KR101584592B1 - Novel saponin compound, the method of preparation thereof, the pharmaceutical compositions, the cosmetic compositions, and the health food composition is a composition having that as active ingredient - Google Patents

Abstract

The present invention relates to a hwanggi extract having excellent antiinflammatory activity and a foodstuff and a pharmaceutical composition containing the same as an effective ingredient. The saponin compound derived from the Hwanggi extract of the present invention is excellent in the inhibitory activity against nitrogen monoxide which causes inflammation, and is a natural-derived extract which has been used for a long time, and can be used safely and effectively since it has no cytotoxicity.

Description

TECHNICAL FIELD The present invention relates to a novel saponin compound, a process for producing the same, a pharmaceutical composition containing the same as an active ingredient, a cosmetic composition and a health functional food composition composition having that active ingredient}

More particularly, the present invention relates to a novel saponin compound, an anti-inflammatory active ingredient, a method for producing the same, and a pharmaceutical composition, a cosmetic composition and a health functional food composition containing the same as an active ingredient. .

The present invention relates to a pharmaceutical composition for the treatment of inflammation, which comprises a saponin extract of yellow mugwort. More particularly, the present invention relates to a pharmaceutical composition for treating inflammation comprising saponin extracted from plants as an active ingredient.

Inflammation is known to be caused by various causes, but recent studies have shown that a large amount of nitrogen monoxide released by inductive NN synthase (iNOS) is an important factor causing inflammation [Nadler EP, et al., Semin. Pediatr. Surg., Vol 8, p 148 ~ 154, 1999]. It is also known that the release of an appropriate amount of nitrogen monoxide protects the organism from pathogens, but an inappropriately large amount of released nitrogen monoxide is known to cause inflammation and inflammation related diseases [Nadler E.P., et al., Semin. Pediatr. Surg., Vol 8, p 148 ~ 154, 1999].

Nitric oxide (NO), a very unstable gas, is known to be an important macrophage mediator that kills pathogens and tumor cells in the human body and regulates the immune system [Brightbill HD, et al., Science, 285, p732-736 ]. However, when a large amount of nitrogen monoxide is produced, there is a side effect that kills host cells and causes inflammation. If macrophages continue to produce excess nitrogen monoxide even after the death of the pathogen, they cause shock or severe inflammatory disease after sepsis and cause autoimmune disease [Hobbs A.J., et al., Annu. Rev. Pharmacol. Toxicol., 39, p191-220, 1999]. Therefore, the generation of mononuclear nitric oxide needs to be well controlled. In other words, when the production of nitrogen monoxide is insufficient, it is a problem in the role of macrophages as mediators, but excessive production may cause inflammation or side effects. Therefore, the amount of nitrogen monoxide generated in iNOS needs to be appropriately regulated.

Induction of iNOS in tissues leads to continuous production of high concentrations of nitrogen monoxide, which, when inappropriately overproduced, causes inflammatory effects such as vascular swelling and cytotoxicity. Therefore, in order to treat inflammation caused by nitrogen monoxide, it is necessary to appropriately control or suppress the amount of nitrogen monoxide produced in iNOS.

Currently, anti-inflammatory drugs include dexamethasone and cortisone using corticosteroids. However, they act as anti-inflammatory drugs, but they are toxic and cause side effects such as edema. In addition, they sometimes fail to act selectively for the cause of inflammation and cause severe immunosuppression [Check W.A. and Kaliner M. A., Am. Rev. Respir. Dis., 141, pp. 44-51. 1990]. Thus, among anti-inflammatory agents for corticosteroids, anti-inflammatory compositions comprising an extract of Corynebacterium glutinum as an active ingredient are provided in the patent publication 10-2013-0094145 (published on Aug. 23, 2013).

In addition, steroid drugs using corticosteroids, which are currently used as anti-inflammatory drugs, have severe side effects such as edema. Therefore, non-steroidal medicines with no side effects are urgently required.

