KR20130064908A - Expression inhibitors of inos protein using dendropanax morbifera lev. leaf and the extracting method thereof - Google Patents

Abstract

PURPOSE: An inhibitor of iNOS expression using Dendropanax morbifera Lev. leaves and a method for extracting the same are provided to obtain the inhibitor which effectively suppresses iNOS. CONSTITUTION: An inhibitor of iNOS expression using Dendropanax morbifera Lev. leaves contains a Dendropanax morbifera Lev. leaf extract. A tablet, a capsule, and a syrup contain the inhibitor. A method for extracting the inhibitor comprises: a step of extracting Dendropanax morbifera Lev. leaves with hydrous alcohol, and concentrating(S101); and a step of fractioning the concentrate with a non-polar solvent and preparing a fraction(S103). [Reference numerals] (AA) Start; (BB) End; (S101) Concentration step of manufacturing concentrate by extracting Dendropanax morbifera Lev. leaves with hydrous alcohol; (S103) Fraction manufacturing step of fractionating the concentrate obtained from the concentration step using a non-polar solvent

Description

Inhibitors of nitric oxide synthase (IINOS) protein expression using the leaves of Hilchi chinensis and method for extracting the same [EXPRESSION INHIBITORS OF iNOS PROTEIN USING Dendropanax morbifera Lev. LEAF AND THE EXTRACTING METHOD THEREOF}

The present invention relates to a nitric oxide synthase protein expression inhibitor using a yellow lacquer tree leaf and a method for extracting the same, and more particularly, to nitric oxide using a yellow lacquer tree leaf excellent effect of inhibiting the expression of nitric oxide synthase protein which is an inflammatory disease trigger. It relates to a synthase protein expression inhibitor and its extraction method.

The present invention relates to a nitric oxide synthase protein expression inhibitor using a yellow lacquer tree leaf and a method for extracting the same, and more particularly, to nitric oxide using a yellow lacquer tree leaf excellent effect of inhibiting the expression of nitric oxide synthase protein which is an inflammatory disease causing factor. It relates to a synthase protein expression inhibitor and a method for producing the same.

Inflammatory reactions involve damage to tissues (cells) or external infectious agents (bacteria, fungi, viruses, various types of allergens), which involve various enzymes and immune cells in local blood vessels and body fluids. A series of complex physiological reactions, including substance secretion, fluid infiltration, cell migration, and tissue destruction, and external symptoms such as erythema and edema, fever and pain. In normal cases, the inflammatory response removes external infectious agents and regenerates damaged tissues to restore life function.However, if the inflammatory response occurs excessively or continuously due to the absence of antigens or internal substances, the main pathology of the disease (sensitizing disease) , Chronic inflammation) and also become a disorder in the treatment process such as blood transfusion, drug administration, and organ transplantation.

In fact, the inflammatory response factors present in vivo include nitric oxide (NO), including inhibition of tumor necrosis factor (TNF) -α production, interleukin (IL) -3 and interleukin-12 mRNA expression, and prostaglandin (PGE2). ), By inhibiting the production of reactive oxygen species (ROS) has the effect of regulating the function of macrophage (Macrophage) involved in immune regulation, and natural herbal extracts useful for the prevention and treatment of diseases caused by immune response It has been attracting attention.

Nitric Oxide (NO) is an inorganic low molecular radical produced by nitric oxide synthase (NOS) (Feldman PL and Griffith OW; Chem. Eng. News., 12, pp 26-38, 1993). This enzyme is composed of constitutive NOS (cNOS), which is responsible for the production of nitric oxide for normal physiological functions such as neurotransmitter function, blood coagulation and blood pressure control, and induced nitric oxide synthase induced under special circumstances. Separated by. General nitric oxide synthase (cNOS) is further divided into neuronal nitric oxide synthase (neuronal NOS, nNOS) and endothelial nitric oxide synthase (endothelial NOS, eNOS). They are dependent on calcium (Ca2 +) and calmodulin (CaM) and produce a small amount of nitric oxide in a short time by stimulation, whereas inducible nitric oxide synthase (iNOS) is independent of calcium and normally Although not activated, its expression is induced by stimulation of liposaccharide (LPS), interferon-γ (Interferon-γ, INF-γ), and cancer necrosis factor-α (TNF-α). , Producing large amounts of nitric oxide for a long time (Mayer B. and John M .; FEBS Lett, 288, pp187-191, 1991).

