RU2317077C2 - Receptor ligand activated by peroxisome proliferator and method for its preparing - Google Patents

Abstract

FIELD: organic chemistry, medicine, pharmacy.

SUBSTANCE: invention relates to drugs and in particular provides a receptor ligand activated by peroxisome proliferator γ (PPAR γ), and a composition comprising the indicated ligand used for aim in improving insulin-resistance state or for prevention and/or improving the insulin-resistance syndrome state. Invention relates to a receptor ligand activated by peroxisome proliferator that comprises prenylflavonoid, chalcone derivative, flavonol derivative and their salt, glycoside and/or esterified form. Also, invention relates to composition comprising abovementioned ligand, a vegetable extract containing abovementioned ligand, and to a novel compound (dehydroglyasperin D) represented by the following formula:

.

EFFECT: improved preparing method, valuable medicinal properties of ligand.

33 cl, 26 tbl, 13 ex

Description

Field of Invention

The present invention relates to a ligand of a receptor activated by a peroxisome proliferator, a method for preparing it and a composition for improving the state of insulin resistance or for preventing and / or improving the state of insulin resistance syndrome (type 2 diabetes, hyperinsulinemia, dyslipidemia, obesity, hypertension and a disease characterized by arteriosclerosis) peroxisome proliferator activated receptor ligand as an active ingredient.

Prior art

The peroxisome proliferator activated receptor (PPAR) is a ligand-dependent transcription regulator that belongs to the intranuclear receptor family. This family is defined as a family of transcription regulators involved in the regulation of the expression of a group of genes that provides lipid metabolism. In mammals, 3 subtypes are known, namely PPARα, PPARδ (PPARβ, NUC-1, FAAR) and PPARγ, with PPARα expressed mainly in the liver, and PPARδ expressed everywhere. There are 2 isoforms for PPAR γ, namely PPAR γ1 and PPARγ2, moreover, PPARγ1 is expressed not only in adipose tissue, but also in the organs of the immune system, adrenal glands and small intestine. PPARγ2 is expressed specifically in adipose tissue and is the main regulator that regulates the differentiation and maturation of adipocytes (Teruo Kawada, "Igaku No Ayumi", 184, 519-523, 1998).

Synthetic compounds including thiazolidine derivatives such as troglitazone, pioglitazone and rosiglitazone are known as PPARγ ligands. As PPARγ ligands, natural compounds are known, including arachidonic acid metabolites, such as 15-deoxy-Δ ^12,14- prostaglandin J ₂ and Δ ^12,14- prostaglandin J ₂ , unsaturated fatty acids such as ω-3 polyvalent unsaturated fatty acid, α-linolenic acid, eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) and eicosanoids such as 9-hydroxyoctadecadienoic acid and 13-hydroxyoctadecadienoic acid (J. Auwerx, Diabetologia, 42 , 1033-1049) , 1999). Japanese publication Kokai 2000-355538 describes a C _10-26 conjugated unsaturated fatty acid having a conjugated triene structure or conjugated tetraenic structure. With regard to flavonoids, it is reported that krizin and apigenin, as derivatives of flavon, and kempferol, as a derivative of flavonol, are PPARγ ligands. (YC Liang, et al., FEBS Letters, 496 , 12-18, 2001). Flavonoid is a substance widely distributed in plants, and it is known to have antioxidant properties. However, it is not known whether the prenyl flavonoid contained only in specific plants is a PPARγ ligand.

The agonistic activity of thiazolidine derivatives as a PPARγ ligand correlates with its hypoglycemic effect, therefore, such derivatives attract attention in relation to their state-improving effect on insulin resistance and are being developed as agents that improve the state of insulin resistance for use in type 2 diabetes mellitus (insulin- independent diabetes mellitus: NIDDM). That is, a thiazolidine derivative, which is a PPAR ligand, activates PPAR, causing an increase in the number of normally functioning small adipocytes differentiated from preadipocytes. Accordingly, the number of hypertrophied adipocytes in which TNF-α (TNFα) and free fatty acids are excessively produced and secreted, contributing to insulin resistance, decreases due to apoptosis. As a result, the state of insulin resistance is improved (A. Okuno, et al., Journal of Clinical Investigation, 101, 1354-1361, 1998). The PPARγ ligand improves insulin resistance and is thus also effective in preventing and / or improving insulin resistance syndrome, including not only type 2 diabetes mellitus, but also hyperinsulinemia, dyslipidemia, obesity (in particular visceral obesity), hypertension and arteriosclerosis (RA DeFzo & E. Ferrannini, Diabetes Care, 14, 173-194, 1991). Pathological conditions of the same kind as insulin resistance syndrome include Syndrome X (GM Reaven, Diabetes, 37 , 1595-1607, 1988), the deadly quartet (NM Kaplan, Archives of Internal Medicine, 149 , 1514-1520, 1989) and the syndrome visceral obesity (Y. Matsuzawa, Diabetes / Metabolism Reviews, 13 , 3-13, 1997).

The PPARγ ligand is also effective in preventing and / or improving inflammatory and cancerous conditions due to its ability to inhibit the production of inflammatory cytokines (C. Jiang, et al., Nature, 391 , 82-86, 1998) and induce apoptosis, thus inhibiting the growth of cancer cells (Y. Tsubouchi, et al., Biochemical and Biophysical Research Communications, 270 , 400-405, 2000).

SUMMARY OF THE INVENTION

In light of the above, the PPARγ ligand has an effect on improving the state of insulin resistance and preventing and / or improving the state of insulin resistance syndrome, for example type 2 diabetes mellitus, hyperinsulinemia, dyslipidemia, obesity (in particular visceral obesity), hypertension and arteriosclerosis. An object of the present invention is to provide a PPAR ligand found in natural materials, a method for more easily and efficiently producing the like, and a composition for improving insulin resistance or for preventing and / or improving insulin resistance syndrome containing the ligand as an active ingredient.

The inventors of the present invention searched for a substance having PPARγ ligand activity in natural materials and as a result found that an extract of a plant, such as licorice, had PPARγ ligand activity. The authors of the present invention intensively studied its active ingredient and as a result found that a specific component of the extract has PPARγ ligand activity. In addition, the inventors of the present invention have found that an organic solvent, preferably a fatty acid ester or a water-soluble organic solvent, especially a water-soluble organic solvent, can be used to efficiently extract such a component from licorice to carry out the present invention.

That is, the first aspect of the present invention relates to a ligand of a peroxisome proliferator-activated receptor ligand, which includes a prenyl flavonoid, a chalcone derivative other than prenyl flavonoids, a flavonol derivative other than prenyl flavonoids, and a salt thereof, a glycoside and / or an esterified pharmaceutical form suitable for or a food or drink.

A second aspect of the present invention relates to a plant extract, which comprises a peroxisome proliferator activated receptor ligand comprising a prenyl flavonoid, a chalcone derivative other than prenyl flavonoids, a flavonol derivative other than prenyl flavonoids, and a salt thereof, a glycoside and / or an esterified pharmaceutical form suitable for drug or food product or drink.

A third aspect of the present invention relates to a composition for improving insulin resistance or for preventing and / or improving insulin resistance syndrome, which includes, as an active ingredient, a prenyl flavonoid having the ability to bind to the ligand-binding portion of a peroxisome proliferator proliferator derivative, a chalcone derivative other than from prenyl flavonoids, a derivative of flavonol other than prenyl flavonoids, and their salt, glycoside and / or esterified form, when acceptable as a pharmaceutical or food product or drink.

A fourth aspect of the present invention relates to a composition for the prevention and / or improvement of inflammatory and cancerous processes, which comprises as an active ingredient a prenyl flavonoid having the ability to bind to a ligand-binding site of a peroxis proliferator-activated receptor, a chalcone derivative other than prenyl flavonoids, a flavonol derivative other than prenylflavonoids and their salt, glycoside and / or esterified form acceptable as a pharmaceutical preparation or a food or drink.

A fifth aspect of the present invention relates to a method for producing an extract described above, which comprises extracting an extract from licorice having a ratio of bark region (shell) to a total surface area of 30% or more.

A sixth aspect of the present invention relates to a method for producing the extract described above, wherein extraction from licorice is carried out using a fatty acid ester or a water-soluble organic solvent.

A seventh aspect of the present invention relates to a method for producing an extract described above, which comprises extracting from licorice using an organic solvent having a water content of 30% by volume or less.

DETAILED DESCRIPTION OF THE INVENTION

Embodiments of the present invention are described in detail below.

A PPAR ligand (preferably a PPARγ ligand) is a compound having a PPAR ligand activity (preferably a PPARγ ligand activity), i.e., a compound having the ability to bind to a PPAR ligand binding site (preferably PPARγ). The activity of the PPARγ ligand can be measured, for example, by reporter analysis, where the binding of a compound to a fusion protein of the PPARγ and GAL4 ligand-binding region is evaluated in luciferase expression units (BM Forman, et al., Cell, 83 , 803-812, 1995), competitive binding analysis using a protein having a PPARγ ligand binding site (SA Kliewer, et al., Cell, 83 , 813-819, 1995), etc. Typically, the activity of a sample in these analyzes is compared to that of a control solvent, and a sample having a higher activity than that of a control solvent, as well as having indicators that the activity is dose-dependent, was regarded as having "PPARγ ligand activity".

The PPAR ligand, preferably the PPARγ ligand, of the present invention is at least one compound selected from the group consisting of prenyl flavonoid, a chalcone derivative other than prenyl flavonoids, a flavonol derivative other than prenyl flavonoids and their salt, glycoside and / or esterified a form acceptable as a pharmaceutical preparation or food product or beverage.

A salt acceptable as a pharmaceutical preparation or food or drink includes, but is not limited to, for example, sodium salts, potassium salts, magnesium salts, calcium salts, etc.

A glycoside suitable as a pharmaceutical preparation or food product or beverage includes, but is not limited to, for example, glycosides of monosaccharides such as glucose and galactose, glycosides of disaccharides or oligosaccharides, etc.

The esterified substances acceptable as a pharmaceutical preparation or food product or drink include, but are not limited to, for example, acetates, propionates, phosphates, sulfates, etc.

The composition of the present invention, comprising at least one species selected from these compounds as an active ingredient, improves the state of insulin resistance and, therefore, is useful as a means to prevent and / or improve the state of insulin resistance syndrome, including diabetes mellitus.

The prenyl flavonoid, which is the PPAR ligand of the present invention, is not particularly limited, but is preferably at least one compound selected from the group consisting of 3-arylcoumarin derivatives, isoflav-3-ene derivatives, isoflavan derivatives, isoflavanone derivatives, derivatives flavonol, flavanone derivatives, chalcone derivatives and dibenzoylmethane derivatives.

