KR20030003669A - Antagonistic Compositions Against Dioxin-like Compounds Comprising Extract of Angelica koreana Maxim - Google Patents

Abstract

PURPOSE: An antagonistic composition containing as a main component an extract of Angelicae Koreanae Radix which can remove toxicity of dioxin or a dioxin-like compound is provided. The composition has lower toxicity than synthetic medicines and shows antagonistic action against a dioxin-like compound while effectively reducing toxicity of the dioxin-like material, therefore it can be used as food additives as well as drugs. CONSTITUTION: The pharmaceutical composition for prophylaxis and treatment of diseases caused by toxicity of a dioxin-like material contains a pharmaceutically effective amount of an Angelicae Koreanae Radix extract and a pharmaceutically acceptable carrier. The dioxin-like compound is selected from the group consisting of PCDD, PCDF and PCB.

Description

Antagonistic Compositions Against Dioxin-like Compounds Comprising Extract of Angelica koreana Maxim}

The present invention relates to an antagonistic composition for a dioxin-like substance having an active extract, and a pharmaceutical composition and a health food composition using the active extract, and more particularly, to a dioxin-like substance in an environmental endocrine disruptor. A composition for inhibiting intracellular action and a pharmaceutical composition and a health food composition for the treatment or prevention of diseases caused by dioxin analogs.

Endocrine disruptors are defined by the U.S. Environmental Protection Agency (EPA) and are exogenous that interfere with the production, release, transport, metabolism, binding, action, or excretion of natural hormones in the body responsible for maintaining and controlling homeostasis. Say a substance. Endocrine disruptors can be roughly classified into drugs, vegetable endocrine disruptors and environmental endocrine disruptors, and the most problematic of these is the environmental endocrine disruptors called environmental hormones. About 100 environmental endocrine disruptors are currently known, most representative of which is dioxin, PCB, DDT, heavy metals, plastic plasticizers, and the like.

The environmental endocrine disruptors can be roughly classified according to their mechanism of action as follows: (1) Diethylstilbestrol (DES), which mimics natural hormones and induces cellular responses such as natural hormones; (2) blockade of the receptor to which the natural hormone binds to inhibit the action of the natural hormone, for example DDT; (3) triggers a series of abnormal cellular responses by binding to protein receptors independent of the endocrine system, such as dioxin; And (4) heavy metals such as lead, for example by increasing or decreasing the synthesis, storage, etc. of natural hormones indirectly without binding to receptors.

Among the endocrine disruptors described above, dioxin is currently receiving the most attention. The term "dioxin" is generally used in the media and is used as an abbreviation for 2,3,7,8-tetrachlorodibenzo-p-dioxin (hereinafter referred to as "TCDD"). However, TCDD is a kind of polychlorinated dibenzo-p-dioxin (hereinafter referred to as "PCDD") family, which has 75 congeners depending on the position and number of chlorine atoms. Biologically, TCDD is known as PCDD with the strongest toxicity.

On the other hand, other aromatic hydrocarbons are known which have biological properties in common with TCDD, such as polychlorinated dibenzofuran (PCDFs) family and polychlorinated biphenyls (PCBs) family. Accordingly, the term "dioxin" as used herein refers to all compounds included in the PCDD, and "dioxin-like substance" includes the above-described PCDD, PCDFs and PCBs, and exhibits the same cellular effect as PCDD. Means.

Dioxins are carcinogenic substances that have been reported to cause serious diseases such as reproductive and developmental disturbances, damage to the immune system and interference with hormonal regulation (Yang, JH et al., Carcinogenesis . 20: 13-18 (1999). ), Lee, YW et al., Toxicol. Lett., 102-103: 29-83 (1998). According to the US Environmental Protection Agency (EPA), statistics on human adverse effects from exposure to dioxins and dioxin-like substances show that sperm counts in men are reduced to 50% and that the incidence of testicular cancer is about three times greater than 50 years ago. Prostate cancer has been reported to be more than doubled. In addition, due to dioxins and the like, endometrial hyperplasia and an increase in the incidence of breast cancer in women have been confirmed unlike the past.