Disclosure of Invention Technical Problem [8] The present invention relates to an extract of Hwanggi extract for solving the problems of the prior art, and it has been found that an active ingredient containing a novel saponin compound or a pharmaceutically acceptable salt thereof has excellent antiinflammatory activity. That is, the object of the present invention is to provide a natural extract, a pharmaceutical composition containing it as an active ingredient, a cosmetic composition and a health functional food composition.

In order to solve the above-mentioned problems, the present invention relates to a saponin compound, wherein the saponin compound of the present invention comprises a compound represented by the following formula (1).

In one preferred embodiment of the present invention, the saponin compound may be characterized in that it is derived from an Hwanggi extract.

In addition, the present invention can produce an anti-inflammatory active ingredient of the extract containing the saponin compound as described above.

In one preferred embodiment of the present invention, the anti-inflammatory activity of the anti-inflammatory activity of the present invention is characterized by having an anti-inflammatory activity by controlling the amount of nitrogen monoxide produced in iNOS.

Further, another aspect of the present invention relates to a method for producing the above-described antiinflammatory active saponin compound, which comprises preparing a first extract from Hwanggi, dividing it with ethyl acetate and water, and concentrating it under reduced pressure to prepare a second extract Purified and separated by chromatography to produce an anti-inflammatory active saponin compound.

In another preferred embodiment of the present invention, the method for preparing an anti-inflammatory active saponin compound of the present invention comprises the steps of: obtaining an extract from a solution containing a mixture of sulfur and solvent; Separating the primary extract with ethyl acetate and water to prepare a partition extract from the ethyl acetate organic layer solution, and concentrating the partition extract to obtain a secondary extract; And separating the second extract by chromatography and purifying the second extract.

In a preferred another embodiment of the present invention, first the extract of preparing an extract and produced by performing the extraction process, once to three times, the solvents include alcohols and combinations of water, C ₁ ~ C ₄ And a mixed solvent of water and an organic solvent.

In another preferred embodiment of the present invention, the extract extracted with the water and / or alcohol solvent may be extracted at room temperature and concentrated under reduced pressure to prepare a first extract.

In another preferred embodiment of the present invention, the step of extracting the step of obtaining the secondary extract may comprise using the ethyl acetate and water in a volume ratio of 1.5 to 2.5: 1.

In another preferred embodiment of the present invention, the step of subjecting the step of preparing the secondary extract to reduced pressure may be carried out at a temperature of 35 to 60 DEG C and a pressure of 80 to 100 atm.

In another preferred embodiment of the present invention, the purifying step comprises: separating the second extract by silica gel column chromatography to obtain a first separated product; And separating the primary separation product by reversed phase silica gel column chromatography to obtain purified white mugwort extract.

In another preferred embodiment of the present invention, the saponin compound obtained from the re-separated and purified Hwanggi extract is analyzed by IR spectroscopy, NMR measurement, atomic impact mass spectrometry, nonlinearity measurement and melting point measurement Process can be performed.

Another aspect of the present invention is to provide a pharmaceutical composition, a cosmetic composition and a health functional food composition having an anti-inflammatory effect, comprising the saponin compound or its pharmaceutically acceptable salt separated by extraction and fractionation as the active ingredient, can do.

In another preferred embodiment of the present invention, the active ingredient of the saponin compound may be formulated into an anti-inflammatory pharmaceutical composition which can be administered in an amount of 1 to 1000 mg per 1 kg of body weight per day, divided into several times to several times.

In another preferred embodiment of the present invention, the active ingredient of the saponin compound may be prepared by a conventional method to produce an anti-inflammatory cosmetic composition.

In another preferred embodiment of the present invention, the active ingredient of the saponin compound may be prepared by a conventional method to produce an anti-inflammatory health functional food composition.

Hereinafter, terms used in this specification will be briefly described.

"Inflammation" of the present invention may mean a pathological state of an abscess formed by the infiltration of an external infectious agent (bacteria, fungi, viruses, various kinds of allergens), and "inflammatory disease" It means a disease accompanied by inflammation.

&Quot; Prevention "of the present invention means all actions that inhibit or delay the onset of inflammatory disease by administering the composition, and" treatment "means that symptoms caused by an inflammatory disease Means any act that improves or benefits.