The prenyl flavonoid referenced herein is defined as a compound having at least one of the following (A) and (B) as the side chain of a flavonoid: (A) a prenyl group, or (B) a structure having 6- a member cycle in which the prenyl group is bonded to a neighboring hydroxyl group to form [-CH = CHC (CH ₃ ) ₂ O-]. The term "flavonoid" is a generic term referring to a group of substances having 2 phenyl groups bonded to each other via 3 carbon atoms, and examples thereof include flavon, flavonol, flavanone, flavanonol, isoflavone, isoflavonol, isoflavanone, isoflavan, isoflavone 3-ene, 3-arylcoumarin, chalcone, dihydrochalcone, dibenzoylmethane, cumestan, pterocarpan, catechin, anthocyanidin, etc.

Derivatives of 3-arylcoumarin, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the general formula (1). And, for example, the compounds shown in Table 1 may be mentioned, such as glycicumarin, glycyrin, gancaonin W, gliasperin L, canzonol W, etc. Derivatives thereof, such as salts, glycosides, and esterified substances may also be mentioned. Among them, glycicoumarin and glycyrin are more preferred.

In the formula, at least one of R1-R7 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH═CHC (CH ₃ ) ₂ O— and the other R groups are H , OH or OCH ₃ .

Table 1 R1 R2 R3 R4 R5 R6 R7 3-arylcoumarin (s) Och ₃ prenil IT N IT N IT glycicumarin Och ₃ prenil Och ₃ N IT N IT glycyrin Och ₃ N IT N IT prenil IT gankaonin W Och ₃ N IT N IT -CH = CHC (CH ₃ ) ₂ O- gliasperin L N N -OC (CH ₃ ) ₂ CH = CH- IT N IT canzonol W N N -OC (CH ₃ ) ₂ CH = CH- IT N Och ₃ RL-P N N IT N -OC (CH ₃ ) ₂ CH = CH- IT RL-U

Derivatives of isoflav-3-ene, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (2). For example, the compounds shown in table 2, such as dehydrogliasperin C, dehydrogliasperin D, glabrene and RL-S. Derivatives thereof, such as salts, glycosides and esterified substances, may also be mentioned. Among them, dehydrogliasperin C, dehydrogliasperin D and glabrene are preferred. Dehydrogliasperin D is a new compound first discovered in the present invention.

In the formula, at least one of R1-R7 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

table 2 R1 R2 R3 R4 R5 R6 R7 Isoflav-3-en (s) Och ₃ prenil IT N IT N IT dehydrogliasperin C Och ₃ prenil Och ₃ N IT N IT dehydrogliasperin D N N IT N -OC (CH ₃ ) ₂ CH = CH- IT glazed N N -OC (CH ₃ ) ₂ CH = CH- IT N IT RL-S

Derivatives of isoflavan, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (3). For example, the compounds shown in table 3, such as gliasperin C, gliasperin D, gliasperin I, licoricidin and licorizoflavan A. Their derivatives, such as salts, glycosides and esterified substances may also be mentioned. Among them, gliasperin D, glabridin, gisaglabridin B, 4'-O-methylglabridine and 3'-hydroxy-4'-O-methylglabridine are more preferred.

In the formula, at least one of R1-R7 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

Table 3 R1 R2 R3 R4 R5 R6 R7 Isoflavan (s) Och ₃ prenil IT N IT N IT gliasperin C Och ₃ prenil Och ₃ N IT N IT gliasperin D Och ₃ prenil IT N Och ₃ N IT gliasperin I Och ₃ prenil IT N IT prenil IT licoricidin Och ₃ prenil Och ₃ N IT prenil IT licorizoflavan A Och ₃ prenil Och ₃ N -OC (CH ₃ ) ₂ CH = CH- IT canzonol I N prenil IT N -OC (CH ₃ ) ₂ CH = CH- IT gliinflanin I N -CH = CHC (CH ₃ ) ₂ O- N IT N Och ₃ gankaonin x Och ₃ -CH = CHC (CH ₃ ) ₂ O- N IT prenil Och ₃ canzonol N Och ₃ -CH = CHC (CH ₃ ) ₂ O- N -OC (CH ₃ ) ₂ CH = CH- Och ₃ cansonol J N -CH = CHC (CH ₃ ) ₂ O- N -OC (CH ₃ ) ₂ CH = CH- N Glyinflanin J N N IT N IT prenil Och ₃ gankaonin Z Och ₃ N IT N IT prenil Och ₃ canzonol R N N IT prenil IT prenil IT canzonol X N N -OC (CH ₃ ) ₂ CH = CH- IT N IT glabridin N N -OC (CH ₃ ) ₂ CH = CH- IT N Och ₃ 4'-O-methylglabridine N N -OC (CH ₃ ) ₂ CH = CH- IT IT Och ₃ 3'-hydroxy-4'-O-methylglabridine N N -OC (CH ₃ ) ₂ CH = CH- IT Och ₃ IT 3'-O-methylglabridine N N -OC (CH ₃ ) ₂ CH = CH- Och ₃ IT Och ₃ gliasperin H N N -OC (CH ₃ ) ₂ CH = CH- IT prenil IT gispaglabridin A N N -OC (CH ₃ ) ₂ CH = CH- IT -CH = CHC (CH ₃ ) ₂ O- gispaglabridin B N N -OC (CH ₃ ) ₂ CH = CH- Och ₃ -CH = CHC (CH ₃ ) ₂ O- methylgispaglabridin B N N -OC (CH ₃ ) ₂ CH = CH- -OC (CH ₃ ) ₂ CH = CH- N Glyinflanin K N N IT N -OC (CH ₃ ) ₂ CH = CH- Och ₃ gankaonin Y N N IT N IT -CH = CHC (CH ₃ ) ₂ O- phaseollinisoflavan N N IT prenil IT -CH = CHC (CH ₃ ) ₂ O- 8-prenyl-phaseollinisoflavan

Derivatives of isoflavanone, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (4). For example, the compounds shown in table 4, such as gliasperin B, gliasperin K, canzonol G, 3'-prenyl-kieviton and gliasperin F. Also derivatives thereof such as salts, glycosides and esterified substances may be mentioned. Among them, gliasperin B and glycyrrhizoflavanone are more preferred.

In the formula, at least one of R1-R8 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH═CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

Table 4 R1 R2 R3 R4 R5 R6 R7 R8 Isoflavonone (s) IT prenil Och ₃ N IT N IT N gliasperin b IT prenil Och ₃ N IT N Och ₃ N gliasperin K IT prenil Och ₃ N IT prenil IT N canzonol G IT N IT prenil IT prenil IT N 3'-prenylkieviton (3 ', 8-diprenyl-dalbergioidine) IT N IT N -OC (CH ₃ ) ₂ CH = CH- IT N gliasperin F IT N IT prenil -OC (CH ₃ ) ₂ CH = CH- IT N gliasperin J Och ₃ N IT N N IT -CH = CHC (CH ₃ ) ₂ O- glycyrrhizoflavanone Och ₃ N IT N IT -CH = CHC (CH ₃ ) ₂ O- N gliasperin M IT N IT N IT -CH = CHC (CH ₃ ) ₂ O- N lycoisoflavanone N N -OC (CH ₃ ) ₂ CH = CH- IT N IT N RL-Q N N -OC (CH _{3) 2} CH = CH- IT N Och ₃ N RL-R IT prenil IT N IT prenil IT N Untitled

Derivatives of isoflavone, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (5). For example, the compounds shown in table 5, such as viteon, gankonin A, gankonin B, gankonin G and gankonin N. Their derivatives, such as salts, glycosides and esterified substances can also be mentioned. Among them, glyuralline B, lupivitone, semilicoisoflavone B and glycyrrhizoflavone are more preferred.

In the formula, at least one of R1-R9 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH═CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

Table 5 R1 R2 R3 R4 R5 R6 R7 R8 R9 Isoflavonone (s) IT prenil IT N N N IT N N viteon IT prenil IT N N N Och ₃ N N gankaonin A IT prenil IT N N IT Och ₃ N N gankaonin B IT prenil Och ₃ N N N IT N N gankaonin G IT prenil IT N IT N Och ₃ N N gankaonin N IT prenil Och ₃ N IT N IT N N 7-O-methylluteon IT prenil Och ₃ N IT prenil IT N N canzonol K IT prenil IT prenil -OS (CH ₃ ) ₂ CH = CH- IT N N canzonol L IT prenil IT N N IT IT prenil N isoangustone A IT prenil IT N N IT -OS (CH ₃ ) ₂ CH = CH- N gankaonin H IT N IT prenil N N IT N N lupiviteon IT N IT prenil N IT IT N N gankaonin L IT N IT prenil N N Och ₃ N N gankaonin M IT N IT prenil N IT IT prenil N gliurallin B IT N IT prenil -OS (CH ₃ ) ₂ CH = CH- IT N N gliasperin N N N IT N IT prenil IT N N euricarpine A IT N IT N IT prenil IT N N lycoisoflavone A IT N IT N IT -CH = CHC (CH ₃ ) ₂ O- N N lycoisoflavone B N N IT N IT -CH = CHC (CH ₃ ) ₂ O- N N glabron IT N IT N N IT IT prenil N glycyrrizoflavone Och ₃ N IT N N IT IT prenil N glizoflavon IT N IT N N IT -OC (CH _{3) 2} CH = CH- N semilicoisoflavone B N N IT N IT N IT prenil Och ₃ glycorinone N N IT N IT N Och ₃ prenil Och ₃ licorinon

Derivatives of flavonol, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (6). For example, the compounds shown in table 6, such as lycoflavonol, gankaonin P, topazolin, gliasperin A and isolicoflavonol. Derivatives thereof, such as salts, glycosides and esterified substances, may also be mentioned. Among them, more preferred are isolicoflavonol, lycoflavonol, and topazoline.

In the formula, R1 represents OH or OCH ₃ , at least one of R2-R9 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHC (CH ₃ ) ₂ O—, and the other R groups are H, OH or OCH _3.

Table 6 R1 R2 R3 R4 R5 R6 R7 R8 R9 Isoflavonone (s) IT IT prenil IT N N N IT N lycoflavonol IT IT prenil IT N N IT IT N gankaonin P Och ₃ IT prenil IT N N N IT N topazoline IT IT prenil IT N N prenil IT N gliasperin A IT IT N IT N N prenil IT N Isolicoflavonol IT IT N IT prenil N N N N gllepidotin A IT N IT IT N prenil IT IT N neouralinol IT IT N IT N N IT IT prenil uralenol Och ₃ IT N IT N N IT IT prenil uralenol-3-O-methyl ether Och ₃ IT IT N N N IT IT prenil Uralan

Derivatives of flavanone, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (7). For example, the compounds shown in Table 7, such as 6-prenylpinocembrin, 6-prenylnaringenin, 6-prelenyriodiktiol, sinoflavanone B and paratocarpine L. Derivatives thereof, such as salts, glycosides and esterified substances may also be mentioned. Among them, glabrol is more preferred.

