KR20150014299A - Preparation and composition of natural vegetables having anti-inflammatory activity - Google Patents

Abstract

The present invention relates to a method for preparing a composition with anti-inflammatory activity using natural vegetables and to the composition thereof and, more specifically, to a method for preparing a composition with anti-inflammatory activity using natural vegetables comprising: a precipitation step of washing Phellodendron amurense bark, clove, Halls honeysuckle, Morus alba L., rose water, Glycyrrhiza uralensis Fischer, Foremost mugwort, Peony root, and Sweet-flag, and precipitating the same in 15-20% ethanol for 20-30 minutes in order for an active extract to be easily extracted; an extraction step of dumping the ethanol from the precipitation step and extracting the active extract by respectively mixing the Phellodendron amurense bark, clove, Halls honeysuckle, Morus alba L., rose water, Glycyrrhiza uralensis Fischer, Foremost mugwort, Peony root, and Sweet-flag with water and heating the same; a distillation step of mixing each active extract extracted in the extraction step and distilling the same at 110~120°C for 4-5 hours; and a filtration step of inputting the extract obtained from the distillation step into a filter. The method for preparing the composition with anti-inflammatory activity using natural vegetables and the composition thereof according to the present invention use the natural vegetables as raw materials, thereby having anti-inflammatory activity through thermal-extraction and distillation processes, and being harmless for human body.

Description

TECHNICAL FIELD [0001] The present invention relates to a method for preparing an anti-inflammatory active ingredient using a natural plant material,

The present invention relates to a method for preparing an anti-inflammatory active ingredient using a natural plant material and a composition thereof, and more particularly to a method for producing an anti-inflammatory active ingredient using a natural plant material, The present invention relates to a method for preparing an anti-inflammatory active ingredient using a natural plant material having an anti-inflammatory activity and a composition thereof.

Generally, inflammation is a generic term for diseases found in many things such as infectious diseases (tuberculosis, syphilis, etc.) as well as inflammation such as otitis media or pleuritis, and most diseases of daily life This belongs to this.

This inflammation is one of the defense reactions of biopsies against certain stimuli. It is a local protective protection reaction against the damage of the biotissue, and the blood component escapes into the tissue through the blood vessel wall. The chemicals involved in this process are histamines, leukotrienes, kinis and prostaglandins. Histamine acts to bring more blood and lymph to the injured area, and Kinin relaxes muscle contraction to smooth blood flow through the capillary, making the pain felt. In addition, prostaglandin is synthesized when leukocytes enter cells, causing pain and heat.

Recently, factors causing inflammation have been increasing due to an increase in social stress, a sudden change in environmental pollution and food culture, and an increase in the elderly population due to aging. As a result, inflammatory diseases such as allergies, asthma, inflammatory bowel disease or arthritis Is also increasing.

For example, Korean Patent Laid-Open No. 10-2012-0051928 (published May 23, 2012) discloses acorn extract having anti-inflammatory activity and a pharmaceutical composition containing the extract, and Korean Patent Laid- 0049043 (published May 16, 2012) relates to an anti-inflammatory composition containing a mussel hydrolyzate as an active ingredient.

As such, inflammation causes many diseases, so efforts to develop functional foods and medical materials have been actively pursued by exploring materials having a function of suppressing inflammation.

However, these inflammation-inhibiting biomaterials depend on imports. For example, arthritis is a non-steroidal anti-inflammatory agent, anti-rheumatic agent, steroid agent and analgesic agent, which can relieve the joint pain or stiffness but does not inhibit disease progression, It can cause side effects in the gastrointestinal or cardiovascular system.

Therefore, it is urgently required to develop a natural anti-inflammatory biomaterial which is safe and does not cause side effects to the human body and has excellent anti-inflammatory effect.

Korean Patent Laid-Open No. 10-2012-0051928 (published on May 23, 2012) Korean Patent Publication No. 10-2012-0051928 (May 16, 2012)

SUMMARY OF THE INVENTION The present invention has been made in view of the above problems, and an object of the present invention is to provide an anti-inflammatory activity effect by heat extraction and distillation using a natural vegetable material as a raw material, A method for producing an inflammatory active composition and a composition thereof.

