceutical composition or a composition for food and drink.

In addition, although the composition for adjusting intestinal bacteria according to the above aspect is not limited, the intestinal bacterium is A. muchiniphila and P. goldsteinii is preferred.

In addition, although the composition for adjusting intestinal bacteria according to the above aspect is not limited, A. muchiniphila and P. It is preferable to increase the abundance ratio of at least one of goldsteinii.

An anti-obesity composition according to another aspect of the present invention contains isoliquiritigenin (ILG), and at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). It contains a licorice extract containing seeds as an active ingredient.

Moreover, in this aspect, although not limited, it is preferably a pharmaceutical composition or a food and drink composition.

An anti-diabetic composition according to another aspect of the present invention contains isoliquiritigenin (ILG) and at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). It contains a licorice extract containing seeds as an active ingredient.

Moreover, in this aspect, although not limited, it is preferably a pharmaceutical composition or a food and drink composition.

In addition, a composition for controlling NLRP3 inflammasome activity according to another aspect of the present invention contains isoliquiritigenin (ILG), and further liquiritigenin (L), isoliquiritin (IL) and liquiritigenin (LG) It contains as an active ingredient a licorice extract containing at least one of

Moreover, in this aspect, although not limited, it is preferably a pharmaceutical composition or a food and drink composition.

As described above, according to the present invention, it is possible to provide a composition that provides new effects of the licorice extract and the compounds contained therein and exerts new medicinal effects.

FIG. 4 is a diagram showing changes in body weight in each mouse group. FIG. 3 is a diagram showing food intake in each mouse group. FIG. 12 shows the liver weight of each mouse group at 12 weeks after the start of feeding. FIG. 10 is a diagram showing the epididymal cell composition weight of each mouse group at 12 weeks after the start of feeding. FIG. 4 shows the results of insulin resistance tests in each mouse group. FIG. 10 shows the results of plasma insulin concentration in each mouse group 12 weeks after the start of the experiment. FIG. 10 is a diagram showing the confirmation results of intestinal bacteria in each mouse group by 16S-rRNA analysis. FIG. 4 is a diagram showing analysis results of intestinal bacteria at the Species level. FIG. 2 shows the results of Western Blot expression analysis of Claudin-1 in large intestine tissue. FIG. 10 shows the expression level of Claudin-1 in FIG. 9. FIG. FIG. 2 shows the results of bacterial flora analysis for Akkermansia muciniphila and Parabacteroides goldsteinii. FIG. 10 is a diagram showing analysis results of feces of each mouse group two weeks after feeding. It is a figure which shows the result of having analyzed the abundance ratio of Akkermansia muciniphila and Parabacteroides goldsteinii in intestine. FIG. 8 shows the amount of visceral fat in mice 8 weeks after transplantation. It is a figure which shows the change of the body weight of each mouse|mouth. Macrophage-like cell line J774. FIG. 2 shows the results of measuring the effect on IL-1β production from A1. FIG. 2 shows the results of measuring the effect on IL-1β production from visceral adipose tissue.

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, embodiments of the present invention will be described in detail. However, the present invention can be embodied in many different forms, and is not limited only to the specific exemplifications in the following embodiments and examples.

(Method for producing licorice extract)
The method for producing a licorice extract according to the present embodiment (hereinafter referred to as "this method") has a step of removing glycyrrhizic acid from licorice roots.

First, this method has a step of removing glycyrrhizic acid as described above. This step makes it possible to reduce the concentration of glycyrrhizic acid in the licorice extract.

Here, the raw material "Glycyrrhiza" is, as described above, the scientific name of G. glabra, G. uralensis, G. echinata, G.; inflata, G. yunnanensis, G. eurycarpa, G.; aspera, or G. It is a plant called pallidiflora (Japanese name: licorice, English name: Licorice), and is a perennial herb that is widely distributed in the northeastern, northern and northwestern parts of China, Mongolia, Siberia, and the like. Glycyrrhiza is characterized by a large rhizome and underground stems that run in all directions, and a long taproot that reaches 1 to 2 m.

