KR101866372B1 - Pharmaceutical composition for preventing or treating obesity, comprising a compounds derived from an extract or fraction or Juglans mandshurica - Google Patents

Abstract

The present invention relates to a novel compound isolated from a spruce tree extract or a fraction thereof, and more particularly to a pharmaceutical composition for preventing and treating obesity containing the above compound as an active ingredient. It is a newly identified compound that a total of 14 active compounds isolated from the spruce extract of the present invention or fractions thereof, of which 8 compounds are novel compounds and two compounds can be separated from the spruce tree extract. The total of 14 compounds of the present invention can be used effectively as an active ingredient of anti-obesity agent because it does not significantly exhibit toxicity to cells and can significantly increase lipolysis effect in adipocytes.

Description

[0001] The present invention relates to a pharmaceutical composition for the prevention and treatment of obesity, which contains, as an active ingredient, a compound which is isolated from a spruce tree extract or a fraction thereof,

The present invention relates to a novel compound isolated from a spruce tree extract or a fraction thereof, and more particularly to a pharmaceutical composition for preventing and treating obesity containing the above compound as an active ingredient.

WHO defines obesity as a state of accumulation of fat that is ideal or excessive in location as a risk to health. Obesity is viewed as a major causative factor for a variety of chronic diseases such as diabetes, cardiovascular disease or cancer. In 2014, more than 1.9 billion people over the age of 18 are expected to be overweight. The criteria for overweight were defined as having a BMI of 25 or more, but over 600 million people were reported to have a BMI of over 30. Thus, obesity has been reported to be costly and expensive to treat and prevent worldwide, costing more than $ 200 million per year in the United States. However, there are not many drugs currently approved and sold as anti-obesity drugs. Thus, attempts have been made to increase the biocompatibility of anti-obesity agents using natural products or compounds derived therefrom for the treatment of obesity.

Spruce trees ( Juglans mandshurica Maxim.) is a medicinal plant also known as Manchurian walnut. It lives mainly in the north central part of Korea or the northeast part of China, and is used as herbal medicine. In one room, the bark is picked from spring to fall, and it is said to be dried, and it has effects such as convergence, fever, and clearing the eyes, and it is prescribed to enteritis, diarrhea, In addition, it is reported that it can be used for treatment of skin diseases, cancer and gastritis.

Studies have been made on useful active ingredients contained in the spiny woods, and reports on substances extracted from roots and bark are known. However, the compounds contained in the fruit of the spruce tree were not actively studied.

Specifically, Korean Patent Laid-Open Publication No. 10-2004-0058736 (Patent Document 001) discloses an active compound isolated from an extract of P. bark and its use as an anti-complement, and Korean Patent Registration No. 10-1292837 002) discloses the use of an active compound for the isolation of an extract of P. falciparum and its use for improving skin wrinkles. In addition, it has been known that the active compound isolated from the extract of P. bark extract inhibits HIV-1 reverse transcriptase and Rnase activity (Non-Patent Document 008).

Regarding the anti-obesity effect of spruce tree extract, it has been known that water extract of spruce tree can exhibit anti-obesity effect by controlling the activity of pancreatic lipase (Non-Patent Document 009). However, there have been no reports on other active ingredients contained in the extracts of P. thunbergii, and it is expected that the novel compounds isolated therefrom will exhibit various physiological activities, particularly anti-obesity activity.

Accordingly, the present inventors have made efforts to provide an anti-obesity agent derived from an extract of a natural product, and as a result, a total of 14 compounds were isolated from the methanol extract of P. falciparum and its active fraction. As a result of the structural analysis, it was confirmed that eight out of 14 compounds were novel compounds, and that the two compounds were structurally analyzed to be able to be isolated from P. gut extract as a known compound. In addition, the present inventors have found that the total of 14 compounds do not exhibit cytotoxicity to adipocytes and significantly increase the lipolytic effect of adipocytes. Therefore, the compounds of the present invention can be used as an effective ingredient of a pharmaceutical composition for the prevention and treatment of obesity The present invention has been completed.

KR 10-20074-0058736 A1 (July 7, 2004) KR 10-1292837 B1 (2013.07.29)

 Sy, L.-K., Brown, G. D., 1999. Coniferaldehyde derivatives from tissue culture of Artemisia annua and Tanacetum parthenium. Phytochemistry 50, 781-785.  Kouno, I., Yanagida, Y., Shimono, S., Shintomi, M., Yang, C.-S., 1992.  Wu, T.-S., Yeh, J.-H., Wu, P.-L., 1995. The heartwood constituents of Tetradium glabrifolium. Phytochemistry 40, 121-124.  Miki, K., Takehara, T., Sasaya, T., Sakakibara, A., 1980. Lignans of Larix leptolepis. Phytochemistry 19, 449-453. (Non Patent Patent 005) Li, Gao, et al. "Four new diarylheptanoids from the roots of Juglans mandshurica." Chemical and pharmaceutical bulletin 51.3 (2003): 262-264.  Liu, Lijuan, et al. "New. ALPHA.-Tetralonyl Glucosides from the Fruit of Juglans mandshurica." Chemical and pharmaceutical bulletin 52.5 (2004): 566-569.  Yao, Da Lei, et al. "Chemical constituents from the leaves of Juglans mandshurica." Archives of pharmacal research 38.4 (2015): 480-484.  Min, Byung-Sun, et al. "Inhibition of human immunodeficiency virus type 1 reverse transcriptase and ribonuclease H activities by constituents of Juglans mandshurica." Chemical and pharmaceutical bulletin 48.2 (2000): 194-200.  Han, Likun, et al. "Inhibitory effects of compounds from Juglans mandshurica on pancreatic lipase." Journal of Natural Medicines 61.2 (2007): 184-186.

Thus, the present inventors isolated 14 compounds from the methanol extract of P. vellum and the active fractions thereof. As a result of structural analysis, eight compounds out of the total 14 compounds were novel compounds, and two compounds were known compounds Can be separated from the spruce tree extract. In addition, the inventors of the present invention have confirmed that the total of 14 compounds do not exhibit cytotoxicity to adipocytes and significantly increase the lipid degradation effect of adipocytes, thereby completing the present invention.

It is therefore an object of the present invention to provide novel active compounds which are isolated from the spruce extracts or fractions thereof.

It is still another object of the present invention to provide a pharmaceutical composition for preventing and treating obesity, which comprises the above compound as an active ingredient.

Still another object of the present invention is to provide a health functional food for preventing and improving obesity comprising the above-mentioned compound as an active ingredient.

In order to achieve the above object, the present invention provides a compound represented by any one of the following formulas (1) to (3), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

(Wherein, R ₁ is hydrogen (-H), hydroxy (-OH), oxo group (= O), alkoxy, C _1- alcohols, C _1-4 alkyl, C _1-4 of C _{1-4 And} the benzene ring may be unsubstituted or substituted with one of a hydroxyl group (-OH) or an oxo group (= O);

R ₂ and R ₃ are each independently selected from the group consisting of -H, -OH, C _1-4 alkyl, C _1-4 alcohols, C _1-4 alkoxy, C _1-4 alkenes, C _1-4 alkenes Or C _1-4 aldehyde),

(2)

(Wherein, R ₁ to R ₃ are each independently selected from hydrogen (-H), hydroxy (-OH), oxo group (= O), an alcohol, C _1-4 alkyl, C _1-4 of C _1-4 C _1-4 alkenes, C _1-4 alkenes or C _1-4 aldehydes), or

(3)

.

.

The present invention also provides a process for the preparation of said compounds, which comprises the following steps i) to iii):

i) extracting the spruce tree with water, C ₁ -C ₂ lower alcohol or a mixed solvent thereof;

ii) obtaining an ethyl acetate or chloroform fraction of the extract obtained in step i); And

iii) subjecting the fraction obtained in step ii) to column chromatography to obtain a separated compound.

In one preferred embodiment of the present invention, the compound may be a compound represented by any one of the following Chemical Formulas 3 to 10:

(3)

,

,

[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),

[Chemical Formula 5]

(Wherein R is either H or OCH ₃ , and the stereochemical arrangement is any one of 8 R and 8 S )

[Chemical Formula 6]

Wherein R ₁ and R ₂ are independently or alternatively H, CH ₃ or OCH ₃ and the relative stereochemistry is either threo or erythro ,

(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 9]

, 및

, And

[Chemical formula 10]

.

.

In a preferred embodiment of the present invention, the compound may be isolated from a spruce tree extract or a fraction thereof.

In a preferred embodiment of the present invention, the extract may be an extract obtained by extracting the spruce tree with water, a C ₁ to C ₄ lower alcohol or a mixture thereof as a solvent, and the lower alcohol may be methanol or ethanol .

In a preferred embodiment of the present invention, the fraction may be a fraction prepared by additionally adding a solvent to the spruce extract, and the solvent may be water, chloroform, ethyl acetate.

The present invention also relates to a pharmaceutical composition for the prevention and treatment of obesity comprising as an active ingredient a compound represented by any one of the following formulas (1) to (3), an optical isomer thereof or a pharmaceutically acceptable salt thereof Lt; / RTI >

In addition, the present invention relates to a health functional food for obesity prevention and improvement, which comprises, as an active ingredient, a compound represented by any one of the following Chemical Formulas 1 to 3, an optical isomer thereof, or a pharmaceutically acceptable salt thereof Lt; / RTI >

[Chemical Formula 1]

(Wherein, R ₁ is hydrogen (-H), hydroxy (-OH), oxo group (= O), alkoxy, C _1- alcohols, C _1-4 alkyl, C _1-4 of C _{1-4 And} the benzene ring may be unsubstituted or substituted with one of a hydroxyl group (-OH) or an oxo group (= O);

R ₂ and R ₃ are each independently selected from the group consisting of -H, -OH, C _1-4 alkyl, C _1-4 alcohols, C _1-4 alkoxy, C _1-4 alkenes, C _1-4 alkenes Or C _1-4 aldehyde),

(2)

(Wherein, R ₁ to R ₃ are each independently selected from hydrogen (-H), hydroxy (-OH), oxo group (= O), an alcohol, C _1-4 alkyl, C _1-4 of C _1-4 C _1-4 alkenes, C _1-4 alkenes or C _1-4 aldehydes), or

(3)

.