Excessive nitric oxide produced by nitric oxide synthase (iNOS) causes severe sepsis in patients with septicemia due to vascular dilatation, increased inflammation and stimulation of biosynthesis of inflammatory mediators such as prostaglandins through activation of COX-2, It is involved in the development and deepening of brain diseases caused by neuronal cell death. In other words, enzymes related to the biosynthesis of nitric oxide synthase (NOS) and prostaglandins (PGs) play an important role in mediating the inflammatory response, which is why L-arginine NOS, an enzyme that produces NO from, and COX (cyclooxygenase), an enzyme involved in synthesizing prostaglandins from arachidonic acid, are the main targets in blocking inflammation. In summary, transcriptional inflammatory factors, including iNOS and COX-2, which enhance the degree of NO and PGs as the cause of inflammation, are characterized by various sclerosis, Parkinson's disease, Alzheimer's disease (AD), and It is associated with the pathogenesis of chronic diseases, including colon cancer.

In addition, according to recent studies, there are several types of NOS, such as bNOS (brain NOS) present in the brain, nNOS (neuronal NOS) present in the nervous system, and eNOS (endothelial NOS) present in the vascular system. It is expressed at a certain level, and a small amount of NO produced by them plays an important role in maintaining normal homeostasis, such as inducing neurotransmission and vasodilation, whereas it is induced by various cytokines or external stimulants. NO, which is excessively produced by nitric oxide synthase, is known to cause cytotoxicity and various inflammatory reactions. Chronic inflammation has been associated with an increase in nitric oxide synthase activity (Miller MJ et al., Mediators of inflammation). , 4, pp 387-396, 1995: Appleton L. et al., Adv. Pharmacol., 35, pp 27-28, 1996).

Macrophages, on the other hand, are derived from blood mononuclear cells (leukocytes) and are a special kind of tissue cells. Macrophages remove antigeny debris or kill pathogenic microorganisms and collect into lymphocyte cells for antigenic action (Kinne et al., 2000). Therefore, macrophage activity plays an important role in effective internal immune system. When the body is subject to pathogenic stimuli or damage, macrophages can produce a variety of proinflammatory cytokines (TNF-α, interleukin-1, -2), chemokines responsible for signaling between cells, and inflammatory molecules. Releases nitric oxide (NO), reactive oxygen species (ROS) and PGE2, and also upregulates the surface level of glycoproteins (co-stimulatory molecules CD80 and CD86, adhesion molecules) (Bresnihan, 1999; Burmester et al. , 1997; Gracie et al., 1999).

However, the properties of these macrophages depend on the magnitude of the host's immune response. Macrophages derived from inflammatory mediators become more plentiful in the blood and cause inflammatory diseases such as septic shock, rheumatoid arthritis, and atherosclerosis (Gracie et al., 1999; Michaelsson et al., 1995; Stuhlmuller et al., 2000). To prevent damage. Therefore, whether the disease recovers from severe conditions depends on the regulation of macrophage function.

In addition, from the point of view of external substances causing inflammation-producing factors such as nitric oxide (NO), when treated with antibiotics under bacterial infection, LPS released from the extracellular membrane while killing bacteria causes endotoxin shock. Severe destruction in the process will not neutralize excessively generated LPS (Dunn, DL, Surg. Clin. North. Am., 74, 1994: Giroir, BP, Crit. Care Med., 21, 1993: Yamaguchi , Y. et al., J. Reticuloendothel. Soc., 409, 1982). Therefore, the anti-inflammatory effect can be seen by reducing the action of LPS by drug treatment, which results in the inhibition of nitric oxide (NO) production produced by LPS.