Furthermore, the "quasi-drug product" of the present invention refers to a drug, a drug or the like which is used for treating, alleviating, treating or preventing a disease of a human or an animal, An article which is not an appliance or a machine and which is similar to a product used for sterilization, insecticide and similar uses for the prevention of infectious diseases and which is intended to diagnose, treat, alleviate, Means an article, except machinery, machinery or equipment, which is not an apparatus, machine or apparatus, used for the purpose of causing pharmacological effects on the structure and function of humans or animals;

The pharmaceutical natural composition for the prevention and treatment of inflammatory diseases containing a novel compound isolated from the group consisting of the radish extract and the radish fraction according to the present invention has an activity of inhibiting the production of NO, which is one of the inflammation inducing substances, The side effects can be significantly reduced as compared with the therapeutic agents for inflammatory diseases.

Fig. 1 shows ¹ H NMR results of NMR data measured in Experimental Example 1. Fig.
Fig. 2 shows ¹³ C NMR results of NMR data measured in Experimental Example 1. Fig.
Figure 3 shows the result of high resolution mass spectrometry (HR-FAB / MS) data.
4 is a graph showing the inhibitory activity against nitrogen monoxide according to the concentration of the novel saponin compound measured in Experimental Example 2. FIG.
5 is a graph showing cell viability according to the concentration of the novel saponin compound measured in Experimental Example 3. FIG.

Hereinafter, the present invention will be described in more detail.

As described above, there are few natural extracts having anti-inflammatory activity and solving the problems of conventional synthetic pharmaceutical compositions.

The present invention has been made to solve the above-mentioned problems by producing a saponin compound represented by the following formula (1) or a pharmaceutically acceptable salt thereof. Accordingly, the saponin compound can provide a pharmaceutical composition for the prevention or treatment of inflammatory diseases, and is free from cytotoxicity, thereby providing a pharmaceutical composition which is free from the human body.

[Chemical Formula 1]

The present invention provides a pharmaceutical composition for preventing or treating inflammatory diseases, which comprises the compound represented by the formula (1) or a pharmaceutically acceptable salt thereof.

The compound represented by the general formula (1) can be obtained by general chemical synthesis, separated from natural products, or sold, and preferably those separated from natural products can be used. More preferably, Can be used.

The present invention also relates to an anti-inflammatory active ingredient of the present invention, which comprises a compound represented by the general formula (1).

The salt of the compound represented by the formula (1) may be an acid addition salt formed by a pharmaceutically acceptable free acid, and the free acid may be an organic acid or an inorganic acid. Examples of the inorganic acid include hydrochloric acid, bromine Sulfuric acid, sulfuric acid, phosphoric acid and the like can be used. As the organic acid, citric acid, acetic acid, maleic acid, fumaric acid, gluconic acid, metal sulfonic acid, acetic acid, glycolic acid, succinic acid, tartaric acid, Lactic acid, galacturonic acid, embic acid, glutamic acid, citric acid, and spartanic acid.

The addition salt according to the present invention can be obtained by a conventional method, that is, dissolving the compound represented by Formula 1 in a water-miscible organic solvent such as acetone, methanol, ethanol or acetonitrile, adding an equivalent amount or an excess amount of an organic acid, Or by precipitation or crystallization after adding an aqueous solution, or by evaporating a solvent or an excessive amount of acid, and then drying or precipitating the salt by suction filtration.

The present invention includes not only compounds represented by the above formula (1) and pharmaceutically acceptable salts thereof, but also solvates, hydrates and stereoisomers which exhibit the same effects as those which can be prepared therefrom.

In addition, the saponin compound of the present invention inhibits nitrogen monoxide, which is one of the inflammation inducing factors, to have antiinflammatory activity, and has an effect of inhibiting the inflammation induced by nitrogen monoxide.

A method for producing an anti-inflammatory active saponin compound derived from the present invention is described below.

The first extract was prepared from the anti-inflammatory active extract of the present invention, and the extract was partitioned with ethyl acetate and water and concentrated under reduced pressure to prepare a second extract. The extract was purified and separated by chromatography to prepare an anti-inflammatory active saponin compound can do.

