KR102234567B1 - New C-p-hydroxybenzylflavonol glycosides from Pumpkin's Tendrile and Their Uses - Google Patents

Abstract

The present invention relates to a novel compound derived from a natural product, and more specifically, to a novel compound derived from amber tendril having antioxidant activity and/or immune enhancing activity, and a use thereof.

Description

New C-p-hydroxybenzylflavonol glycosides from Pumpkin's Tendrile and Their Uses}

The present invention relates to a novel compound derived from a natural product, and more specifically, to a novel compound derived from amber tendril having antioxidant activity and/or immune enhancing activity, and a use thereof.

Pumpkin (Cucurbita spp.) is an annual vine plant belonging to the Cucurbita or Cucuraceae family, and is mainly cultivated in Asia, including the Americas, Korea and Japan. Largely, pumpkins are divided into oriental pumpkins ( Cucurbita moschata Duch.), western pumpkins ( Cucurbita maxima Duch.), and pepper pumpkins ( Cucurbita pep o L.). In Korea, the fruits, leaves, seeds, and stems of oriental pumpkins can be used for food, but only the fruits and seeds of Western pumpkins are used for food. In Korea, fruits, leaves, and seeds are mainly consumed, but stems are not used for food. Pumpkin has been reported in people with weak stomachs, patients in recovery phase, alleviation of edema, and anti-obesity effects. In addition, pumpkin has been consumed a lot when cooking after childbirth.

On the other hand, pumpkin tendril, a by-product of pumpkin, is a tendril that winds around other objects while the pumpkin tendril stretches out, and recently, it has been reported that it has the effect of preventing osteoporosis and preventing inflammation (Hana Jang et al., Korean J. Food Preserv. , 24, 1122~ 1128, 2017) and revealed the existence of rutin as an active ingredient, but research on the ingredients contained in pumpkin tendrils is insufficient.

Korean Patent Registration No. 10-1775071

The present inventors completed the present invention by purifying and separating a novel compound contained in pumpkin tendrils through a number of studies.

Accordingly, it is an object of the present invention not only to confirm that various physiologically active substances are contained in pumpkin tendrils, but also to provide a novel compound having a new structure that was not known at all before.

Another object of the present invention is to provide a food composition and a health functional food using the antioxidant activity and/or immunity enhancing activity of a novel compound isolated and structured from pumpkin tendrils.

The object of the present invention is not limited to the above-mentioned object, and even if not explicitly mentioned, the object of the invention that one of ordinary skill in the art can recognize from the description of the detailed description of the invention to be described later may naturally be included. .

In order to achieve the object of the present invention described above, the present invention provides a novel compound represented by the following formula (1).

[Formula 1]

Here, R1 is a hydrogen atom, an oxygen atom or a hydroxy group,

,

또는

이다.R2 is

,

or

to be.

In a preferred embodiment, the compound has at least one of antioxidant activity and immune enhancing activity.

In a preferred embodiment, the compound is extracted and isolated from amber tendril.

In a preferred embodiment, the compound is 8-carbon-para-hydrocybenzylquacertin 3-oxy-lutinoside, 8-carbon-para-hydrocybenzylquacertin 3-oxy-beta-di-glucopyrano Side, 7-carbon-para-hydrocybenzylchemferol 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di-galatopyranoside, 8-carbon-para-hydrocybenzylchemferol 3-oxy-lutinoside, 8-carbon-para-hydrocybenzylisoramnetin 3-oxy-lutinoside, 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyra It is any one or more selected from the group consisting of nosyl (1→6)-beta-di-galatopyranoside.

In addition, the present invention is to obtain a pumpkin tendril extract; Concentrating the pumpkin tendril extract under reduced pressure; Obtaining an ethyl acetate fraction through the solvent fractionation of the concentrate; And purifying the ethyl acetate fraction by HPLC using Amberlite XAD-2 and ODS column chromatography and reverse phase column.

In a preferred embodiment, the step of obtaining the pumpkin tendril extract is performed by pulverizing the pumpkin tendril and extracting hot water.

In addition, the present invention provides an antioxidant food composition comprising the above-described compound or a food pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a food composition for enhancing immunity comprising the above-described compound or a food pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides an antioxidant health functional food comprising the above-described compound or a food pharmaceutically acceptable salt thereof as an active ingredient.

In addition, the present invention provides a health functional food for enhancing immunity comprising the above-described compound or a food pharmaceutically acceptable salt thereof as an active ingredient.

According to the present invention described above, there is provided a novel compound derived from amber tendril and a method for separating the same, wherein the novel compound has antioxidant activity and/or immunity enhancing activity, and a composition comprising the same as an active ingredient is used for antioxidant and/or immune enhancing activity. It can be usefully used as a food material.

These technical effects of the present invention are not limited only to the ranges mentioned above, and even if not explicitly mentioned, the effects of the invention that can be recognized by those of ordinary skill in the art from the description of specific details for the implementation of the invention described later are also Of course it is included.

