JP4100623B2 - Composition of herbal extract with nicotine and dioxin detoxification - Google Patents

Description

  The present invention relates to a composition composed of a herbal extract having an in vivo detoxifying action of typical modern toxic substances such as nicotine and dioxin.

  Toxic substances related to the human body include externally ingested drugs and food-derived toxic substances, microorganisms, polluted air, water, peroxides and free radicals generated during metabolic processes in the body, Toxins produced by harmful microorganisms in the intestines. As harmful substances generated by the interaction between the external environment and the body, the natural frequency of the human body is interfered by microwaves and ultrasonic waves, electromagnetic waves, noise, ultraviolet rays, radiation, etc., leading to abnormalities in the nervous system and endocrine secretion. It is known to cause abnormalities in the system and affect metabolism by mutagenesis of cells and generation of harmful substances (Kwon, YT Environmental contamination and human exposure by dioxin-like compounds. The Research Institute of Engineering Technology. 15: 851-867 (1998)). In this way, as the modern environment is extensively polluted with various harmful substances, the immunity of the human body declines and the chances of enjoying healthy life are decreasing, but we often come in contact with them. The most representative harmful substances that can be mentioned are smoking-related substances and environmental hormones generated during waste incineration.

  Nicotine, which is a typical harmful substance in tobacco, is nitrosamine-4- (methylnitrosamino) -1- (3-pyridyl) -1-butanone {nitrosamine-4 (methylnitrosamino), a substance that induces lung cancer. ) -1- (3-pyridyl) -1-butanone (NNK)}, a powerful environmental toxin. Symptoms of acute toxicity of nicotine induce pupil dilation, corneal opacity, vomiting, difficulty breathing, increased secretions in saliva and respiratory organs, decreased heart rate, etc. Systemic diseases (Brunneman, KD, Prokopezyk, B., Djordjevic, MV and Hoffmann, D. Formation and analysis of tobacco-specific N-nitrosamines. J. Natl. Cancer Inst. 89: 868-73 (1996); Carmella, SG, Borukhova, A., Akerkar, SA and Hecht, SS Analysis of human urine for pyridine-N-oxidemetabolites of 4- (methylnitrosamino) -1- (3-pyridyl) -1-butanone, a tobacco-specific lung Carcinogen. Cancer Epidemiol Biomarkers Prev. 6: 113-20 (1997); and Wynder, EL, And Muscat, JE The changing epidemiology of smoking and lung cancer histology. Environ Health Perspect. 103 Suppl 8: 143-148 (1995)) .

  On the other hand, dioxin is a substance that induces reproduction, neurotoxicity, and carcinogenesis. It is the worst poisonous and deleterious substance made by humans. It is toxic by the action of halogenated aromatic hydrocarbons and is the most powerful. 2,3,7,8-tetrachloro-dibenzo-p-dioxin (TCDD), known as a dioxin, is the main component of defoliants. Known as endocrine disrupting compounds, or environmental hormones (Poland, A. And Knutson, JC 2, 3, 7, 8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons; examination of the mechanism of Toxicol Ann. Rev. Pharmacol. Toxico 22: 517-554 (1982)). TCDD is a polyhalogenated aromatic hydrocarbon produced during pulp bleaching, forest fires, garbage incineration, automobile exhaust, smoking and other combustion processes, and has a half-life in the body of 11 years. Pyrolysis at 800-850 ° C. TCDD is absorbed into the body through the gastrointestinal tract, respiratory organs, and skin, and then passes through the cell membrane to help aromatic hydrocarbon receptors and heat shock proteins present in the cytoplasm. To reach the cell nucleus and act on DNA in the nucleus to generate a new enzyme, causing toxic effects such as weight loss, immune function suppression, induction of skin toxicity, birth defects, abnormal liver function and lipid metabolism, carcinogenesis, etc. Ibid. 26: 371-399 (1986); and Gelboin, HV Benzo (a) pyrene metabolism, Safe, SH Comparative toxicology and mechanism of action of polychlorinate dibenzo-p-dioxins and dibenzofurans. activation and carcinogenesis: role and regulation of mixed-function oxidases and felated enzymes. Physiol, Rev. 60: 1107-1166 (1980)).