To date, nonsteroidal antiinflammatory drugs (NSAIDs) have been the most widely used in inflammatory diseases and cannot be denied that they have made a significant contribution in terms of their effectiveness. In addition, most non-narcotic analgesics (such as aspirin) also have anti-inflammatory effects and have been used appropriately for the treatment of acute and chronic inflammatory diseases. However, due to side effects, it is difficult to use in the long term, and as a result, the seriousness of the side effects is so high that the development of new or improved therapies is indispensable and urgent. In connection with the problem of overcoming these side effects, efforts have recently been made to find the active ingredient in natural products that are used in the private sector.

Therefore, there is a need for the development of a drug that effectively inhibits the expression of iNOS that intensifies inflammation and even causes brain diseases and develops a safe inflammatory disease treatment without side effects. As a result, natural herbal medicine-derived materials may be an important material that can meet such demands.

Hwangchil-tree (Dendropanax morbifera Lev.) Is an evergreen broad-leaved arboreous tree that grows only in the southwestern coastal area of Jeollanam-do and Jeju Island.黄 漆) is called.

Hwangchil has been used as a rare paint that emits the golden color of emperor's armor, helmets, and other metal ornaments since the Three Kingdoms.It is a Korean envoy, written in the Goryeo Dynasty, and Guilin History, Guilin, and Haedong History. It is written on the back, and it remains in the Tangbu History Book Book District, which is a special product of Baekje.

However, not only Hwangchil tree extract but also Hwangchil tree is effective in eliminating heat, detoxifying alcohol, treating eye and jaundice, treating burns, and leprosy, and it is known to be harmless to humans (Lee Si-jin, Herb, Chinese Moonshine Book, 1590). It has been known to exhibit sedation and tonic action (research of unique global agricultural commodities (Hwang Chil-pyeon), Jeollanam-do, 1996). In addition, it is known that the leaf extract fraction has anti-cancer activity and slightly weaker antioxidant activity than α-tocopherol (Patent Publication No. 2000-0004499, Park Ho-geun et al., 1998), but it has been used for thousands of years. Considering the value, the results of pharmacological activity are very poor.

An object of the present invention is an inhibitor of the expression of nitric oxide synthase protein using a yellow lacquer tree leaf extract or a method of extracting the extract from the yellow lacquer tree leaf extract or the yellow chile leaf extract, which is excellent in inhibiting the expression of nitric oxide synthase protein which is an inflammatory disease causing factor. To provide.

An object of the present invention is achieved by providing an inhibitor of nitric oxide synthase protein expression using the hwangchil tree leaves, characterized in that consisting of the hwangchil tree leaves extract.

In addition, an object of the present invention can also be achieved by providing an inhibitor of nitric oxide synthase protein expression using the hwangchil leaves, characterized in that consisting of a fraction of the hwangchil leaves extract.

According to a preferred feature of the present invention, the inhibitor of nitric oxide synthase protein expression using the yellow lacquer tree leaf will further comprise a pharmaceutically acceptable inert carrier.

According to a further preferred feature of the invention, the inert carrier is to be made of one or more selected from the group consisting of lactose, starch and pregelatinized corn starch.

According to a more preferred feature of the present invention, the nitrogen oxide synthase protein expression inhibitor using the hwangchil leaves further comprises at least one additive selected from the group consisting of diluents, flavors, solubilizers, lubricants, suspensions, binders and disintegrants It shall be included.

In addition, the object of the present invention can also be achieved by providing a tablet characterized in that it contains a nitric oxide synthase protein expression inhibitor using the yellow lacquer tree leaves.

In addition, an object of the present invention can also be achieved by providing a capsule, characterized in that containing the nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves.

In addition, an object of the present invention can also be achieved by providing a syrup that is characterized by containing nitric oxide synthase protein expression inhibitor using the hwangchil leaves.