1 is a diagram showing the separation and purification process of new compounds 1 to 6 having the structural formula of Formula 1 according to an embodiment of the present invention.
2 is an 8-carbon-para-hydrocybenzylquacertin 3-oxy-lutinoside (8- C - p -hydroxybenzylquercetin 3-), which is compound 1 of the novel compounds having the structural formula of Formula 1 according to an embodiment of the present invention. O- rutinoside) of ¹ H- ¹ H-COSY correlation (bold line), gHMBC correlation (arrow) and structural formula are shown.
FIG. 3 is an 8-carbon-para-hydrocybenzylquacertin 3-oxy-beta-di-glucopyranoside (8- C- ), which is compound 2 of the novel compounds having the structural formula of Formula 1 according to an embodiment of the present invention. p- hydroxybenzylquercetin 3- O- β-D-glucopyranoside) ¹ H- ¹ H-COSY correlation (bold line), gHMBC correlation (arrow) and structural formula are shown.
FIG. 4 is a 7-carbon-para-hydrocybenzylcamperol 3-oxy-alpha-L-rhamnopyranosyl (1→6)- which is a compound 3 of a novel compound having the structural formula of Formula 1 according to an embodiment of the present invention. beta-D-gala topira furnace side [8- C - p -hydroxybenzylkeampferol 3- O -α-L-rhamnopyranosyl (1 → 6) -β-D-galactopyranoside] 1 H- 1 H-COSY correlation (bold line in the ), gHMBC correlation (arrow) and structural formula.
5 is a new compound having the structural formula of Formula 1 according to an embodiment of the present invention, compound 4 , 7-(para-hydroxybenzyl)chemferol 3-oxy-lutinoside [7-( p-hydroxybenzyl)kaempferol 3 -O- rutinoside] of ¹ H- ¹ H-COSY correlation (bold line), gHMBC correlation (arrow) and structural formula are shown.
6 is an 8-carbon-para-hydrocybenzylisoramnetin 3-oxy-lutinoside (8-C - p -hydroxybenzylisorhamnetin 3), which is compound 5 of the novel compounds having the structural formula of Formula 1 according to an embodiment of the present invention. - O ¹ H- ¹ H-COSY correlation -rutinoside) (thick line), gHMBC shows the correlation (arrows) and the structural formula.
7 is 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl (1→6), which is compound 6 of the novel compounds having the structural formula of Formula 1 according to an embodiment of the present invention. -beta-D-gala topira furnace side - ¹ H- ¹ H-COSY correlation of [8- C p -hydroxybenzyl isorhamnetin 3- O -α-L-rhamnopyranosyl (1 → 6) -β-D-galactopyranoside] ( Bold line), gHMBC correlation (arrow), and structural formula.
8 shows the DPPH free radical scavenging effect of compounds having the structural formula of Formula 1 isolated from amber tendril according to an embodiment of the present invention.
9 shows the results of evaluating the cytotoxicity of compounds having the structural formula of Formula 1 isolated from pumpkin tendrils according to an embodiment of the present invention.
10A and 10B show the results of inducing the production of cytokines IL-6 (10a) and TNF-α (10b) in cells by compounds having the structural formula of Formula 1 isolated from pumpkin tendrils according to an embodiment of the present invention. This is the graph shown.

The terms used in the present invention are only used to describe specific embodiments, and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the presence of features, numbers, steps, actions, components, parts, or a combination thereof described in the description of the invention, but one or more other It is to be understood that it does not preclude the presence or addition of features, numbers, steps, actions, components, parts, or combinations thereof.

Terms such as first and second may be used to describe various constituent elements, but the constituent elements should not be limited by the terms. These terms are only used for the purpose of distinguishing one component from another component. For example, without departing from the scope of the present invention, a first element may be referred to as a second element, and similarly, a second element may be referred to as a first element.

Unless otherwise defined, all terms used herein including technical or scientific terms have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms as defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and should not be interpreted as an ideal or excessively formal meaning unless explicitly defined in the present invention. Does not.

In interpreting the constituent elements, it is interpreted as including an error range even if there is no explicit description. In particular, when the terms "about", "substantially" and the like of degree are used, it may be interpreted as being used in or close to the numerical value when manufacturing and material tolerances specific to the stated meaning are presented. .

In the case of a description of a temporal relationship, for example,'after','following','after','before', etc. This includes cases that are not continuous unless' is used.

Hereinafter, the technical configuration of the present invention will be described in detail with reference to the accompanying drawings and preferred embodiments.

However, the present invention is not limited to the embodiments described herein and may be embodied in other forms. The same reference numerals used to describe the present invention throughout the specification denote the same elements.

The technical feature of the present invention lies in the isolation of a novel compound that was not previously known to be contained in the amber tendril, confirming the structure, as well as confirming their exact physiological activity.

That is, the present invention is a composition comprising a novel compound of Formula 1 having a new structure that was not previously known at all and confirming its activity, as well as through the antioxidant activity and/or immunity enhancing activity of the new compound as an active ingredient. This is because silver can be usefully used as a food material for antioxidant and/or immune enhancing activity. Because.

Therefore, the novel compound of the present invention has a structural formula represented by the following formula (1).

[Formula 1]

Here, R1 is a hydrogen atom, an oxygen atom or a hydroxy group,

,

또는

이다.R2 is

,

or

to be.

The novel compound having the structure as shown in Formula 1 has at least one of antioxidant activity and immune enhancing activity. In particular, a novel compound having a structure as shown in Formula 1 can be extracted and separated from amber tendril, but may also be synthesized by a known method.

Among the new compound group belonging to Formula 1, the most preferred compounds may be compounds 1 to 6 represented by the following Formulas 2 to 7.

Compound 1 has the structural formula shown in Formula 2 below, and was named as 8-carbon-para-hydroxybenzylquercetin 3-oxy- lutinoside (8-C - p -hydroxybenzylquercetin 3- O- rutinoside).

[Formula 2]

Compound 2 has the structural formula shown in Formula 3 below, 8-carbon-para-hydroxybenzyl quercetin 3-oxy-beta-di-glucopyranoside (8- C - p- hydroxybenzyl quercetin 3- O-β- D-glucopyranoside).

[Formula 3]

Compound 3 has the structural formula shown in Formula 4 below, 7-carbon-para-hydrocybenzylchemferol 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di-galatopyranoside [8 -C - p -hydroxybenzylkeampferol 3- O -α-L-rhamnopyranosyl(1→6)-β-D-galactopyranoside].

[Formula 4]

Compound 4 has the structural formula shown in Formula 5 below, and was named as 8-carbon-para-hydrocybenzylchemferol 3-oxy-lutinoside (8-C - p -hydroxybenzylkaempferol 3- O-rutinoside).

[Formula 5]

Compound 5 has the following structural formula (6), and was named as 8-carbon-para-hydroxybenzylisoramnetin 3-oxy- lutinoside (8-C - p -hydroxybenzylisorhamnetin 3- O- rutinoside).

[Formula 6]

Compound 6 has the structural formula shown in Formula 7 below, 8-carbon-para-hydroxybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di-galatopyranoside[ 8- C - p- hydroxybenzylisorhamnetin 3- O- α-L-rhamnopyranosyl(1→6)-β-D-galactopyranoside].

[Formula 7]

In the present invention, the novel compound having the structure of Chemical Formula 1 is obtained by obtaining a pumpkin tendril extract; Concentrating the pumpkin tendril extract under reduced pressure; Obtaining an ethyl acetate fraction through the solvent fractionation of the concentrate; And purifying the ethyl acetate fraction by HPLC using Amberlite XAD-2 and ODS column chromatography and reverse phase column. Here, the step of obtaining the pumpkin tendril extract may be performed by pulverizing the pumpkin tendril and extracting hot water.

Meanwhile, the compounds of the present invention represented by Formula 1 may be prepared as pharmaceutically acceptable salts and solvates according to conventional methods in the art.