In the formula, at least one of R1-R8 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH═CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

Table 7 R1 R2 R3 R4 R5 R6 R7 R8 Flavanon (s) IT prenil IT N N N N N 6-prenylpinocembrin IT prenil IT N N N IT N 6-prenylnaringenin IT prenil IT N N IT IT N 6-prenyleriodikthiol IT prenil IT prenil N N N N sinoflavanone B IT prenil IT N N prenil IT N paratocarpine L N N IT prenil N N N N ovaliflavanone B N N IT prenil N N IT N isobavin IT N IT prenil N N N N glabranin IT N IT prenil N N IT N Soforaflavanone B IT N IT prenil N IT IT N 8-prenyleriodikthiol IT N IT prenil N Och ₃ IT N exiguaflavanone K N N IT prenil N prenil IT N glabrol IT N IT prenil N prenil IT N eucrestaflavanone A IT N IT prenil N IT IT prenil gankaonin E IT N -OC (CH _{3) 2} CH = CH- N N IT N citflavanone N N -OS (CH ₃ ) ₂ CH = CH- N prenil IT N shinflavanon N N -OS (CH ₃ ) ₂ CH = CH- N -CH = CHC (CH ₃ ) ₂ O- N xambiuna IT N IT N N prenil IT N lycoflavanone IT N IT N N IT IT prenil Sigmoidin B (Uralenin) IT N IT N N N -OS (CH ₃ ) ₂ CH = CH- sigmoidin C N prenil IT prenil N N IT N Untitled

Chalcone derivatives, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (8). For example, the compounds shown in table 8, such as lycochalcon C, lycochalcon D, glyinflanin G, canzonol B and canzonol C. Also derivatives thereof such as salts, glycosides and esterified substances may be mentioned. Among them, lycochalcon C. is more preferred.

In the formula, at least one of R1-R9 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH═CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

Table 8 R1 R2 R3 R4 R5 R6 R7 R8 R9 Chalcon (s) Och ₃ prenil IT N N N IT N N Lycochalcon C Och ₃ IT IT N N prenil IT N N Lycochalcon D N -CH = CHC (CH ₃ ) ₂ O- IT IT -CH = CHC (CH ₃ ) ₂ O- N N Glyinflanin G N -CH = CHC (CH ₃ ) ₂ O- N IT N IT N N canzonol B N prenil IT N N N IT prenil IT canzonol C N N IT N IT N Och ₃ prenil N abalone N prenil IT N IT prenil IT N N Untitled N IT -CH = CHC (CH ₃ ) ₂ O- IT prenil IT N N Untitled N -CH = CHC (CH ₃ ) ₂ O- IT N N IT prenil IT Untitled

Derivatives of dibenzoylmethane, which are the prenyl flavonoids used in the present invention, are not particularly limited, but their preferred examples include compounds represented by the following general formula (9). For example, the compounds shown in table 9, such as glycyrrhione A, glycyrrhione B, glycyrrhione C, glyinflanin B and glyinflanin D. Their derivatives such as salts, glycosides and esterified substances may also be mentioned. Among them, glycyrrhione A and glycyrrhione C are more preferred.

In the formula, at least one of R1-R7 is a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H OH or OCH ₃ .

Table 9 R1 R2 R3 R4 R5 R6 R7 Dibenzoylmethane (s) IT N IT prenil N prenil IT glycyrdione A (glyinflanin A) IT N IT prenil N -CH = CHC (CH ₃ ) ₂ O- glycyrdione B IT N -OC (CH _{3) 2} CH = CH- N prenil IT glycyrdione C (glyinflanin C) IT N -OC (CH _{3) 2} CH = CH- N N IT Glyinflanin B IT N -OC (CH _{3) 2} CH = CH- N -CH = CHC (CH ₃ ) ₂ O- Glyinflanin D IT N IT N N prenil IT canzonol A IT N IT prenil N N IT 5'-prenyl-lycodion

Chalcone derivatives, excluding prenyl flavonoids, used in the present invention are not particularly limited, and their preferred examples include echinatin, isoquiritigenin, lycochalcon A and lycochalcon B. Their derivatives such as salts, glycosides and esterified substances may also be mentioned.

Derivatives of flavonol, excluding the prenyl flavonoids used in the present invention, are not particularly limited, and their preferred examples include campferol 3-O-methyl ester. Derivatives thereof, such as salts, glycosides and esterified substances, may also be mentioned.

For the names of the compounds can be made to post T. Hiraga & K. Kajiyama (Meiji Pharmaceutical University Bulletin, 27, 9-57, 1997) and post T. Nomura & T. Fukai (Fortschritte der Chemie organischer Naturstoffe, 73, 1-158 , 1998).

The PPAR ligand of the present invention can be any compound obtained from natural sources, such as plants, chemically synthesized or biosynthesized in cultured cells, etc., if the compound is the aforementioned compound, but the compound obtained from natural sources that a person can eat is preferred . The method for producing such compounds is not particularly limited and the compounds can be obtained from the licorice family Leguminosae, which a person can eat, or from other plants. The other plants mentioned above are not particularly limited provided that the plants contain a PPAR ligand, preferably a PPARγ ligand, and in addition to plants of the Leguminosae family, other families can be mentioned, such as the Moraceae family, Eucommiceae family, Cannabaceae family and Urticaceae family.

An example of a method for producing the PPAR ligand of the present invention from licorice is described below, but the starting material is not necessarily limited to licorice.

The licorice used may be a plant of the genus Glycyrrhiza of the Leguminosae family and examples thereof include Glycyrrhiza uralensis Fisch. et DC, G. Inflata BAT., G. Glabra L., G. Glabra L. var. glandu rifera Regel et Herder, G. echinata L., G. pallidiflora Maxim and other plants of the same genus (Leguminosae). Such plants are grown, for example, in Xin Jiang, northeast China, northwest China, Mongolia, Russia, Afghanistan, Iran, Turkey, and others. Among them, those having a higher content of the active ingredient of the PPAR ligand, such as G. uralensis (growing areas: north-eastern China, north-western China, Mongolia, Xin Jiang, etc.), G. glabra (growing areas: Russia, Afghanistan, Iran, Turkey, etc.) and G. inflata (regions cultivation: Xin Jiang et al.), and G. uralensis is particularly preferred for use.

The present invention preferably uses the root, rhizome, or stolon of licorice mentioned above, and is used in the form of a finely ground product, ground product, cut product, or periderm of the aforementioned licorice. A finely ground product refers to a powdered or nearly powdered product, and the ground product refers to the product in the form of threads, fibers or cotton wool (for example, a product having a length of about 5-10 mm). The cut product is obtained by slicing a root, rhizome or stolon of licorice (for example, thin round slices having a length of usually about 1-10 cm, preferably about 1-5 cm, more preferably about 2-4 cm and a thickness usually about 3 cm or less, preferably about 2 cm or less, more preferably about 1 cm or less), usually cylindrical or substantially cylindrical in shape. Finely ground product, ground product and cut product can be obtained using conventional equipment. The finely ground product can be obtained in the form of a powdery or almost powdery product using a Masscolloider mill, a grinding mortar, etc., and the ground product can be obtained using a grinding hammer mill or the like. The cut product can be obtained by cutting the root or other parts of the licorice into pieces having the aforementioned length using a conventional cutting machine.

The composition and content of the active ingredient of licorice varies to some extent depending on its type, cultivation area, harvest time, etc. Therefore, it is preferable to use one in which a large amount of the active ingredient is established by preliminary experiment.

Extraction methods include methods that include extraction of licorice with an organic solvent. However, the residue from aqueous extraction obtained by extraction of licorice with water, or the dry matter of the residue can also be subjected to extraction with an organic solvent.

The organic solvent used herein is not particularly limited, but is preferably a safe solvent used to create and process pharmaceuticals, foods, food additives, and the like. Examples thereof include acetone, a monohydroxy alcohol having 1-4 carbon atoms, glycerin, fatty acid esters, diethyl ether, cyclohexane, carbon dioxide, propylene glycol, etc., and two or more of these solvents may be used as a mixture. Among them, preferred solvents are fatty acid esters, such as ethyl acetate, or water-soluble organic solvents, such as a monohydroxy alcohol having 1-4 carbon atoms, acetone, propylene glycol, glycerin, etc., and particularly preferably water-soluble organic solvents.

After extraction, it is preferable to increase the content of the active ingredient in the extract by purification, such as absorption by activated carbon, resin or the like, and / or fractionation. The substance obtained by extraction can be subjected to silica gel column chromatography, ODS, ion exchange resin, etc., where the target compound can be concentrated, fractionated or isolated.

It is clear that the target compound can be obtained by other chemical synthesis.

Below, a method for extracting licorice with a fatty acid ester or a water-soluble organic solvent is described in more detail as a preferred method for producing licorice extract containing PPAR ligand in accordance with the present invention. Licorice extract isolated using a fatty acid ester or a water-soluble organic solvent tends to exhibit better PPAR ligand activity, and a water-soluble organic solvent is particularly preferred for use since the active substance, the PPAR ligand, can be more efficiently prepared.

When the target substance is extracted from the licorice with a fatty acid ester or a water-soluble organic solvent, the finely divided product, the crushed product, the cut product or the licorice periderm as such may be extracted, but as described above, the pre-extracted licorice residue obtained by preliminary extraction from licorice with another solvent (for example, water, an aqueous solution of alkali, etc.), can be extracted with a fatty acid ester or a water solution an organic solvent to remove impurities, etc.

After pre-extraction from the licorice with another solvent (water, an aqueous solution of alkali, etc.), the pre-extract can be divided into the extract and the residue of the extracted licorice by the usual separation technique (e.g. pressure filtration, vacuum filtration, filter press, centrifugation, sedimentation, etc.) to obtain the residue of previously extracted licorice. In this separation procedure, if necessary, commonly used filter aid and / or adsorbent substances, such as activated carbon and activated clay, can be used.

The pre-extracted residue thus obtained is used when extracting a fatty acid ester or a water-soluble organic solvent as such or after removing partially or completely another solvent used (water, an aqueous solution of alkali, etc.) by a conventional drying method (for example, drying in a stationary state, drying with stirring, mixed drying, fluidized drying, thermal drying, spray drying, freeze-drying, concentration by washing azhivaniem et al.).