According to an aspect of the present invention for achieving the above object, the first invention provides a method for cleaning a yellowish white, a clove, a rhododendron, a mulberry, a rosemary, a licorice, a wormwood, a peony and an iris, Precipitation in 15 to 20% ethanol for 20 to 30 minutes; An extraction step of discarding the ethanol in the precipitation step and extracting the effective extract solution by mixing and heating the precipitated yellowish white, clove, rhododendron, mulberry, rosewater, licorice, wormwood, peony and iris respectively with water; A distillation step of mixing the respective effective extracts extracted in the extraction step and distilling at a temperature of 110 to 120 DEG C for 4 to 5 hours; And a filtration step in which the extract obtained in the distillation step is put into a filter and filtered 2 to 4 times.

In a second aspect of the present invention, in the first invention, in the precipitation step, the sedimentation step comprises 550 to 600 parts by weight of cling, 250 to 300 parts by weight of rhizome, 350 to 400 parts by weight of mulberry, 150 to 200 parts by weight of rosemary, 50-100 parts by weight of licorice, 50-100 parts by weight of licorice, 150-200 parts by weight of mugwort, 50-100 parts by weight of peony and 50-100 parts by weight of iris.

According to a third aspect of the present invention, in the first aspect of the present invention, in the extraction step, 300 parts by weight of water is mixed with 100 parts by weight of yellowish white, 1000 parts by weight of water is mixed with 550 to 600 parts by weight of clay, 500 parts by weight of water were mixed, 800 parts by weight of water was mixed with 350 to 400 parts by weight of mulberry, 400 parts by weight of water was mixed with 150-200 parts by weight of rosin, 300 parts by weight of water was added to 50-100 parts by weight of licorice 400 parts by weight of water is mixed with 150-200 parts by weight of mugwort, 300 parts by weight of water is mixed with 50-100 parts by weight of peony powder, and 300 parts by weight of water is mixed with 50-100 parts by weight of iris .

In a fourth aspect of the present invention, in the first invention, in the distillation step, 200 parts by weight of the effective extract of clove, 100 parts by weight of the effective extract of Phalaenopsis, 200 parts by weight of the effective extract of mulberry, 200 parts by weight of rosewater 100 parts by weight of the extract, 50 parts by weight of the effective extract of licorice, 100 parts by weight of the effective extract of mugwort, 100 parts by weight of the effective extract of peony root and 100 parts by weight of the effective extract of iris.

The fifth invention is characterized by the composition according to the first invention to the fourth invention.

The method for producing an anti-inflammatory active composition using a natural plant material according to the present invention and its composition have an anti-inflammatory activity effect through heat extraction and distillation using a natural plant material as a raw material, and are harmless to the human body.

1 is a flowchart showing a method for preparing an anti-inflammatory active composition using a natural plant material according to an embodiment of the present invention,
2 is a graph showing the cytotoxicity of BO2 against RAW 264.7 cells,
3 is a graph showing inhibitory effect of BO2 on NO secretion in LPS-stimulated RAW 264.7 cells,
4 is a graph showing the inhibitory effect of BO2 on PGE2 secretion in LPS-stimulated RAW 264.7 cells,
FIGS. 5A and 5B are graphs showing the inhibitory effect of BO2 on TNF-.alpha. And IL-6 secretion in LPS-stimulated RAW 264.7 cells,
FIG. 6 is a graph showing the inhibitory effects of intracellular iNOS and COX-2 on BO2 pretreatment,
FIGS. 7A and 7B are graphs showing the densitometric analysis of iNOS and OCX-2 expression,
8 is a graph showing the effect of inhibiting MAPK activation by BO2 pre-treatment,
9A to 9C are graphs showing the densitometric analysis of MAPK activation inhibition,
10 is a graph showing the inhibitory effect of NF-kB activation by BO2 pretreatment,
FIGS. 11A and 11B are graphs showing the densitometric analysis of inhibition of NF-kB activation,
12A to 12C are graphs showing the effect of suppressing secretion of cytokines in blood in LPS-administered mice.

Hereinafter, a method for preparing an anti-inflammatory active composition using a natural plant material according to the present invention and a composition thereof will be described in detail with reference to the accompanying drawings.

1 is a flowchart illustrating a method for preparing an anti-inflammatory active ingredient using a natural plant material according to an embodiment of the present invention.