As the pharmaceutically acceptable salt, an acid addition salt formed by a pharmaceutically acceptable free acid is useful. Acid addition salts are prepared by conventional methods, for example, dissolving the compound in an excess aqueous acid solution, and precipitating the salt using a water-miscible organic solvent such as methanol, ethanol, acetone or acetonitrile. The same molar amount of the compound and an acid or alcohol (eg glycol monomethyl ether) in water may be heated and then the mixture may be evaporated to dryness, or the precipitated salt may be suction filtered.

In addition, a pharmaceutically acceptable metal salt can be made using a base. The alkali metal or alkaline earth metal salt is obtained, for example, by dissolving the compound in an excess alkali metal hydroxide or alkaline earth metal hydroxide solution, filtering the undissolved compound salt, and evaporating and drying the filtrate. In this case, as the metal salt, it is particularly pharmaceutically suitable to prepare sodium, potassium or calcium salt, and the corresponding silver salt is obtained by reacting an alkali metal or alkaline earth metal salt with an appropriate silver salt (eg, silver nitrate).

Accordingly, pharmaceutically acceptable salts of the compound having the structure of Formula 1 include acidic or basic salts that may exist in the compound having the structure of Formula 1, unless otherwise indicated. For example, pharmaceutically acceptable salts include sodium, calcium and potassium salts of the hydroxy group, and other pharmaceutically acceptable salts of the amino group include hydrobromide, sulfate, hydrogen sulfate, phosphate, hydrogen phosphate, and dihydrogen There are phosphate, acetate, succinate, citrate, tartrate, lactate, mandelate, methanesulfonate (mesylate) and p-toluenesulfonate (tosylate) salts, and methods or processes for preparing salts known in the art. It can be manufactured through.

In addition, the food composition of the present invention contains a compound having the structure of Formula 1 and/or a pharmaceutically acceptable salt thereof as an active ingredient. If necessary, not only can it contain various flavoring agents or natural carbohydrates, etc. as an additional ingredient like a normal food composition, but also flavoring agents such as various nutrients, vitamins, minerals (electrolytes), synthetic flavoring agents and natural flavoring agents. , Colorants and thickeners (cheese, chocolate, etc.), pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonates used in carbonated beverages, etc. It may contain. In addition, the food composition of the present invention may contain flesh for the production of natural fruit juice and fruit juice beverages and vegetable beverages.

Here, examples of the above-described natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose, sucrose, and the like; And polysaccharides, for example, common sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. The above-described flavoring agent can be advantageously used as a natural flavoring agent (taumatin), stevia extract (eg, rebaudioside A, glycyrrhizin, etc.), and synthetic flavoring agents (saccharin, aspartame, etc.).

The food composition of the present invention can be formulated and used as a functional food or added to various foods. Foods to which the composition of the present invention can be added include, for example, beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gums, candy, ice cream, alcoholic beverages, vitamin complexes and health supplements, etc. There is this.

The compound having the structure of Formula 1 or a pharmaceutically acceptable salt thereof, which is the active ingredient of the present invention, is a material extracted from a medicinal crop and has almost no side effects such as chemicals, so it is intended to provide functionality for antioxidant or immunity enhancement. It can be used with confidence even when taking it for a long time.

That is, the food composition of the present invention is related to excessive production of free radicals through strong antioxidant activity. It can be usefully used as a functional food composition for the prevention or improvement of immunity, and as a functional food composition for the prevention or improvement of diseases caused by weakening of the immune system such as autoimmune diseases or strengthening of the immune system after chemotherapy through immunity enhancing activity. It can be usefully used.

In addition, the health functional food for antioxidant or immunity enhancement of the present invention comprises a compound having the structure of Formula 1 or a pharmaceutically acceptable salt thereof as an active ingredient. The health functional food of the present invention can be manufactured and processed in the form of tablets, capsules, powders, granules, liquids, pills, etc. for the purpose of antioxidant or immunity enhancement.

In the present invention, the term "health functional food" refers to a food manufactured and processed using raw materials or ingredients having useful functions for the human body according to the Health Functional Food Act No.6727, and nutrients for the structure and function of the human body It refers to ingestion for the purpose of controlling or obtaining beneficial effects for health purposes such as physiological effects. The health functional food of the present invention may contain ordinary food additives, and whether it is suitable as a food additive is determined according to the general rules and general test methods of food additives approved by the Food and Drug Administration, unless otherwise specified. It is judged according to the standards and standards.

Items listed in the "Food Additives Code" include, for example, chemical compounds such as ketones, glycine, calcium citrate, nicotinic acid, and cinnamic acid; Natural additives such as reduced pigment, licorice extract, crystalline cellulose, high color pigment, and guar gum; Mixed preparations, such as a sodium L-glutamate preparation, an alkali additive for noodles, a preservative preparation, and a tar color preparation, etc. are mentioned.

For example, the health functional food in the form of a tablet is a mixture of a compound having the structure of Formula 1 or a pharmaceutically acceptable salt thereof, which is an active ingredient of the present invention, with an excipient, a binder, a disintegrant, and other additives. After granulating by the method of, compression molding by putting a lubricant or the like, or directly compression molding the mixture. In addition, the health functional food in the form of a tablet may contain a mating agent or the like, if necessary.

Among the health functional foods in the form of capsules, the hard capsules are prepared by filling a conventional hard capsule with a mixture of a compound having the structure of Formula 1, the active ingredient of the present invention, or a pharmaceutically acceptable salt thereof, with an additive such as an excipient. It may be prepared, and the soft capsule may be prepared by filling a mixture of a compound having the structure of Formula 1 or a pharmaceutically acceptable salt thereof with an additive such as an excipient in a capsule base such as gelatin. The soft capsule may contain a plasticizer such as glycerin or sorbitol, a colorant, a preservative, and the like, if necessary.

The cyclic health functional food is a compound having the structure of Formula 1, which is an active ingredient of the present invention, or a mixture of pharmaceutically acceptable salts thereof and excipients, binders, disintegrants, etc., formed by a conventionally known method. It can be prepared by using, and if necessary, it can be coated with a white sugar or other coating agent, or the surface can be coated with a substance such as starch or talc.

The health functional food in the form of granules is a mixture of a compound having the structure of Formula 1, which is an active ingredient of the present invention, or a pharmaceutically acceptable salt thereof, and a mixture of an excipient, a binder, a disintegrant, etc. It can be prepared by, and if necessary, may contain a flavoring agent, a flavoring agent, and the like.