On the other hand, the most important route that dioxin is introduced into the human body is food, because dioxin is fat-soluble, it is introduced into the human body when ingesting commercial foods, such as meat and milk containing a large amount of lipid components. In addition, since dioxin is hydrophobic, it does not dissolve in water but accumulates in fish and the like, and shows a similar pattern in animals, and it is introduced into the upper layer according to the food chain of the ecosystem, and its effect is shown throughout the ecosystem.

In vivo studies on the mechanism of action of toxic substances, including dioxins, are being conducted in various tissue cells, including animal experiments. In particular, the various routes accessible such as the mechanism that causes the intermediate material, and the oxidative stress of the gene and the cell signaling pathway associated with induction function, the immune system of the genes associated with metabolic oxidation in the in vivo and in vitro centered around the stem cell ought.

On the other hand, studies on the changes in the expression of target genes and the biological activity of intermediates involved in the signaling system mediated by TCDD have been actively reported. Jeong et al. (Jeong, HG, and Lee, SS, Cancer Lett ., 138: 131-137 (1997)) suggest that alpha-Hederin is induced by TCDD in rat Hepa-Ic1c7 cells. It has been reported that it inhibits the expression of cypla-1, and estradiol inhibits the binding of the aryl hydrocarbon (Ah) receptor to the dioxin-response element, or xenobiotic response element, It has been reported to reduce some of the physiological effects of TCDD (Jeong, HG and Lee, SS, Cancer Lett., 133: 177-184 (1998)).

Yang et al. (Yang, JH et al., Carcinogenesis ., 20: 13-18 (1999)) describe TGF-β, plasminogen activator inhibitors using methods such as RT-PCR in cells transformed by TCDD. By exploring changes in growth regulators such as -2 and TNF-α, it has been reported that dysregulation of key growth regulator genes in cells may be involved in the transformation of TCDD-induced cells. Lee et al. (Lee, YW et al, Toxicol. Lett., (1998)) reported that 2-amino-3-methylimidazo [4,5-f] quinoline is immunosuppressed by down-regulating protein kinase C. Reported inhibitory effect.

Park et al. (Park, R. et al., Biochem Biophys Res Commun ., 253: 577-581 (1998)) examined the effect of TCDD on the molecular interactions of attorter complexes located in tyrosine kinase signaling systems. Induction of cyp1a-1 gene expression of benzimidazole by the mediation of, Ah receptor, ARNT, and xenobiotic response element (XRE) (Backlund, M. et al., Eur. J. Biochem., 261: 66- 71 (1999).

One of the important research flows related to the recent TCDD is reports suggesting that the toxic effects of TCDD in hepatocytes are closely related to the oxidative stress-induced mechanisms, such as Radijendirane et al. (1649-1655 (1999)) reported that TCDD-mediated CYP1A-1 gene augmented oxidative stress and induced NAD (P) H: quinone oxidoreductase I gene through an antioxidant site, Shertzer et al. Shuttzer, HG et al., Biochem. Reported.

These changes in various biological systems due to dioxin have been demonstrated that dioxin acts on the Ah receptor, causing binding to XRE, thereby inducing the expression of genes containing the XRE response site up or down (Telakowoski-). Hopkins, CA et al.,Proc. Natl. Acad. Sci., 85: 1000-1004 (1988); Denison, M.S. et al.,J. Biol. Chem, 263: 17221-17224 (1988); Kamath, A. B. et al.,Toxicol. Appl. Pharmacol.,142: 367-377 (1997); McConkey, D. J. et al.,Science,242: 256-259 (1988); McConkey, D. J. and Orrenius, S.,Biochem. Biophys. Res. Commun. 160: 1003-1008 (1989).

PCBs also enter the body through a pathway similar to dioxins, which have been shown to present risks by inhibiting the uptake of neurotransmitters dopamine, glutamate, GABA and serotonin and showing affinity for sex hormone-binding globulin in the blood (Mariussen E, Fonnum F Toxicology 21: 11-21 (2001) and Jury HH, Zacharewski TR, Hammond GL J Steroid Biochem Mol Biol 15: 167-76 (2000). In addition, it has been reported that the structural effects of PCB homologues lead to differences in the induction effect of the CYP1A1 gene in the liver and the use of dioxin's action pathway through the Ah receptor (Van der Burght AS et al., Chemosphere 41: 1697-). 708 (2000), van der Burght AS et al., Toxicol. Appl. Pharmacol. 15: 13-23 (1999), Tilson HA, Kodavanti PR Neurotoxicology 19: 517-25 (1998).