In this method, the part of the licorice used is not limited as long as the desired active ingredient can be obtained. The licorice without the skin (skinless licorice) may be used, but the root is preferable from the viewpoint of obtaining a more effective extract.

In addition, here, the method for removing glycyrrhizic acid is not limited as long as it can remove glycyrrhizic acid. Removal by precipitation at , removal by metal chloride, or the like can be used.

In addition, the content of glycyrrhizic acid to be removed here is preferably 1% by weight or less, more preferably 0.5% by weight or less, when the sufficiently dried extract is 100% by weight. is. Assuming that the total weight of the extract before general removal is 100% by weight, it generally contains 5.0% by weight or more of glycyrrhizic acid, but it is preferable to keep it below the above range.

Removal of glycyrrhizic acid is preferably performed on an extract obtained from licorice. The method for obtaining an extract from licorice is not particularly limited, and a general method can be adopted. A method of obtaining a precipitate and then concentrating and drying the filtrate after filtration can be employed.

The licorice extract according to the present embodiment (hereinafter referred to as "the present extract") can be obtained by using the extract obtained by the above method as it is as the licorice extract. It may be concentrated after removing glycyrrhizic acid, dried, or dried and dissolved again in a solvent such as water.

(manufactured licorice extract)
As described above, this extract is obtained by the above method, and specifically contains isoliquiritigenin (ILG) as an essential component, and further contains liquiritin (L), isoliquiritin (IL) and It contains at least one of liquiritigenin (LG), and more preferably contains at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG) in addition to the above essential ingredients. be.

In this extract, isoliquiritigenin (ILG) is an essential component of this extract and is a compound represented by the following formula.

In addition, in the present extract, isoliquiritigenin (ILG) is not limited as long as the present extract can obtain the desired effect, but it should be 0.1% by weight or more and 30% by weight or less. is preferred, more preferably 1% by weight or more and 20% by weight or less, and even more preferably 3% by weight or more and 15% by weight or less.

In addition, liquiritin (L) is selectively contained in this extract, but is a compound capable of exhibiting the same effects as isoliquiritigenin, and is represented by the following formula.

In addition, in the present extract, liquiritin (L) is not limited as long as the present extract can obtain the desired effect, but when it contains liquiritin (L), it is preferably 10% by weight or less. , more preferably 5% by weight or less.

In addition, isoliquiritin (IL) is selectively contained in this extract, but is a compound capable of exhibiting the same effect as isoliquiritigenin (ILG), and is represented by the following formula: is.

In the present extract, isoliquiritin (IL) is not limited as long as the present extract can obtain the desired effect, but when it contains isoliquiritin (IL), it is preferably 15% by weight or less. , more preferably 10% by weight or less, and still more preferably 5% by weight or less.

Liquiritigenin (LG) is selectively contained in this extract, but it is a compound that can exhibit the same effect as isoliquiritigenin (ILG), and is represented by the following formula: is.

In addition, in the present extract, the content of liquiritigenin (LG) is not limited as long as the present extract can obtain the desired effect, but when it contains liquiritigenin (LG), it is preferably 30% by weight or less. , more preferably 25% by weight or less, and still more preferably 20% by weight or less.

In this extract, the concentrations of isoliquiritigenin (ILG), etc. are as described above. Tigenin (ILG) is preferably 0.1% by weight or more and 30% by weight or less, more preferably 1% by weight or more and 20% by weight or less, still more preferably 3% by weight or more and 15% by weight or less, Liquiritin ( L) is preferably 10% by weight or less, more preferably 5% by weight or less, and isoliquiritin (IL) is preferably 15% by weight or less, more preferably 10% by weight or less, and still more preferably 5% by weight. % by weight or less, and Liquiritigenin (LG) is preferably 30% by weight or less, more preferably 25% by weight or less, and even more preferably 20% by weight or less.