The health functional food containing the compound having the structure of Formula 1 or a pharmaceutically acceptable salt thereof of the present invention as an active ingredient has excellent antioxidant and/or immunity enhancing effects, such as excessive production of active oxygen or weakening of the immune system. It is effective in preventing or improving diseases caused by. The health functional food may be beverages, meat, chocolate, foods, confectionery, pizza, ramen, other noodles, gum, candy, ice cream, alcoholic beverages, vitamin complexes, and health supplements.

The food composition or health functional food of the present invention may contain a compound having the structure of Formula 1 or a pharmaceutically acceptable salt thereof in a concentration of 0.001 to 1000 μM, and may be included in an amount of 0.1 to 95% by weight based on the total weight of the composition. I can.

Example 1

1. Extraction and solvent fraction

Pumpkin tendrils were dried and sold in Damyang-gun, Jeollanam-do, and 300 g of dried squash tendrils were pulverized, and then 9 L of hot water of 90°C or higher was added and extracted for 15 minutes. After filtration with a nonwoven fabric, 6 L of hot water of 90° C. or higher was added to the obtained residue, followed by extraction and filtering in the same manner. The obtained extraction filtrate was concentrated under reduced pressure at 45° C. using a vacuum concentrator (A-3S, Eyela, Tokyo, Japan) equipped with an aspirator (CCA-1110, Eyela, Tokyo, Japan). The obtained concentrate was repeated three times with hexane, ethyl acetate, and saturated butanol, 3 L each, and the solvent was sequentially fractionated.

2. Component separation

As shown in FIG. 1, Amberlite XAD-2 column chromatography was performed on 2.27 g of an ethyl acetate fraction obtained by solvent fractionation of the hot water extract of amber tendrils. That is, the ethyl acetate fraction _{was dissolved in H 2} O and then injected into XAD-2 (521 μm, Supelco, USA) column (3x60 cm), H ₂ O/acetonitrile (MeCN) = 8:2, It was eluted by 800 mL each with a solvent of 6:4, 4:6, and 0:10 (v/v). The obtained fractions were spotted on a silica gel TLC plate (silica gel 60 F254, 0.25 mm thickness, Merck) and developed with n -BuOH/acetic acid/H ₂ O=4:1:1 (v/v) solvent. After the analysis was performed, a 1% cerium IV sulfate ethanol solution was sprayed on the developed TLC plate to determine the distribution trend of the compounds and grouped. Subsequently, a relatively high purity and high amount of PT-E (H ₂ O/MeCN = 6:4, v/v, 599.1 mg) was dissolved in 25% MeCN, and then injected into an ODS column (2x55 cm). %, 30%, 35%, and 40% MeCN were eluted with solvents (200 mL each) and fractionated. The obtained fraction PT-E5 (30% MeCN, 31 mg) was subjected to HPLC by applying an ODS column (Spherisorb S5 ODS2, 10x250 mm) and a 23% MeCN solvent (5.0 ml/min) to obtain a peak of t _{R 15.3 min.} A single compound having (Compound 1 , 15.6 mg, yellow powder) was isolated, and 7-(para-hydroxybenzyl)quacertin 3-oxy-lutinoside [7-(p -hydroxybenzyl)quercetin 3- O-rutinoside ].

Example 2

The fraction PT-E6 was subjected to ODS-HPLC in the same manner as described in Example 1 except for the 25% MeCN solvent (7.0 ml/min), and a single compound having a peak of t _R 12.6 min (Compound 2 , 3.9 mg , Yellow powder), a single compound with a peak of t _R 13.4 min (compound 3 , 7.6 mg, yellow powder), a single compound with a peak of t _R 14.5 min (compound 4 , 8.7 mg, yellow powder), and t _R A mixed compound having a peak of 16.2 min (Compounds 5 and 6 , 17.7 mg, yellow powder) was isolated. Compounds isolated and purified from fraction PT-E6 were 8-carbon-para-hydroxybenzylquacertin 3-oxy-beta-di-glucopyranoside (8- C - p -hydroxybenzyl)quercetin 3- O-β- D-glucopyranoside) (Compound 2 ), 7-carbon-para-hydrocybenzylchemferol 3-oxy-alpha-L-rhamnopyranoside (1→6)-beta-di-galatopyranoside[8- C- p- hydroxybenzylkeampferol 3- O -α-L-rhamnopyranosyl(1→6)-β-D-galactopyranoside] (Compound 3 ), 8-carbon-para-hydrocybenzylchemferol 3-oxy-lutinoside (8- C - p -hydroxybenzylkaempferol 3- O- rutinoside) (compound 4 ), 8-carbon-para- hydrocybenzylisoramnetin 3-oxy-lutinoside (8-C - p- hydroxybenzyl isorhamnetin 3- O- rutinoside) (Compound 5 ), and 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di-galatopyranoside [8- C - p- hydroxybenzylisorhamnetin 3- O -α-L-rhamnopyranosyl (1→6)-β-D-galactopyranoside] (compound 6 ).

Experimental Example 1

The structure of compound 1 was analyzed as follows, and the analysis results are shown in Table 1 and FIG. 2.

Compound 1 HR-ESI-MS (negative ) a m / z peak from the molecular spectrum 715.1879 [MH] of ^- was observed, it was found that the molecular formula of this compound is C ₃₄ H ₃₆ O ₁₇ (molecular weight 716). And from the ¹ H-NMR (600 MHz, CD ₃ OD) spectrum, four kinds of methine proton signals attributed to quacertin, a flavonol, [δ 7.72 (1H, d, J = 2.1 Hz, H-2'), 6.84 (1H, d, J = 9.0 Hz, H-5'), 7.46 (1H, dd, J = 9.0, 2.1 Hz, H-6') and 6.31 (1H, s, H-6)] and p − Three kinds of proton signals belonging to the hydroxylbenzyl group [δ 4.05 (2H, s, H-1''), 7.07 (2H, d, J = 8.4 Hz, H-3'' and H-7''), and 6.64 (2H, d, J = 8.4 Hz, H-4'' and H-6'')] were observed. In addition, two kinds of anomeric proton signals belong to two kinds of sugars, glucose and rhamnose [δ 5.08 (1H, d, J = 7.8 Hz, H-1``') and 4.52 (1H, d, J = 1.2). Hz, H-1''')] and other proton signals [δ 3.87-3.18, 1.12 (3H, d, J = 6.6 Hz, H-6'''')] were observed.

^{In the 13} C-NMR (125 MHz) spectrum, a total of 34 carbon signals including one carbonyl carbon signal [δ 179.8 (C-4)] were observed, which is the HR-ESI-MS and ¹ H-NMR spectra. It was consistent with the results of Trump's analysis.