Fatty acid esters or water-soluble organic solvents that can preferably be used in this extraction method include, but are not limited to, acetates such as ethyl acetate, monohydric alcohols having 1-4 carbon atoms, acetone, propylene glycol, glycerol, and mixtures thereof. Among them, acetates such as ethyl acetate, monohydric alcohols having 1-4 carbon atoms, acetone or mixtures thereof are preferable, and ethyl acetate, ethanol, acetone or mixtures thereof are more preferred. In particular, ethanol is preferred in view of the safety of the solvent for the human body. To reduce pollution with water-soluble impurities such as glycyrrhizin described below, monohydric alcohols having 2-4 carbon atoms, fatty acid esters and acetone are preferred, fatty acid esters and acetone are particularly preferred, acetates and acetone are more preferred, and still more preferred are ethyl acetate and acetone. Obviously, other solvents can coexist in such a range so as not to have side effects.

In the present invention, it is important to increase the content of the active ingredient in the licorice extract, along with a decrease in the content of water-soluble impurities such as glycyrrhizin.

Glycyrrhizin is also used as a pharmaceutical, but it has side effects such as high blood pressure, cardiovascular disease and hydremia (referred to by Harders, H. & Rausch-Stroomann, JG, Munch. Med. Wschr. 95 , 580 (1953)), hypokalemia, decreased plasma renin activity, symptoms of pseudo-aldosteronism and relaxing limb paralysis (referred to by Conn, JW, Rovner, DR & Cohen, EL, J. Am. Med. Assoc. 205 , 492 (1968) ), decreased secretion of aldosterone in the urine, swelling, headache and drowsiness (referred to by Epstein, MT, et al., Brit. Med. J., 1 , 488 (1977)). Glycyrrhizin is 150 times sweeter than sucrose. Glycyrrhizin contamination inhibits the various effects of licorice extract and causes side effects and strong sweetness, thus causing inconvenience when used in food products, etc.

To increase the content of the active ingredient in the resulting licorice extract and at the same time reduce the content of water-soluble impurities, such as glycyrrhizin, the water content present during extraction with the organic solvent is preferably reduced.

To reduce the water content, it is important to use licorice, dried to the maximum degree, as well as the use of an anhydrous organic solvent.

However, it should be noted that licorice is a plant and conventional drying in the sun is not always sufficient to obtain sufficiently dried licorice, therefore, it becomes necessary to use drying machines, etc. This represents a serious weakness in production on a commercial scale.

Even with an anhydrous organic solvent, it is difficult to completely prevent the organic solvent from being contaminated with water from the used licorice and the working atmosphere, and thereby return the organic solvent to the cycle due to the increased water content. Since the organic solvent is difficult to return to the cycle as such, the solvent is thrown away or the necessary special and expensive cleaning agents and methods are used to obtain an anhydrous solvent, which leads to an increase in production costs.

Accordingly, if high quality licorice extract with a high content of active ingredient and a lower content of water-soluble impurities can be obtained quite easily and inexpensively, significant advantages can be expected.

As a result of intensive research, the present inventors have found that it is preferable to use licorice in a form such that the ratio of the area of the crust (shells) to the total surface area thereof is high, usually 30% or more, preferably 50% or more, more preferably 70 % or more, even more preferably 80% or more, even more preferably 90% or more (especially a cut product or licorice periderm (including a product mainly containing the periderm)), in order to obtain extra KTA high quality licorice. In addition, the inventors have found that when licorice is used in the above form, a high quality licorice extract can be obtained even if a hydrated, water-soluble organic solvent is used.

Among the cut product and the periderm (including a product mainly containing the periderm), the cut product is more preferable for use, since it can be more easily processed, and the ratio of the crust area to the total surface area of the cut product is usually 50% or more, preferably 70 % or more, more preferably 80% or more, and even more preferably 90% or more.

According to this method, the effect can be maximized, especially when using the aforementioned water-soluble organic solvent, preferably a monohydroxy alcohol having 2-4 carbon atoms, more preferably ethanol.

The water content of the organic solvent used is usually about 30% by volume or less, preferably about 20% by volume or less, more preferably about 10% by volume or less, and even more preferably about 8% by volume or less. The lower limit is not particularly limited, but is usually from a practical point of view about 3% by volume, preferably 4% by volume.

Thus, licorice extract having a higher content of the active ingredient and a lower content of water-soluble impurities can preferably be obtained. The glycyrrhizin content in the resulting extract can be minimized, preferably up to 0.5% by volume or less. Even when an inexpensive, hydrated, water-soluble organic solvent is used as the extracting solvent, a high quality extract can be preferably obtained (without special drying of licorice).

Extraction with a fatty acid ester or a water-soluble organic solvent mentioned above can be carried out in accordance with a conventional method and is not particularly limited. The extraction temperature is not particularly limited, and the extraction can be carried out preferably between the pour point and the boiling point in the system, typically from −20 to 100 ° C., usually from 1 to 80 ° C., preferably from 20 to 60 ° C.

The extraction procedure can be carried out, for example, for 0.1 hours or more, preferably 0.2 hours or more, more preferably 0.5 hours or more using a fatty acid ester or a water-soluble organic solvent in, for example, 1-20 -fold excess, preferably 2-10-fold excess, based on the volume of licorice or previously extracted residue. Typically, the time for each extraction is preferably about 1-10 hours. The upper limit is not particularly limited and is about 1 day, but the extraction can be carried out for a longer time. The extraction can be carried out one or several times as necessary, and a suitably mixed solvent can be used. The pressure during extraction is not particularly limited. The extraction is carried out at normal pressure or under pressure (one to several atmospheric pressures), but if desired, can be carried out under reduced pressure. The extraction can be carried out with reflux or under low pressure.

After extraction, the pre-extract can be separated into the extract and the residue of extracted licorice using a conventional separation technique (e.g., pressure filtration, vacuum filtration, centrifugation, sedimentation, etc.) and, if necessary, washed with a solvent to obtain licorice extract. In the separation procedure, if necessary, commonly used filter aid and / or adsorbent substances, such as activated carbon and resin, can be used.

From the licorice extract thus obtained, the solvent used can be removed using a conventional solvent removal technique (e.g., concentration under normal pressure, vacuum concentration, spray drying, freeze drying, freeze concentration), whereby a solvent free licorice extract can be obtained. After extraction, the extract as described above is more preferably purified by adsorption using activated carbon or clay and / or fractionation, thereby increasing the content of the active ingredient in the extract.

In the method of the present invention, a number of the above procedures, especially solvent extraction, or solvent extraction with the following procedures (separation of the extract, removal of the solvent, adsorption with activated carbon, clay or the like and / or fractionation, etc.) are preferably carried out in an oxygen-free atmosphere, such as an inert gas atmosphere, using nitrogen gas, etc. In order to prevent oxidation, the technique can be carried out in the presence of antioxidants such as ascorb other acid, sodium ascorbate, ascorbyl palmitate, ascorbyl stearate, tocopherol, etc.

Licorice extract obtained after removing the solvent is usually brown (e.g., tan to dark brown).

The weight ratio (based on dry weight) of dry licorice extract obtained by treating licorice as such is usually about 0.01 or more.

The glycyrrhizin content of the extract of the present invention, based on dry weight, is usually as low as 0.5% by mass or less, preferably 0.3% by mass or less, more preferably 0.2% by mass or less, still more preferably 0.1% by weight or less. When the glycyrrhizin content of the extract of the present invention is from 0.001 to 0.5% by weight based on dry weight, there is practically no problem of side effects and disadvantages of use.

The glycyrrhizin content in the extract can be determined, for example, by HPLC analysis using commercial glycyrrhizin or glycyrrhizinate as a standard substance or similar analysis. The content of the PPAR ligand in the extract can be determined, for example, by HPLC analysis using the PPAR ligand of the present invention as a standard substance or similar analysis. The PPAR ligand as a standard substance may be a compound isolated from a plant, such as licorice, or a chemically synthesized compound. The total number of PPAR ligand components in the extract can be calculated as the total content of the measured PPAR ligand components.

The total amount of PPAR ligand components in the extract of the present invention, based on dry weight, is usually 0.5% by mass or more, preferably 1% by mass or more, and more preferably 2% by mass or more. In particular, when the PPAR ligand is extracted with a monohydroxy alcohol having 1-4 carbon atoms, its amount is usually 5% by mass or more, preferably 6% by mass or more, more preferably 7% by mass or more, thus showing an effective selection of the active ingredient. The total amount of PPAR ligand components contained in the extract of the present invention, based on dry weight, is usually not more than 50% by weight, but the content can be further increased by adsorption using activated carbon, resin or the like and / or fractionation.

For example, the extract of the present invention preferably includes, for example, at least one species selected from the group consisting of glycicoumarin, glycyrin, dehydrogliasperin C, dehydrogliasperin D, gliasperin B and gliasperin D as the main component in an amount of usually 0.5% by weight or more, preferably 1% by weight or more based on dry weight. More preferably, the extract includes glycicumarin, glycyrin, dehydrogliasperin C, dehydrogliasperin D, gliasperin B and gliasperin D, respectively, in an amount of usually 0.5% by weight or more, preferably 1% by weight or more based on dry weight. These components are found in larger quantities in licorice extract, especially G. uralensis.

In addition, the extract of the present invention preferably includes, for example, at least one species selected from the group consisting of glycyrrhizoflavanone, glycyrrhizoflavone, gliuralline B, semicoisoflavone B and isoquiquitigenin in an amount of usually 0.01% by weight or more, preferably 0 , 02% by weight or more based on dry weight. More preferably, the extract includes glycyrrhizoflavanone, glycyrrhizoflavon, gliuralin B, semicoisoflavon B and isoquirytigenin, respectively, in an amount of usually 0.01% by mass or more, preferably 0.02% by mass or more based on dry weight.

Moreover, the extract of the present invention preferably includes, for example, at least one species selected from the group consisting of glabren, glabridin, glabrol, 3'-hydroxy-4'-O-methylglabridine, 4'-O-methylglabridine and gisaglabridin B in an amount of usually 0.5% by mass or more, preferably 1% by mass or more based on dry weight. More preferably, the extract includes glabren, glabridin, glabrol, 3'-hydroxy-4'-O-methylglabridine, 4'-O-methylglabridine and gisaglabridin B, respectively, in an amount of usually 0.5% by weight or more, preferably 1% by weight or more based on dry weight. These components are found in greater quantities in licorice extract, especially G. glabra.