Specifically, the method for producing an anti-inflammatory active saponin compound of the present invention comprises the steps of: preparing an extract from a solution containing a mixture of a sulfur source and a solvent, and concentrating it under reduced pressure to prepare a first extract; Separating the primary extract with ethyl acetate and water to prepare a partitioned extract from the ethyl acetate organic layer solution, and concentrating the partitioned extract to obtain a secondary extract; And separating the secondary extract by chromatography, followed by purification.

In the step of preparing the primary extract, the solvent is water, C ₁ - alcohols of C ₄ and these may be prepared by extraction with a solvent mixture, preferably water, methanol, ethanol, propanol, or 50% (v / v) aqueous solution of said at least an alcohol, more preferably 70 to 85 % Aqueous methanol solution may be used.

The method for obtaining the above extract is not particularly limited as long as it is a method for obtaining an extract from a natural product, but preferably one or more selected from the group consisting of an ultrasonic extraction method, a filtration method and a reflux extraction method may be used.

The vacuum concentration may be performed using a vacuum decompression concentrator or a vacuum rotary evaporator, but is not limited thereto.

The concentration under reduced pressure may be 35 to 60 ° C, preferably 40 to 50 ° C, more preferably 45 ° C, and the pressure is 80 to 100 atm, preferably 85 to 95 atm , And more preferably 90 atm. If the temperature is lower than 35 ° C during the concentration under reduced pressure, the extraction may not be performed properly during the extraction process. If the temperature exceeds 80 ° C, the concentrate may boil over. Also, when the pressure is less than 80 atm, it may take too long to concentrate, and when it exceeds 100 atm, the concentrate may boil over.

Next, in the step of obtaining a liquid distribution extract using an organic solvent, the organic solvent is preferably selected from the group consisting of C ₁ to C ₅ alcohol, water, hexane, methylene chloride and ethyl acetate May be at least one selected, more preferably water and / or ethyl acetate. The ethyl acetate and water may be extracted at a volume ratio of 1: 3: 1, preferably 1.5: 2.5: 1, more preferably 2: 1. Do. If the volume ratio of ethyl acetate and water is less than 1: 1, the ethyl acetate fraction may not be well separated in the aqueous layer. If the volume ratio is more than 3: 1, the reproducibility of the ethyl acetate fraction is poor, There is a problem in that the amount of the liquid is reduced.

The liquid distribution extract is not particularly limited as long as it is prepared using a conventional liquid distribution extraction method. The liquid dispensing and extraction method is for performing a purification using a chromatographic method in the next step more effectively by removing a large amount of impurities from a polar substance to a non-polar substance contained in the extract, and the large amount of impurities is solubility And the mixture of the solvent and the solvent.

Then extracting the solution separated with ethyl acetate and water, and concentrating the filtrate under reduced pressure to prepare a second extract. The step of purifying is a step of separating the second extract by silica gel column chromatography to obtain a first separated product; And separating the primary separated product by reverse phase silica gel column chromatography to obtain purified white mugwort extract.

The purification method is not particularly limited so long as it is a method of separating and purifying the components, but chromatography can be preferably carried out

Furthermore, the present invention can separate the secondary extracts obtained by concentration under reduced pressure by chromatography, and purify them to obtain an active fraction exhibiting anti-inflammatory activity. The chromatography is preferably performed three to several times using silica gel column chromatography. The active fraction obtained by silica gel column chromatography is further subjected to reversed phase silica gel column chromatography to obtain a more purified purified active fraction. When the chromatography is carried out, a mixed solvent in which chloroform and methanol or methanol and distilled water are mixed can be used as the developing solvent.

Further, the saponin compound obtained by separation and purification by the above chromatography was measured by IR spectroscopy using a Perkin model 599B (Buckinghamshire, England), NMR (Nuclear Magnetic Resonance) was measured with a 400 MHz FT-NMR spectrometer The fast atom bombardment mass spectrum was measured using JMSAX 700 (JEOL), and the non-linearity was measured with a polarimeter P-1020 (JASCO, Tokyo, Japan) ), And the melting point was determined using a Fisher-John's Melting Point Apparatus (Fisher Scientific, Miami, Fla., USA) to analyze the structure of the saponin compounds derived from the Hwanggi extract.