.

It is a newly identified compound that a total of 14 active compounds isolated from the spruce extract of the present invention or fractions thereof, of which 8 compounds are novel compounds and two compounds can be separated from the spruce tree extract. The total of 14 compounds of the present invention can be used effectively as an active ingredient of anti-obesity agent because it does not significantly exhibit toxicity to cells and can significantly increase lipolysis effect in adipocytes. In particular, since the novel compounds of the present invention are biocompatible compounds derived from natural products, they can be safely applied to pharmaceutical compositions or health functional foods and are effective.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 shows the structures of the spruce-derived compounds 1 to 14 of the present invention. FIG.
FIG. 2 is a graph showing the results of the HMBC analysis and COZY (chemical shift correlated spectroscopy) analysis of the heterogeneous nuclei of Compound 1, Compound 3, and Compound 5 to Compound 7 derived from the spruce tree of the present invention .

Hereinafter, the present invention will be described in detail.

As described above, although obesity patients continue to increase, anti-obesity agents that can be safely used in the body have not yet been provided, and compounds isolated from extracts derived from natural products such as spruce trees are used as anti-obesity agents There is a continuing study on fitness for this.

It is a newly identified compound that a total of 14 active compounds isolated from the spruce extract of the present invention or fractions thereof, of which 8 compounds are novel compounds and two compounds can be separated from the spruce tree extract. The total of 14 compounds of the present invention can be used effectively as an active ingredient of anti-obesity agent because it does not significantly exhibit toxicity to cells and can significantly increase lipolysis effect in adipocytes. In particular, since the novel compounds of the present invention are biocompatible compounds derived from natural products, they can be safely applied to pharmaceutical compositions or health functional foods and are effective.

Accordingly, the present invention provides a compound represented by any one of the following formulas (1) to (3), an optical isomer thereof, or a pharmaceutically acceptable salt thereof:

[Chemical Formula 1]

(Wherein, R ₁ is hydrogen (-H), hydroxy (-OH), oxo group (= O), alkoxy, C _1- alcohols, C _1-4 alkyl, C _1-4 of C _{1-4 And} the benzene ring may be unsubstituted or substituted with one of a hydroxyl group (-OH) or an oxo group (= O);

R ₂ and R ₃ are each independently selected from the group consisting of -H, -OH, C _1-4 alkyl, C _1-4 alcohols, C _1-4 alkoxy, C _1-4 alkenes, C _1-4 alkenes Or C _1-4 aldehyde),

(2)

(Wherein, R ₁ to R ₃ are each independently selected from hydrogen (-H), hydroxy (-OH), oxo group (= O), an alcohol, C _1-4 alkyl, C _1-4 of C _1-4 C _1-4 alkenes, C _1-4 alkenes or C _1-4 aldehydes), or

(3)

.

.

The present invention also provides a process for the preparation of said compounds, which comprises the following steps i) to iii):

i) extracting the spruce tree with water, C ₁ -C ₂ lower alcohol or a mixed solvent thereof;

ii) obtaining an ethyl acetate or chloroform fraction of the extract obtained in step i); And

iii) subjecting the fraction obtained in step ii) to column chromatography to obtain a separated compound.

The "compound" of the present invention is preferably, but not limited to, a compound represented by any of the following formulas (3) to (10)

(3)

,

,

[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),

[Chemical Formula 5]

(Wherein R is either H or OCH ₃ , and the stereochemical arrangement is any one of 8 R and 8 S )

[Chemical Formula 6]

Wherein R ₁ and R ₂ are independently or alternatively H, CH ₃ or OCH ₃ and the relative stereochemistry is either threo or erythro ,

(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 9]

, 및

, And

[Chemical formula 10]

.

.

More specifically, the compounds of the present invention

A compound 1 having the structure of Formula 11: 2- (4-formyl-2-methoxyphenoxy) -propan-1,3-diol (2- (4-Formyl-2 -methoxyphenoxy) -propan- 1,3-diol): < RTI ID = 0.0 >

(11)

;

;

Compound 2 having a structure represented by the following Chemical Formula 12: 2- (4-hydroxymethyl-2-methoxyphenoxy) -propan- -1, 3-diol)

[Chemical Formula 12]

,

,

To have the structure of Formula 13], the compound 3 is the three-dimensional arrangement 8 R: (+) - 3- hydroxy-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxy (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) propan-1-

[Chemical Formula 13]

;

;

To the compound 4 has a structure of [formula 14], the relative three-dimensional arrangement is threo: threo-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -propane-1, 3-diol (threo -2- (4-hydroxy- 3- methoxyphenyl) -1- (4-hydroxyphenyl) -propan-1,3-diol) can be:

[Chemical Formula 14]

;

;

To have the structure of Formula 15], the relative three-dimensional arrangement is threo compound 5: -1- (4-hydroxyphenyl) -1-methoxy-2- (4-hydroxy-3-methoxyphenyl) -propan (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -1-methoxy-propan-

[Chemical Formula 15]

;

;

Compound 6 having the structure of the following formula 6: (7S, 8R) -3'-methoxy-4 ', 7-epoxy-8,5'-neo lignan- 7 S, 8 R) -3'- methoxy-4 ', 7-epoxy-8,5'-neolignan-4,9,7'-triol) may be:

(3)

;

;

To have the structure of Formula 7], the relative three-dimensional arrangement, and the threo, the three-dimensional arrangement is S 7, S 8 The compound 7: threo - (7S, 8S) -4,7,9- trihydroxy-1'- acrylic aldehyde -8-O-4'- neo lignans (threo - (7 S, 8 S) -4,7,9-trihydroxy-1'-acrylaldehyde-8- O -4'-neolignan) may be:

(7)

;

;

It has a structure of the above-mentioned [Chemical Formula 7], the relative three-dimensional arrangement and the erythro, the compound 8 is the three-dimensional arrangement 7 R, 8 S: erythro - (7R, 8S) -4,7,9- trihydroxy-1'- Acrylic aldehyde-8-O-4'-neo lignan (erythro- (7R, 8S) -4,7,9-trihydroxy-1'-acrylaldehyde-8-0-4'-neolignan);

To have the structure of Formula 8], and the relative three-dimensional arrangement is threo, compound of the three-dimensional arrangement has 7 S, 8 S 9: threo - (7S, 8S) -3'- methoxy -4,7,9- tree Hydroxy-1'-hydroxypropyl-8-O-4'-neo lignan (threo- (7S, 8S) -3'- methoxy-4,7,9-trihydroxy- 4'-neolignan):

[Chemical Formula 8]

;

;

It has a structure of the above-mentioned [Chemical Formula 8], and the relative three-dimensional arrangement is erythro, the three-dimensional arrangement 7 R, 8 S of compound 10: erythro - (7R, 8S) -3'- methoxy -4,7,9- tree hydroxy-1'-hydroxypropyl -8-O-4'- neo lignans (erythro- (7 R, 8 S ) -3'- methoxy -4,7,9-trihydroxy-1'-hydroxypropyl-8- O -4 ' -neolignan).

Compound 1 to compound 14 of the present invention, Azusa (Juglans mandshurica Maxim.) extract or fractions thereof. The spruce tree may be any of bark, root, leaf, flower or fruit, and more preferably fruit. In addition, the spruce tree can be used without limitation such as cultivated or commercially available.

Preferably, the extract is extracted with water, C ₁ to C ₄ lower alcohols or a mixture thereof as a solvent, and the lower alcohol is preferably ethanol, methanol or butanol.

The extract is preferably, but not exclusively, prepared by a process comprising the steps of:

1) extracting the spruce tree by adding an extraction solvent;

2) filtering the extract of step 1); And

3) Concentrating the filtered extract of step 2) under reduced pressure and drying.

In the above method, the extraction method of the spruce tree extract may be a conventional method in the art such as filtration, hot water extraction, immersion extraction, reflux cooling extraction and ultrasonic extraction.

In the above method, it is preferable to use a vacuum decompression concentrator or a vacuum rotary evaporator for the decompression concentration in step 3), but it is not limited thereto. The drying is preferably performed under reduced pressure, vacuum drying, boiling, spray drying or freeze drying, but not always limited thereto.

Preferably, the fraction is a fraction prepared by addition of an organic solvent to the extract of spruce tree. The organic solvent is preferably one or more selected from the group consisting of chloroform, hexane, ethyl acetate and butanol, Chloroform and / or ethyl acetate is more preferable, but is not limited thereto.

In a specific embodiment of the present invention, we prepared methanol extracts and fractions from the spruce tree fruit, from which a total of 14 compounds 1 to 14 were isolated. As a result of analyzing the structures of the total of 14 compounds of the above-mentioned compounds 1 to 14, it can be seen that the compounds 1 to 8 of the present invention are novel compounds which have no known structure and characteristic in the art, It was confirmed that this compound is a compound which is not known in the art as a compound which can be separated from the spruce tree (Fig. 1, Fig. 2, and Tables 1 to 3).

The compound of the present invention can be used in the form of a pharmaceutically acceptable salt. As the salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts include those derived from inorganic acids such as hydrochloric acid, nitric acid, phosphoric acid, sulfuric acid, hydrobromic acid, hydroiodic acid, nitrous acid or phosphorous acid, and aliphatic mono- and dicarboxylates, phenyl-substituted alkanoates, hydroxyalkanoates, Dioleate, aromatic acid, aliphatic and aromatic sulfonic acids. Such pharmaceutically innocuous salts include, but are not limited to, sulfate, pyrosulfate, bisulfate, sulfite, bisulfite, nitrate, phosphate, monohydrogenphosphate, dihydrogenphosphate, metaphosphate, pyrophosphate chloride, bromide, Butyrate, caprate, heptanoate, propiolate, oxalate, malonate, succinate, succinate, maleic anhydride, maleic anhydride, , Sebacate, fumarate, maleate, butyne-1,4-dioate, hexane-1,6-dioate, benzoate, chlorobenzoate, methylbenzoate, dinitrobenzoate, hydroxybenzoate, Methoxybenzoate, phthalate, terephthalate, benzene sulfonate, toluene sulfonate, chlorobenzene sulfide Propyl sulphonate, naphthalene-1-yne, xylenesulfonate, phenylsulfate, phenylbutyrate, citrate, lactate,? -Hydroxybutyrate, glycolate, maleate, Sulfonate, naphthalene-2-sulfonate or mandelate.