Thus, for more accurate structural analysis, ^{2D-NMR analysis such as gHSQC, 1} H- ¹ H-COSY, and gHMBC was performed. As a result, it was confirmed that compound 1 is a compound composed of quacertin, p- hydroxybenzyl group, and rutinose in which ramose and glucose are bound. In particular, a correlation indicating that an ester bond was formed between δ 5.08 rutinose H-1"' and δ 135.8 quacertin C-3 was observed in gHMBC. In addition, the correlation between the quercetin 3-O- rutinoside C-7 and C-9 of δ 4.05 p -hydroxybenzyl group H-1' from δ 4.05 and 156.0) in gHMBC and the NOE of the compound dissolved in DMSO-d ₆ In the analysis results, correlations were observed between the proton signals between δ 4.05 (H-1''), δ 7.72 (H-2'') and 7.46 (H-6'), respectively, confirming their binding style.

From Table 1 in which the results of this instrumental analysis are presented, Compound 1 was determined as 8-carbon-para-hydroxybenzylquercetin 3-oxy- lutinoside (8-C - p -hydroxybenzylquercetin 3- O- rutinoside), and novel It turned out to be a compound.

Position δ _H ( Int. , Multi. , J in Hz) δ _C 2 - 159.5 3 - 135.8 4 - 179.8 5 - 161.0 6 6.31 (1H, s) 99.6 7 - 163.9 8 - 108.4 9 - 156.0 10 - 105.8 One' - 123.4 2' 7.72 (1H, d, 2.1) 118.2 3' - 145.9 4' - 150.0 5' 6.84 (1H, d, 9.0) 116.1 6' 7.46 (1H, dd, 9.0, 2.1) 123.6 One'' 4.05 (2H, br. s) 28.4 2'' - 133.2 3'', 7'' 7.07 (2H, d, 8.4) 130.4 4'', 6'' 6.64 (2H, d, 8.4) 116.2 5'' - 156.5 One''' 5.08 (1H, d, 7.8) 105.2 2''' 3.46 (1H, dd, 7.8, 7.8) 75.9 3''' 3.41 (1H, dd, 7.8, 7.8) 78.4 4''' 3.29 (1H, dd, 7.8, 7.8) 71.4 5''' 3.30 (1H, m) 77.3 6'''a
6'''b 3.79 (1H, dd, 12.4, 1.2)
3.38 (1H, dd, 12.4, 5.4) 68.6 One'''' 4.52 (1H, d, 1.2) 102.5 2'''' 3.64 (1H, dd, 3.0, 1.2) 72.2 3'''' 3.56 (1H, dd, 9.0, 3.0) 72.4 4'''' 3.28 (1H, dd, 9.0, 9.0) 74.1 5'''' 3.44 (1H, m) 69.8 6'''' 1.12 (3H, d, 6.6) 18.0

^{Table 1 shows 1} H- (600 MHz) and ¹³ of 8-carbon-para-hydroxybenzylquercetin 3-oxy- lutinoside (8-C - p -hydroxybenzylquercetin 3- O- rutinoside) (compound 1 ). C-NMR (150 MHz) data (CD ₃ OD). Further, the 8-carbon-para-dihydro when benzyl query seotin 3-oxy-Rutino side (8-C-3-O p -hydroxybenzylquercetin -rutinoside) ¹ H- ¹ H-COSY correlation (bold lines), the gHMBC The correlation (arrow) and structure are as shown in FIG. 2.

Experimental Example 2

The structure of compound 2 was analyzed as follows, and the analysis results are shown in Table 2 and FIG. 3.

Compound 2 HR-ESI-MS (negative ) a m / z peak from the molecular spectrum 569.1303 [MH] of ^- was observed, it was found that the molecular formula of this compound is C ₂₈ H ₂₄ O ₁₃ (molecular weight 570). And ¹ H- (500 MHz) and ¹³ C-NMR (125 MHz) spectra of compound 2, but is very similar to that of the compound 1, are proton and carbon signal was not observed attributable to rhamnose. Compound 2 was structured in the same manner as in Experimental Example 1.

From Table 2 below, where the results of this instrumental analysis are presented, Compound 2 is 8-carbon-para-hydroxybenzyl quercetin 3-oxy-beta-di-glucopyranoside (8- C - p- hydroxy benzyl quercetin 3- O- It was determined as β-D-glucopyranoside) (compound 2 ), and it turned out to be a new compound.

Table 2 below shows 8-carbon-para-hydrocybenzylquacertin 3-oxy-beta-di-glucopyranoside (8- C - p- hydroxybenzylquercetin 3- O- β-D-glucopyranoside) (Compound 2 ) ¹ H- (500 MHz) and ¹³ C-NMR (125 MHz) data (CD ₃ OD).

Position δ _H ( Int. , Multi. , J in Hz) δ _C 2 - 157.5 3 - 134.1 4 - 178.3 5 - 159.5 6 6.32 (1H, s) 98.0 7 - 162.3 8 - 106.8 9 - 154.4 10 - 104.3 One' - 121.7 2' 7.78 (1H, d, 2.0) 116.5 3' - 144.5 4' - 148.5 5' 6.84 (1H, d, 8.5) 114.5 6' 7.42 (1H, dd, 8.5, 2.0) 121.8 One'' 4.04 (2H, s) 26.8 2'' - 131.6 3'', 7'' 7.07 (2H, d, 8.5) 128.8 4'', 6'' 6.63 (2H, d, 8.5) 114.6 5'' - 155.6 One''' 5.25 (1H, d, 7.5) 103.0 2''' 3.48 (1H, dd, 8.5, 7.5) 74.3 3''' 3.43 (1H, dd, 8.5, 8.5) 76.7 4''' 3.36 (1H, dd, 8.5, 8.5) 69.8 5''' 3.22 (1H, ddd, 12.0, 5.0, 2.5) 77.0 6'''a
6'''b 3.71 (1H, dd, 12.0, 2.5)
3.58 (1H, dd, 12.0, 5.0) 61.1

^{In addition, 1} H- ¹ H- of 8-carbon-para-hydrocybenzylquacertin 3-oxy-beta-di-glucopyranoside (8- C - p- hydroxybenzylquercetin 3- O- β-D-glucopyranoside) The COSY correlation (bold line), gHMBC correlation (arrow) and structure are as shown in FIG. 3.

Experimental Example 3

The structure of compound 3 was analyzed as follows, and the analysis results are shown in Table 3 and FIG. 4.