In addition, the saponin compound can prevent or treat inflammation by controlling the amount of nitrogen monoxide produced in iNOS. Induction of iNOS in tissues leads to continuous production of high concentrations of nitrogen monoxide, which, when inappropriately overproduced, causes inflammatory effects such as vascular swelling and cytotoxicity. Therefore, in order to treat inflammation caused by nitrogen monoxide, it is necessary to appropriately control or suppress the amount of nitrogen monoxide produced in iNOS.

The composition of the present invention can be prepared into a powder, a granule, a tablet, a capsule, a suspension, a syrup, a tablet or the like by mixing with a carrier or a pharmaceutically acceptable carrier, a diluent or the like by a method known in the pharmaceutical field. An external preparation, an injection, or the like. The dose of the active ingredient according to the present invention is appropriately selected depending on the degree of absorption of the active ingredient in the body, the rate of water activation and excretion, the age, sex and condition of the patient, severity of the disease to be treated, The composition may be administered in an amount of 1 to 1000 mg per 1 kg of body weight, once or several times.

In addition, the present invention can provide an anti-inflammatory cosmetic composition comprising a saponin compound derived from an extract of Angelica keiskei koidz. As an active ingredient.

Specific formulations of the cosmetic composition of the present invention include formulations such as skins, toners, lotions, creams, essences, soaps, shampoos, cleansing foams and powders. The cosmetic composition may contain any other components commonly used in cosmetics such as a solubilizer, an antioxidant, a suspending agent, a stabilizer, a blocking agent, a foaming agent, a surfactant, and an emulsifier.

Further, the present invention relates to an anti-inflammatory health functional food comprising a saponin compound derived from a hwanggi extract as an active ingredient. The saponin compound derived from the Hwanggi extract can be prepared by the same method as the pharmaceutical composition. There is no particular limitation on the kind of the food. Examples of the food to which the above substance can be added include all kinds of health foods in a usual sense, such as meat, bread, confectionery, rice cakes, chocolate, noodles, gums, dairy products including ice cream, various soups, . The mixing amount of the horsetail extract can be suitably determined according to the purpose of use thereof.

Generally, the amount of the extract of the health functional food composition may comprise from 0.1 to 95% by weight, preferably from 5 to 95% by weight, of the total food weight. In addition, in the case of long-term ingestion intended for health and hygiene purposes or for the purpose of controlling health, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount exceeding the above range.

Hereinafter, the present invention will be described in more detail with reference to the following Examples. However, these drawings and the following embodiments are only illustrative of the contents and scope of the technical idea of the present invention, and the technical scope of the present invention is not limited or changed. It will be apparent to those skilled in the art that various modifications and variations are possible within the scope of the technical idea of the present invention based on these examples.

[ Example ]

Example  1. Compound separation

Example  1-1: Hwanggi  Extract preparation

The hwanggi cultivated under hydroponic cultivation conditions was harvested for 1 year, and the harvested hwanggi was peeled and dried, followed by pulverization to obtain 10 kg of hwanggi powder sample. The yellowish powder sample was immersed in an aqueous solution of 80% methanol for 24 hours at 25 ° C, and the extract was filtered to obtain a filtrate. The filtrate was concentrated under reduced pressure to obtain 587 g of concentrated extract of reduced pressure 1. Thereafter, the above-mentioned extraction was repeated twice, and the filtrate obtained by repeated extraction was combined and concentrated under reduced pressure to obtain a reduced pressure concentrated extract 2 (800 g).

The depressurized concentrated extracts 1 and 2 were combined to obtain a depressurized concentrated extract of 1.387 g in total.

Example  1-2: Hwanggi  Manufacture of distribution extract

The concentrated extract under reduced pressure obtained in Example 1-1 was partitioned with ethyl acetate (EtOAc, 6 L) and water (3 L), and the water layer was further extracted with butanol (n-BuOH, 5 L). Each layer (ethyl acetate, water, butanol) was concentrated under reduced pressure at a temperature of 45 캜 and a pressure of 90 atm to obtain an ethyl acetate distribution extract (57 g), a butanol distribution extract and a water distribution extract.