The acid addition salt according to the present invention can be obtained by a conventional method, for example, by dissolving any one of the above-mentioned chemical formulas 1 to 3 in an excess amount of an acid aqueous solution, and mixing the salt with a water- For example, by precipitation using methanol, ethanol, acetone or acetonitrile. Or by heating the acid or alcohol in the same amount of the compound and water and then evaporating and drying the mixture or by suction filtration of the precipitated salt.

In addition, bases can be used to make pharmaceutically acceptable metal salts. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess amount of an alkali metal hydroxide or an alkaline earth metal hydroxide solution, filtering the insoluble compound salt, and evaporating and drying the filtrate. At this time, it is preferable for the metal salt to produce sodium, potassium or calcium salt. The corresponding silver salt is also obtained by reacting an alkali metal or alkaline earth metal salt with a suitable salt (such as silver nitrate).

In addition, the compound of any one of the above formulas (1) to (3) of the present invention includes not only pharmaceutically acceptable salts, but also all salts, hydrates and solvates which can be prepared by conventional methods .

The addition salt according to the present invention can be prepared by a conventional method, for example, by reacting a compound of any one of Chemical Formulas 1 to 3 with a water-miscible organic solvent such as acetone, methanol, ethanol, Nitrile, etc., adding an excess amount of an organic acid, or adding an aqueous acid solution of an inorganic acid, followed by precipitation or crystallization. Subsequently, in this mixture, a solvent or an excess acid is evaporated and dried to obtain an additional salt, or the precipitated salt may be produced by suction filtration.

The present invention also relates to a pharmaceutical composition for the prevention and treatment of obesity comprising as an active ingredient a compound represented by any one of the following formulas (1) to (3), an optical isomer thereof or a pharmaceutically acceptable salt thereof Lt; / RTI >

[Chemical Formula 1]

(Wherein, R ₁ is hydrogen (-H), hydroxy (-OH), oxo group (= O), alkoxy, C _1- alcohols, C _1-4 alkyl, C _1-4 of C _{1-4 And} the benzene ring may be unsubstituted or substituted with one of a hydroxyl group (-OH) or an oxo group (= O);

R ₂ and R ₃ are each independently selected from the group consisting of -H, -OH, C _1-4 alkyl, C _1-4 alcohols, C _1-4 alkoxy, C _1-4 alkenes, C _1-4 alkenes Or C _1-4 aldehyde),

(2)

(Wherein, R ₁ to R ₃ are each independently selected from hydrogen (-H), hydroxy (-OH), oxo group (= O), an alcohol, C _1-4 alkyl, C _1-4 of C _1-4 C _1-4 alkenes, C _1-4 alkenes or C _1-4 aldehydes), or

(3)

.

.

The "compound" of the present invention is preferably, but not limited to, a compound represented by any of the following formulas (3) to (10)

(3)

,

,

[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),

[Chemical Formula 5]

(Wherein R is either H or OCH ₃ , and the stereochemical arrangement is any one of 8 R and 8 S )

[Chemical Formula 6]

Wherein R ₁ and R ₂ are independently or alternatively H, CH ₃ or OCH ₃ and the relative stereochemistry is either threo or erythro ,

(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 9]

, 및

, And

[Chemical formula 10]

.

.

More specifically, the compounds of the present invention

A compound 1 having the structure of Formula 11: 2- (4-formyl-2-methoxyphenoxy) -propan-1,3-diol (2- (4-Formyl-2 -methoxyphenoxy) -propan- 1,3-diol): < RTI ID = 0.0 >

(11)

;

;

Compound 2 having a structure represented by the following Chemical Formula 12: 2- (4-hydroxymethyl-2-methoxyphenoxy) -propan- -1, 3-diol)

[Chemical Formula 12]

,

,

To have the structure of Formula 13], the compound 3 is the three-dimensional arrangement 8 R: (+) - 3- hydroxy-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxy (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) propan-1-

[Chemical Formula 13]

;

;

To the compound 4 has a structure of [formula 14], the relative three-dimensional arrangement is threo: threo-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -propane-1, 3-diol (threo -2- (4-hydroxy- 3- methoxyphenyl) -1- (4-hydroxyphenyl) -propan-1,3-diol) can be:

[Chemical Formula 14]

;

;

To have the structure of Formula 15], the relative three-dimensional arrangement is threo compound 5: -1- (4-hydroxyphenyl) -1-methoxy-2- (4-hydroxy-3-methoxyphenyl) -propan (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -1-methoxy-propan-

[Chemical Formula 15]

;

;

Compound 6 having the structure of the following formula 6: (7S, 8R) -3'-methoxy-4 ', 7-epoxy-8,5'-neo lignan- 7 S, 8 R) -3'- methoxy-4 ', 7-epoxy-8,5'-neolignan-4,9,7'-triol) may be:

(3)

;

;

To have the structure of Formula 7], the relative three-dimensional arrangement, and the threo, the three-dimensional arrangement is S 7, S 8 The compound 7: threo - (7S, 8S) -4,7,9- trihydroxy-1'- acrylic aldehyde -8-O-4'- neo lignans (threo - (7 S, 8 S) -4,7,9-trihydroxy-1'-acrylaldehyde-8- O -4'-neolignan) may be:

(7)

;

;

It has a structure of the above-mentioned [Chemical Formula 7], the relative three-dimensional arrangement and the erythro, the compound 8 is the three-dimensional arrangement 7 R, 8 S: erythro - (7R, 8S) -4,7,9- trihydroxy-1'- Acrylic aldehyde-8-O-4'-neo lignan (erythro- (7R, 8S) -4,7,9-trihydroxy-1'-acrylaldehyde-8-0-4'-neolignan);

To have the structure of Formula 8], and the relative three-dimensional arrangement is threo, compound of the three-dimensional arrangement has 7 S, 8 S 9: threo - (7S, 8S) -3'- methoxy -4,7,9- tree Hydroxy-1'-hydroxypropyl-8-O-4'-neo lignan (threo- (7S, 8S) -3'- methoxy-4,7,9-trihydroxy- 4'-neolignan):

[Chemical Formula 8]

;

;

It has a structure of the above-mentioned [Chemical Formula 8], and the relative three-dimensional arrangement is erythro, the three-dimensional arrangement 7 R, 8 S of compound 10: erythro - (7R, 8S) -3'- methoxy -4,7,9- tree hydroxy-1'-hydroxypropyl -8-O-4'- neo lignans (erythro- (7 R, 8 S ) -3'- methoxy -4,7,9-trihydroxy-1'-hydroxypropyl-8- O -4 '-neolignan);

Compound 11 having the structure of the following formula (9): (2-glyceryl) -O-coniferaldehyde)

[Chemical Formula 9]

;

;

Compounds having the structure of the following formula 12: 2- [4- (3-Hydroxypropyl) -2-methoxyphenoxy] -1,3-propanediol ) -2-methoxyphenoxy] -1,3-propanediol)

[Chemical formula 10]

;

;

To have the structure of Formula 16], a compound the three-dimensional arrangement 13 is 8 S: (-) - morpholine EVO B be ((- -) evofolin B) are:

[Chemical Formula 16]

; 및

; And

To the compound 14 has the structure of Formula 17], the relative three-dimensional arrangement is erythro: 1,2-bis- (4-hydroxy-3-methoxyphenyl) -propane-1,3-diol (1,2- bis- (4-hydroxy-3-67 methoxyphenyl) -propane-1,3-diol).

[Chemical Formula 17]

.

.

Compound 1 to compound 14 of the present invention, Azusa (Juglans mandshurica Maxim.) extract or fractions thereof.

Preferably, the extract is extracted with water, C ₁ to C ₄ lower alcohols or a mixture thereof as a solvent, and the lower alcohol is preferably ethanol, methanol or butanol.

Preferably, the fraction is a fraction prepared by addition of an organic solvent to the extract of spruce tree. The organic solvent is preferably one or more selected from the group consisting of chloroform, hexane, ethyl acetate and butanol, Chloroform and / or ethyl acetate is more preferable, but is not limited thereto.

In another specific example of the present invention, the present inventors confirmed anti-obesity effects on the compounds 1 to 14 isolated from the spruce tree extract or fractions thereof. As a result, the compounds 1 to 14 showed a toxicity to the cells (Table 4). In addition, the lipid-degrading effect in adipocytes can be significantly increased without being expressed as adipocytes.

Therefore, the compounds 1 to 14 of the present invention can be effectively used as an active ingredient of an anti-obesity agent.

When the composition of the present invention is used as a medicine, a pharmaceutical composition containing, as an active ingredient, any one or more selected from the group consisting of the compounds 1 to 14 of the present invention, or a pharmaceutically acceptable salt thereof, May be formulated and administered in a variety of oral or parenteral dosage forms such as, but not limited to,

The compositions of the present invention may be of various oral or parenteral formulations. When formulating the composition, one or more buffers (e.g., saline or PBS), antioxidants, bacteriostats, chelating agents (e.g., EDTA or glutathione), fillers, extenders, binders, adjuvants Aluminum hydroxide), suspending agents, thickening agents, disintegrating agents or surfactants, diluents or excipients.