On the other hand, as described above, the molecular level research on the toxicity of dioxin and dioxin-like substances are actively progressed, but studies on antagonists capable of inhibiting their in vivo activity have been insufficient.

Throughout this specification, a number of articles are referenced and their citations are indicated in parentheses. The disclosure of the cited article is incorporated herein by reference in its entirety, so that the level of the technical field to which the present invention belongs and the contents of the present invention are more clearly explained.

Accordingly, it is an object of the present invention to provide an antagonistic composition for dioxin analogs that inhibits the intracellular action of dioxin analogs.

Another object of the present invention to provide a pharmaceutical composition for the treatment or prevention of diseases caused by the toxicity of dioxin-like substances.

It is another object of the present invention to provide an antagonistic health food composition against dioxin analogs.

1 is a genetic map of TRAS-XRE used in the present invention; And

Figure 2 is a graph showing the antagonistic action of dioxins analogues of the active extract of the present invention.

According to one aspect of the present invention, the present invention provides a dioxin analogue that uses active extract as an active ingredient and inhibits the intracellular action of dioxin analogue by interfering with the action of XRE related signaling system of dioxin analogue. It provides a pharmaceutical composition.

The present inventors have tried to find a substance that can treat or prevent the toxicity of dioxins or dioxin-like substances. As a result, extracts of Angelica koreana Maxim, which is conventionally used as a herbal medicine, can effectively remove the toxicity of dioxins or dioxin-like substances. It was confirmed that there is.

The active ingredient used in the present invention is essential oils such as limonene, o-cresol, and alpha-pinene, and isoimperatoine and oxype. Coumarins such as oxypeucedanin, osthol and the like are contained as main components, and they have been used in herbal medicine as analgesics, antibacterial agents and antiperspirants.

Antagonist compositions for dioxin analogs of the present invention are particularly useful for inhibiting the intracellular action of dioxin analogs belonging to PCDD, PCDF, PCB, more preferably dioxin analogs belonging to PCDD and PCB, most preferred. Preferably, it is useful for inhibiting the intracellular action of TCDD, which has the highest intracellular toxicity.

The antagonistic action of dioxin analogs of the active extracts of the present invention is shown by inhibiting the action of dioxin analogs on XRE related signal transduction, as is clearly illustrated in the following examples.

According to another aspect of the invention, the invention features another pharmaceutical composition for the treatment or prevention of diseases caused by the toxicity of dioxin-like substances comprising a pharmaceutically effective amount of active extract and a pharmaceutically acceptable carrier.

Pharmaceutical compositions of the present invention comprising a pharmaceutically effective amount of an active extract are useful for the treatment or prevention of diseases caused by dioxin analogs. The disease caused by the dioxin-like substance is very complex, sperm count, testicular cancer, prostate cancer, endometrial hyperplasia, breast cancer, hepatotoxicity, decreased immune function, hyperlipidemia, subesophageal urethra, latent, birth defects, vascular damage, Hepatocellular carcinoma, hypertrophy of the liver, adenofibrosis, weight loss, hair loss, oral edema, bleeding bleeding and gastric mucosal ulcers (Carter et al., Science 188: 738 (1975); Fourth Annual Report on Carcinogens , NTP 85-002 : 170, 185 (1985)).

On the other hand, the pharmaceutical composition comprising the active extract of the present invention, as described above, exhibits an antagonistic action on the dioxin-like substance at the molecular level, it can be used as an effective and original treatment or prevention agent.

Pharmaceutically acceptable carriers included in the pharmaceutical compositions of the present invention are those commonly used in the preparation, such as lactose, dextrose, sucrose, sorbitol, mannitol, starch, acacia rubber, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil, and the like It doesn't happen. In addition to the above components, the pharmaceutical composition of the present invention may further include a lubricant, a humectant, a sweetener, a flavoring agent, an emulsifier, a suspending agent, a preservative, and the like.

The pharmaceutical composition of the present invention can be administered orally or parenterally, in the case of parenteral administration, it can be administered by intravenous infusion, subcutaneous infusion, intramuscular infusion, etc., preferably by intravenous infusion.