In addition, the object of the present invention is the oxidation step using the hwangchil wood leaves, characterized in that the extract comprises the step of extracting the extract from the Hwangchil wood with a hydrous alcohol to produce a concentrate and the fraction obtained by fractionating the concentrate obtained in the concentration step with a non-polar solvent It can also be achieved by providing a method for extracting nitrogenase enzyme expression inhibitors.

According to a preferred feature of the present invention, the step of concentrating is prepared by applying hot-water or reflux for 5 to 24 hours at a temperature of 40 to 100 ℃ the sulfuric acid leaves to the water-containing alcohol to produce the yellow-chilewood leaf extract, The supernatant of the filtrate obtained by filtration after cooling to 10 to 40 ° C. is separated, and then the separated supernatant is concentrated by evaporation.

According to a more preferred feature of the present invention, the concentration step is to put the hwangchil wood leaves in the extraction solvent, and chilled for 5 to 7 days at a temperature of 4 to 20 ℃ to produce the hwangchil wood leaf extract, After filtration, the filtered filtrate is concentrated by evaporation.

According to a more preferable feature of the present invention, the nonpolar solvent is an alkane nonpolar solvent having 4 to 10 carbon atoms.

According to a still further preferred feature of the present invention, the nonpolar solvent is comprised of at least one selected from the group consisting of normal hexane, chloroform and ethyl acetate.

Inhibitor of nitric oxide synthase protein expression using the hwangchil tree leaves according to the present invention and the extraction method thereof is an inhibitor of the expression of nitric oxide synthase protein using the hwangchil tree leaves excellent in inhibiting the expression of nitric oxide synthase protein which is an inflammatory disease causing factor Excellent effect provided.

1 is a flow chart showing a method for extracting nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves according to the present invention.
Figure 2 is a graph showing the effect of inhibiting nitric oxide production of nitric oxide synthase protein expression inhibitor using the hwangchil tree according to the present invention.

Hereinafter, preferred embodiments of the present invention and physical properties of the respective components will be described in detail with reference to the accompanying drawings. However, the present invention is not limited thereto, And this does not mean that the technical idea and scope of the present invention are limited.

Nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves according to the present invention consists of a hwangchil tree leaf extract, or a fraction of the hwangchil tree leaf extract.

In addition, the inhibitor of nitric oxide synthase protein expression using the yellow lacquer tree leaf may further include a pharmaceutically acceptable inert carrier.

The inert carrier serves to formulate the nitric oxide synthase protein expression inhibitor using the yellow lacquer leaf into a pharmaceutical preparation in solid, semi-solid or liquid form suitable for oral administration, and can be suitably used for this purpose. The inert carrier may be a solid or a liquid, preferably consisting of one or more selected from the group consisting of lactose, starch and pregelatinized corn starch.

In addition, the nitric oxide synthase protein expression inhibitor using the yellow lacquer tree leaf may further include one or more additives selected from the group consisting of diluents, flavors, solubilizers, lubricants, suspensions, binders and disintegrants.

In addition, the nitric oxide synthase protein expression inhibitor using the yellow lacquer tree leaf extract or a yellow lacquer tree leaf extract fraction as described above may be used in addition to tablets, capsules and syrups.

The tablets were sieved nitric oxide synthase protein expression inhibitor using the yellow lacquer tree leaves, mixed with lactose, starch and pregelatinized corn starch, granulated into powder by the addition of purified water and dried granules It is prepared by mixing with magnesium stearate and compressing, wherein the preferred ingredient and content of the tablet is 5.0 mg of nitric oxide synthase protein expression inhibitor using lacquer leaves, lactose BP 150.0 mg, starch BP 30.0 mg, pregelatinized corn starch 15.0 mg BP and 1.0 mg magnesium stearate.

In addition, the capsule is prepared by sieving the nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves and then mixed with excipients and filled into gelatin capsules, wherein the preferred component and content of the capsule is the 5.0 mg of the nitric oxide synthase protein expression inhibitor used, 100.0 mg of starch (1500), and 1.0 mg of magnesium stearate BP.