In addition, the extract of the present invention preferably includes, for example, at least one species selected from the group consisting of lycochalcone A, lycochalcone B, lycochalcone C, glycyrrhione A and glycyrrhione C in an amount of usually 0.5% by weight or more preferably 1% by weight or more based on dry weight. More preferably, the extract more preferably includes lycochalcon A, lycochalcon B, lycochalcon C, glycyrdione A and glycyrdione C, respectively, in an amount of usually 0.5% by mass or more, preferably 1% by mass or more based on dry weight. These components are found in greater quantities in licorice extract, especially G. inflata.

The composition of the present invention to improve insulin resistance or to prevent and / or improve the condition of insulin resistance syndrome and the composition of the present invention to prevent and / or improve inflammation or cancer are compositions comprising the PPAR ligand of the present invention. That is, they are compositions that include at least one compound selected from the group consisting of prenyl flavonoid, a chalcone derivative other than prenyl flavonoids, a flavonol derivative other than prenyl flavanides, and their salts, glycosides and esterified substances, acceptable as a pharmaceutical preparation or food product or drink.

As described above, the prenylflavonoid is not particularly limited, but is preferably at least one compound selected from the group consisting of a 3-arylcoumarin derivative, an isoflav-3-ene derivative, an isoflavan derivative, an isoflavanone derivative, an isoflavone derivative, an isoflavanol derivative, a flavanone derivative, a chalcone derivative and a dibenzoylmethane derivative.

The above compound may be a pure compound or a semi-purified or untreated compound such that it does not contain impurities unsuitable for pharmaceutical preparations or food products. In this case, the extract may be used as such or may be further purified. The compound is not limited in its form and can be used, for example, in the form of food products or drinks, such as health food products (specialized health products, nutritious functional food products), healthy food products and auxiliary nutritious food products, pharmaceutical preparations and pseudo-drugs.

The composition of the present invention can be administered to humans as well as to pets or domestic animals, such as dogs, cats, cows, horses, pigs, chickens, sheep, goats, mice and rats. For use as a food or drink, it may be swallowed directly or may be presented as easily swallowed products, such as capsules, tablets, granules, powders, etc., using known carriers, adjuvants or the like for oral administration. Additionally, a substance for the composition other than the PPAR ligand of the present invention may be suitably added and mixed by conventional methods. Such a substance is not particularly limited, and, for example, excipients, disintegrants, lubricants, binding agents, antioxidants, coloring agents, aggregation inhibitors, adsorption activators, stabilizers, etc. may be mentioned. The amount of PPAR ligand of the present invention in a product thus created may be 0.1-100% by weight, preferably 10-90% by weight. In addition, it can be mixed with raw materials for all types of foods or drinks, for example, confectionery such as chewing gum, chocolate, sweets, jellies, cookies and crackers; frozen sweets such as ice cream and frozen sweets; drinks such as tea, soft drinks, nutritious drinks and drinks for cosmetic purposes; noodles such as Japanese wheat noodles, Chinese noodles, spaghetti and instant noodles; fish paste products such as minced and steamed fish ( kamaboko) , fish sausages ( chikuva ) and minced meat ( hannpen ); seasonings such as gravy, mayonnaise and sauces; oily products such as margarine, butter and lettuce oil; bakery products, ham, soups, baked products, frozen foods and so on. When consuming compositions of such products or drinks, the recommended daily intake for an adult is 0.1-3000 mg / kg, more preferably 1-300 mg / kg, as the PPAR ligand of the present invention. Such compositions can also be used as food for pets and domestic animals or as animal feed, and the recommended daily intake for such use is preferably 0.1-3000 mg / kg as the PPAR ligand of the present invention.

For use as a pharmaceutical product, the dosage form is not particularly limited and includes capsules, tablets, granules, powders, injectable preparations, suppositories, plasters, etc. Such dosage forms may be prepared from suitably formulated pharmaceutically acceptable substances for pharmaceutical preparations, such as excipients, disintegrants, lubricants, binders, antioxidants, coloring agents, aggregation inhibitors, adsorption activators, solubilizers and stabilizers. The daily dosage of such compositions for an adult is 0.1-3000 mg / kg, preferably 1-300 mg / kg in the form of the PPAR ligand of the present invention, this dose can be administered in one dose or in several separate doses throughout the day. The composition can also be used as a pharmaceutical preparation for pets and domestic animals, and the daily dosage for such use is preferably 0.1-3000 mg / kg as the PPAR ligand of the present invention.

When the composition of the present invention is presented in the form of tablets, capsules or powders and when the above extract is contained in the form of a PPAR ligand, the composition preferably comprises an extract in an amount of 0.1-1000 mg per tablet.

The PPAR ligand of the present invention is an extract component obtained from a plant or a compound associated with it, and is believed to have low toxicity. The extract obtained according to the present invention can also be added to foods for which sweetness or the like is undesirable. It is highly stable when compared with the highly unsaturated fatty acids, which are usually considered PPARγ ligands, and is the best in terms of form suitable for food products and pharmaceutical compositions.

Best Mode for Carrying Out the Invention

Below the present invention is described in more detail by reference to examples, but the range of the present invention is not limited to these examples.

(Example 1) Extraction and isolation of the compound from licorice (1)

The finely ground product (1.2 kg) of licorice (G. uralensis) was immersed in 5.5 L of ethyl acetate and extracted for 7 days at room temperature with shading. Then the mixture was filtered to obtain an extract. The extract was concentrated under reduced pressure to remove the solvent, and 74.0 g of extract was obtained. The resulting extract was subjected to silica gel column chromatography (1500 ml) to obtain a fraction eluted with chloroform: 19: 1 methanol (v / v). The eluting fraction was concentrated under reduced pressure to remove the solvent, and 55.4 g of the crude fraction was obtained. This untreated fraction was purified by repeating column chromatography on silica gel, column chromatography on ODS silica gel, high performance liquid chromatography equipped with an ODS column, gel filtration column chromatography and fractional thin layer chromatography. Thus, compound 1 (225 mg), compound 2 (80.7 mg), compound 3 (19.6 mg), compound 4 (22.2 mg), compound 5 (3.9 mg), compound 6 ( 72.8 mg), compound 7 (28.3 mg), compound 8 (12.0 mg), compound 9 (6.8 mg), compound 10 (8.5 mg), compound 11 (58.7 mg) and compound 12 (10.2 mg).

As a result of the structural analysis, compounds 1-3 and 5-12 are known compounds and are respectively identified as the following: compound 1 is glycumarin, compound 2 is glycyrin, compound 3 is glyuralline B, compound 5 is echinatin, compound 6 is is isolicoflavonol, compound 7 is dehydrogliasperin C, compound 8 is gliasperin B, compound 9 is glycyrrhizoflavanone, compound 10 is lupivitone, compound 11 is gliasperin D and compound 12 is semilicoisoflavone B. In addition, for structural data identification, the compound, spectral data described by S. Demizu, et al., Chemical and Pharmaceutical Bulletin, 36, 3474-3479 ( 1988) refer to compound 1, Kinoshita, et al., Chemical and Pharmaceutical Bulletin, 26, 1350140 (1978) refer to compound 2, M. Shibano, et al., Heterocycles, 45, 2053-2060 (1997) refer to compounds 3 and 7, K. Kajiyama, et al., Phytochemistry, 31, 3229-3232 (1992) refer to compound 5, T. Hatano, et al., Chemical and Pharmaceutical Bulletin, 36, 2090-2097 (1988) refer to compounds 6 and 9, L. Zeng, et a l., Heterocycles, 34, 575-587 (1992) refer to compounds 8 and 11, Y. Hashidoko, et al., Agricultural and Biological Chemistry, 50, 1797-1807 (1986) refer to compound 10 and F. Kiuchi, et al., Heterocycles, 31, 629-636 (1990) refer to compound 12.

Compound 4 is a new compound, and its structure has been defined as 3- (2 ', 4'-dihydroxyphenyl) -6- (3``, 3''- dimethylallyl) -5,7-dimethoxy-2H-chromene using spectral spatial centric NMR analysis ( ¹ H- ¹ H COSY, HMQC, HMBC, PHNOESY), it was called dehydrogliasperin D.

The characteristics and spectral data of compound 4 were as follows:

Dehydrogliasperin D, brown powder, C ₂₂ H ₂₄ O ₅ .

EI-MS m / z: 368.1610 [M] + (calculated value; C ₂₂ H ₂₄ O ₅ 368.1624).

UVmax (methanol) nm: 330 (logε = 4.25), 243 cor. (logε = 4.21).

IR (KBr tablet) cm ^-1 : 3375, 2929, 1613, 1516, 1458, 1308, 1198, 1164, 1114, 1092, 1024, 978, 837.

¹ H-NMR (dimethyldisulfoxide) ppm: 7.06 (1H, d, J = 8.4 Hz, H-6 '), 6.68 (1H, s, H-4), 6.34 ( 1H, d, J = 2.3 Hz, H-3 '), 6.33 (1H, s, H-8), 2.26 (1-H, dd, J = 8.4, 2.3 Hz , H-5 '), 5.09 (1H, br t, J = 6.9 Hz, H-10), 4.90 (2H, s, H-2), 3.75 (3H, s, C-7-OMe), 3.67 (3H, s, C-7-OMe), 3.18 (2H, brd, J = 6.7 Hz, H-9), 1.71 (3H, s, Me-13), 1.63 (2H, s, Me-12).

¹³ C-NMR (dimethyldisulfoxide) ppm: 67.9 (C-2), 128.8 (C-3), 114.7 (C-4), 110.4 (C-4a), 154, 9 (C-5), 115.6 (C-6), 158.0 (C-7), 95.8 (C-8), 153.2 (C-8a), 22.6 (C-9 ), 124.1 (C-10), 130.4 (C-11), 26.0 (C-12), 18.1 (C-13), 116.8 (C-1 '), 156, 7 (C-2 '), 103.3 (C-3'), 158.7 (C-4 '), 107.4 (C-5'), 129.2 (C-6 '), 62, 3 (C-5-OMe), 56.3 (C-7-Me)

(Example 2) Extraction and isolation of the compound and licorice (2)

In the same procedure as in Example 1, compounds 13-17 were obtained by extraction of the finely divided licorice product (G. uralensis) with ethanol for various chromatography. As a result of structural analysis, the following compounds were identified; Compound 13 was isoliquirithigenin, Compound 14 was Kempferol 3-O-methyl ester, Compound 15 was lycoflavonol, Compound 16 was topazoline and Compound 17 was glycyrrhizoflavone.

(Example 3) Extraction and isolation of the compound from licorice (3)

In the same procedure as in Examples 1 and 2, Compounds 18-23 and Compounds 24-28 were obtained by extracting finely divided licorice product (G. glabra) and finely divided licorice product (G. inflata) with ethanol, respectively. As a result of structural analysis, the following compounds were identified; Compound 18 was glabridin, Compound 19 was glabren, Compound 20 was gisaglabridin B, Compound 21 was 4'-O-methylglabridine, Compound 22 was 3'-hydroxy-4'-O-methylglabridine, Compound 23 was glabrol, compound 24 was lycochalcon A, compound 25 was lycochalcon B, compound 26 was lycochalcon C, compound 27 was glycyrrhione A and compound 28 was glycyrdione C.