Chromatographic analysis of the components of the above-mentioned distribution extracts revealed that the ethyl acetate distribution extract contained a large amount of substances which were confirmed to be saponins rather than the butanol distribution extract and the water distribution extract. Thus, the compounds were then separated in the ethyl acetate distribution extract.

Example  1-3: Isolation of compounds

The ethyl acetate fraction obtained in Example 1-2 was sequentially subjected to silica gel column chromatography and reverse phase chromatography to separate and purify the substance exhibiting anti-inflammatory activity. The silica gel was suspended in dichloromethane, methanol and water in a developing solvent, filled in a glass column, and then the ethyl acetate fraction obtained in Example 1-2 was loaded and eluted. At this time, methanol and water solvent system were used as developing solvent in a ratio of 1: 1.8 parts by weight. As a result of the chromatography, 23 eluted fractions were obtained and Thin layer chromatography (hereinafter referred to as TLC) was carried out to confirm the distribution of the substance to be processed, and a similar substance was collected. As a result, it was found that the fraction showing the greatest anti-inflammatory activity was the 21st fraction, and the obtained amount was 140 mg. This was further subjected to reversed phase silica gel column chromatography using methanol and water solvent system in a ratio of 3: 1 by weight. And 12 mg of the final active substance was isolated and purified by elution with a mixed solvent.

Experimental Example  1. Structure analysis of compounds

For the structural analysis of the final active material obtained in Example 1-3, a 400 MHz FT-NMR spectrometer (Varian Inova AS 400, Palo Alto, Calif.) Was used for one-dimensional and two-dimensional NMR, , The structure of the compound was analyzed by measuring using pyridine-d5 (Merck, Germany) reagent. ¹ H NMR and NMR data of ¹³ C-NMR are shown in FIG. 1 and FIG. 2, respectively. Through the above structural analysis, it was confirmed that the compound was a compound represented by the following formula (1).

[Chemical Formula 1]

1) Properties: White solid form

2) ¹ H-NMR (400 MHz, pyridine-d5,?) And ¹³ C-NMR (100 MHz, pyridine-

3) Analysis results

Compound 1 (white solid) showed absorption of hydroxyl group (3,433 cm ^-1 ) and absorption of carbonyl (1,724 cm ^-1 ) in the IR spectrum and molecular weight was 811.4777 [MH ] ^- . ≪ ^/ RTI >

J = 8.0, 8.0, 5.2 Hz), 3.53 (ddd, J = 9.5, 9.5, 4.5 Hz) in the ¹ H-NMR (400 MHz, pyridine- ), 3.41 (dd, J = 10.5, 2.2 Hz), 3.23 (dd, J = 11.3, 4.0 Hz) and 6 methyl groups (δH 0.95, 1.28, 1.38, 1.41, 1.43, and 1.78 ) And one acetal group (δH 2.03) were observed. The carbon signal was also observed at 19.9 ppm, 16.6 ppm, 18.6 ppm, 25.7 ppm, 26.5 ppm and 28.3 ppm, and one acetyl carbon (δ C 21.2) was observed. In addition, a cyclopropyl-ring system showing germinal coupling was observed with δH 0.53 (1H, d, J = 4.0 Hz, H-19a) and δH 0.07 (1H, d, J = 4.0 Hz, H- Methylene (methylene) group was present. Two anomeric hydrogen signals observed from the sugar molecule were observed [delta H 4.77 (J = 7.6 Hz), and 4.96 (J = 7.2 Hz)]. The types of bound sugars were determined by comparing the chemical shifts of carbon NMR with those of β-xylose and β-glucose. The HMBC experiment was conducted to confirm the binding sites of the two sugars. The signal of the first anomer proton, 4.77, confirmed the signal and cross peak of 89.0, confirming that the first xylose sugar binds to position 3 of the aglycon. The binding position of the second sugar was confirmed by confirming the 79.1 carbon signal and the cross peak of the proton signal of 4.96 and binding to the 6th of the aglycon. And the ester carbon signal (170.0 ppm) showed two proton signals (δH 5.52, H-2 ') and cross peak of Xylose and the binding site was determined. As a result of synthesis of these results, it was found that a novel compound which has not been reported so far is 3-O-? - (2'-O-acetyl) -D-xylopyranosyl-6-0-? -D-glucopyranosyl- , 24α, 25-tetrahydroxy-9,19-cyclolanostane, and named agroastragaloside V.