As shown in FIG. 1, the present invention relates to an anti-inflammatory active composition using a natural plant material having an anti-inflammatory activity effect using a natural plant material such as yellowtail, clove, rhododendron, mulberry, rose water, licorice, And a manufacturing method thereof.

The method for preparing an anti-inflammatory active ingredient using a natural plant material comprises four steps including a precipitation step (S110), an extraction step (S120), a distillation step (S130) and a filtration step (S140).

The precipitation step (S110) cleans the yellowish white, cloves, rhododendron, mulberry, rosemary, licorice, wormwood, peony and iris and precipitates in 15 to 20% ethanol for 20 to 30 minutes to easily extract the effective extract . In addition, the ethanol can be used as a spirit.

In this case, the precipitation step (S110) comprises 550 to 600 parts by weight of clove, 250 to 300 parts by weight of rhizome, 350 to 400 parts by weight of mulberry, 150 to 200 parts by weight of rosin, 50 to 100 parts by weight of licorice, 50-100 parts by weight of licorice, 150-200 parts by weight of mugwort, 50-100 parts by weight of peony and 50-100 parts by weight of iris.

In the precipitation step (S110), the yellow, white, clover, mulberry, rosemary, licorice, mugwort, peony, and iris may be cut into appropriate size before the ethanol is added.

The extracting step S120 is a step of discarding the ethanol in the precipitation step S110 and mixing and heating the precipitated yellowish white, clove, mulberry, mulberry, rosewater, licorice, Respectively.

At this time, the extraction step (S120) may be a step of mixing 300 parts by weight of water with respect to 100 parts by weight of yellowish white, and heating the mixture at a temperature of 110 to 120 DEG C for 1 to 2 hours to extract 150 parts by weight of the effective extract.

In the extracting step (S120), 1000 parts by weight of water may be mixed with 550 to 600 parts by weight of the cling, and the mixture may be heated at 110 to 120 DEG C for 4 to 5 hours to extract 600 parts by weight of the effective extract.

In the extracting step (S120), 500 parts by weight of water may be mixed with 250 to 300 parts by weight of the vinegars, and the mixture may be heated at 110 to 120 DEG C for 1 to 2 hours to extract 200 parts by weight of the effective extract .

In the extracting step (S120), 800 parts by weight of water is mixed with 350 to 400 parts by weight of mulberry, and the mixture is heated at 110 to 120 DEG C for 3 to 4 hours to extract 400 parts by weight of the effective extract.

In the extracting step (S120), 400 parts by weight of water may be mixed with 150-200 parts by weight of rosin, and the mixture may be heated at 110-120 캜 for 1 to 2 hours to extract 200 parts by weight of the effective extract.

In the extracting step (S120), 300 parts by weight of water may be mixed with 50-100 parts by weight of licorice and heated at 110-120 ° C for 1-2 hours to extract 150 parts by weight of the effective extract.

In the extracting step (S120), 400 parts by weight of water may be added to 150-200 parts by weight of mugwort, and the mixture may be heated at 110-120 ° C for 1-2 hours to extract 200 parts by weight of the effective extract.

In the extracting step (S120), 300 parts by weight of water may be mixed with 50-100 parts by weight of peanuts, and the mixture may be heated at 110-120 ° C for 1 to 2 hours to extract 150 parts by weight of the effective extract.

In the extraction step (S120), 300 parts by weight of water may be mixed with 50 to 100 parts by weight of iris, and the mixture may be heated at 110 to 120 캜 for 1 to 2 hours to extract 150 parts by weight of the effective extract.

The distillation step (S130) is a step of mixing the respective effective extracts extracted in the extraction step (S120) and distilling at a temperature of 110 to 120 DEG C for 4 to 5 hours.

At this time, each of the effective extractives to be mixed in the distillation step (S130) comprises 200 parts by weight of an effective extract of clove, 50 parts by weight of an effective extract of yellowish white, 200 parts by weight of an effective extract of mulberry, 100 parts by weight of an effective extract of rosemary extract, 50 parts by weight of an effective extract of licorice, 100 parts by weight of an effective extract of mugwort, 100 parts by weight of an effective extract of peony root and 100 parts by weight of an effective extract of iris.

In the filtration step (S140), the extract obtained in the distillation step (S130) is put into a filter and filtered 2 to 4 times.