In addition, the above compounds contained in this extract are efficiently extracted from licorice, and as will be apparent from the examples described later, the desired effects are exhibited by using a predetermined amount of the above compounds. be able to. On the other hand, by using a predetermined amount of the present extract itself, it is possible to exhibit the desired effect, and in many cases the effect is higher than when the compound is used alone. However, it is almost impractical to directly specify all the components in this extract at the time of filing the present patent application, and it is practical to specify them by the production method. Specifically, this extract is produced by a process in which glycyrrhizic acid is removed, contains isoliquiritigenin (ILG), and further contains liquiritin (L), isoliquiritin (IL) and It can be said that it contains at least one selected from Liquiritigenin (LG).

As described above, the present extract contains at least the predetermined compound, and can be extracted and produced by the present method described above.

As described above, this extract and the compounds contained therein (hereinafter also referred to as "this extract, etc.") have an intestinal bacterium-regulating action, an intestinal barrier function-enhancing action, an anti-obesity action, an anti-diabetic action, and an NLRP3 inflamma A composition containing the present extract, etc. (hereinafter referred to as "the present composition"), which exhibits a regulating effect on the activity of the soma, is a composition for regulating intestinal bacteria, a composition for enhancing intestinal barrier function, an anti-obesity composition, It can be used as an antidiabetic composition and a composition for controlling NLRP3 inflammasome activity.

In addition, the present composition is not limited in its form as long as it can exhibit each of the above actions, and can take the form of, for example, a pharmaceutical composition and a composition for food and drink.

When the present composition is a pharmaceutical composition, the pharmaceutical is preferably a pharmaceutical for humans or a quasi-drug, but may be a pharmaceutical for animals.

In addition, when the composition is a pharmaceutical composition, its administration method and its form are not limited as long as it can exhibit its effect, and tablets, capsules, powders, granules containing the above extract etc. as an active ingredient It may be in the form of internal medicines such as preparations, solutions, suspensions and syrups, external medicines such as poultices and ointments, and injections such as liquids. That is, the present composition can be administered orally, injected, sublingually, cutaneously, and the like.

In addition, when the present composition is a pharmaceutical composition, excipients, stabilizers, Preservatives, buffering agents, suspending agents, emulsifiers, coloring agents, flavoring agents, viscosity modifiers, binders, tensioning agents, solubilizers, preservatives, antioxidants, coagulants, coating agents, etc. Ingredients other than active ingredients, so-called pharmaceutical additives, can be included. Of course, solvents such as water and physiological saline, vitamins, minerals, fragrances, etc., can also be added to the extent that the quality and effect of the present composition are not changed.

Moreover, when this composition is a composition for eating and drinking, it includes so-called foods and beverages. Food and drink are preferably edible for humans, but may be edible for animals.

In addition, when the present composition is a composition for food and drink, its form is not particularly limited, and may be, for example, a food, a beverage, a seasoning, a supplement, a food with function claims, or a food for specified health uses. Examples of foods include sweets such as candy, candies, gums, and chocolates, and examples of beverages include soft drinks, carbonated drinks, nutritional drinks, fruit drinks, lactic acid drinks, and the like. . In the case of functional foods, supplements, and foods for specified health uses, tablets, capsules, drink preparations, and the like can be in various forms like the above pharmaceuticals.

Also, when the present composition is a food composition, as in the case of the pharmaceutical composition, there is no particular limitation as long as it can exhibit various effects, and excipients, stabilizers, preservatives, buffers , Suspending agents, emulsifiers, coloring agents, flavoring agents, viscosity modifiers, binders, tensioning agents, solubilizers, preservatives, antioxidants, coagulants, coating agents, etc. , so-called food additives. Of course, solvents such as water and physiological saline, vitamins, minerals, fragrances, etc., can also be added to the extent that the quality and effect of the present composition are not changed.

(Composition for adjusting intestinal bacteria)
As described above, this extract can be used as a composition for regulating intestinal bacteria. That is, the composition for adjusting intestinal bacteria according to the present embodiment contains licorice extract as an active ingredient, and the extract contains isoliquiritigenin (ILG) as an essential ingredient, liquiritin (L), Contains at least one of isoliquiritin (IL) and liquiritigenin (LG). In this case, it is preferable to contain at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG).