Solid formulations for oral administration include tablets, pills, powders, granules, capsules and the like, which may contain at least one excipient, such as starch (cornstarch, wheat starch, rice starch, potatoes Starch and the like), calcium carbonate, sucrose, lactose, dextrose, sorbitol, mannitol, xylitol, erythritol maltitol, cellulose, methylcellulose, sodium carboxymethylcellulose and hydroxypropylmethylcellulose - It is prepared by mixing cellulose or gelatin. For example, tablets or tablets may be obtained by combining the active ingredient with a solid excipient, then milling it, adding suitable auxiliaries, and processing the mixture into granules.

In addition to simple excipients, lubricants such as magnesium stearate, talc, and the like may also be used. Liquid preparations for oral administration include suspensions, solutions, emulsions or syrups. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances or preservatives are included . In addition, crosslinked polyvinylpyrrolidone, agar, alginic acid, or sodium alginate may optionally be added as a disintegrant, and may further include an anticoagulant, a lubricant, a wetting agent, a flavoring agent, an emulsifying agent and an antiseptic agent .

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations or suppositories. Examples of non-aqueous solvents and suspensions include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate, and the like. As a base for suppositories, witepsol, macrogol, tween 61, cacao paper, laurin, glycerol, gelatin and the like can be used.

The composition of the present invention may be administered orally or parenterally, and may be administered externally for parenteral administration; Intraperitoneal, rectal, intravenous, intramuscular, subcutaneous, intrauterine, or intracerebral injection; Percutaneous administration agents; Or in the form of a nasal inhaler, according to methods known in the art.

In the case of such injections, they must be sterilized and protected against contamination of microorganisms such as bacteria and fungi. Examples of suitable carriers for injectables include, but are not limited to, solvents or dispersion media containing water, ethanol, polyols (e.g., glycerol, propylene glycol and liquid polyethylene glycol, etc.), mixtures thereof and / or vegetable oils . More preferably, suitable carriers include isotonic solutions such as Hanks' solution, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, or sterile water for injection, 10% ethanol, 40% propylene glycol and 5% dextrose Etc. may be used. In order to protect the injection from microbial contamination, various antibacterial and antifungal agents such as parabens, chlorobutanol, phenol, sorbic acid, thimerosal and the like may be further included. In addition, the injections may in most cases additionally include isotonic agents, such as sugars or sodium chloride.

Examples of transdermal dosage forms include ointments, creams, lotions, gels, solutions for external use, pastes, liniments, and air lozenges. In the above, transdermal administration means that the pharmaceutical composition is locally administered to the skin, so that an effective amount of the active ingredient contained in the pharmaceutical composition is delivered into the skin.

In the case of an inhalation dosage form, the compounds used according to the invention can be formulated into a pressurized pack or a pressurized pack using a suitable propellant, for example dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, carbon dioxide or other suitable gases. It can be conveniently delivered in the form of an aerosol spray from a nebulizer. In the case of a pressurized aerosol, the dosage unit may be determined by providing a valve that delivers a metered amount. For example, gelatin capsules and cartridges for use in an inhaler or insufflator may be formulated to contain a compound, and a powder mixture of a suitable powder base such as lactose or starch. Formulations for parenteral administration are described in Remington's Pharmaceutical Science, 15th Edition, 1975. Mack Publishing Company, Easton, Pennsylvania 18042, Chapter 87: Blaug, Seymour, commonly known in all pharmaceutical chemistries.

The composition of the present invention is administered in a pharmaceutically effective amount. In the present invention, " pharmaceutically effective amount " means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level will depend on the type of disease, severity, , Sensitivity to the drug, time of administration, route of administration and rate of release, duration of treatment, factors including co-administered drugs, and other factors well known in the medical arts. The composition of the present invention can be administered as an individual therapeutic agent or in combination with other therapeutic agents, and can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. That is, the total effective amount of the composition of the present invention may be administered to a patient in a single dose and administered by a fractionated treatment protocol administered over a long period in multiple doses . It is important to take into account all of the above factors and to administer the amount in which the maximum effect can be obtained in a minimal amount without side effects, which can be easily determined by those skilled in the art.

The dosage of the pharmaceutical composition of the present invention varies depending on the patient's body weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. The daily dose is preferably such that it is preferably administered in an amount of 0.01 to 50 mg, more preferably 0.1 to 30 mg per kg of body weight per day on the basis of the compound during parenteral administration, and when administered orally, May be administered in one to several divided doses so as to be preferably administered in an amount of 0.01 to 100 mg, more preferably 0.01 to 10 mg per kg of body weight per day. However, the dosage may be varied depending on the route of administration, the severity of obesity, sex, weight, age, etc. Therefore, the dosage is not limited to the scope of the present invention by any means.

The composition of the present invention may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers.

The pharmaceutical composition of the present invention can also be provided as a formulation of an external preparation. When the pharmaceutical composition for preventing and treating obesity of the present invention is used as an external preparation for skin, it may further contain at least one selected from the group consisting of fatty substances, organic solvents, solubilizers, thickeners and gelling agents, softeners, antioxidants, suspending agents, stabilizers, foaming agents, Surfactants, water, ionic emulsifiers, nonionic emulsifiers, fillers, sequestering agents, chelating agents, preservatives, vitamins, blocking agents, wetting agents, essential oils, dyes, pigments, hydrophilic active agents, Such as lipid vesicles, and any other ingredients conventionally used in dermatologic external preparations. The components can also be introduced in amounts commonly used in the field of dermatology.

When the pharmaceutical composition for preventing and treating obesity of the present invention is provided as an external preparation for skin, it may be a formulation such as ointments, patches, gels, creams or sprays.

Further, the present invention relates to a health functional food for improving obesity prevention, which comprises, as an active ingredient, a compound represented by any one of the following Chemical Formulas 1 to 3, an optical isomer thereof, or a pharmaceutically acceptable salt thereof: to provide:

[Chemical Formula 1]

(Wherein, R ₁ is hydrogen (-H), hydroxy (-OH), oxo group (= O), alkoxy, C _1- alcohols, C _1-4 alkyl, C _1-4 of C _{1-4 And} the benzene ring may be unsubstituted or substituted with one of a hydroxyl group (-OH) or an oxo group (= O);

R ₂ and R ₃ are each independently selected from the group consisting of -H, -OH, C _1-4 alkyl, C _1-4 alcohols, C _1-4 alkoxy, C _1-4 alkenes, C _1-4 alkenes Or C _1-4 aldehyde),

(2)

(Wherein, R ₁ to R ₃ are each independently selected from hydrogen (-H), hydroxy (-OH), oxo group (= O), an alcohol, C _1-4 alkyl, C _1-4 of C _1-4 C _1-4 alkenes, C _1-4 alkenes or C _1-4 aldehydes), or

(3)

.

.

The "compound" of the present invention is preferably, but not limited to, a compound represented by any of the following formulas (3) to (10)

(3)

,

,

[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),

[Chemical Formula 5]

(Wherein R is either H or OCH ₃ , and the stereochemical arrangement is any one of 8 R and 8 S )

[Chemical Formula 6]

Wherein R ₁ and R ₂ are independently or alternatively H, CH ₃ or OCH ₃ and the relative stereochemistry is either threo or erythro ,

(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),

[Chemical Formula 9]

, 및

, And

[Chemical formula 10]

.

.

More specifically, the compounds of the present invention

A compound 1 having the structure of Formula 11: 2- (4-formyl-2-methoxyphenoxy) -propan-1,3-diol (2- (4-Formyl-2 -methoxyphenoxy) -propan- 1,3-diol): < RTI ID = 0.0 >

(11)

;

;

Compound 2 having a structure represented by the following Chemical Formula 12: 2- (4-hydroxymethyl-2-methoxyphenoxy) -propan- -1, 3-diol)

[Chemical Formula 12]

,

,

To have the structure of Formula 13], the compound 3 is the three-dimensional arrangement 8 R: (+) - 3- hydroxy-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxy (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) propan-1-

[Chemical Formula 13]

;

;

To the compound 4 has a structure of [formula 14], the relative three-dimensional arrangement is threo: threo-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -propane-1, 3-diol (threo -2- (4-hydroxy- 3- methoxyphenyl) -1- (4-hydroxyphenyl) -propan-1,3-diol) can be:

[Chemical Formula 14]

;

;

To have the structure of Formula 15], the relative three-dimensional arrangement is threo compound 5: -1- (4-hydroxyphenyl) -1-methoxy-2- (4-hydroxy-3-methoxyphenyl) -propan (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -1-methoxy-propan-

[Chemical Formula 15]

;

;

Compound 6 having the structure of the following formula 6: (7S, 8R) -3'-methoxy-4 ', 7-epoxy-8,5'-neo lignan- 7 S, 8 R) -3'- methoxy-4 ', 7-epoxy-8,5'-neolignan-4,9,7'-triol) may be:

(3)

;

;

To have the structure of Formula 7], the relative three-dimensional arrangement, and the threo, the three-dimensional arrangement is S 7, S 8 The compound 7: threo - (7S, 8S) -4,7,9- trihydroxy-1'- acrylic aldehyde -8-O-4'- neo lignans (threo - (7 S, 8 S) -4,7,9-trihydroxy-1'-acrylaldehyde-8- O -4'-neolignan) may be:

(7)

;

;

It has a structure of the above-mentioned [Chemical Formula 7], the relative three-dimensional arrangement and the erythro, the compound 8 is the three-dimensional arrangement 7 R, 8 S: erythro - (7R, 8S) -4,7,9- trihydroxy-1'- Acrylic aldehyde-8-O-4'-neo lignan (erythro- (7R, 8S) -4,7,9-trihydroxy-1'-acrylaldehyde-8-0-4'-neolignan);

To have the structure of Formula 8], and the relative three-dimensional arrangement is threo, compound of the three-dimensional arrangement has 7 S, 8 S 9: threo - (7S, 8S) -3'- methoxy -4,7,9- tree Hydroxy-1'-hydroxypropyl-8-O-4'-neo lignan (threo- (7S, 8S) -3'- methoxy-4,7,9-trihydroxy- 4'-neolignan):

[Chemical Formula 8]

;

;

It has a structure of the above-mentioned [Chemical Formula 8], and the relative three-dimensional arrangement is erythro, the three-dimensional arrangement 7 R, 8 S of compound 10: erythro - (7R, 8S) -3'- methoxy -4,7,9- tree hydroxy-1'-hydroxypropyl -8-O-4'- neo lignans (erythro- (7 R, 8 S ) -3'- methoxy -4,7,9-trihydroxy-1'-hydroxypropyl-8- O -4 '-neolignan);

Compound 11 having the structure of the following formula (9): (2-glyceryl) -O-coniferaldehyde)

[Chemical Formula 9]

;

;

Compounds having the structure of the following formula 12: 2- [4- (3-Hydroxypropyl) -2-methoxyphenoxy] -1,3-propanediol ) -2-methoxyphenoxy] -1,3-propanediol)

[Chemical formula 10]

;

;

To have the structure of Formula 16], a compound the three-dimensional arrangement 13 is 8 S: (-) - morpholine EVO B be ((- -) evofolin B) are:

[Chemical Formula 16]

; 및

; And

To the compound 14 has the structure of Formula 17], the relative three-dimensional arrangement is erythro: 1,2-bis- (4-hydroxy-3-methoxyphenyl) -propane-1,3-diol (1,2- bis- (4-hydroxy-3-67 methoxyphenyl) -propane-1,3-diol).