  In the study on the reduction of toxicity to nicotine and TCDD, green tea extract has the effect of suppressing the occurrence of lung cancer in mice in which lung cancer has been induced, and has the effect of suppressing the occurrence of lung cancer by NNK. (Chung. FL The prevention of lung cancer induced by a tobacco-specific carcinogen in rodents by green and black Tea. Proc. Soc. Exp. Viol. Med. 220: 244-248 (1999); and Fujiki, H., Suganuma, M., Okabe, S., Sueoka N., Komori, A., Sueoka, E., Kozu, T., Tada, Y., Suga , K., Imai, K. and Nakachi, K. Cancer inhibition by green tea. Mutat. Res. 18: 307-310 (1998)).

  It is also known that green tea extract and plant extract mixture containing green tea extract increase nicotine degradability (Kyung, YJ and Lee, DH Enhanced conversion to cotinine from nicotine by green tea extract. Korean J. Environmental 19: 147-153 (2000); and Kim, JH, Lee, DH, Cho, HJ and Jeong, JM Effects of phyto-extract mixture on the nicotine decomposition. J. Fd Hyg. Safety. 17: 8-14 (2002)). The biochemical reaction of TCDD and similar compounds is related to the increased synthesis of cytochrome P450 complex present in liver microsomes, and the mechanism of toxic expression of TCDD is influenced by oxidative stress. (Kitchin, KT and Woods, JS 2,3,7,8-Tetrachlorodibenzo-p-dioxin effect on hepatic microsomal cytochromes p448-mediated activities. Toxicol. Appl. Phamacol. 47: 537-546 (1979); and Mohammadpour, H., Murray, WJ and Stohs, SJ 2,3,7,8-Tetrachloro-dibenzo-p-dioxin (2 , 3, 7, 8-TCDD) -induced lipid peroxidation in genetically responsive and non-responsive mice. Arch. Environ. Contam. Toxicol. 17: 645-650 (1988)).

  The inventors of the present application, in particular, are effective herbal medicines that can detoxify the above-mentioned typical representative toxic substances among the medicinal plants known to have a detoxifying function in traditional Chinese medicine books such as Tokankan and folk remedies. As a result of diligent research to find out, it was found that a mixed composition of several specific medicinal plant extracts can significantly reduce the toxicity of dioxins such as nicotine in the body and TCDD in the body, thus completing the present invention. .

The present invention has been made in view of the above problems, and an object of the present invention is to provide a composition comprising a herbal extract having an in vivo detoxifying action of typical representative toxic substances such as nicotine and dioxin. To do.

  Another object of the present invention is to provide a method for producing the composition.

In order to achieve the above object, according to the first aspect of the present invention, there is provided a mixed composition of herbal extracts containing black beans, koei kan and gold and silver flowers, which have a body nicotine and / or dioxin degradation effect. Features.

  According to the second aspect of the present invention, the mixture of herbal extracts of black beans, koei kan and gold and silver flowers, which has a body nicotine and / or dioxin degrading effect, is selected from the group consisting of Hanatsusei, Seiko and licorice, or It is characterized by providing a composition further comprising a further herbal extract.

According to a third aspect of the present invention, there is provided a method for producing a mixture composition of herbal extracts according to the first and second aspects of the present invention.
According to the 4th aspect of this invention, the foodstuff containing the composition by the 1st and 2nd aspect of this invention is provided, It is characterized by the above-mentioned.

  The composition according to the present invention is a typical toxic substance that can be easily obtained from herbal medicines that are easily available in the vicinity, can be easily manufactured from herbal medicines that are harmless to the human body, and causes serious damage to the health of modern people without causing side effects even when taken in an overdose It has an excellent effect on reducing in vivo toxicity of nicotine and dioxin.

  First, the effect of each herbal medicine constituting the composition according to the present invention will be briefly described based on matters described in traditional folk remedies or Chinese medicine related books.

According to Tokankan, Honsotsuna, etc., black beans (Glycine max Merrill), that is, black beans, have long been known to be effective in detoxification, live blood, jaundice, edema, numbness, diuresis and the like. Therefore, in countries such as Korea where rice is the staple food, rice is cooked with black beans, or soaked in vinegar and then eaten as is, or steamed with other cereals to make flour and ingredients for healthy food It is a food that is widely consumed due to its effectiveness, such as use as However, we never expected that black beans themselves would have a direct superior effect on nicotine or dioxin detoxification.