Compound 3 HR-ESI-MS (negative ) a m / z peak from the molecular spectrum 699.1921 [MH] of ^- was observed, it was found that the molecular formula of this compound is C ₃₄ H ₃₄ O ₁₆ (molecular weight 700). And ¹ H- (600 MHz) and ¹³ C-NMR (150 MHz) spectra of compound 3 is very similar to that of the compound 1. In the case of compound 3 ^{, in the 1} H-NMR spectrum, δ 7.94 (2H, d, J = 9.0 Hz, H-2'' and H-6'') and 6.84 (2H, d, J = 9.0 Hz, H-3''AndH-5''), and δ 114.6 (C-2'' and C-6'') and 131.1 (C-2'' and C-6'') in the ^{13 C-NMR (125 MHz) spectrum.} From the observation of compound 3 is of compound 1 It was confirmed that it was camperol instead of quacerol. Compound 3 was structured in the same manner as in Experimental Example 1.

From Table 3 below, where the MS and NMR analysis results are presented, compound 3 is 7-carbon-para-hydrocybenzylchemferol 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di-galatopyrano It was determined as a side [8- C - p -hydroxybenzylkeampferol 3- O- (α-L-rhamnopyr anosyl(1→6)-β-D-galactopyranoside] (Compound 3 ), and it was found to be a new compound.

Table 3 shows 7-carbon-para-hydroxybenzylkeampferol 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di- galatopyranoside[8-C - p- hydroxybenzylkeampferol 3- O -(α-L-rhamnopyranosyl(1→6)-β-D-galactopyranoside] (Compound 3 ) ¹ H- (600 MHz) and ¹³ C-NMR (150 MHz) data (CD ₃ OD).

In addition, 7-carbon-para-hydroxybenzylkeampferol 3-oxy-alpha-L-rhamnopyranosyl(1→6)-beta-di- galatopyranoside[8-C - p- hydroxybenzylkeampferol 3- O- (α-L-rhamnopyranosyl (1 → 6) -β-D-galactopyranoside] 1 H- 1 H-COSY correlation (bold lines), gHMBC correlation (arrows) and the structure is as shown in Fig.

Position δ _H ( Int. , Multi. , J in Hz) δ _C 2 - 157.9 3 - 134.3 4 - 178.4 5 - 159.5 6 6.33 (1H, s) 98.1 7 - 162.3 8 - 106.6 9 - 154.5 10 - 104.2 One' - 121.3 2', 6' 7.94 (2H, d, 9.0) 131.1 3', 5' 6.84 (2H, d, 9.0) 114.6 ^a 4' - 160.1 One'' 4.04 (2H, m) 26.8 2'' - 131.5 3'', 7'' 7.04 (2H, d, 8.5) 128.7 4'', 6'' 6.66 (2H, d, 8.5) 114.6 ^a 5'' - 155.0 One''' 5.02 (1H, d, 7.5) 104.4 2''' 3.79 (1H, dd, 8.0, 7.5) 71.6 3''' 3.55 (1H, dd, 8.0, 2.5) 73.7 4''' 3.76 (1H, dd, 3.5, 2.5) 68.7 5''' 3.62 (1H, m) 73.8 6'''a
6'''b 3.74 (1H, dd, 11.5, 6.0)
3.36 (1H, dd, 11.5, 6.5) 65.8 One'''' 4.52 (1H, d, 1.5) 100.4 2'''' 3.61 (1H, dd, 3.0, 1.5) 70.6 3'''' 3.52 (1H, dd, 9.0, 3.0) 70.9 4'''' 3.30 (1H, dd, 9.0, 9.0) 72.5 5'''' 3.53 (1H, m) 68.3 6'''' 1.20 (1H, d, 6.5) 16.6

Experimental Example 4

The structure of compound 4 was analyzed as follows, and the analysis results are shown in Table 4 and FIG. 5.

Compound 4 HR-ESI-MS (negative ) a m / z peak from the molecular spectrum 699.1918 [MH] of ^- was observed, it was found that the molecular formula of this compound is C ₃₄ H ₃₄ O ₁₆ (molecular weight 700). And ¹ H- (600 MHz) and ¹³ C-NMR (150 MHz) spectra of compound 4 but is very similar to that of the compound 3, instead of galactose proton and carbon signal that is attributed to the glucose were observed. Compound 4 was structured in the same manner as in Experimental Example 1.

From Table 4 below, where the MS and NMR analysis results are presented, compound 4 was determined as 8-carbon-para- hydrocybenzylchemferol 3-oxy-lutinoside (8-C - p -hydroxybenzyl kaempferol 3- O- rutinoside) And turned out to be a new compound.

^{Table 4 shows 1} H- (600 MHz) and ¹³ C of 8-carbon-para- hydrocybenzylchemferol 3-oxy-lutinoside (8-C - p -hydroxybenzylkaempferol 3- O- rutinoside) (compound 4 ) -NMR (150 MHz) data (CD ₃ OD).

Further, the 8-carbon-para-dihydro when benzyl Chem Ferrol 3-oxy-Rutino side (8-C - p-3-O hydroxybenzylkaempferol -rutinoside] ¹ H- ¹ H-COSY correlation (coarse (Compound 4) Line), gHMBC correlation (arrow) and structure are as shown in FIG. 5.

Position δ _H ( Int. , Multi. , J in Hz) δ _C 2 - 157.9 3 - 134.1 4 - 178.2 5 - 159.5 6 6.33 (1H, s) 98.1 7 - 162.3 8 - 106.6 9 - 154.5 10 - 104.3 One' - 121.4 2', 6' 7.93 (2H, d, 9.0) 131.0 3', 5' 6.85 (2H, d, 9.0) 114.7 4' - 160.0 One'' 4.05 (2H, m) 26.8 2'' - 131.6 3'', 7'' 7.05 (2H, d, 8.5) 128.7 4'', 6'' 6.66 (2H, d, 8.5) 114.6 5'' - 155.0 One''' 5.12 (1H, d, 7.5) 103.4 2''' 3.44 (1H, dd, 7.5, 7.5) 74.4 3''' 3.41 (1H, dd, 7.5, 8.0) 76.8 4''' 3.28 (1H, dd, 8.0, 8.0) 69.9 5''' 3.34 (1H, m) 75.8 6'''a
6'''b 3.80 (1H, dd, 11.0, 1.5)
3.39 (1H, dd, 11.0, 6.0) 65.8 One'''' 4.52 (1H, d, 1.5) 101.0 2'''' 3.65 (1H, dd, 3.5, 1.5) 70.7 3'''' 3.55 (1H, dd, 9.5, 3.5) 70.9 4'''' 3.30 (1H, dd, 9.5, 9.5) 72.5 5'''' 3.47 (1H, m) 68.3 6'''' 1.13 (1H, d, 6.0) 16.5

Experimental Example 5

The structures of compounds 5 and 6 were analyzed as follows, and the analysis results are shown in Tables 5 and 6, and Tables 6 and 7 respectively.