In addition, the syrup is dissolved in sucrose in 500 ml of purified water, and dissolved in carboxymethyl cellulose sodium in 400 ml of purified water in a separate container, and then the purified water in which the white sugar is dissolved and purified water in which methyl cellulose sodium is dissolved are mixed. After dissolving by adding propylparaben and adding ethanol, purified water is added so that the total solution has a volume of 1000 ml, and it is prepared by suspending an inhibitor of nitric oxide synthase protein expression using the yellow lacquer leaves. , Preferred components and contents of the syrup are 5.0g of nitric oxide synthase protein expression inhibitor using sulfur lacquer leaves, 637.5g per white, sodium carboxymethylcellulose 2.0g, 0.28g methylparaben, 0.12g propylparaben, 20ml, purified water The remaining amount is.

In addition, the extraction method of the nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves according to the present invention is concentrated step (S101) and the concentrate obtained in the concentration step (S101) to extract the hwangchil tree leaves with a hydrous alcohol Fraction preparation step of fractionating with a non-polar solvent (S103).

The concentration step (S101) is the concentration step is to put the yellow lacquer leaves in water-containing alcohol and hot water or reflux extraction for 5 to 24 hours at a temperature of 40 to 100 ℃ to prepare the yellow lacquer leaves extract, After cooling to 10 to 40 ° C., the supernatant of the filtrate obtained by filtration is separated, and then the separated supernatant is concentrated by evaporation, or the leaves of the yellow lacquer tree are placed in an extraction solvent, and 5 to 5 ° C. at a temperature of 4 to 20 ° C. After cooling for 7 days to prepare the Hwangchil-tree leaf extract, and after filtering the Hwangchil-tree leaf extract, the filtered filtrate is concentrated by evaporation, in order to increase the yield of the active ingredient contained in the concentrate step of the concentration step ( It is preferable to use a method of repeating S101) three times to combine the supernatants obtained in each concentration step into a final concentrate.

In the two concentration steps (S101) as described above, a hydrous alcohol is used as an extraction solvent, which can be extracted using water in addition to the hydrous alcohol, and the alcohol used in the hydrous alcohol has a concentration of 10 to 100% methanol or Ethanol can be used.

The concentrate obtained through the concentration step (S101) may be obtained in a liquid state with fluidity, but may be obtained in a lyophilized powder state at -80 ° C using a lyophilization method.

The fraction production step (S103) is a step of fractionating the concentrate obtained in the concentration step (S101) with a non-polar solvent, the concentrate obtained in the concentration step (S101) is made by separating into a non-polar solvent, the fraction of the fraction In order to increase the yield, the liquid or dried powder concentrate was extracted and filtered three times with a non-polar solvent, and repeated three times, and the filtrates obtained in each extraction and filtration step were combined to obtain a final fraction. It is preferable to use a collecting method.

At this time, the nonpolar solvent is not particularly limited as long as it can fractionate the concentrate obtained in the concentration step (S101), but it is preferable to use an alkane nonpolar solvent having 4 to 10 carbon atoms, and normal hexane, chloroform and ethyl acetate. It is more preferred to use one or more selected from the group consisting of:

When two or more non-polar solvents are used in the fraction production step (S103), the fractions are processed using normal hexane, and the same fractionation process as the fraction production step (S103) is applied to the remaining filtrate, but the fraction is processed by applying chloroform. In addition, the same fractionation process as in the step of producing the fraction (S103) is applied to the remaining filtrate after fractionation using chloroform, but fractionation is performed using ethyl acetate.

Hereinafter, the method of extracting the nitrogen oxide synthase protein expression inhibitor using the hwangchil tree leaves according to the present invention and the effect of inhibiting the expression of the nitrogen oxide synthase protein expression of the extracted extract will be described with reference to Examples.