Suitable dosages of the pharmaceutical compositions of the invention vary depending on factors such as the formulation method, mode of administration, age, weight, sex, morbidity, condition of food, time of administration, route of administration, rate of excretion and response to reaction, Usually a skilled practitioner can easily determine and prescribe a dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, in the case of oral administration of the pharmaceutical composition of the present invention, a suitable dosage is to administer 10-400 ml of the distilled water extract of active water once a day on an adult basis.

The pharmaceutical compositions of the present invention may be prepared in unit dose form by formulating with a pharmaceutically acceptable carrier and / or excipient according to methods which can be easily carried out by those skilled in the art. Or may be prepared by incorporating into a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or an aqueous medium, or may be in the form of extracts, powders, granules, tablets or capsules, and may further include a dispersant or stabilizer.

According to another aspect of the present invention, the present invention provides a health food composition comprising an active extract. The food of the present invention may include ingredients that are commonly added during food production. For example, when prepared as a drink, in addition to the active extract of the present invention, citric acid, liquid fructose, sugar, glucose, acetic acid, malic acid, fruit juice, tofu extract, jujube extract, licorice extract may be further included. Considering the environment that is constantly exposed to endocrine disruptors, the food of the present invention is very useful for the prevention and treatment of diseases caused by endocrine disruptors.

Since the composition of the present invention not only effectively reduces the toxicity of dioxin-like substances, but also contains active extract, which is a natural product that has been used as a medicament in the past, as an active ingredient, and exhibits particularly antagonistic action against dioxin-like substances, Side effects on the human body are extremely less than chemical synthetic medicines.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are only for illustrating the present invention in more detail, it is to those skilled in the art that the scope of the present invention is not limited by these examples in accordance with the gist of the present invention. Will be self-evident.

Example

Example material

Samples and reagents used in the examples were used as molecular biology grade products from restriction enzymes from NEB and various reagents from SIGMA, unless otherwise noted in the examples. Kangheol purchased the dried products in the shade at Gyeongdong Market, located in Jongno-gu, Seoul, Korea and used them to manufacture extracts.

Example 1 Preparation of Extracts of Actives

After adding 4 ml of tertiary distilled water per 1 g of dried active water, the mixture was distilled at 121 ° C. for 30 minutes and then centrifuged at 4000 × g at room temperature, and a supernatant was used as an extract of the present invention.

Example 2: Cell Culture

All cell lines used in the examples were cultured and maintained under conditions of 5% CO ₂ and 37 ° C., with penicillin (100 units / ml, Gibco BRL) and streptomycin (100 μg / ml, Gibco BRL) antibiotics and 10 Dulbecco's modified Eagle's medium (DMEM) containing% fetal bovine serum (Gibco BRL) was used.

Cell lines used in the present invention, HeLa cell line and HepG2 cell line is purchased from ATCC.

Example 3: Measurement of Cellular Activity of TCDD and PCB

I. Construction of TRAS-XRE and TRAS-mXRE Reporter Vectors

A TRAS-XRE reporter vector was constructed that contains three overlapping base sequences of the XRE primitives known to bind the dioxin (or dioxin analog) -receptor complex:

First, in order to obtain BGH pA, f1 ori, neomycin resistance gene, ColE1 and ampicillin resistance gene, pcDNA3 (Invitrogen) was cut with Bgl II (Promega) and Xba I (Promega) and the luciferase gene was cut. To obtain, pGL3-basic vector (Promega) was digested with Bgl II (Promega) and Xba I (Promega) to obtain fragments. The two fragments obtained by cleavage were linked with T4 DNA ligase (Promega).

Separately, DNA sequences corresponding to multiple cloning sites (see SEQ ID NO: 1) were designed and submitted to Genotech (Korea) for synthesis. Subsequently, the synthesized multiple cloning site was inserted at positions Bgl II and Hin dIII of the vector intermediate to prepare a TRAS-promoter-basic vector. Subsequently, only a small portion of the promoter (see SEQ ID NO: 2) except for the enhancer region of the known early early promoter (Genbank accession number: k03104) of hCMV was commissioned from Genotech, and Bam was placed on the 5 'side. HI linker and Hin dIII linker was added to the 3 'side. Then, the TRAS-promoter-basic vector was digested with Bam HI (Promega) and Hin dIII (Promega), and the produced CMV minimal promoter was linked with T4 DNA ligase (Promega) to TRAS-enhancer- A basic vector was produced.