Tables 10-13 show the structural formulas of compounds 1-28.

Table 10 Connection number Connection Name Structural formula Compound 1 Glycicumarin

Compound 2 Glycyrin

Compound 3 Gliurallin B

Compound 4 Dehydrogliasperin D

Compound 5 Echinatin

Compound 6 Isolicoflavonol

Compound 7 Dehydrogliasperin C

Compound 8 Gliasperin B

Table 11 Connection number Connection Name Structural formula Compound 9 Glycyrrizoflavanone

Compound 10 Lupiviteon

Compound 11 Gliasperin D

Compound 12 Semilicoisoflavone B

Compound 13 Isoliquitritigenin

Compound 14 Kempferol 3-O-methyl ester

Compound 15 Lycoflavonol

 Table 12 Connection number Connection Name Structural formula Compound 16 Topazoline

Compound 17 Glycyrrizoflavone

Compound 18 Glabridin

Compound 19 Glabren

Compound 20 Gisaglabridin B

Compound 21 4'-O-methylglabridine

Compound 22 3'-hydroxy-4'-O-methylglabridine

Table 13 Connection number Connection Name Structural formula Compound 23 Glabrol

Compound 24 Lycochalcon A

Compound 25 Lycochalcon B

Compound 26 Lycochalcon C

Compound 27 Glycyrdione A

Compound 28 Glycyrdione C

(Example 4) Measurement of PPARγ Ligand Activity

CV-1 cells (cultured cells obtained from the kidney of a male African green macaque) were inoculated into a 96-well culture plate at a concentration of 6 × 10 ³ cells / well and incubated at 37 ° C. in 5% CO ₂ for 24 hours. The medium used was DMEM (Dulbecco's Modified Eagle's Medium: GIBCO) containing 10% FCS (FBS) (fetal calf serum), 10 ml / L solution of penicillin and streptomycin (containing 5000 IU / ml and 5000 μg / ml, respectively: GIBCO ) and 37 mg / L ascorbic acid (Wako Chemical Industries, Ltd). Cells were washed with OPTI-MEM (GIBCO) and then transfected with pM-mPPARγ and 4xUASg-luc using Lipofect Amine Plus (GIBCO). PM-mPPARγ is a plasmid for the expression of a chimeric protein comprising the GAL4 yeast transcription factor gene (amino acid sequence 1-147) and the associated mouse gene of the PPARγ ligand binding site (amino acid sequence 174-475). 4xUASg-luc is a reporter plasmid comprising four GAL4 response sequences (UASg) inserted in the region upstream of the luciferase gene. About 24 hours after transfection, the medium was changed to a medium containing a sample, and the cells were cultured for 24 hours (n = 4). The medium containing the sample was obtained by dissolving the sample in dimethyl sulfoxide (DMSO) in the amount of 1/1000 based on the medium, whereas for the control group, DMSO in the amount of 1/1000 based on the medium was used instead of the sample. After the cells were washed with Ca- and Mg-containing phosphate buffered saline (PBS), LucLite (Packard) was added to them and luciferase luminescence intensity was measured using a Topcount (Packard) selective scintillation / luminescence counter.

In the control group, pM (a plasmid from which the PPARγ ligand binding site was deleted) was evaluated in place of pM-mPPARγ in the same manner as in the study group. For each sample, the ratio (test group / control group) of the average luminescence intensity of the test group (n = 4) to that of the control group was calculated, and the specific activity relative to the untreated control activity was considered as the PPARγ ligand activity of the sample.

Tables 14-17 show the results of measurements of the activity of the PPARγ ligand of compounds 1-28 obtained in examples 1-3. As comparative compounds, glycyrrhizin (glycyrrhizic acid, Wako Pure Chemical Industries, Ltd.), glycyrrhetinic acid (Wako Pure Chemical Industries, Ltd.) and quercetin (Sigma) were measured by their PPARγ ligand activity.

Table 14 Added concentration PPARγ ligand activity Raw Control (DMSO) 0.1% 1.00 Troglitazone 0.5 μmol 2.17 ± 0.08 1 μmol 3.36 ± 0.24 2 μmol 4.60 ± 0.27 Compound 1
(glycicumarin) 2 μg / ml (5.4 μmol) 2.03 5 μg / ml (13.6 μmol) 2.79 10 μg / ml (27.2 μmol) 3.82 Compound 2
(glycyrin) 2 μg / ml (5.2 μmol) 2.81 5 μg / ml (13.1 μmol) 3.34 10 μg / ml (26.2 μmol) 3.67 Compound 3
(gliuralline B) 2 μg / ml (4.7 μmol) 1.28 5 μg / ml (11.8 μmol) 1.96 10 μg / ml (23.7 μmol) 2.73 Compound 4
(dehydrogliasperin D) 2 μg / ml (5.4 μmol) 2.95 5 μg / ml (13.6 μmol) 3.72 10 μg / ml (27.2 μmol) 4.00 Compound 5
(echinatin) 2 μg / ml (7.4 μmol) 1.45 5 μg / ml (18.5 μmol) 2,35 10 μg / ml (37.0 μmol) 2.49

Table 15 Added concentration PPAR Ligand Activity Raw Control (DMSO) 0.1% 1.00 Troglitazone 0.5 μmol 2.44 ± 0.10 1 μmol 3.99 ± 0.24 2 μmol 5.74 ± 0.31 Compound 6
(Isolicoflavonol) 2 μg / ml (5.6 μmol) 1.73 5 μg / ml (14.1 μmol) 2,50 10 μg / ml (28.2 μmol) 3.82 Compound 7
(dehydrogliasperin C) 2 μg / ml (5.6 μmol) 2.90 5 μg / ml (14.1 μmol) 3.73 10 μg / ml (28.2 μmol) 4.41 Compound 8
(gliasperin B) 2 μg / ml (5.4 μmol) 2.60 5 μg / ml (13.5 μmol) 3.53 10 μg / ml (27.0 μmol) 3.02 Compound 9
(glycyrrhizoflavanone) 2 μg / ml (5.4 μmol) 2.01 5 μg / ml (13.6 μmol) 3.45 10 μg / ml (27.2 μmol) 4.68 Compound 10
(lupivitone) 2 μg / ml (5.9 μmol) 1,67 5 μg / ml (14.8 μmol) 2,53 10 μg / ml (29.6 μmol) 3.75 Compound 11
(gliasperin D) 2 μg / ml (5.4 μmol) 3.22 5 μg / ml (13.5 μmol) 3.28 10 μg / ml (27.0 μmol) 4.06 Compound 12
(semilicoisoflavone B) 2 μg / ml (5.7 μmol) 1.86 5 μg / ml (14.2 μmol) 2.66 10 μg / ml (28.4 μmol) 3.15 Glycyrrhizin 2 μg / ml (2.4 μmol) 1.21 5 μg / ml (6.1 μmol) 1.10 10 μg / ml (12.2 μmol) 0.75 Glycyrrhetinic acid 2 μg / ml (4.2 μmol) 0.97 5 μg / ml (10.6 μmol) 0.95 10 μg / ml (21.2 μmol) 0.97 Quercetin 2 μg / ml (5.9 μmol) 1.15 5 μg / ml (14.8 μmol) 1.72 10 μg / ml (29.6 μmol) 1.70

Table 16 Added concentration PPAR Ligand Activity Raw Control (DMSO) 0.1% 1.00 Troglitazone 0.5 μmol 2.24 ± 0.56 1 μmol 3.29 ± 0.60 2 μmol 4.80 ± 1.30 Compound 13
(isoliquiritigenin) 2 μg / ml (7.8 μmol) 1.47 5 μg / ml (19.5 μmol) 2.71 10 μg / ml (39.0 μmol) 3.39 Compound 14
(campferol-3-O-methyl ether) 2 μg / ml (6.7 μmol) 1.86 5 μg / ml (16.7 μmol) 2,37 10 μg / ml (33.3 μmol) 3.47 Compound 15
(lycoflavonol) 2 μg / ml (5.6 μmol) 1.44 5 μg / ml (14.1 μmol) 2.27 10 μg / ml (28.2 μmol) 4.07 Compound 16
(topazoline) 2 μg / ml (5.4 μmol) 2,56 5 μg / ml (13.6 μmol) 3.64 10 μg / ml (27.1 μmol) 5.84 Compound 17
(glycyrrhizoflavone) 2 μg / ml (5.6 μmol) 1.78 5 μg / ml (14.1 μmol) 3.05 10 μg / ml (28.2 μmol) 3.20 Table 17 Added concentration PPAR Ligand Activity Raw Control (DMSO) 0.1% 1.00 Troglitazone 0.5 μmol 2.05 ± 0.35 1 μmol 3.30 ± 0.43 2 μmol 5.66 ± 1.06 Compound 18
(glabridin) 2 μg / ml (6.2 μmol) 1,64 5 μg / ml (15.4 μmol) 2.15 10 μg / ml (30.8 μmol) 1.98 Compound 19
(glazed) 2 μg / ml (6.2 μmol) 1.78 5 μg / ml (15.5 μmol) 3.44 10 μg / ml (31.0 μmol) 2.89 Compound 20
(gispaglabridin B) 2 μg / ml (5.1 μmol) 1,04 5 μg / ml (12.8 μmol) 1.45 10 μg / ml (25.6 μmol) 1.94 Compound 21
(4'-O-methylglabridine) 2 μg / ml (5.9 μmol) 1.25 5 μg / ml (14.8 μmol) 1.55 10 μg / ml (29.6 μmol) 2.13 Compound 22
(3'-hydroxyl-4'-O-methylglabridine) 2 μg / ml (5.6 μmol) 1,61 5 μg / ml (14.1 μmol) 2.46 10 μg / ml (28.2 μmol) 1,69 Compound 23
(glabrol) 2 μg / ml (5.1 μmol) 1.85 5 μg / ml (12.7 μmol) 3.48 10 μg / ml (25.5 μmol) 3.74 Compound 24
(lycochalcon A) 2 μg / ml (5.9 μmol) 4.02 5 μg / ml (14.8 μmol) 4.14 10 μg / ml (29.6 μmol) 4.16 Compound 25
(lycochalcon B) 2 μg / ml (7.0 μmol) 1.27 5 μg / ml (17.5 μmol) 1.93 10 μg / ml (34.9 μmol) 1,61 Compound 26
(lycochalcon C) 2 μg / ml (5.9 μmol) 1.84 5 μg / ml (14.8 μmol) 2.20 10 μg / ml (29.6 μmol) 3.71 Compound 27
(glycyrdione A) 2 μg / ml (4.9 μmol) 1.44 5 μg / ml (12.2 μmol) 2.49 10 μg / ml (24.5 μmol) 3.05 Compound 28
(glycyrdione C) 2 μg / ml (4.9 μmol) 0.96 5 μg / ml (12.3 μmol) 1.48 10 μg / ml (24.6 μmol) 2.04

The above PPARγ ligand activity of each compound was compared with that of the positive control, troglitazone (Sankyo Co., Ltd.). As a result, it was found that compounds 1-19, 21-24, 26 and 27 have a higher PPARγ ligand activity than the positive control containing 0.5 μmol of troglitazone. It was also found that compounds 20, 25, and 28 have PPARγ ligand activity, although it is slightly weaker than that of the positive control containing 0.5 μmol of troglitazone. However, glycyrrhizin, which is a hydrophilic compound and the main component of licorice, glycyrrhetinic acid (a derivative of glycyrrhizin in which the carbohydrate residue is hydrolyzed) and quercetin (a flavonol derivative of compound 6, in which the OH group replaces the prenyl group), did not have PPARγ ligand activity. The fact that quercetin does not have PPARγ ligand activity is consistent with information in the literature (Liang, YC, et al., FEBS Letters, 496 , 12-18, 2001).