In addition, the compounds contained in the present extract can be separated from the extract, exhibit their effects as synthesized or isolated compounds, and can be used as a composition for regulating intestinal bacteria. That is, the composition for adjusting intestinal bacteria according to the present embodiment contains at least one of liquiritin (L) and isoliquiritigenin (ILG) as an essential component, and further contains isoliquiritin (IL) and liquiritigenin (LG). It contains at least one of them as an active ingredient. In this case, it is more preferable to contain at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG) as active ingredients.

In the present embodiment, the composition for adjusting intestinal bacteria is a composition capable of adjusting the abundance ratio of intestinal bacteria. You can adjust the ratio. Types of intestinal bacteria include, but are not limited to, A. muchiniphila and P. goldsteinii has been confirmed to be effective, and at least one of these targets is preferable.

(Composition for enhancing intestinal barrier function)
In addition, the compounds contained in the present extract can be separated from the extract, exhibit their effects as synthesized or isolated compounds, and can be used as a composition for enhancing intestinal barrier function. That is, the composition for adjusting intestinal bacteria according to the present embodiment contains isoliquiritigenin (ILG) as an essential active ingredient. Furthermore, it preferably contains at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG) as an active ingredient. In this case, more preferably, at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG) are contained as active ingredients.

(Antiobesity composition)
As described above, this extract can be used as an anti-obesity composition. That is, the anti-obesity composition according to the present embodiment contains licorice extract as an active ingredient, and the extract contains isoliquiritigenin (ILG) as an essential ingredient, liquiritin (L), isoliquiritin ( IL) and at least one of Liquiritigenin (LG). In this case, it is preferable to contain at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG).

(Antidiabetic composition)
As described above, this extract can be used as an anti-diabetic composition. That is, the anti-diabetic composition according to the present embodiment contains licorice extract as an active ingredient, and the extract contains isoliquiritigenin (ILG) as an essential ingredient, liquiritin (L), isoliquiritin ( IL) and at least one of Liquiritigenin (LG). In this case, it is preferable to contain at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG).

(Composition for controlling NLRP3 inflammasome activity)
In addition, the licorice extract according to the present embodiment can also be used as a composition for controlling NLRP3 inflammasome activity. More specifically, the composition for controlling NLRP3 inflammasome activity according to the present embodiment contains licorice extract as an active ingredient, and the extract contains isoliquiritigenin (ILG) as an essential ingredient. Furthermore, it contains as an active ingredient a licorice extract containing at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). In this case, it is preferable to contain at least two of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG).

As described above, this extract exhibits intestinal bacteria regulating action, intestinal barrier function enhancing action, anti-obesity action, anti-diabetic action, and NLRP3 inflammasome activity regulating action, and provides new effects of licorice extract. It can exert its medicinal effect.

Here, the licorice extract according to the above embodiment was actually produced and its various effects were confirmed. A specific description will be given below.

125 L of ammonia water was added to 30 kg of licorice for extraction, and sulfuric acid was added thereto to obtain 3 kg of precipitate. Next, 80% by weight ethanol and activated carbon were added to 3 kg of the precipitate, followed by filtration. Ammonia water was added to the filtrate, the filtrate was filtered again, and the filtrate was concentrated and dried to obtain 1127 g of an extract.

Next, from 1127 g of the extract obtained above, using synthetic adsorption resins XAD-7HP and HP-20, glycyrrhizic acid and its analogue compounds are removed, concentrated and dried to obtain glycyrrhizic acid-removed licorice. 46.8 g of extract were obtained. The licorice extract was quantitatively analyzed by HPLC, and the results were as follows.

(Administration to mice)
Six 10-week-old male experimental mice (C57BL/6) were grouped, and each group was fed (1) normal diet (ND), (2) high-fat diet (HFD), and (3). High-fat diet (HFD), water for (1) and (2), licorice extract (GE-1) for (3), orally administered for 12 weeks using a gastric tube (5 times a week) The body weight, food intake, liver weight, epididymal cell tissue weight, insulin resistance test, and plasma insulin concentration were examined for each of them. The dose of the licorice extract was set to 4 mg/animal per day.