Then, a DNA sequence was synthesized to additionally form a Bam HI linker at 5 'and a Bgl II linker at 3' in addition to the DNA sequence of XRE, and this was repeated three times to finally synthesize the DNA sequence set forth in SEQ ID NO: 3 ). Subsequently, the TRAS-enhancer-basic vector was digested with Bam HI (Promega Co.), and the DNA sequence of SEQ ID NO: 3 synthesized at that position was finally linked by using T4 DNA ligase (Promega Co.). Vectors were produced. Gene map of TRAS-XRE is shown in FIG.

On the other hand, TRAS-mXRE to confirm the specificity of the dioxin-receptor to the XRE site, first modified the XRE by a site-directed mutagenesis process, the DNA of SEQ ID NO: 4 with three overlapping mXRE Base sequences were synthesized (Gennotex). After that, the manufacturing process of TRAS-mXRE is the same as that of TRAS-XRE.

Ⅱ. Transformation of Animal Cell Lines

The TRAS-XRE reporter vector prepared above was transformed into HepG2 cells by the lipofectin method in the following manner: 2 μl lipofectin (Gibco BRL Co., Ltd.) was mixed with 100 μl serum-free and antibiotic-free DMEM, After incubation at room temperature for 30 minutes, 1 μg of the reporter vector was mixed with 100 μl serum-free and antibiotic-free DMEM. Subsequently, incubated at room temperature for 15 minutes, 1 ml of 800 μl serum-free and antibiotic-free DMEM was added, and then sprinkled onto HepG2 cells washed twice with PBS. After 6 hours, DMEM was replaced with medium supplemented with 10% fetal calf serum and penicillin (100 units / ml, Gibco BRL) and streptomycin (100 μg / ml, Gibco BRL) antibiotics. Incubated at 37 ° C. for a defined time.

III. Construction of TRAS-XRE HepG2 and TRAS-mXRE HepG2

From one day after the HepG2 cell line was transformed by the method described in II), the cells were grown in DMEM of selective medium containing neomycin (400 µg / ml, Gibco BRL). Two days after transformation, cells were divided by 1/10 (subculture). Cells were then grown for 20 more days in the selection medium described above and then grown separately from each neomycin-resistant colony.

The HepG2 cells thus separated were named "TRAS-XRE HepG2" cell lines, and were deposited with the Korea Cell Line Research Foundation on October 16, 2000, and were given accession number KCLRF-BP-00034. Meanwhile, the HepG2 cell line transformed with TRAS-mXRE was named "TRAS-mXRE HepG2".

Ⅳ. Effect of TCDD or PCB on Expression of TRAS-XRE Reporter Vector

TCND 10 nM and PCB 32 homologues 3.2 ng / ml final concentration (PCB homolog mix; Accustandard, C-SCA-06) or TCDD 5 nM and 32 PCBs in TRAS-XRE HepG2 Each homologue was treated with a mixture of 1.6 ng / ml final concentration and incubated at 37 ° C. for 24 h. Cells were then harvested to normalize transfection efficiency and then luciferase activity was determined as follows: First, the medium in the dish was removed, washed with 1X PBS and lysed buffer (1% Triton X-100). 25 μl glycylglycine, pH 7.8; 15 mM MgSO ₄ ; 4 mM EGTA; and 1 mM DTT) were added and incubated at room temperature for 10 minutes. The dish was then added to a tube to which 360 μl assay buffer (25 mM glycylglycine, pH 7.8; 15 mM potassium phosphate, pH 7.8; 15 mM MgSO ₄ ; 4 mM EGTA; 2 mM ATP; and 1 mM DTT) was added. 100 μl of the solution was added, 200 μl luciferin was added, and fluorescence was measured on a luminometer (Turner Designs Instrument, TD 2020). The result is shown in FIG.