≪ Example 1 >

100 g of yellow chile leaves were cut into suitable sizes, and put into 500 ml of methanol solution (methanol: water = 7: 3) with a concentration of 70%, and warmed for 12 hours at a temperature of 95 to 100 ° C. in a water bath equipped with a reflux cooling device. Or reflux extraction, the extracted extract is cooled to a temperature of 30 ℃ and then filtered three times to take the supernatant by filtration with filter paper, and then the supernatant obtained through three iterations are combined, and the combined supernatant is a rotary evaporator After depressurizing with a rotary evaporator, water and methanol were completely evaporated to extract the yellow lacquer leaf extract.

<Example 2>

100 g of yellow chile leaves are chopped to a suitable size and poured into 500 ml of 20% methanol aqueous solution (methanol: water = 1: 4) and cooled at room temperature for 3 days, followed by filtration with multiple layers of gauze to obtain a supernatant solution. After the repetition, the supernatants obtained through the three repetitions were combined, the supernatant was decompressed with a rotary evaporator to completely evaporate water and methanol to extract the yellow lacquer leaves, and the extracted yellow lacquer leaves. The extract was dissolved in distilled water to prepare an aqueous solution of yellow chile leaf extract.

<Example 3>

The aqueous solution of the Hwangchil-tree leaf extract obtained in Example 2 was added to a separatory filter, and extracted at room temperature with hexane for 1 day, followed by filtration of several layers of filter paper and taking a supernatant three times. The filtrates were combined and the solvent was evaporated completely under reduced pressure using a rotary evaporator to fractionate the yellow lacquer leaf extract.

<Example 4>

After the fractionation process is completed through the process of Example 3, the remaining filtrate is added to the separatory filter, extracted with chloroform for 1 day at room temperature, filtered through several layers of filter paper, and the supernatant is taken three times. The filtrates obtained through the above were combined and the solvent was evaporated completely under reduced pressure using a rotary evaporator to fractionate the yellow lacquer leaves.

<Example 5>

After the fractionation process is completed through the procedure of Example 4, the remaining filtrate is added to the separatory filter, extracted with ethyl acetate for 1 day at room temperature, filtered through several layers of filter paper to take a supernatant, and then repeated three times. The filtrates obtained through the procedure were combined and the solvent was evaporated completely under reduced pressure using a rotary evaporator to fractionate the yellow lacquer leaves.

&Lt; Comparative Example 1 &

After the fractionation process is completed through the procedure of Example 5, the remaining filtrate is added to the separatory filter, and extracted at room temperature with butanol for 1 day, and then filtered through several layers of filter paper to take a supernatant three times. The filtrates obtained through the repetition process were combined and the solvent was evaporated completely under reduced pressure using a rotary evaporator to fractionate the yellow lacquer leaf extract.

Comparative Example 2

The filtrate (water layer) remaining after the fractionation process was completed through the procedure of Comparative Example 6 was concentrated.

<Experimental Example 1>

Example 1 to 5 and Comparative Examples 1 to 2 obtained by measuring the effect of inhibiting the nitric oxide leaf extracts, fractions of the hwangchil wood leaf extract and filtrate produced nitric oxide production in Figure 2 below.

In Figure 2 below, veh is a vehicle (control), LPS (Lipopolysaccharide) is a lipopolysaccharide, DPL 001 (Example 1), 002 (Example 2) is a water-containing ethanol extract of the leaves of Hwangchil, DPL_H is an example Fraction 3, DPL_C is the fraction of Example 4, DPL_EA is the fraction of Example 5, DPL_Bu is the fraction of Comparative Example 1, and DPL_H2O is the filtrate of Comparative Example 2.

The physiological activity of the samples prepared in Examples 1 to 5 and Comparative Examples 1 to 2 was evaluated as an inhibitory effect of nitric oxide (NO) production in macrophages activated with lipopolysaccharide (LPS). RAW 264.7 cells, which are mouse macrophage lines, were treated with 1 μg / ml LPS in the presence of samples and incubated for 20 hours.