(Weight and anti-obesity action)
FIG. 1 shows the results of body weight changes for the above mouse groups. The horizontal axis indicates the elapsed time (weeks) from the start of administration, and the vertical axis indicates body weight (g). In addition, FIG. 2 shows the food intake per day in each mouse group.

As shown in these figures, even with the same amount of food intake, the body weight increased over time in the high-fat diet (HFD) feeding group, while the licorice extract (GE-1) was administered orally. It was confirmed that administration could suppress the weight gain. That is, the anti-obesity action of the licorice extract (GE-1) could be confirmed.

In addition, the liver weight and epididymal fat composition weight of mice at 12 weeks after the start of feeding were also confirmed in the same manner. The results are shown in FIGS. 3 and 4, respectively.

In this result also, when only the high-fat diet was given, a significant increase in liver weight and epididymal fat composition weight was confirmed, whereas in the mouse group orally administered with licorice extract, these increases were suppressed. I confirmed that it is possible.

As described above, in this example, it was confirmed that the licorice extract having the composition of this example has an effect as an anti-obesity composition.

(Anti-diabetic action)
Next, FIG. 5 shows the results of an insulin resistance test. In this figure, 0.75 IU/kg of insulin was intraperitoneally administered to each mouse group at 11 weeks after the start of feeding, after fasting for 2 hours, and after administration, 30 minutes, 60 minutes, and 120 minutes. It is a measurement of blood sugar level.

According to the results shown in this figure, insulin-induced blood glucose level reduction by insulin was slight, whereas blood glucose level reduction was large in mice orally administered with licorice extract. That is, insulin resistance induced by high-fat diet was ameliorated by oral administration of licorice extract.

Further, FIG. 6 shows the plasma insulin concentration results 12 weeks after the start of the experiment. Plasma insulin concentrations at the time of dissection were measured by ELISA.

The results in this figure show that the plasma insulin concentration is higher in the group of mice fed the high-fat diet than in the group of mice fed the normal diet, but the increase in insulin concentration is suppressed in the mice group orally administered with licorice extract. confirmed that it is possible. It was confirmed that by using the licorice extract according to the present example as an active ingredient, it was possible to exhibit an effect as an anti-diabetic composition.

(Composition for adjusting intestinal bacteria)
Next, in order to confirm the effect of isoliquilitigenin (ILG) according to this example, the same mice as described above were used to confirm intestinal bacteria. Intestinal bacteria were confirmed by 16S-rRNA analysis for mouse intestinal bacteria. In this example, (1) normal diet (ND), (2) normal diet + isolitigenin (ND + ILG), (3) high-fat diet (HFD), (4) high-fat diet + isolitic Validation was performed in groups of mice fed one of four diets of iritigenin (HFD+ILG). This analysis result is shown in FIG. This figure shows the analysis results at the Phylum (phylum) level for each mouse group 1 to 4 weeks after the start of feeding. Here, ILG was administered by mixing 0.5% with high-fat diet.

According to the analysis results shown in this figure, more than 70% of Bacteroidetes and Firmicutes are present in the intestine. It was confirmed that this value can be increased by feeding isoliquiritigenin (ILG) on either a diet (ND) or a high-fat diet (HFD).

Next, in the above, analysis was performed at the Species level, proceeding further from the Phylum (phylum). This result is shown in FIG.

According to this figure, it was confirmed that although Akkermansia muciniphila of Verrucomicrobia phylum decreased by starting a high-fat diet, it turned to increase by mixing isoliquiritigenin (ILG). In addition, it was found that Parabacteroides goldsteinii of Bacteroidetes was the bacterium whose abundance ratio increased most by feeding isoliquiritigenin (ILG).

From the above results, isoliquiritigenin (ILG) is a composition for adjusting the intestinal bacteria that can regulate the abundance ratio of intestinal bacteria, and more specifically, the abundance ratio of Akkermansia muciniphila and Parabacteroides goldsteinii. It was confirmed that the

In addition, as a detailed analysis, the expression of intestinal barrier function molecules was analyzed. In this analysis, in the same manner as the 16S-rRNA analysis described above, groups of fed mice were used, and 5 mice in each group 6 weeks after the start of feeding were analyzed by Western Blot of Claudin-1 in large intestine tissue. These are the results of expression analysis. The results are shown in FIGS. 9 and 10. FIG. FIG. 9 shows the results of Western Blot, and FIG. 10 quantifies the expression level (ratio to β-actin).