Experimental Example  2. Anti-inflammatory activity measurement

The amount of NO produced from RAW 264.7 cells was determined using the Griess reagent as the form of NO2- present in the cell culture. The compound obtained in Example 1-3 was added to 0.1% dimethyl sulfoxide (DMSO) to prepare a sample solution having concentrations of 1.5625 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM or 50 μM. Then, the cells were equally divided into 96-well plates at a density of 1 × 10 ⁵ cells / well and cultured for 24 hours. Then, RAW 264.7 cells were cultured at various concentrations (1.5625 μM , 3.125 [mu] M, 6.25 [mu] M, 12.5 [mu] M, 25 [mu] M or 50 [mu] M. 100 μl of the cell culture supernatant and 100 μl of the Griess reagent were mixed and reacted on a 96-well plate for 10 minutes, and the absorbance was measured at 540 nm. The measured nitric oxide is shown in FIG.

The sample solutions of various concentrations were prepared by adding the compound represented by the formula 1 prepared in Example 1-3 to 0.1% dimethyl sulfoxide at 1.5625, 3.125, 6.25, 12.5, 25 or 50 μM Respectively.

The Griess reagent was added to 1% (w / v) sulfanilamide with 5% (v / v) phosphoric acid and 0.1% (w / v) naphthylethylenediamine- HCl). As a result of examining the effect of inhibiting nitric oxide on the cell culture treated with the saponin compound, the NO production inhibitory activity was confirmed to be about 30 to 35% at the concentration of 1.56 μM, and the NO production inhibitory activity at the concentration of 3.12 μM was about 65 to 70% (See Fig. 4). ≪ RTI ID = 0.0 >

Experimental Example  3. Measurement of cytotoxic activity of compounds

In order to measure the cytotoxic activity of the compound represented by the formula (1) obtained in Example 1-3, the cell viability was measured.

The compound obtained in Example 1-3 was added to 0.1% dimethylsulfoxide to prepare a sample solution having concentrations of 1.56, 3.125, 6.25, 12.5, 25, 50 or 100 μM. The cells were then equally divided into 96-well plates at a density of 1 × 10 ⁵ cells / well and cultured for 24 hours. Then, the cells were cultured at various concentrations (1.56 μM, 3.125 μM, 6.25 μM, 12.5 μM, 25 μM, or 50 μM) was added to the medium. After 1 hour of the addition, 1 占 퐂 / ml of lipopolysaccharide (LPS) was added to only one group. After 1 hour, MTT (Microculture Tetrazolium) reagent was added and the mixture was allowed to stand for 12 hours, and the supernatant was removed to obtain a precipitate. Then, 100 μl of dimethyl sulfoxide (DMSO) was added to the precipitate, and the formazan formed in the precipitate was dissolved to prepare a solution. After 30 minutes of the DMSO addition, the absorbance of the solution was measured at 540 nm.

The survival rate of the cells was measured by the ratio of surviving cells in the group treated with the compound represented by the formula (1) and the control group in which no treatment was carried out according to the following formula (1). The measured cell viability is shown in Fig.

As can be seen in FIG. 5, it was confirmed that the cell viability of the test group treated with the compound represented by Formula 1 was not decreased from 6.25 μM to 12.5 μM as compared with the cell survival rate of the control group that was not treated with anything. Thus, it was confirmed that the compound represented by Formula 1 prepared in Example 1-3 had no cytotoxicity. (See Fig. 5)

[ Manufacturing example ]

Manufacturing example  1: Preparation of pharmaceutical composition

The preparation examples of the pharmaceutical compositions comprising the compound represented by the formula (1) (hereinafter referred to as " compound 1 ") prepared in Example 1-3 of the present invention will be described, but the present invention is not limited thereto But rather just to explain it specifically.