The extracts (hereinafter referred to as BO2) extracted from natural plant materials such as yellowish white, clove, rhododendron, mulberry, rosewater, licorice, mugwort, peony and iris by the disclosed manufacturing method are natural components without toxicity, It is harmless and has no side effects.

[Experimental Example 1]

Name of material Additive amount (g) Water Addition Amount (ml) Heating temperature (℃) Heating time (h) Extract
Recovery (ml) cloves 600 1,000 110-120 5 600 Rhinoceros 300 500 110-120 2 200 Yellowish white 100 300 110-120 2 150 mulberry tree 400 800 110-120 4 400 Roses 200 400 110-120 2 200 licorice 100 300 110-120 2 150 Mugwort 200 400 110-120 2 200 Cure 100 300 110-120 2 150 calamus 100 300 110-120 2 150

The anti-inflammatory active composition using the natural plant material of the present invention was tested according to Table 1 above.

2 is a graph showing the cytotoxicity of BO2 against RAW 264.7 cells.

As a result of measuring the cytotoxicity of BO2 by MTT, BO2 was found to be a safe concentration that does not significantly affect cell density up to 10 times dilution.

FIG. 3 is a graph showing the inhibitory effect of BO2 on NO 2 secretion in LPS stimulated RAW 264.7 cells, FIG. 4 is a graph showing inhibitory effect of BO2 on PGE2 secretion in LPS stimulated RAW 264.7 cells, and FIGS. Lt; RTI ID = 0.0 > IL-6 < / RTI >

As a result of measuring the inhibitory effect of BO2 on the secretion of NO, one of the inflammatory mediators, NO2 production was observed in RAW 264.7 cells stimulated with LPS by BO2 pretreatment from 10 times dilution to 50 times dilution which did not show cytotoxicity An inhibitory effect was observed.

In addition, PGE2 is an important inflammatory mediator that is secreted from macrophages stimulated by LPS. As a result of the measurement of the effect on PGE2 production, it was confirmed that PGE2 production is also easily inhibited by BO2 pretreatment.

TNF-α and IL-6, which are important inflammatory cytokines secreted from macrophages in inflammatory reactions, were investigated for the inhibitory activity of BO2 on the secretion of these cytokines. As a result, it was confirmed that both TNF-α and IL-6 were easily inhibited by pretreatment of BO2 as in NO.

These results suggest that BO2 inhibits the secretion of inflammatory cytokines as well as NO and PGE2 in the inflammatory response of macrophages by LPS.

FIG. 6 is a graph showing the inhibitory effect of intracellular iNOS and COX-2 expression by BO2 pretreatment, and FIGS. 7A and 7B are graphs showing the densitometric analysis of iNOS and OCX-2 expression.

iNOS is a NO biosynthetic enzyme induced by stimulation of inflammatory response. The mechanism of action of inhibition of NO secretion can be analyzed by examining the expression level of this enzyme. In addition, COX-2 is an enzyme that produces PGE2, one of the inflammatory mediators, which is also increased by LPS stimulation. As a result of intracellular expression of the iNOS and COX-2 enzymes, intracellular expression of the iNOS and COX-2 enzymes was markedly inhibited by pretreatment of BO2, and the inhibitory effect was due to the concentration of BO2.

In addition, in order to quantitatively observe the results of Western blotting, the inhibition of the expression of these enzymes was also confirmed by a densitometer analysis.

FIG. 8 is a graph showing the effect of suppressing MAPK activation by pretreatment with BO2, FIGS. 9A to 9C are graphs showing the densitometric analysis of inhibition of MAPK activation, and FIG. 10 is a graph showing inhibitory effect of NF- , And Figs. 11A to 11B are graphs showing the densitometric analysis for suppression of NF-kB activation.

When macrophages cause inflammation by LPS stimulation, inflammation is induced through various intracellular signaling pathways. In particular, MAPKs such as p38, JNK, and ERK are very important signal transduction molecules in these inflammatory responses, and inflammation is induced by phosphorylation of these molecules.

In the group treated with BO2, phosphorylation of all three molecules was markedly suppressed, and it was confirmed that the anti - inflammatory activity of the Korean extract was related to the inhibition of activation of MAPK.