Compound 5 and 6 HR-ESI-MS (negative ) a m / z molecular weight peaks from the spectrum 729.2036 [MH] of ^- was observed, molecular formula of this compound was found to be C ₃₅ H ₃₈ O ₁₇ (molecular weight 730). In the ¹ H-NMR (600 MHz) spectrum of compounds 5 and 6 , it was confirmed that two kinds of compounds were mixed at a ratio of 9:1, and among them, a large amount of compounds was present as compound 5 and in trace amounts. The present compound was designated as compound 6.

Position δ _H ( Int. , Multi. , J in Hz) δ _C 2 - 157.3 3 - 134.1 4 - 178.1 5 - 159.5 6 6.31 (1H, s) 98.1 7 - 162.4 8 - 106.3 9 - 154.5 10 - 104.3 One' - 121.6 2' 7.78 (1H, d, 2.0) 112.7 3' - 146.9 4' - 149.4 5' 6.85 (1H, d, 8.5) 114.6 6' 7.53 (1H, dd, 8.5, 2.0) 123.0 -OCH ₃ 3.82 (3H, s) 55.3 One'' 4.05 (2H, m) 26.7 2'' - 131.3 3'', 7'' 7.04 (2H, d, 8.0) 128.6 4'', 6'' 6.67 (2H, d, 8.0) 114.7 5'' - 155.0 One''' 5.22 (1H, d, 7.5) 103.3 2''' 3.49 (1H, dd, 7.5, 7.5) 74.5 3''' 3.46 (1H, dd, 7.5, 8.0) 76.8 4''' 3.29 (1H, dd, 8.0, 8.0) 70.0 5''' 3.38 (1H, m) 75.9 6'''a
6'''b 3.81 (1H, dd, 11.5, 1.5)
3.42 (1H, dd, 11.0, 6.0) 67.0 One'''' 4.54 (1H, d, 1.5) 101.0 2'''' 3.64 (1H, dd, 3.5, 1.5) 70.6 3'''' 3.52 (1H, dd, 9.5, 3.5) 70.9 4'''' 3.27 (1H, dd, 9.5, 9.5) 72.5 5'''' 3.40 (1H, m) 68.3 6'''' 1.11 (1H, d, 6.0) 16.5

¹ H- (600 MHz) and ¹³ C-NMR (150 MHz) spectra of compound 5 were very similar to those of the compound 1, where the proton and carbon signal (δ 3.82 and 55.3), attributable to the methoxyl group observed Became. Compound 5 was structured in the same manner as in Experimental Example 1. From Table 5 below, where the MS and NMR analysis results are presented, compound 5 is 8-carbon-para- hydrocybenzylisoramnetin 3-oxy-lutinoside (8-C - p- hydroxybenzyl isorhamnetin 3- O- rutinoside). It was determined, and it turned out to be a new compound.

^{Table 5 shows 1} H- (600 MHz) of 8-carbon-para- hydrocybenzylisoramnetin 3-oxy-lutinoside (8-C - p -hydroxybenzylisorhamnetin 3- O- rutinoside) (compound 5) and ¹³ C-NMR (150 MHz) data (CD ₃ OD).

Further, the 8-carbon-para-dihydro when benzyl is soram netin ^3-oxy-1 H- ¹ H correlation of the p-COSY -hydroxybenzylisorhamnetin 3-O -rutinoside] (Compound 5) (- Rutino side (8-C Bold line), and gHMBC correlation (arrow) and structure are as shown in FIG.

In the case of compound 6 , galactose instead of glucose of compound 5 ^{was observed in 1} H- (600 MHz) and ¹³ C-NMR (150 MHz) spectra. Compound 6 was structured in the same manner as in Experimental Example 1. From Table 6 below, where the MS and NMR analysis results are presented, compound 6 is 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di-galatopyra It was determined as a noside [8- C - p -hydroxybenzylisorhamnetin 3- O- (α-L-rhamnopyranosyl(1→6)-β-D-galactopyranoside] and proved to be a new compound.

Position δ _H ( Int. , Multi. , J in Hz) δ _C 2 - 157.3 3 - 134.1 4 - 178.3 5 - 159.5 6 6.31 (1H, s) 98.1 7 - 162.4 8 - 106.3 9 - 154.5 10 - 104.3 One' - 121.6 2' 7.85 (1H, d, 2.0) 112.8 3' - 147.0 4' - 149.4 5' 6.84 (1H, d, 8.5) 114.5 6' 7.48 (1H, dd, 8.5, 2.0) 122.9 -OCH ₃ 3.84 (3H, s) 55.4 One'' 4.05 (2H, m) 26.7 2'' - 131.3 3'', 7'' 7.03 (2H, d, 8.0) 128.7 4'', 6'' 6.65 (2H, d, 8.0) 114.7 5'' - 155.0 One''' 5.20 (1H, d, 7.5) 104.4 2''' 3.83 (1H, dd, 8.0, 7.5) 71.7 3''' 3.54 (1H, dd, 8.0, 2.5) 73.6 4''' 3.75 (1H, dd, 3.5, 2.5) 68.6 5''' 3.66 (1H, m) 73.9 6'''a
6'''b 3.76 (1H, dd, 11.5, 6.0)
3.36 (1H, dd, 11.5, 6.5) 65.8 One'''' 4.53 (1H, d, 1.5) 100.4 2'''' 3.61 (1H, dd, 3.0, 1.5) 70.6 3'''' 3.52 (1H, dd, 9.0, 3.0) 70.9 4'''' 3.30 (1H, dd, 9.0, 9.0) 72.5 5'''' 3.53 (1H, m) 68.3 6'''' 1.19 (1H, d, 6.5) 16.6

Table 6 shows 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl (1→6)-beta-di- galatopyranoside [8-C - p- hydroxybenzylisorhamnetin 3- O- (α-L-rhamnopyranosyl(1→6)-β-D-galactopyranoside] (compound 6 ) ¹ H- (600 MHz) and ¹³ C-NMR (150 MHz) data (CD ₃ OD).

In addition, 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl(1→6)-beta-di- galatopyranoside[8-C - p- hydroxybenzylisorhamnetin 3- O to (α-L-rhamnopyranosyl (1 → 6) -β-D-galactopyranoside] 1 H- 1 H-COSY correlation (bold lines), gHMBC correlation (arrows) and the structure of Figure 7 (compound 6) As shown.