As can be seen in Figure 2, when treated with TCDD in the TRAS-XRE HepG2 5 times compared to the untreated control, about 1.8 times when treated with PCB, and about when treated with a mixture of TCDD and PCB It exhibited about 3.8-fold increased luciferase activity. These results indicate that TCDD or PCB binds to the Ah receptor and stimulates the Ah receptor, which in turn causes the TCDD-Ah receptor to bind to Arnt (Aryl hydrocarbon receptor nuclear translocator), and the resulting complex binds to XRE to activate the transcriptional regulator. This is because it triggers the expression of luciferase. On the other hand, the PCB than TCDD triggers the expression of less luciferase, it can be seen that TCDD is the most potent toxicant and PCB can also be toxic, which is consistent with the existing reports, Evidence confirms the accuracy of the analysis system. On the other hand, no increase in luciferase activity was observed when TRAS-mXRE HepG2 was treated with TCDD or PCB (data is not described herein).

As a result, as disclosed in the above-mentioned known documents, TCDD or PCB triggers a signaling system that specifically acts on the XRE sequence, thereby confirming that pathological conditions such as cytotoxicity such as TCDD or PCB are caused.

Example 4: Confirmation of the antagonism of dioxins or PCBs of the active extract of the present invention

TCDD 10 nM and PCB 32 homologues 3.2 ng / ml final concentration (PCB homolog mix; Accustandard, C-SCA-06) or TCDD 5 nM and 32 PCBs in TRAS-XRE HepG2 cells constructed in Example 3 Each mixture of final homologs of 1.6 ng / ml was treated at 37 ° C. for 24 hours, and then 50 μl of the active extract obtained in Example 1 was added and incubated at 37 ° C. for 24 hours. Subsequently, the treated cells were harvested to normalize transformation efficiency, and then the luciferase activity was measured in the same manner as in Example 3, and the results are shown in FIG. 2.

As can be seen in Figure 2, the active extract of the present invention can be seen that the effect of inducing luciferase expression by TCDD or PCB to the level of the untreated control. On the other hand, it can be seen that the luciferase expression is not affected at all in the experimental example treated only active extract, the offset effect of the activity extract is based on the antagonism of the action of TCDD or PCB.

In addition, the cellular action by TCDD or PCB is due to the triggering of the signaling system acting on the XRE as disclosed in the above-mentioned literature, and the analysis system employed in this embodiment is a reporter depending only on the binding to the XRE. Since the expression level of the gene (luciferase gene) is measured, the above results can be seen that the antagonistic action of the active extract is achieved by inhibiting the triggering of the XRE-associated signaling system by TCDD or PCB.

Example 5: Confirm specific antagonistic action of the active extract of the present invention

I. Creation of Reporter Vector

A TRAS-p53 vector having a site where a protein encoded by p53, an important gene related to cell cycle regulation and death, functions was constructed in the same manner as in TRAS-XRE of Example 3, except that p53 was encoded instead of the XRE sequence. The site where the protein acts (see SEQ ID NO: 5) was synthesized and made by Genotech.

The TRAS-NF-κB vector having a site where NF-κB is known to play an important role in various stress and signaling systems in cells is prepared in the same manner as in TRAS-XRE of Example 3, but instead of the XRE sequence. The site where NF-κB acts (see SEQ ID NO: 6) was synthesized (Genotec).

In addition, a TRAS-PRE vector having PRE, a site where progesterone, an endocrine-related hormone in the cell functions, was prepared in the same manner as in TRAS-XRE of Example 3, but in place of the XRE sequence of PRE (see SEQ ID NO: 7). Was synthesized (genotech).

Ⅱ. Confirmation of specificity of active extract

In the same manner as described in Example 3, each vector was introduced into the HepG2 cells in the same amount (1 μg), and then divided into the experimental group and the control group, and 50 μl of the active extract of Example 1 was added to the experimental group, respectively. , Incubated at 37 ℃ for 24 hours. Subsequently, luciferase activity was measured in the same manner as in Example 3, and the results are shown in FIG. 2.

As can be seen in Figure 2, the active extract of the present invention appeared to have little effect on the binding site of the p53 protein, the site where NF-κB acts, and PRE. In particular, NF-κB is an important factor in inducing various physiological phenomena by mediating the stress delivered to cells, and sudden activity change may have unexpected effects on the cells, so the active extract does not affect the activity of NF-κB. Has an important meaning. In addition, PRE is a site where progesterone acts as a hormone, and the results of the experiment indicate that it does not inhibit the system of homeostasis due to the natural role of the hormone, and the active extract of the present invention is also an excellent active ingredient. It can be shown that it can be used.