In contrast, the activation of NF-kB by NF-kB is important for the inflammation induction of macrophages by LPS stimulation. NF-kB is associated with I-kB in normal state, When -kB is phosphorylated, NF-kB is converted to a free state and a potential in the nucleus is generated. Therefore, activation of NF-kB can be measured through phosphorylation of I-kB.

When BO2 was pretreated before LPS stimulation, it was confirmed that phosphorylation of I-kB by LPS was inhibited depending on the concentration. In addition, it was confirmed that the process of decomposition in cytoplasm, which is liberated from NF-kB as I-kB is phosphorylated, is inhibited by treatment with BO2.

These results suggest that the anti-inflammatory activity of BO2 inhibits the activation of MAPK and NF-kB, inhibits the production of NO and PGE2 by inhibiting the expression of COX-2 and iNOS, and inhibits inflammatory cytokines It can be seen that a series of mechanisms occurs.

12A to 12C are graphs showing the effect of suppressing secretion of cytokines in blood in LPS-administered mice.

The in vivo model of the anti - inflammatory effect of BO2 identified in vitro was examined. When Balb / c mice were intravenously injected with LPS at 400 ug / mouse, an experimental model with a peak serum inflammatory cytokine level was used after 2 hours.

BO2 was diluted to 1/3 of the original volume. The solution was administered once per day for 3 days, and LPS was administered. After 2 hours, serum was collected and analyzed by ELISA kit for TNF-α, IL-6 and IL- Lt; RTI ID = 0.0 > IL-1 < / RTI > As a result, the blood levels of TNF-α and IL-6 decreased very little in BO2-treated mice, but the plasma levels of IL-1β were markedly decreased by BO2 administration.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the invention as defined by the appended claims. It is obvious that you can do it.

Claims (5)

A precipitation step (S110) in which the precipitate is precipitated in ethanol at 15 to 20% for 20 to 30 minutes so as to wash out yellowish white, clove, rhododendron, mulberry, rosewater, licorice, mugwort, peony and iris and extract the effective extract easily;
(S120) in which the ethanol is discarded in the precipitation step (S110) and the effective extract is extracted by mixing and heating the precipitated yellowish white, clove, mulberry, mulberry, rosemary, licorice, ;
A distillation step (S130) of mixing the respective effective extracts extracted in the extraction step (S120) and distilling at a temperature of 110 to 120 DEG C for 4 to 5 hours; And
A filtration step (S140) in which the extract obtained in the distillation step (S130) is put into a filter and filtered 2 to 4 times;
The method of claim 1 or 2,
The method of claim 1, wherein the depositing step (Sl 10)
The present invention relates to a method for producing a fermented soybean curd, which comprises 550 to 600 parts by weight of clove, 250 to 300 parts by weight of rhizome, 350 to 400 parts by weight of mulberry, 150 to 200 parts by weight of rosin, 50 to 100 parts by weight of licorice, And 50 to 100 parts by weight of iris are precipitated.
2. The method according to claim 1, wherein the extracting step (S120)
300 parts by weight of water was mixed with 100 parts by weight of yellowish white,
1000 parts by weight of water is mixed with 550 to 600 parts by weight of the cling,
500 parts by weight of water was mixed with 250 to 300 parts by weight of the vinegar,
800 parts by weight of water was mixed with 350 to 400 parts by weight of mulberry,
400 parts by weight of water was mixed with 150-200 parts by weight of the rosin,
300 parts by weight of water was mixed with 50 to 100 parts by weight of licorice,
400 parts by weight of water was mixed with 150-200 parts by weight of mugwort,
300 parts by weight of water is mixed with 50 to 100 parts by weight of peony powder,
And 300 parts by weight of water is mixed with 50-100 parts by weight of iris.
2. The method according to claim 1, wherein the distillation step (S130)
200 parts by weight of the effective extract of Clove, 100 parts by weight of the effective extract of Phloem clover, 200 parts by weight of the effective extract of Mulberry, 100 parts by weight of the effective extract of rosewater, 50 parts by weight of the effective extract of licorice, , 100 parts by weight of an effective extract of Peony root, and 100 parts by weight of an effective extract of Peony root are mixed.
An anti-inflammatory active composition using a natural plant material, which is characterized in that it is composed of any one of claims 1 to 4.

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