Experimental Example 6

Free radical scavenging of 1,1-diphenyl-2-picrylhydrazyl (DPPH, Sigma-Aldrich, St. Louis, MO, USA) for the antioxidant activity of the compounds 1 to 3 , 5 and 6 obtained in Examples 1 and 2 Measurements were made as follows using the activation method, and the results are shown in FIG. 8. A solution (final concentration 10 μM) and 990 μL of DPPH ethanol solution (final 100 μM DPPH concentration) were mixed with a vortex mixer. After reacting the mixed solution for 10 minutes in a dark room at room temperature, absorbance was measured at 517 nm (UV/VIS Spectrophotometer, V-550, Jasco, Tokyo, Japan). In addition, quercetin was used as a positive control. The antioxidant activity of the compounds isolated from amber tendril showed the degree of decrease in the absorbance compared to that of the DPPH ethanol solution to which the sample solution was not added.

From FIG. 8, all of the compounds isolated from amber tendril showed the activity of scavenging DPPH free radicals. Among them, it was found that Compound 1 and Compound 2 exhibited almost similar DPPH free radical scavenging activity to the positive control quercetin.

Experimental Example 7

Cytotoxicity of Compounds 1 to 3 , 5 and 6 obtained in Examples 1 and 2 was tested as follows, and the results are shown in FIG. 9.

Bone marrow was isolated from femur and tibia C57BL/6 male 8-week-old mice. IMDM medium containing 10% FBS, 1% Penicillin/Streptomycin (P/S), Sodium pyruvate, and MEM Non-Essential Amino Acids (NEAA) was used, and 30% of L929 cell culture was added to differentiate into macrophages. . After 3 days, a medium containing L929 cell culture was added, and _{cultured at 37°C and 5% CO 2 for} 6 days (Bone marrow-derived macrophages, BMDMs). Compounds (1~3, 5,6) isolated from the hot water extract of Pumpkin tendril were treated at a concentration of 250 μM. After 24 hours, the cell supernatant was harvested and inflammatory cytokines IL-6 and TNF-α were measured by Enzyme-Linked ImmunoSorbent Assay (ELISA). ^{BMDMs were dispensed to a 96 well plate at 1×10 4} /well on the 6th day of culture, and cultured for 24 hours, and then the MTT method was performed to evaluate the toxicity to cells of the single-compound hot water extract of Zucchini. After treating the hot water extract of amber hand for 24 hours, the MTT solution was added and the light was blocked for 4 hours at _{37°C and 5% CO 2 incubator.} When a purple, water-insoluble formazan was formed, it was dissolved by adding DMSO. Absorbance was measured at a wavelength of 570 nm and graphed as a relative quantitative value of the control.

From FIG. 9, it was confirmed that there was no s-cell toxicity by the compound isolated from the hot water extract of pumpkin tendril. In particular, it was found that Compound 3 and Compound 5 + 6 induced 30% cell proliferation at a concentration of 250 μM.

Experimental Example 8

The immunity enhancing activity of the compounds 1 to 3 , 5 and 6 obtained in Examples 1 and 2 was tested as follows, and the results are shown in FIGS. 10A and 10B.

BMDMs were dispensed to a 48 well plate at 2×10 ⁵ /well and incubated for 24 hours, and then the compounds ( 1 ~ 3 , 5 , 6 ) isolated from the hot water extract of Zucchini tendril were treated with 250 μM each and cultured for 24 hours. The amount of secretion of IL-6 and TNF-α in the cell supernatant was measured using an ELISA kit. ELISA was performed by referring to the protocol of the kit.

As shown in Figs. 10a and 10b, all of the compounds isolated from amber tendril induced the production of IL-6 and TNF-α, and in particular, were produced the highest by compound 3. Through this, it was found that the five compounds are immune enhancing substances as they induce cytokine production.

As shown in the above-described experimental results, the compound of the present invention or a pharmacologically acceptable salt thereof is effective as long as the novel compound derived from amberson having the structural formula as in Formula 1 of the present invention has antioxidant activity and/or immunity enhancing activity. It can be seen that when a food is prepared by including it as an ingredient, a functional food having antioxidant activity and immunity enhancing activity can be prepared.

Although the present invention has been shown and described with a preferred embodiment as described above, it is not limited to the above-described embodiment, and within the scope not departing from the spirit of the present invention, to those of ordinary skill in the art. Various changes and modifications will be possible.

Claims (10)

A new compound represented by the following formula (1).
[Formula 1]

Here, R1 is a hydrogen atom, an oxygen atom or a hydroxy group,
R2 is

,

or

to be.
The method of claim 1,
The compound is a novel compound, characterized in that it has at least one of antioxidant activity and immune enhancing activity.

The method of claim 1,
The compound is a novel compound, characterized in that the extraction and separation from pumpkin tendrils.
The method of claim 1,
The compound is 8-carbon-para-hydrocybenzylquacertin 3-oxy-lutinoside, 8-carbon-para-hydrocybenzylquacertin 3-oxy-beta-di-glucopyranoside, 7-carbon- Para-hydrocybenzylcamperol 3-oxy-alpha-L-rhamnopyranosyl(1→6)-beta-di-galatopyranoside, 8-carbon-para-hydrocybenzylcamperol 3-oxy-lutino Side, 8-carbon-para-hydrocybenzylisoramnetin 3-oxy-lutinoside, 8-carbon-para-hydrocybenzylisoramretin 3-oxy-alpha-L-rhamnopyranosyl (1→6) -A new compound, characterized in that at least one selected from the group consisting of beta-di-galatopyranoside.
Obtaining a pumpkin tendril extract;
Concentrating the pumpkin tendril extract under reduced pressure;
Obtaining an ethyl acetate fraction through the solvent fractionation of the concentrate; And
Separation method of a novel compound derived from amber tendril comprising the step of purifying the ethyl acetate fraction by HPLC using Amberlite XAD-2 and ODS column chromatography and reverse phase column.
The method of claim 5,
The step of obtaining the amber tendril extract is performed by pulverizing the amber tendril and then extracting hot water.
An antioxidant food composition comprising the compound of claim 1 as an active ingredient.
A food composition for enhancing immunity comprising the compound of claim 1 as an active ingredient.
An antioxidant health functional food comprising the compound of claim 1 as an active ingredient.
A health functional food for enhancing immunity comprising the compound of claim 1 as an active ingredient.

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