Manufacturing example  1-1. Sanje  Produce

Compound 1 2 g

Lactose 1 g

The above components were mixed and packed in airtight bags to prepare powders.

Manufacturing example  1-2. Manufacture of tablets

Compound 1 100 mg

Corn starch 100 mg

Lactose 100 mg

2 mg of magnesium stearate

After mixing the above components, tablets were prepared by tableting according to a conventional method for producing tablets.

Manufacturing example  1-3. Preparation of capsules

Compound 1 100 mg

Corn starch 100 mg

100 mg of milk

2 mg of magnesium stearate

After mixing the above components, the capsules were filled in gelatin capsules according to the conventional preparation method of capsules.

Manufacturing example  1-4. Manufacture of rings

Compound 1 1 g

Lactose 1.5 g

Glycerin 1 g

0.5 g of xylitol

After mixing the above components, they were prepared so as to be 4 g per one ring according to a conventional method.

Manufacturing example  1-5. Manufacture of granules

Compound 1 150 mg

Soybean extract 50 mg

200 mg of glucose

600 mg of starch

After mixing the above components, 100 mg of 30% ethanol was added and the mixture was dried at 60 캜 to form granules, which were then filled in a capsule.

Manufacturing example  2: Preparation of cream containing Compound 1

The atopic cream containing the compound represented by the formula (1) (hereinafter referred to as "compound 1") prepared in Example 1-3 was prepared by mixing in the following amounts.

Compound 1 3 ml

4 ml of glycerin monostearate

4.5 ml of cetearyl alcohol

Stearic acid 3 ml

Wax 1.5 ml

Glycerin 3 ml

Squalane 8 ml

Betaine 6 ml

0.5 ml of sodium hypaloronate

Hardened vegetable oil 2 ml

Polysorbate 60 3 ml

Add 100 ml of distilled water

Although the composition ratio of the ingredients suitable for the formulation of the cosmetic composition is mixed and prepared as a preferable preparation example, the compounding ratio may be arbitrarily varied depending on the regional or national preference such as the demand level, the demanded country, and the use purpose.

Claims (12)

A saponin compound comprising a compound represented by the following formula (1);
[Chemical Formula 1]

2. The saponin compound according to claim 1, wherein the saponin is derived from an extract from a hull. delete delete A pharmaceutical composition for preventing or treating inflammation, which comprises the saponin compound of claim 1 as an active ingredient. An anti-inflammatory cosmetic composition comprising the saponin compound of claim 1 as an active ingredient. An anti-inflammatory health functional food composition comprising the saponin compound of claim 1 as an active ingredient. Extracting the extract from a solution containing a mixture of a sulfur source and a solvent, and concentrating it under reduced pressure to prepare a first extract;
Separating the primary extract with ethyl acetate and water to prepare a partitioned extract from the ethyl acetate organic layer solution, and concentrating the partitioned extract to obtain a secondary extract; And
Separating the second extract by chromatography and purifying it to obtain a saponin compound represented by Formula 1 below;
: ≪ / RTI > a method for producing an anti-inflammatory active saponin compound,
[Chemical Formula 1]

The method according to claim 8, wherein the extract of the step of preparing the first extract is prepared by performing the extraction process one to three times, and the solvent is selected from water, a C ₁ to C ₄ alcohol system and a mixed solvent thereof Wherein the anti-inflammatory activity is at least one kind selected from the group consisting of: 9. The method according to claim 8, wherein the step of obtaining the second extract comprises using the ethyl acetate and water in a volume ratio of 1.5 to 2.5: 1. 9. The method according to claim 8, wherein the step of preparing the second extract is carried out at a temperature of 35 to 60 DEG C and a pressure of 80 to 100 atm. The method of claim 8, wherein the purifying step
Separating the second extract by silica gel column chromatography to obtain a first separated product; And
Separating the primary product by reverse phase silica gel column chromatography to obtain a purified saponin compound represented by Formula 1;
≪ / RTI > wherein the anti-inflammatory activity is selected from the group consisting of:

Images