[Chemical Formula 17]

.

.

Compound 1 to compound 14 of the present invention, Azusa (Juglans mandshurica Maxim.) extract or fractions thereof.

Preferably, the extract is extracted with water, C ₁ to C ₄ lower alcohols or a mixture thereof as a solvent, and the lower alcohol is preferably ethanol, methanol or butanol.

Preferably, the fraction is a fraction prepared by addition of an organic solvent to the extract of spruce tree. The organic solvent is preferably one or more selected from the group consisting of chloroform, hexane, ethyl acetate and butanol, Chloroform and / or ethyl acetate is more preferable, but is not limited thereto.

The total of 14 compounds of the present invention can be used as an effective ingredient of a health functional food for prevention and improvement of obesity since it does not significantly exhibit toxicity to cells and can significantly increase the lipid degradation effect in adipocytes.

The food composition according to the present invention can be prepared in various forms according to a conventional method known in the art. Common foods include but are not limited to beverages (including alcoholic beverages), fruits and processed foods (eg, canned fruits, jam, maamalade, etc.), fish, meat and processed foods (Eg butter, chewing), edible vegetable oil, margarine (such as corn oil, etc.), breads and noodles (eg udon, buckwheat noodles, ramen noodles, spaghetti, macaroni, etc.), juice, various drinks, cookies, , Vegetable protein, retort food, frozen food, various kinds of seasoning (e.g., miso, soy sauce, sauce, etc.). The nutritional supplement may be prepared by adding the compound of the present invention to capsules, tablets, rings and the like. The health functional food is not limited thereto. For example, the compound of the present invention itself may be prepared in the form of tea, juice, and drink to be liquefied, granulated, encapsulated, can do. In order to use the compound of the present invention in the form of a food additive, it may be prepared in the form of powder or concentrate. In addition, the compound of the present invention can be mixed with a known active ingredient known to have an effect of preventing and / or improving obesity in the form of a composition.

When the compound of the present invention is used as a health drink, the health beverage composition may contain various flavors or natural carbohydrates as an additional ingredient such as ordinary beverages. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose; Polysaccharides such as dextrin, cyclodextrin; Xylitol, sorbitol, erythritol, and the like. Sweeteners include natural sweeteners such as tau Martin and stevia extract; Synthetic sweetening agents such as saccharin and aspartame, and the like can be used. The ratio of the natural carbohydrate is generally about 0.01 to 0.04 g, preferably about 0.02 to 0.03 g per 100 mL of the composition of the present invention.

In addition, the compound of the present invention may be contained as an active ingredient of a food composition for prevention and improvement of obesity. The amount thereof is not particularly limited as long as it is effective for achieving an obesity-preventing and improving action, It is preferably 0.01 to 100% by weight. The food composition of the present invention can be prepared together with other active ingredients known to be effective in compositions for preventing and improving obesity together with the compounds.

In addition to the above, the health food of the present invention may contain various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acid, protective colloid thickener, pH adjusting agent, Glycerin, an alcohol or a carbonating agent, and the like. In addition, the health food of the present invention may contain natural fruit juice, fruit juice drink, or pulp for the production of vegetable drinks. These components may be used independently or in combination. The proportion of such additives is not critical, but is generally selected in the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the composition of the present invention.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these examples are for illustrative purposes only and that the scope of the present invention is not construed as being limited by these examples.

Spruce tree extract and its From the fraction  Isolation of active compounds

≪ 1-1 > Preparation of Methanol Extract and Fractions of Spruce Tree

To isolate the compounds of the present invention, extracts of spruce wood were prepared.

Specifically, P. thunbergii was purchased from the Kyungdong Pharmaceutical Market in Seoul, Korea in 2014. 10 kg of the obtained pine nut was immersed in 5 L, ultrasonically extracted for 4 hours, and the extract was filtered. The solvent obtained by filtration was concentrated under reduced pressure in vacuum to obtain 200.0 g of spruce methanol extract.

The obtained methanol extract of spruce wood was suspended in distilled water, and an equal amount of chloroform was added thereto. The mixture was shaken to obtain a chloroform fraction, which was concentrated under reduced pressure to obtain 17.5 g of a chloroform fraction (JM1). Ethyl acetate was further added to the distilled water and shaken to obtain an ethyl acetate fraction, which was concentrated under reduced pressure to obtain 45.7 g of a fraction (JM2). Then, finally, the remaining aqueous layer was concentrated under reduced pressure to obtain 78.6 g of a water fraction (JM3).

<1-2> Spruce tree extract and its From the fraction  Isolation of active ingredient

The useful active components were separated from the methanol extract of spruce wood and its chloroform, ethyl acetate and water fractions. The chromatographic equipment used to separate the active ingredients is as follows. For separation HPLC, an AGILENT 1200 HPLC system was used. Column chromatography used a silica-gel column (Kieselgel 60, 70-230 mesh and 230-400 mesh, Merck) or YMC RP-18 resin (30-50 μm, Fujisilisa Chemical Ltd.). In the thin film chromatography (TLC) run, silica-gel 60 F254 TLC plates (0.25 mm, Merck) and RP-18 F254S plates (0.25 mm, Merck) were used.

Specifically, 17.5 g of the chloroform fraction (JM1) obtained in the above Example <1-1> was separated by chromatography on a silica gel column. At this time, the mobile phase solvent was concentration-graded to hexane: acetone at a ratio of 1: 1 (v / v) at 20: 1 (v / v) to obtain a total of five fractions. The fractions were JM1A (3.4 g), JM1B 4.1 g), JM1C (2.2 g), JM1D (1.2 g) and JM1E (2.5 g).

Of these, the JM1C fraction was loaded on a silica gel column, and 5 small fractions were eluted using a mobile phase solvent in which chloroform: methanol was mixed at a ratio of 5: 1 (v / v), and each of the JM1C1 (0.4 g), JM1C2 (0.3 g), JM1C3 (0.8 g), JM1C4 (0.8 g) and JM1C5 (0.3 g). Then, given the re-flow the JM1C2 small fraction, loaded on a HPLC equipped with a J'sphere ODS H-80 column (250 ㎜ × 20 ㎜), and a 22% aqueous solution of MeCN at a flow rate of 3 ㎖ / min and finally the compound 1 (4.3 mg), compound 11 (5.1 mg) and compound 13 (4.0 mg). Further, the above JM1C3 sub-fraction was loaded onto HPLC under the same conditions as above to give Compound 3 (6.3 mg), Compound 6 (3.7 mg), Compound 9 (3.5 mg) and Compound 10 (5.2 mg). Further, the JM1C4 sub-fraction was loaded onto HPLC under the same conditions as above to obtain Compound 7 (2.7 mg), Compound 8 (7.3 mg) and Compound 14 (10.3 mg). Further, the JM1C5 fraction was loaded onto HPLC equipped with a J'sphere ODS H-80 column (250 mm x 20 mm), and a 15% aqueous MeCN solution was flowed at a flow rate of 3 ml / min to finally obtain Compound 4 5.7 mg) and compound 12 (8.3 mg).

Further, 45.7 g of the ethyl acetate fraction (JM2) obtained in the above Example <1-1> was separated by chromatography on a silica gel column. At this time, the mobile phase solvent was concentration-graded so that the chloroform: methanol was 1: 1 (v / v) at 20: 1 (v / v) to obtain a total of five fractions. JM2A (7.0 g), JM2B 6.3 g), JM2C (5.8 g), JM2D (4.3 g) and JM2E (8.2 g).

The JM2C fraction was loaded on a YMC RP-18 column and eluted with a mobile phase solvent mixture of acetone: water 1: 1 (v / v) to yield 1.6 g of JM2Cl JM2C2 (2.3 g). Then, given the re-flow the JM2C1 small fraction, loaded on a HPLC equipped with a J'sphere ODS H-80 column (250 ㎜ × 20 ㎜), and a 15% aqueous solution of MeCN at a flow rate of 3 ㎖ / min and finally the compound 2 (2.3 mg) and compound 5 (3.3 mg).

As a result, as shown in Fig. 1, a total of 14 compounds were isolated from the methanol extract of spruce and fractions thereof. Of these, a total of eight compounds of Compound 1 to Compound 8 are novel compounds, and a total of six compounds of Compound 9 to Compound 14 were known compounds. In addition, a total of four compounds of compounds 11 to 14 in known compounds were known to be able to separate from spruce tree extracts through previous studies, and compounds 9 and 10 could be isolated from spruce extracts Lt; / RTI &gt;

Structural characterization of isolated compounds

Experimental equipments and devices used to isolate the compounds isolated from the spruce extracts and to identify their structural characteristics are as follows. Chemical shifts were analyzed in parts per million (ppm) using TMS. NMR spectra were analyzed using hydrogen and carbon, and analyzed using an Agilent 400-MR-NMR spectrophotometer at 400 MHz and 100 MHz, respectively. The results were analyzed using the MestReNova ver.6.0.2 program. HS-ESI-MS spectral analysis was obtained using an AGILENT 6550 iFunnel Q-TOF LC / MS system. Optical rotations of the compounds were measured using a Jasco DIP-370 automatic polarimeter.