The concentration of nitric oxide released to the culture system was measured in the form of nitrite by the Greries method (0.1% N- (1-naphthyl) -ethylenediamine dihydrogen dissolved in 1% sulfanylamide + 5% phosphoric acid). Chloride) (Green, LC; Wagner, DA; Glogowski, J .; Skipper, PL; Wishnok, JS; Tannenbaum, SR Anal. Biochem. 1982, 126, 131-138). 100uL of the culture solution and 100uL of Griess reagent were mixed and allowed to stand at room temperature for 10 minutes, and then absorbance was measured using a microplate reader at 540 nm. A standard calibration curve was prepared using sodium nitrite.

As shown in Table 2 below, the nitric oxide synthase protein expression inhibitor extracted by the present invention exhibits excellent physical properties for inhibiting nitric oxide production, and in particular, the nitric oxide production inhibitory effect of the fractions fractionated through Example 4 is most effective. It can be seen that it is excellent.

S101: concentration step
S103: fraction production step

Claims (13)

Inhibitor of the nitric oxide synthase protein expression using the yellow lacquer tree leaf, characterized in that consisting of the yellow lacquer tree leaf extract.
Nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves, characterized in that consisting of a fraction of the hwangchil tree leaves extract.
The method according to claim 1 or 2,
Inhibitor of nitric oxide synthase protein expression using the hwangchil tree leaves, Nitrogen oxide synthase protein expression inhibitor using the hwangchil wood leaf characterized in that it further comprises a pharmaceutically acceptable inert carrier.
The method according to claim 3,
The inert carrier is a lactose, starch and nitric oxide synthase protein expression inhibitor using hwangchil tree, characterized in that consisting of at least one selected from the group consisting of pregelatinized corn starch. The method according to claim 1 or 2,
Nitrogen oxide synthase protein expression inhibitor using the hwangchil tree leaves further comprises one or more additives selected from the group consisting of diluents, flavors, solubilizers, lubricants, suspensions, binders and disintegrants Nitric oxide synthase protein expression inhibitor using.
Tablet comprising the inhibitor of nitric oxide synthase protein expression using the yellow-leaf tree according to any one of claims 1 to 4.
A capsule comprising a nitric oxide synthase protein expression inhibitor using a yellow lacquer tree leaf according to any one of claims 1 to 4.
Syrup, characterized in that it contains a nitric oxide synthase protein expression inhibitor using the hwangchil tree leaves according to any one of claims 1 to 4.
Concentrating step of producing a concentrate by extracting the hwangchil tree leaves with a hydrous alcohol; And
A method for extracting a nitric oxide synthase protein expression inhibitor using a hwangchil tree, characterized in that consisting of; fractions manufacturing step of fractionating the concentrate obtained in the concentration step with a non-polar solvent.
The method according to claim 9,
In the concentration step, the yellow lacquer tree leaf was added to the water-containing alcohol, and hot water or reflux extraction was performed for 5 to 24 hours at a temperature of 40 to 100 ° C. to prepare the yellow lacquer tree leaf extract. After separating the supernatant of the filtrate obtained by filtration afterwards, the extracted supernatant is concentrated by evaporation.
The method according to claim 9,
The concentration step is to put the yellow lacquer leaves in the extraction solvent, and to cool for 5 to 7 days at a temperature of 4 to 20 ℃ to prepare the yellow lacquer leaves extract, and after filtering the yellow lacquer leaves extract, the filtered filtrate Extraction method of nitric oxide synthase protein expression inhibitor using the hwangchil tree, characterized in that the evaporation concentrated.
The method according to claim 9,
The nonpolar solvent is a method for extracting a nitrogen oxide synthase protein expression inhibitor using a yellow lacquer tree, characterized in that the alkanes nonpolar solvent having 4 to 10 carbon atoms.
The method according to claim 9,
The non-polar solvent is a method for extracting the nitric oxide synthase protein expression inhibitor using the hwangchil tree, characterized in that at least one selected from the group consisting of normal hexane, chloroform and ethyl acetate.

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