(Example 5) Effect on a Type 2 Diabetes Model in Mice

Using KK-Ay mice that are genetically obese and type 2 diabetes, the effects of Compound 2 on diabetes mellitus were evaluated. As a positive control, pioglitazone, i.e., means for improving the state of insulin resistance / means for treating type 2 diabetes, was used.

KK-Ay mice (females, 15 weeks old) were divided into 3 groups, each of which consisted of 5 mice, and they were freely given food that included only the main diet, i.e., powdered CE-2 (CLEA Japan, Inc.) for the non-supplementing group (control group), a food comprising the main diet and pioglitazone, the food group including the main diet and compound 2, the group receiving compound 2. For pioglitazone, ACTOS 30 tablets (containing 30 mg pioglitazone tablet, Takeda Chemical Industries, Ltd.) was ground in an agate mortar and added a basic diet, so that the content of pioglitazone in the final feed was 0.02%. Compound 2 was prepared using the same procedure as in Example 1, and was added to the main diet so that the content of compound 2 in the final feed was 0.1%. A day before administration and 4 days after administration, a small amount of blood was taken from the tail vein of mice and blood glucose was measured using a simple glucose meter Glutest Ace (Sanwa Kagaku Kenkyusho Co., Ltd.). Table 18 shows the results.

After the mixed feed was given for 1 week, the mice were fasted overnight, and then they were tested for glucose tolerance. That is, 20 mg / kg of pioglitazone or 100 mg / kg of compound 2 dissolved in a 0.5% sodium carboxymethyl cellulose solution (CMC-Na (CMC-Na)) was forcibly administered orally to hungry mice. The group to which the drug was not administered (control group) was injected with a 0.5% solution of CMC-Na in an amount of 5 ml / kg. 30 minutes after administration, mice were given 2 g / kg of a 40% glucose solution. Before the sugar load and 30 minutes, 1 hour and 2 hours after the sugar load, a small amount of blood was taken from the tail vein of mice and blood glucose was measured in them using a simple glucose meter Glutest Ace (Sanwa Kagaku Kenkyusho Co., Ltd.) . Table 19 shows the results.

Table 18 Blood glucose level (mg / dl) The group to which the drug was not administered (control group) Pioglitazone group The group receiving compound 2 (glycyrin) Before introduction 476 ± 22 486 ± 26 474 ± 27 4 days after administration 420 ± 14 191 ± 6 ** 278 ± 14 ** ** (p <0.01)

Table 19 Blood glucose level (mg / dl) The group to which the drug was not administered (control group) Pioglitazone group The group receiving compound 2 (glycyrin) Before loading sugar 80 ± 2 89 ± 3 78 ± 4 30 minutes after sugar loading 355 ± 18 158 ± 11 ** 225 ± 27 ** 1 hour after sugar load 199 ± 11 136 ± 5 * 129 ± 8 * 2 hours after sugar loading 100 ± 4 106 ± 4 100 ± 3 * (p <0.05), ** (p <0.01)

As can be seen from table 18, the blood glucose level (mean ± CO, n = 5) of mice from the group receiving compound 2 was statistically significantly reduced in the same way as that of mice from the group receiving pioglitazone, on the fourth day after administration . As can be seen from table 19, compound 2, like pioglitazone, statistically significantly inhibited the rapid increase in blood glucose after loading with sugar. These results confirm that compound 2 has a decreasing effect and an inhibitory effect on the increase in blood glucose levels, therefore, it was suggested that this compound has an action that improves the state of insulin resistance, like pioglitazone.

(Example 6)

To a finely ground product (500 g) of licorice (G. uralensis) (Kaneka Sun Spice Co., Ltd) was added 5 kg of ethanol (99.5% (v)). The mixture was extracted at 25 ° C. for 5 hours, and the residue was filtered to obtain an extract. The solvent was removed from the obtained extract under reduced pressure to obtain 45 g of licorice extract having a tan to dark brown color. This licorice extract showed good PPARγ ligand activity. Table 20 shows the content of ingredients having PPARγ ligand activity and glycyrrhizin contained in this licorice extract.

In addition, the content of these active ingredients and glycyrrhizin was obtained by HPLC analysis under the following conditions.

(Conditions for the analysis of glycyrrhizin)

Speaker; J'sphere ODS-H80 (YMC Co., Ltd.) 4.6 mm (inner diameter) × 250 mm (length); column temperature: 40 ° C; mobile phase: acetonitrile / 10 mmol of phosphoric acid solution = 33/67 (v / v); flow rate: 1 ml / min; detection wavelength: 254 nm; retention time: 27.1 minutes

(Conditions for analysis of active ingredient)

Speaker; J'sphere ODS-H80 (YMC Co., Ltd.) 4.6 mm (inner diameter) × 250 mm (length); column temperature: 40 ° C. Mobile phase: a gradient in which the ratio of acetonitrile relative to 10 mmol of phosphoric acid solution is maintained at 35% up to 15 min after the start of the analysis, then increases to 70% at a constant rate from 15 min to 65 min, and then maintained at 70% period from 65 minutes to 70 minutes. Flow rate: 1 ml / min; Detection wavelength: 254 nm. The retention time for each component is as follows: 5.2 min for lycochalcone B, 13.3 min for glycyrrhizoflavanone, 14.7 min for isoliquiritigenin, 32.7 min for glycicoumarin, 34.2 min for seven-isoflavone B, 35.9 min for glabrena, 36.3 min for glycyrrhizoflavone, 37.2 min for dehydrogliasperin C, 39.1 min for lycochalcone C, 40.2 min for lycochalcone A, 44.7 min for glabridin, 48.1 min for glycyrin, 51.1 min for gliasperin B, 51.8 min for glabrol, 52.7 min for gliasperin D, 53.4 min for glycyrdione A, 53.5 min for 3'-hydroxyl-4'-O-methylglabridine, 55.2 min for dehydrogum liasperin D, 58.7 min for gliuralline B, 59.8 min for 4'-O-methylglabridine, 63.0 min for glycyrdione C, and 74.1 min for gisaglabridin B.

Table 20 Connection Name Content (wt.%) Glycyrrhizin 0.28 Glycyrrizoflavanone 0.15 Isoliquitritigenin 0.19 Glycicumarin 2,55 Semilicoisoflavone B 0.45 Glycyrrizoflavone 0.50 Dehydrogliasperin C 2.82 Glycyrin 1.46 Gliasperin B 1.12 Gliasperin D 2.44 Dehydrogliasperin D 1.39 Gliurallin B 0,03 Active component: total 13.10%

(Example 7)

Licorice extract (18 g) was obtained by the same procedure as in Example 6, except that extraction was carried out using acetone instead of ethanol. The obtained licorice extract showed good PPARγ ligand activity. The contents of the active ingredients and glycyrrhizin contained in such licorice extract were evaluated in the same manner as in Example 6, and table 21 shows the results.

Table 21 Connection Name Content (wt.%) Glycyrrhizin 0.06 Glycyrrizoflavanone 0.19 Isoliquitritigenin 0.16 Glycicumarin 3,50 Semilicoisoflavone B 0.01 Glycyrrizoflavone 0.69 Dehydrogliasperin C 4.09 Glycyrin 1.91 Gliasperin B 1.40 Gliasperin D 3.38 Dehydrogliasperin D 1.82 Gliurallin B 0,03 Active component: total 17.18%

(Example 8)

Licorice extract (18 g) was obtained by the same procedure as in Example 6, except that extraction was carried out using ethyl acetate instead of ethanol. The obtained licorice extract showed good PPARγ ligand activity. The content of active ingredients and glycyrrhizin contained in such licorice extract was evaluated in the same manner as in Example 6, and table 22 shows the results.

Table 22 Connection Name Content (wt.%) Glycyrrhizin 0.05 Glycyrrizoflavanone 0.18 Isoliquitritigenin 0.09 Glycicumarin 3.44 Semilicoisoflavone B 0.62 Glycyrrizoflavone 0.70 Dehydrogliasperin C 4.09 Glycyrin 1.96 Gliasperin B 1,52 Gliasperin D 3.41 Dehydrogliasperin D 1.93 Gliurallin B 0,03 Active component: total 17.97%

(Comparative example 1)

Licorice extract was obtained by the same procedure as in Example 6, however, extraction was carried out using water instead of ethanol. The obtained licorice extract did not exhibit PPARγ ligand activity.

(Example 9)

To a finely ground product (500 g) of licorice (G. uralnsis) (Kaneka Sun Spice Co., Ltd) was added 5 kg of water, and the mixture was extracted at 60 ° C. for 1 day. Then the residue was filtered and dried under reduced pressure. To the obtained pre-extracted residue was added 2.5 kg of ethanol (99.5% v / v) and then was extracted at 25 ° C. for 5 hours, and the residue was filtered to obtain an extract. The solvent was removed from the obtained extract under reduced pressure to obtain 47 g of licorice extract having a tan to dark brown color. The obtained licorice extract showed good PPARγ ligand activity. Table 23 shows the content of the active ingredients of the PPARγ ligand and glycyrrhizin contained in this licorice extract.