According to the results of this figure, the expression level of Claudin-1, which is a barrier function molecule, is decreased by feeding a high-fat diet (HFD), while administration of isoliquiritigenin (ILG) reduces the expression of Claudin-1. It was confirmed that the expression level of -1 was increased, that is, the intestinal barrier function was restored.

On the other hand, next, the effect of liquiritin (L) in regulating intestinal bacteria was compared. Specifically, (1) normal diet (ND), (2) high-fat diet (HFD), (3) high-fat diet + isoliquiritigenin (HFD + ILG), (4) high-fat diet + liquiritin (HFD + L) ), 2 weeks after the start of feeding, the feces of 4 mice in each mouse group were analyzed for bacterial flora, especially for Akkermansia muciniphila and Parabacteroides goldsteinii. The results are shown in FIG.

According to the above results, feeding a high-fat diet (HFD) decreases the abundance of both Akkermansia muciniphila and Parabacteroides goldsteinii. However, it was confirmed that by feeding isoliquiritigenin (ILG) or liquiritin (L), the decrease in the abundance ratio of both bacteria due to HFD was restored and the abundance ratio was greatly increased.

Regarding whether or not the same effect can be obtained with regard to licorice extract, (1) normal diet (ND), (2) high-fat diet (HFD), (3) high-fat diet + 4 mg of the licorice extract / animal (HFD + GE-1 4 mg), (4) high fat diet + the above licorice extract 10 mg / animal (HFD + GE-1 10 mg). It was confirmed whether there was an increase in the abundance ratio of muciniphila. This result is shown in FIG. These results are the analysis results of feces 2 weeks after feeding, and the licorice extract was confirmed separately for 4 mg/animal and 10 mg/animal.

As a result, it was confirmed that there was an increase in the abundance of Akkermansia muciniphila in mice orally administered with the licorice extract (GE-1).

In addition, how much effect on Akkermansia muciniphila and Parabacteroides goldsteinii by transplanting the feces of mice in the high-fat diet + isoliquiritigenin (HFD + ILG) group of mice to mice in the high-fat diet (HFD) group of mice? I checked if there is any. Specifically, a cocktail of antibiotics was administered to a mouse group 4 weeks after the start of feeding with a high-fat diet (HFD) for 4 days, and a recovery period of 3 days was provided. transplanted. Then, after administering a high-fat diet (HFD) for 8 weeks, the abundance ratio of Akkermansia muciniphila and Parabacteroides goldsteinii in the intestine was analyzed. The results are shown in FIG.

As a result, it was confirmed that the abundance ratio of Akkermansia muciniphila and Parabacteroides goldsteinii can be increased by transplanting the feces of the mouse group to which isoliquiritigenin was administered.

Furthermore, as confirmation of this, changes in visceral fat weight and body weight were also confirmed. FIG. 14 shows the visceral fat of the mice 8 weeks after transplantation, and FIG. 15 shows changes in body weight.

From the above results, it was confirmed that both visceral fat weight and body weight were reduced by transplanting feces.

(NLRP3 inflammasome activity control action)
Here, we also confirmed the NLRP3 inflammasome activity control action of the licorice extract. Here, the effect in vitro was confirmed prior to administration to mice. Specifically, the macrophage-like cell line J774. After priming the A1 cell line with LPS 1 μg/mL for 3 hours, washing the cells, adding licorice extract and reacting for 30 minutes, adding Nigericin 10 μM, IL-1β produced in the culture supernatant was measured by the ELISA method. The results are shown in FIG.