<2-1> Compound 1: 2- (4-Formyl-2-methoxyphenoxy) -propan-1,3- diol)

(11)

.

.

Compound 1 of Formula 11 is in the form of a white amorphous powder, and the result of the structural analysis is as follows: _{^{[α] 20 D: + 10.9}} (MeOH, c = 0.2); C ₁₁ H ₁₄ O ₅ , HR-ESI-MS m / z : 227.0912 [M + H] ^{+ (} calcd for C ₁₁ H ₁₅ O ₅ , 227.0919); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 1 below.

^{It was} confirmed through the ¹ H NMR spectrum that the three aromatic protons were located at δ _H 7.46 (s), 7.23 (d, J = 8.2 Hz) and 7.50 (d, J = 8.2 Hz), and one methoxyproton Was located at δ _H 3.89 (3H, s), confirming that one aldehyde proton was located at δ _H 9.79 (s). ^It was confirmed through ¹³ C NMR and DEPT spectra that a total of 10 carbons were contained, containing one aldehyde, three quaternary carbons, four methines, one methylene and one methoxy carbon (Table 1). NMR results of Compound 1 were similar to (2-glyceryl) -O- coniferaldehyde (Non-Patent Document 001), and it was confirmed that the compound had a structure showing only difference in the aldehyde group of C-4. H-7 (δ _H 9.79) and C-3 (δ _c 111.53) / C-4 (δ _c 131.92) / C-5 through the HMBC analysis of heterogeneous nuclei lt; / RTI &gt; _c &lt; RTI ID = _0.0 &gt; 127.26). &lt; / RTI &gt; Methoxy protons (δ _H 3.89) from C- from C-2 (δ _c 151.89) via HMBC analysis between, located methoxy group in C-2 position, oxy-methoxy proton _{H-2 '(δ H 4.50} ) 1 (delta _c 154.97), it was confirmed that the C-1 position was glycerol.

<2-2> Compound 2: 2- (4-hydroxymethyl-2-methoxyphenoxy) -propane-1,3-diol diol)

[Chemical Formula 12]

.

.

The compound 2 of the above formula (12) is in the form of a white amorphous powder, and the result of the structural analysis is as follows: _{^{[α] 20 D: + 11.9}} (MeOH, c = 0.2); C ₁₁ H ₁₆ O ₅ , HR-ESI-MS m / z : 251.0886 [M + Na] ^{+ (} calcd for C ₁₁ H ₁₆ O ₅ Na, 251.0895); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 1 below.

^{Through the 1} H NMR and ¹³ C NMR spectra, the structure of compound 2 was confirmed to have a structure similar to that of compound 1 except that the aldehyde group was substituted with a hydroxymethylene group (Table 1). HMBC analysis between oxymethylene (δ _H 4.52) and C-3 (δ _c 112.62) / C-4 (δ _c 137.31) / C-5 (δ _c 120.71) showed that oxymethylene was located at the C-4 position Respectively.

<2-3> Compound 3: (+) - 3-hydroxy-2- (4-hydroxy-3-methoxyphenyl) - 3-hydroxy-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) propan-

[Chemical Formula 13]

.

.

Compound 3 having the structure of Formula 13 and having a stereoselectivity of 8 S is in the form of a white amorphous powder. _{^{[α] 20 D: +21.7 (}} MeOH, c = 0.2); C ₁₆ H ₁₆ O ₅ , HR-ESI-MS m / z : 289.1073 [M + H] ^{+ (} calcd for C ₁₆ H ₁₇ O ₅ , 289.1076); ¹ H NMR (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) The results are shown in Table 1 below.

¹ H NMR δ _H 6.75 over the spectral (d, J = 8.2 ㎐) , 6.76 (d, J = 8.2 ㎐), 6.81 (d, J = 8.8 ㎐), 6.90 (s) and 7.94 (d, J = 8.8 ㎐ _H 3.84 (3H, s), there is one methoxy group in δ _H 4.76 (dd, J = 5.2, 8.8 ㎐), δ _H 3.72 , One ABX and one AA'BB 'system with one oxymethylene at 4.26 (dd, J = 8.8, 10.7 ㎐), J = 5.2, 10.7 ㎐. ^{Through 13} C NMR and DEPT spectra, it was confirmed that compound 3 had a total of 14 carbons, with one carbonyl group, four quaternary carbons, six methines, one oxymethylene and one methoxy carbon The carbonyl was located at the C-7 position in the HMBC analysis between (δ _H 7.94) / H-8 (δ _H 4.76) / H-9 (δ _H 3.72) and C-7 (δ _c 199.70) , And it was confirmed that the oxymethylene group was located at the C-8 position in the HMBC analysis between H-8 (δ _H 4.76) and C-9 (δ _C 65.50). In addition, HMBC analysis from methoxyproton (delta _H 3.84) to C-3 '(delta _C 149.27) confirmed that the methoxy group was located at the C-3' position. The chemical shift correlated spectroscopy (COZ) analysis, which is a two-dimensional NMR analysis, confirmed the position of the oxymethylene group as a spin system of -CH-CH ₂ - was produced (FIG. 1).

<2-4> Compound 4: threo-2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -propane-1,3-diol (threo-2- (4- hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -propane-1,3-diol)

[Chemical Formula 14]

.

.

Compound 4 having the structure of the above formula (14) and having a relative stereochemistry of threo was obtained as a racemic compound in the form of a white amorphous powder in an optically inactive state. The results of the structural analysis are as follows; [?] ²⁰ _D : +0 (MeOH, c = 0.2); C ₁₆ H ₁₈ O ₅ , HR-ESI-MS m / z : 289.1080 [MH] ^{- (} calcd for C ₁₆ H ₁₇ O ₅ , 289.1076); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 2 below.

^{Through the 1} H NMR and ¹³ C NMR spectra, it was confirmed that the structure of the compound 4 had a structure similar to that of the compound 3 except that the carbonyl group at the C-7 position was substituted with a hydroxy group. HMBC analysis of H-2 (隆_H 7.03) / H-8 (隆_H 2.96) / H-9 (隆_H 3.68) to C-7 (隆_c 75.55) Location. The configuration between C-2 and C-3 was determined to be threo through the fact that the coupling constant between H-2 and H-3 in compound 4 was relatively large J = 7.8 Hz.

2-5 Compound 5: 2- (4-hydroxy-3-methoxyphenyl) -1- (4-hydroxyphenyl) -1-methoxy- -3-methoxyphenyl) -1- (4-hydroxyphenyl) -1-methoxy-propan-

[Chemical Formula 15]

.

.

Compound 5 having the structure of Formula 15 and having a relative stereoselectivity of threo is in the form of a white amorphous powder. [?] ²⁰ _D : +0 (MeOH, c = 0.2); C ₁₇ H ₂₀ O ₅ , HR-ESI-MS m / z : 303.1229 [MH] ^{- (} calcd for C ₁₇ H ₁₉ O ₅ , 303.1232); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 2 below.

^{Through the 1} H NMR and ¹³ C NMR spectra, it was confirmed that the structure of the compound 5 had a structure similar to that of the compound 4 except that the structure further had a methoxy group. The HMBC analysis between methoxyproton (δ _H 3.15) and C-7 (δ _c 87.33) confirmed the presence of the methoxy group at the C-7 position. Further, it was determined that the stereoconfiguration was threo through the fact that the coupling constant between H-2 and H-3 in Compound 5 was relatively large J = 8.8 Hz.

<2-6> Compound 6: (7S, 8R) -3'--methoxy-4 ', 7-epoxy -8,5'- neo lignans -4,9,7'- triol ((7 S, 8 R ) -3'-methoxy-4 ', 7-epoxy-8,5'-neoLignan-4,9,7'-triol)

(3)

.

.

Compound 6 of the above formula 3 is a colorless oil, and the result of the structural analysis is as follows; [?] ²⁰ _D : +12.8 (MeOH, c = 0.2); CD ( c = 2 x 10 ^-4 , MeOH)? (Mm) +0.32 (281); C ₁₇ H ₁₈ O ₅ , HR-ESI-MS m / z : 301.1075 [MH] ^{- (} calcd for C ₁₇ H ₁₇ O ₅ , 301.1076); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 2 below.

¹ H NMR through the spectrum, compound 6, AA'BB 'get the spin pattern, δ _H 6.77 the aromatic ring in (d, J = 8.4 ㎐) and 7.20 (d, J = 8.4 ㎐ ) and, 6.89 (s) And 6.91 (s) with two meta aromatic protons; [delta] _H 3.84 (3H, s), there is one methoxyproton; [delta] _H 3.49 (d, J = 6.2 Hz) with one methine proton;隆_H 5.54 (d, J = 6.2 Hz) with one oxymetin proton; And two oxymethylene protons were located at δ _H 3.77 (m) and 3.84 (dd, J = 5.3, 10.7 ㎐) and δ _H 4.54 (s). ^{Through the 13} C NMR and DEPT spectra, it was confirmed that the compound 6 contained 6 quaternary carbons, 6 methine, 2 oxymethylene, and one methoxy carbon, and was composed of a total of 15 carbons (Table 2). The NMR analysis of Compound 6 showed that when compared to the NMR analysis of the previously known compound 2- (4-hydroxy-3-methoxyphenyl) -3-methyl-5-hydroxymethyl-7-methoxycoumaran, Similar results were observed except that no methoxy groups were present. The methoxy group was located at the C-3 position in the HMBC analysis from methoxyproton (delta _H 3.84) to C-3 '(delta _c 145.37). HMBC analysis between H-2 '(δ _H 6.89) / H-6' (δ _H 6.91) and C-7 '(δ _c 65.41) and H-9 (δ _H 3.77 and 3.84) and C-7 1 and C-8 positions were found through the HMBC analysis between the C-1 and C-8 (δ _c 89.04) / C-8 (δ _c 55.32) / C-5 '(δ _c 129.90) ). The relative stereochemistry of H-7 and H-8 was determined to be threo through their binding constant J = 6.2 Hz. Through the appearance of a positive cotton effect at 281 nm (Δε +0.32), absolute configuration of the C-7 and C-8 of the compound 6 was determined to be 7 S, and R 8.