Table 23 Connection Name Content (wt.%) Glycyrrhizin 0.50 Glycyrrizoflavanone 0.18 Isoliquitritigenin 1.93 Glycicumarin 2.10 Semilicoisoflavone B 0.40 Glycyrrizoflavone 0.44 Dehydrogliasperin C 1.19 Glycyrin 1.44 Gliasperin B 1.19 Gliasperin D 1,59 Dehydrogliasperin D 1,03 Gliurallin B 0,03 Active component: total 11.52%

(Example 10)

Licorice extracts (25.8 g and 29.5 g) were obtained by extracting cut products (the ratio of the total surface area of the bark is about 80% of the total surface area of the licorice) licorice (G. uralensis) 99.5% (v) and 95% (v) ethanol (water content was 0.5 and 5.0% (v), respectively), respectively, using the same procedure as in Example 6. The obtained licorice extracts showed good PPARγ ligand activity. Table 24 shows the content of ingredients exhibiting the activity of PPARγ and glycyrrhizin ligands contained in these licorice extracts.

Table 24 The ethanol water content during extraction 0.5% by volume 5.0% by volume Connection Name Content (% by weight) Content (% by weight) Glycyrrhizin 0.02 0.26 Glycyrrizoflavanone 0.14 0,03 Isoliquitritigenin 0,03 0.05 Glycicumarin 3.12 2.85 Semilicoisoflavone B 0.42 0.41 Glycyrrizoflavone 0.65 0.54 Dehydrogliasperin C 7.53 4,50 Glycyrin 1.95 1.92 Gliasperin B 3.92 3.62 Gliasperin D 6.48 4.23 Dehydrogliasperin D 4.43 3.63 Gliurallin B 0.12 0.11 Active component: total 28.79% 21.89%

(Example 11)

Licorice extract was obtained from the finely divided licorice product (G. glabra) by the same procedure as in Example 6. The contents of the active ingredients and glycyrrhizin contained in such licorice extract were evaluated in the same manner as in Example 6 and table 25 results are shown.

Table 25 Connection Name Content (wt.%) Glycyrrhizin 0.27 Glycyrrizoflavanone 0.09 Isoliquitritigenin 0.12 Glabren 3.14 Glabridin 22.08 Glabrol 4.33 3'-hydroxyl-4'-O-methylglabridine 2.19 4'-O-methylglabridine 3.03 Gisaglabridin B 1.14 Active component: total 36.12%

(Example 12)

Licorice extract was obtained from finely divided licorice product (G. inflata) by the same procedure as in Example 6. The contents of the active ingredients and glycyrrhizin contained in such licorice extract were evaluated in the same manner as in Example 6 and table 26 results are shown.

Table 26 Connection Name Content (wt.%) Glycyrrhizin 0.17 Isoliquitritigenin 0.19 Lycochalcon B 2,57 Lycochalcon C 3.61 Lycochalcon A 20.34 Glycyrdione A 1.91 Glycyrdione C 0.94 Active component: total 29.56%

(Example 13)

Corn starch, lactose, carboxymethyl cellulose and magnesium stearate were mixed with licorice extract obtained in Example 6. In addition, an aqueous solution of polyvinylpyrrolidone was added to them as a binding agent, and the mixture was granulated by conventional methods. Talc was added to the product, mixed and pressed into tablets having the following composition.

Licorice extract 10 parts by weight Corn starch 25 parts by weight Lactose 15 parts by weight Carboxymethyl cellulose 10 parts by weight Magnesium stearate 3 parts by weight Polyvinylpyrrolidone 5 parts by weight Talc 10 parts by weight

Industrial applicability

According to the present invention, a ligand of a peroxisome proliferator activated receptor (PPAR) and a composition containing it can be prepared simply and efficiently. The composition of the present invention is useful for improving the state of insulin resistance or as a means for preventing and / or improving the state of insulin resistance syndrome.

Claims (33)

1. The ligand of the receptor activated by the peroxisome γ proliferator, which includes at least one compound selected from the group consisting of prenylflavonoid, which is a 3-arylcoumarin derivative of the general formula (1):

where at least one of R1-R7 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring comprising —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; isoflav-3-ene derivative of the general formula (2)

where at least one of R1-R7 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring comprising —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; isoflavan derivative of the general formula (3)

where at least one of R1-R7 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring comprising —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; isoflavanone derivative of the general formula (4)

where at least one of R1-R8 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring comprising —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; isoflavone derivative of the general formula (5)

where at least one of R1-R9 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring, SNA comprising -CH = (CH _{3) 2} O-, and the other R-groups are H, OH or OCH _3; flavonol derivative of the general formula (6)

where R1 represents OH or OCH ₃ , at least one of R2-R9 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHS (CH ₃ ) ₂ O—, and other R groups are H, OH or OCH ₃ ; flavanone derivative of the general formula (7)

where at least one of R1-R8 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; chalcone derivative of the general formula (8)

where at least one of R1-R9 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring comprising —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; or dibenzoylmethane derivative of the general formula (9)

where at least one of R1-R7 represents a prenyl group or structure, where 2 adjacent R groups form a 6-membered ring including —CH = CHC (CH ₃ ) ₂ O— and the other R groups are H, OH or OCH ₃ ; a chalcone derivative other than prenyl flavonoids, which is echinatin, isoliquirithigenin, lycochalcon A or lycochalcon B; and a flavonol derivative other than prenyl flavonoids, which is kempferol 3-O-methyl ester. 2. The ligand of the receptor activated by the peroxisome proliferator γ, according to claim 1, where the 3-arylcoumarin derivative represented by the general formula (1) is glycicumarin or glycyrin. 3. The peroxisis γ proliferator activated receptor ligand according to claim 1, wherein the isoflav-3-ene derivative represented by the general formula (2) is dehydrogliasperin D, dehydrogliasperin C, or glabrene. 4. The peroxisis γ proliferator-activated receptor ligand according to claim 1, wherein the isoflavan derivative represented by the general formula (3) is gliasperin D, glabridine, gisaglabridin B, 4'-O-methylglabridine or 3'-hydroxy-4'- O-methylglabridine. 5. The ligand of the receptor activated by the peroxisome proliferator γ, according to claim 1, where the isoflavanone derivative represented by the general formula (4) is gliasperin B or glycyrrhizoflavanone. 6. The ligand of the receptor activated by the peroxisome proliferator γ, according to claim 1, where the isoflavone derivative represented by the general formula (5) is gliuralline B, lupivitone, semilicoisoflavone B or glycyrrhizoflavone. 7. The ligand of the receptor activated by the peroxisome γ proliferator according to claim 1, wherein the flavonol derivative represented by the general formula (6) is isolicoflavonol, lycoflavonol or topazoline. 8. The ligand of the receptor activated by the peroxisome γ proliferator according to claim 1, wherein the flavanone derivative represented by the general formula (7) is glabrol. 9. The ligand of the receptor activated by the peroxisome proliferator γ, according to claim 1, where the chalcone derivative represented by the general formula (8) is lycochalcon C. 10. The ligand of the receptor activated by the peroxisome γ proliferator according to claim 1, wherein the dibenzoylmethane derivative represented by the general formula (9) is glycyrrhione A or glycyrrhione C. 11. The compound represented by formula

12. An extract obtained from licorice to improve insulin resistance or to prevent and / or improve insulin resistance syndrome, which includes a ligand of receptor activated by peroxisome proliferator γ according to claim 1, where the extract is obtained by extraction with an organic solvent, and the glycyrrhizin content is not more than than 0.5% by weight on a dry weight basis. 13. The extract according to item 12, which includes at least 5% by weight of the ligand of the receptor activated by the peroxisome proliferator γ, by dry weight. 14. The extract according to item 12, which includes at least one compound selected from the group consisting of glycycoumarin, glycyrin, dehydrogliasperin C, dehydrogliasperin D, gliasperin B and gliasperin D in an amount of not less than 0.5% by weight by dry weight. 15. The extract according to item 12, which includes glycicoumarin, glycyrin, dehydrogliasperin C, dehydrogliasperin D, gliasperin B and gliasperin D in an amount of not less than 0.5% by weight of dry weight. 16. The extract according to claim 12, which includes at least one compound selected from the group consisting of glycyrrhizoflavanone, glycyrrhizoflavone, gliuralline B, semilicoisoflavone B and isoliquiritigenin in an amount of not less than 0.01% by weight on a dry weight basis. 17. The extract according to item 12, which includes glycyrrhizoflavanone, glycyrrhizoflavon, gliuralin B, semilicoisoflavon B and isoliquiritigenin in an amount of not less than 0.01% by weight of dry weight. 18. The extract according to item 12, which includes at least one compound selected from the group consisting of glabren, glabridin, glabrol, 3'-hydroxy-4'-O-methylglabridine, 4'-O-methylglabridine and gisaglabridin In an amount of not less than 0.5% by weight of dry weight. 19. The extract according to item 12, which includes glabren, glabridin, glabrol, 3'-hydroxy-4'-O-methylglabridine, 4'-O-methylglabridine and gisaglabridin B in an amount of not less than 0.5% by weight dry weight. 20. The extract according to item 12, which includes at least one compound selected from the group consisting of lycochalcone A, lycochalcone B, lycochalcone C, glycyrrhione A and glycyrrhione C in an amount of not less than 0.5% by weight dry weight. 21. The extract according to item 12, which includes lycochalcon A, lycochalcon B, lycochalcon C, glycyrrhione A and glycyrrhidion C in an amount of not less than 0.5% by weight of dry weight. 22. The extract according to item 12, where the content of glycyrrhizin is 0.001-0.5% by weight by dry weight. 23. The extract of claim 12, wherein the licorice is at least one compound selected from the group consisting of Glycyrrhiza uralensis, Glycyrrhiza glabra, Glycyrrhiza inflata and Glycyrrhiza echinata. 24. The extract of claim 12, wherein the licorice is Glycyrrhiza uralensis. 25. The extract of claim 12, wherein the licorice is Glycyrrhiza glabra. 26. The extract of claim 12, wherein the licorice is Glycyrrhiza inflata. 27. A composition for improving the state of insulin resistance or for preventing and / or improving the condition of insulin resistance syndrome, which comprises as an active ingredient an effective amount of a ligand receptor activated by the peroxisome proliferator y according to claim 1. 28. The composition according to item 27, where the insulin resistance syndrome is at least one symptom or disease selected from the group consisting of hyperinsulinemia, diabetes mellitus, dyslipidemia, obesity, hypertension and arteriosclerosis. 29. The composition according to p, where the symptom or disease is diabetes. 30. The composition according to item 27, which is used as part of a food product or drink. 31. The composition according to item 27, which is used in the form of a pharmaceutical preparation. 32. The composition according to item 27, which is used for pets or pets. 33. The composition according to item 27, which is presented in the form of tablets, capsules, granules or powders.