In addition, the NLRP3 inflammasome activity-regulating action of the licorice extract was also confirmed in vivo. Specifically, 10-week-old male mice were fed (1) normal diet (ND), (2) high-fat diet (HFD), and (3) high-fat diet (HFD), ( 1) For (2), water was orally administered, and for (3), licorice extract (GE-1) was orally administered for 12 weeks using a gastric tube (5 times a week). Each epididymal fat was isolated, and the minced epididymal fat was cultured ex vivo to measure the effect on IL-1β production by ELISA. It has been reported that NLRP3 inflammasome is activated by palmitic acid and glucose in visceral adipose tissue in diabetes. The results are shown in FIG.

Summary As described above, according to this example, it was possible to confirm the intestinal bacterium-regulating action, intestinal barrier function-enhancing action, anti-obesity action, anti-diabetic action, and NLRP3 inflammasome activity-regulating action of this extract and this compound. rice field.

The present invention provides a composition containing a licorice extract, an intestinal bacterium-regulating composition, an intestinal barrier function-enhancing composition, an anti-obesity composition, an anti-diabetic composition, and an NLRP3 inflammasome containing the same. It has industrial applicability as a composition for controlling activity.

Claims (21)

A method for producing a licorice extract, comprising a step of removing glycyrrhizic acid. A method for producing a licorice extract, comprising a step of removing the glycyrrhizic acid so that the concentration of the glycyrrhizic acid is 1% by weight or less. A method for producing a licorice extract, comprising a step of removing the glycyrrhizic acid so that the glycyrrhizic acid concentration is 0.5% by weight or less. containing isoliquiritigenin (ILG), and
A licorice extract containing at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). 5. The liquorice extract according to claim 4, which contains at least two of the liquiritin (L), the isoliquiritin (IL) and the liquiritigenin (LG). 0.1% to 30% by weight of the isoliquiritigenin (ILG), 10% by weight or less of the liquiritin, 15% by weight or less of the isoliquiritin (IL), and 30% by weight of the liquiritigenin (LG) % or less of the licorice extract according to claim 5. A licorice extract produced through a process of removing glycyrrhizic acid, containing isoliquiritigenin (ILG), and at least one selected from liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG) Glycyrrhiza extract containing seeds. 0.1% to 30% by weight of the isoliquiritigenin (ILG), 10% by weight or less of the liquiritin (L), 15% by weight or less of the isoliquiritin (IL), and the liquiritigenin (LG) The licorice extract according to claim 7, containing 30% by weight or less. A composition for enhancing intestinal barrier function, containing isoliquiritigenin (ILG) as an active ingredient. A composition for regulating intestinal bacteria, containing at least one of liquitin (L) and isoliquiritigenin (ILG) as an active ingredient. 11. The composition for regulating intestinal bacteria according to claim 10, further comprising at least one of isoliquiritin (IL) and liquiritigenin (LG) as an active ingredient. A composition for regulating intestinal bacteria, containing a licorice extract as an active ingredient. The composition for regulating intestinal bacteria according to any one of claims 10 to 12, which is a pharmaceutical composition or a composition for food and drink. Intestinal bacteria A. muchiniphila and P. 13. The composition for regulating intestinal bacteria according to any one of claims 10 to 12, wherein the composition is at least one of G. goldsteinii. A. above. muchiniphila and the aforementioned P. 15. The composition for regulating intestinal bacteria according to claim 14, wherein the abundance ratio of at least one of goldsteinii is increased. containing isoliquiritigenin (ILG), and
An anti-obesity composition containing, as an active ingredient, a licorice extract containing at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). 17. The anti-obesity composition according to claim 16, which is a pharmaceutical composition or an eating and drinking composition. containing isoliquiritigenin (ILG), and
An antidiabetic composition containing, as an active ingredient, a licorice extract containing at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). 19. The anti-diabetic composition according to claim 18, which is a pharmaceutical composition or a food or drink composition. containing isoliquiritigenin (ILG), and
A composition for controlling NLRP3 inflammasome activity, containing as an active ingredient a licorice extract containing at least one of liquiritin (L), isoliquiritin (IL) and liquiritigenin (LG). 21. The composition for controlling NLRP3 inflammasome activity according to claim 20, which is a pharmaceutical composition or a food or drink composition.

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