<2-7> Compound 7: threo - (7S, 8S) -4,7,9- trihydroxy-1'-acrylic aldehyde -8-O-4'- neo lignans (threo - (7 S, 8 S ) -4,7,9-trihydroxy-1'-acrylaldehyde -8- O -4'-neolignan)

(7)

.

.

Compound 7 having the structure of Formula 7 and having a relative stereo configuration of threo and having a stereoselectivity of 7 S and 8 S is a colorless oil. _{^{[α] 20 D: -11.2 (}} MeOH, c = 0.2); CD ( c = 2 x 10 ^-4 , MeOH)? (Mm) + 0.40 (230); C ₁₈ H ₁₈ O ₅ , HR-ESI-MS m / z : 315.1232 [M + H] ^{+ (} calcd for C ₁₈ H ₁₉ O ₅ , 315.1232); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 3 below.

^{Through the 1} H NMR spectrum, it was confirmed that Compound 7 had two AA'BB 'spin patterns and two olefin protons, one oxymethylene, two oxymethines and one aldehyde proton (Table 3). The E geometry of the double bond was confirmed through the J _{7'8 '} value of 16.0 Hz. Further, it was confirmed that the compound 7 had threo configuration through the J _7,8 value of 6.6 Hz. Relative stereostructures at C-7 and C-8 positions can be identified by showing the difference in chemical shifts (δ _c 84.43) to the downfield (δ _c 84.43) relative to the threo isomer at the C-8 position. NMR analysis of the compound 7 was previously known in the compounds of threo - when compared with (7 R, 8 R) NMR analysis of the -guaiacylglycerol-β-coniferyl aldehyde ether, with the exception that it has two additional methoxy Similarity was confirmed. The C-4 'position in the HMBC analysis between _{H-7' (δ H 7.63} ) and _{C-8 '(δ c 127.20} ) / C-4' (δ c 128.35) / C-3 '(δ c 131.59) ( 2 E ) -2-pentenal ((2 E ) -2-pentenal) residue (FIG. 1). Through the appearance of a positive cotton effect at 230 nm (Δε +0.40), absolute configuration of the C-7 and C-8 of the compound 7 was determined to be S 7 and S 8.

<2-8> Compound 8: erythro - (7R, 8S) -4,7,9- trihydroxy-1'-acrylic aldehyde -8-O-4'- neo lignans (erythro - (7 R, 8 S ) - 4,7,9-trihydroxy-1'- acrylaldehyde-8- O -4'-neolignan)

(7)

.

.

Compound 8 in which the structure of the formula 7 is the erythro structure and the configuration thereof is 7 R and 8 S is a colorless oil, and the result of the structural analysis is as follows; _{^{[α] 20 D: -31.1 (}} MeOH, c = 0.2); CD ( c = 2 x 10 ^-4 , MeOH)? (Mm) + 0.40 (230); C ₁₈ H ₁₈ O ₅ , HR-ESI-MS m / z : 315.1235 [M + H] ^{+ (} calcd for C ₁₈ H ₁₉ O ₅ , 315.1232); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 3 below.

^{Through the 1} H NMR spectrum, it was confirmed that Compound 8 had two AA'BB 'spin patterns and two olefin protons, one oxymethylene, two oxymethines and one aldehyde proton (Table 3). The E geometry of the double bond was confirmed through the J _{7'8 '} value of 16.0 Hz. Also, it was confirmed that Compound 8 had an erythro configuration with J _7,8 value of 5.7 Hz. Relative stereostructures at C-7 and C-8 positions can be identified by showing the difference in chemical shifts (δ _c 83.58) in the upfield (δ _c 83.58) relative to the erythro isomer at the C-8 position. NMR analysis of the compound 8 is the erythro compounds known in the prior - (7 S, 8 R) -guaiacylglycerol -? - when compared to the NMR analysis of coniferyl aldehyde ether, with the exception that it has two additional methoxy Similarity was confirmed. Through the appearance of a positive cotton effect at 230 nm (Δε +0.387), absolute configuration of the C-7 and C-8 of the compound 8 was determined to be 7 R 8 and S.

<2-9> Compound 9: threo- (7S, 8S) -3'-methoxy-4,7,9-trihydroxy-1'-hydroxypropyl- 8-0-4'- neo lignan - (7 S, 8 S) -3'-methoxy-4,7,9-trihydroxy-1'-hydroxypropyl-8- O -4'-neolignan)

[Chemical Formula 8]

.

.

Compound 9 having the structure of Formula 8 and having a relative stereoselection of threo and a stereoselectivity of 7 S , 8 S is a colorless oil, and the structural analysis result is as follows; _{^{[α] 20 D: -19.1 (}} MeOH, c = 0.2); CD ( c = 2 x 10 ^-4 , MeOH)? (Mm) +0.38 (230); C ₁₉ H ₂₄ O ₆ , HR-ESI-MS m / z : 371.1462 [M + Na] ^{+ (} calcd for C ₁₉ H ₂₄ O ₆ Na, 371.1471); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 3 below.

NMR analysis of the compound 9 is a compound of threo -1- (4-hydroxyphenyl) kkuji previously known to be contained in the fruit of the tree (Broussonetia papyrifera) -2- [4- ( 3-hydroxy-1- propyl) - 2-methoxyphenoxy] -1,3-propanediol, but it was confirmed that there was a difference in that there was no optical activity and contained only one stereostructure. Compound 9 has a J _7,8 value of 6.6 Hz, and Compound 9 is threo , and CD spectral results show a positive cotton effect (Δε +1.25), it was confirmed that the absolute stereostructure at the C-7 and C-8 positions was 7 S , 8 S -.

<2-10> Compound 10: erythro- (7R, 8S) -3'-methoxy-4,7,9-trihydroxy-1'-hydroxypropyl- 8-0-4'- neo lignan - (7 R, 8 S) -3'- methoxy -4,7,9-trihydroxy-1'-hydroxypropyl-8- O -4'-neolignan)

[Chemical Formula 8]

.

.

Compound 10 having the structure of Formula 8 and having a relative stereochemical configuration of erythro and a configuration of 7 R , 8 S is a colorless oil. [?] ²⁰ _D : -22.1 (MeOH, c = 0.2); CD ( c = 2 x 10 ^-4 , MeOH)? (Mm) + 0.40 (230); C ₁₉ H ₂₄ O ₆ , HR-ESI-MS m / z : 371.1492 [MH] ^{- (} calcd for C ₁₉ H ₂₃ O ₆ , 347.1495); ¹ H (CD ₃ OD, 400 MHz) and ¹³ C NMR (CD ₃ OD, 100 MHz) are shown in Table 3 below.

A known compound, NMR analysis of the compound 10 is kkuji previously to have been contained in the fruit of the tree (Broussonetia papyrifera) erythro -1- (4 -hydroxyphenyl) -2- [4- (3-hydroxy-1-propyl) - 2-methoxyphenoxy] -1,3-propanediol, but it was confirmed that there was a difference in that there was no optical activity and contained only one stereostructure. Compound 10 has a J _7,8 value of 5.7 Hz, and compound 10 is erythro , and CD spectral results show a positive cotton effect (Δε +1.30), it was confirmed that the absolute stereostructure at the C-7 and C-8 positions was 7 R and 8 S -.

<2-11> Compound 11: (2-glyceryl) -O-coniferaldehyde (2-glyceryl)

[Chemical Formula 9]

.

.

The compound 11 of the above formula (9) is a compound known to be able to be isolated from the spruce tree (Non Patent Patent 001).

<2-12> Compound 12: 2- [4- (3-Hydroxypropyl) -2-methoxyphenoxy] -1,3-propanediol methoxyphenoxy] -1,3-propanediol)

[Chemical formula 10]

.

.

The compound 12 of the above formula (10) is a compound known to be able to be isolated from P. fir (Non Patent Patent 002).

<2-13> Compound 13: (-) - Ebopholine B ((-) - evofolin B)

[Chemical Formula 16]

.

.

Compound 13 having the structure of the above formula (16) and having a configuration of 8 S is a compound known to be able to be isolated from Pyrrhizae (Non Patent Patent 003).

<2-14> Compound 14: 1,2-bis- (4-hydroxy-3-methoxyphenyl) -propane-1,3-diol ) -propane-1,3-diol)

[Chemical Formula 17]

.

.

Compound 14, which has the structure of Formula 17 and whose relative stereochemistry is erythro , is a compound known to be able to be isolated from P. falciparum (Non Patent Patent 004).