In other words, it can be seen that the active extract of the present invention has little effect on the expression and activity of major genes in cells except XRE. Therefore, it can be seen that the active extract of the present invention is an active ingredient that can antagonize very specifically with respect to the dioxin analogue that binds to the XRE sequence.

Example 6: Confirmation of the antagonism of dioxins of the active extract of the present invention using an animal model

For the active extract of the present invention confirmed that the antagonistic action against dioxin at the cellular level through Example 4, it was confirmed as follows whether or not the antagonism actually appeared in vivo using an animal model.

Experimental animals were male 5-year-old guinea pig rats (BioKorea). The active extract treatment group was six. Rats were administered 1 μg of TCDD per kg once intraperitoneally on test start day or 7 days after test start. The active extract was administered negatively for 28 days at a dose of 0.03-0.3 g / day / kg from the start of the test. After breeding for 28 days, the mice were weighed, anesthetized with ethyl ether, the mice were slaughtered, the testicles were taken out, and weighed. The experimental results are shown in Table 1 below:

division Number of animals Weight (g) Absolute organ weight (mg) Relative organ weight ³⁾ (mg) Left testicle Right testicle Left testicle Right testicle No treatment group 6 451.0 ± 47.22 791.20 ± 133.33 776.30 ± 151.23 175.43 ± 41.20 172.13 ± 46.40 TCDD treatment group 6 393.1 ± 44.37 604.87 ± 102.25 601.98 ± 112.26 153.87 ± 30.08 153.14 ± 34.42 Extract treatment group 6 461.9 ± 62.31 822.90 ± 101.11 836.33 ± 103.75 178.15 ± 40.96 181.06 ± 39.41 Extract + TCDD pre-treatment group ¹⁾ 6 463.5 ± 55.31 828.33 ± 156.55 811.98 ± 189.64 178.71 ± 45.90 175.18 ± 49.90 Extract + TCDD post-treatment group ²⁾ 6 454.1 ± 49.54 830.20 ± 189.76 822.60 ± 187.65 182.82 ± 42.86 181.15 ± 41.20 ¹⁾ from the 7th day after treatment TCDD extract negative group ^2), TCDD-treated group on day 7 from the test start date ³⁾ a relative weight (in terms of values with the weight (100 g basis) of the experimental rats)

As can be seen in Table 1 above, the active extract of the present invention can prevent (refer to TCDD pre-treatment group) and treatment (refer to TCDD post-treatment group) weight loss and testicular weight caused by TCDD. It can be seen that. In other words, the active extract of the present invention can be confirmed that the antagonistic action on dioxin not only at the cellular level but also at the individual level.

Preparation Example: Health food drink

80 ml of the active extract of Example 1, 10 ml of liquid fructose, 0.5 ml of citric acid, and 9.5 ml of licorice extract were mixed to prepare a drink of the present invention.

The present invention provides antagonistic compositions for dioxin analogs that inhibit the intracellular action of dioxin analogs. The present invention also provides a pharmaceutical composition for the treatment or prevention of diseases caused by the toxicity of dioxin-like substances. The present invention also provides an antagonistic health food composition against dioxin analogs. Since the composition of the present invention not only effectively reduces the toxicity of dioxin-like substances, but also includes active extract, which is a natural product that has been conventionally used as a medicament, exhibits an antagonistic action against dioxin-like substances very specifically, Side effects on the human body are extremely less than chemical synthetic medicines, and can be used as food additives as well as medicines.

Claims (6)

An antagonistic composition for a dioxin analogue that uses active extract as an active ingredient and inhibits the dioxin-like intracellular action of the dioxin analogue by interfering with the action of the XRE related signaling system. Pharmaceutically effective amount of active extract; And A pharmaceutical composition for the treatment or prevention of a disease against the toxicity of a dioxin-like substance comprising a pharmaceutically acceptable carrier. Antagonistic health food composition for dioxin-like substances comprising active extract as an active ingredient. The composition of claim 1, wherein said dioxin analog is selected from the group consisting of PCDD, PCDF, and PCB. The composition of claim 4, wherein the dioxin analog is PCDD or PCB. 6. The composition of claim 5, wherein said PCDD is TCDD.