NMR spectra of compounds 1 to 3 location One 2 3 δ _C ^{a, b} δ _H ^{a, c} (J in Hz) δ _C ^{a, b} δ _H ^{a, c} (J in Hz) δ _C ^{a, b} δ _H ^{a, c} (J in Hz) One 154.97 - 174.97 - 130.07 - 2 151.86 - 151.94 - 132.41 7.94 (d, 8.8) 3 111.53 7.46 (s) 112.62 6.70 (s) 116.14 6.81 (d, 8.8) 4 131.92 - 137.31 - 163.70 - 5 127.26 7.50 (d, 8.2) 120.71 6.85 (d, 8.1) 116.14 6.81 (d, 8.8) 6 115.58 7.23 (d, 8.2) 118.81 7.03 (d, 8.1) 132.41 7.94 (d, 8.8) 7 192.87 9.79 (s) 65.01 4.52 (s) 199.70 - 8 56.24 4.76 (dd, 5.2, 8.8) 9 65.50 3.72 (dd, 5.2, 10.7)
4.26 (dd, 8.8, 10.7) One' 61.92 3.79 (m) 61.96 3.73 (d, 4.9) 129.73 - 2' 82.07 4.50 (m) 83.00 4.19 (m) 112.71 6.90 (s) 3 ' 61.92 3.79 (m) 61.96 3.37 (d, 4.9) 149.27 - 4' 146.90 - 5 ' 116.54 6.75 (d, 8.2) 6 ' 122.22 6.76 (d, 8.2) 2-OMe 56.44 3.89 (s) 56.39 3.8 (s) 56.39 3'-OMe 56.36 3.84 (s)

NMR spectra of compounds 4 to 6 location 4 5 6 δ _C ^{a, b} δ _H ^{a, c} (J in Hz) δ _C ^{a, b} δ _H ^{a, c} (J in Hz) δ _C ^{a, b} δ _H ^{a, c} (J in Hz) One 135.68 - 132.70 - 134.09 - 2,6 128.92 7.03 (d, 8.3) 129.91 6.86 (d, 8.4) 128.23 7.20 (d, 8.4) 3,5 115.60 6.72 (d, 8.3) 115.59 6.56 (d, 8.4) 116.25 6.77 (d, 8.4) 4 157.61 - 157.74 - 158.47 - 7 75.55 4.90 (d, 7.8) ^b 87.33 4.27 (d, 8.8) 89.04 5.54 (d, 6.2) 8 56.74 2.96 (m) 56.00 2.96 (m) 55.32 3.49 (d, 6.2) 9 646.49 3.65 (dd, 7.4, 10.8)
3.81 (d, 7.4) 65.10 3.82 (dd, 5.6, 10.7)
4.03 (dd, 5.5, 10.9) 65.02 3.77 (m)
3.84 (dd, 5.3, 10.7) One' 132.41 - 132.03 - 136.14 - 2' 114.32 6.70 (d, 2.1) 114.15 6.46 (d, 2.1) 113.05 6.89 (s) 3 ' 148.38 - 148.30 - 145.37 - 4' 146.11 - 145.90 - 148.59 - 5 ' 115.37 6.72 (d, 8.2) 115.69 6.56 (d, 8.2) 129.90 - 6 ' 123.01 6.64 (dd, 2.1, 8.2) 122.52 6.45 (dd, 2.1, 8.2) 117.16 6.91 (s) 7 ' 65.41 4.54 (s) 7-OMe 56.65 3.15 (s) 3'-OMe 56.25 3.79 (s) 56.26 3.66 (s) 56.65 3.84 (s)

Spruce tree extract and its From the fraction  Identification of lipid degradation effects of isolated compounds

Lipid degradation is the process by which lipid structure is degraded and triglyceride is hydrolyzed to glycerol and three fatty acids. Therefore, in order to confirm the lipolytic activity exhibited by the compound of the present invention, the lipolytic activity was confirmed by the degree of secretion of glycerol into the medium by decomposing the fat in the cells.

Specifically, a C3H10T1 / 2 cell line (ATCC, Manassas, VA, USA), which is an embryonic fibrous cell line derived from a mouse, was cultured in a growth medium in a saturated medium, and 20 ng / ml BMP4 morphogenetic protein 4 (R & D system, Minneapolis, MN, USA), and the medium was replaced with differentiation medium for 3 days to induce differentiation into lipid cells. The differentiation medium was DMEM containing 10% FBS, 1% P / S, 2.5 mM isobutylmethylxanthine, IBMX (Cayman Chemical), 1 M examethasone (Cayman Chemical) and 1 g / ml insulin Medium was used. After induction of differentiation for 3 days, any one of the compounds 1 to 14 of the present invention was replaced with fresh DMEM medium (phenol-red free) containing 1 mM concentration and further cultured for 4 hours. After incubation, the medium was harvested and the level of glycerol released into the medium was determined according to the manufacturer's protocol using free glycerol reagent (Sigma). The reagents were treated and incubated at 37 ° C for 5 minutes, and the absorbance was measured at 540 nm.

In order to confirm the cytotoxicity of the compound of the present invention with respect to the cells, the cell viability was confirmed in the cells treated with the compound. Cell viability was analyzed according to the manufacturer's protocol using Ez-cytox cell viability, proliferation & cytotoxicity assay kit (EZ-1000, Dogen, Daeil Lab, Seoul, Korea). C3H10T1 / 2 cells were induced to differentiate in 96-well plates with the same composition as above, and any one of Compounds 1 to 14 of the present invention was added to a final concentration of 1 [mu] M and cultured for 24 hours. Then, the Ez-cytox reagent was treated and incubated at 37 ° C for 2 hours, and the absorbance was analyzed at 450 nm in a microplate reader.

As a positive control for lipolytic activity, 8-Bromoadenosine 3 ', 5'-cyclic monophosphate (8Br-cAMP), known as an AMP-dependent protein kinase activator, was used instead of the compound of the present invention. In addition, the cell survival rate was determined as a relative value of the cell survival rate of each compound-treated group based on the cell survival rate of the solvent control containing only the compound-free solvent.

As a result, as shown in the following Table 4, it was confirmed that Compounds 1 to 14 of the present invention all exhibited significantly increased lipolytic effect as compared with the solvent control. Compared with the control group showing a glycerol secretion amount of 5.57 ± 0.60 mg / ml in the positive control group, 8Br-cAMP treated group, Compound 1 and Compound 12 of the present invention showed 4.94 ± 0.90 mg / ml and 3.99 ± 0.69 mg / Ml, indicating that the lipolytic activity was significantly increased. In addition, it was confirmed that the compounds 1 to 14 of the present invention did not show significant cytotoxicity against C3H10T1 / 2 cells (Table 4).

Comparison of lipolytic effects of the compounds of the present invention in differentiated adipocytes Experimental group Glycerol (mg / mL) Cell viability (%) Solvent control 0.78 + 0.02 100.0 + - 6.73 One 4.94 ± 0.90 ^a 105.01 + - 10.82 2 1.52 + - 0.67 87.74 + - 7.21 3 0.84 0.16 94.60 + - 4.67 4 3.56 + 0.47 ^a 92.43 + - 1.38 5 0.98 ± 0.01 101.53 + - 9.55 6 0.97 + - 0.12 105.01 + - 10.82 7 1.32 ± 0.28 128.70 ± 9.06 8 1.06 + 0.06 120.97 + - 3.31 9 1.43 + 0.64 107.34 + 1.49 10 1.48 ± 0.63 105.34 ± 1.40 11 0.97 ± 0.01 93.73 + - 0.66 12 3.99 ± 0.69 ^a 91.44 + - 3.30 13 2.55 + - 0.60 93.78 ± 1.73 14 0.95 + 0.03 129.97 ± 3.31 8Br-cAMP 5.57 + - 0.60b 99.3 + - 4.34

Claims (12)

delete A compound represented by any one selected from the group consisting of the following formulas (3), (4), (7), (8), (13) and (15) salt:
(3)

,
[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),
(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),
[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),
[Chemical Formula 13]

, And
[Chemical Formula 15]

.
3. The compound according to claim 2, wherein the compound is isolated from a spruce tree extract or a fraction thereof, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
The compound according to claim 3, wherein the extract is an extract obtained by extracting the spruce tree with water, a C ₁ to C ₄ lower alcohol or a mixture thereof as a solvent. The compound, its optical isomer, Acceptable salts.
4. The compound according to claim 3, wherein the fraction is a fraction prepared by additionally adding a solvent to a spruce extract, or an optical isomer thereof, or a pharmaceutically acceptable salt thereof.
i) extracting the spruce tree with water, C ₁ -C ₂ lower alcohol or a mixed solvent thereof;
ii) obtaining an ethyl acetate or chloroform fraction of the extract obtained in step i); And
iii) subjecting the fraction obtained in step ii) to column chromatography to obtain a separated compound.
delete A pharmaceutical composition for preventing or treating obesity, which comprises, as an active ingredient, a compound represented by any one of the following formulas (3) to (10) or a pharmaceutically acceptable salt thereof:
(3)

,
[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),
[Chemical Formula 5]

(Wherein R is either H or OCH ₃ , and the stereochemical arrangement is any one of 8 R and 8 S )
[Chemical Formula 6]

Wherein R ₁ and R ₂ are both H, R ₁ is H, R ₂ is CH ₃ , or R ₁ is OCH ₃ and R ₂ is H, and the relative stereoconfiguration is threo or erythro , etc.),
(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),
[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),
[Chemical Formula 9]

, And
[Chemical formula 10]

.
9. A pharmaceutical composition for preventing and treating obesity according to claim 8, characterized in that the compound is isolated from a spruce tree extract or a fraction thereof.
delete A health functional food for preventing or improving obesity, which comprises, as an active ingredient, a compound represented by any one of the following formulas (3) to (10) or a pharmaceutically acceptable salt thereof:
(3)

,
[Chemical Formula 4]

(Wherein R is any one of CHO and CH ₂ OH),
[Chemical Formula 5]

(Wherein R is either H or OCH ₃ , and the stereochemical arrangement is any one of 8 R and 8 S )
[Chemical Formula 6]

Wherein R ₁ and R ₂ are H, R ₁ is H, R ₂ is CH ₃ , or R ₁ is OCH ₃ and R ₂ is H, and the relative stereoconfiguration is threo or erythro , &Lt; / RTI &gt;
(7)

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),
[Chemical Formula 8]

(Three-dimensional arrangement of the above formula is a 7 S, 8 S, or R 7, S 8 any one of the relative three-dimensional arrangement is any one of threo or erythro.),
[Chemical Formula 9]

, And
[Chemical formula 10]

.
12. The health functional food for preventing and improving obesity according to claim 11, wherein the compound is isolated from a spruce tree extract or a fraction thereof.

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