Licorice (Glycyrrhiza uralensis) has long been well known as a substance that has almost lacked the story of “licorice in pharmacy” to some extent. As for its specific efficacy, Togakukan etc. is nicknamed because it detoxifies the toxicity of all drugs and expresses its efficacy in harmony with 72 stone drugs and 1200 herbs. It is said that it is "Kunio". In addition, licorice is described as having an excellent effect of detoxifying various poisons together with ginger and jujube, and effective in detoxifying food poisoning, various drug poisoning and anticancer drugs.

On the other hand, there is a report that mixed use of the black bean and licorice extract has a detoxifying and protective effect against carbamate poisoning (Shin, IS, Min, KJ and Kang, HY study on the preventive effect of extract). of Glycyrrhizae Radix and Glycine semen on NAC intoxication. Kor. J. Env. Hlth. Soc. 19: 61-68 (1993)).

The gold-silver flower (Lonicerae flos) is a flower of Shinobi Fuyusaku, and it is used by Tokankan for heat poison, diarrhea, etc., and as an antidote and anti-inflammatory agent for treating colds, vomiting, etc. Has been.

Taraxacum mongolicum, also called 蒲 公 草, refers to the dandelion, and in the eastern ginseng, it turns into a heat poison, cures pressure sores, relieves stiffness, dissolves food poison, It is described that it has an effect to disperse. In addition, the Chinese Medicinal Dictionary has effects of antipyretic, detoxification, diuresis, and lactation, and it is said that it cures inflammation and artifacts and is used to treat the liver and gallbladder. In addition, it is noted that if you take dandelion juice continuously, your hair will become darker and your stomach and bones will be stronger. (Hong, SJ, Choi, EG, Lim, HS, Shon, JB and Jeong SS Effect of herbal dentifrice on dental plaque and gingivitis. J Korean Acad Dental Health. 25: 347-355 ( 2001)).

Saururus chinensis is a Chinese herbology book that treats edema and beriberi, passes large and small stools, cuts heels, clears clogs, breaks up masses in the abdomen, treats things and swelling Then it is written. In addition, the Chinese Pharmacy Dictionary, Chinese Medicine Dictionary, Karakusa, Shonan Yakuhin, and Honso Jinjin also treat edema and beriberi, improve bowel movement, and have antibacterial, detoxification, and anti-inflammatory effects. ing.

Seiko is a herbal medicine well known as a tea that reveals eyes. For Tojokan, Akemiko supplemented the liver and flushed with water to treat headache and nosebleed. For Tojo Pharmacy, healed headaches with liver fever, redness of eyes, pain and tears. It is described that it is used for visual acuity and for visual acuity enhancement. In addition, the medicinal plant dictionary describes that it is effective for diuresis, indigestion, gastrointestinal disease, etc. and improves visual acuity.

On the other hand, in natural folk remedies and traditional Chinese medicine, it is known that the use of both the Saururus chinensis and the Cassia obtusifolia has an excellent effect on diuresis and detoxification (Shin, IS, Min, KJ and Kang, HY A study on the preventive effect of extract of Glycyrrhizae Radix and Glycine semen on NAC intoxication.Kor. J. Env. Hlth. Soc. 19: 61-68 (1993); Hong, SJ, Choi, EG, Lim , HS, Shon, JB and Jeong SS Effect of herbal dentifrice on dental plaque and gingivitis. J Korean Acad Dental Health. 25: 347-355 (2001); , Folk remedies), Kyunghee University of Herbal Medicine, Department of Herbal Sciences, Korean Book Publishing Central Association. 970-971, 981-983 (1994)).

As described above, the inventors of the present invention are the main constituents constituting the invention of herbal medicines conventionally known to have various detoxification and diuretic effects by traditional Chinese medicine books and traditional folk remedies. As a result, the present invention was completed. However, as described above, the general fact that each herbal medicine has a certain degree of detoxification, diuresis, and appropriate anti-inflammatory action and activates metabolism in the body is known. No one easily anticipated that nicotine and dioxin, the representative toxic substances that cause serious health problems in modern society, would have particularly good effects on in-vivo detoxification. In addition, when the extracts of each of the herbal medicines are used alone, a significant detoxification effect as in the composition according to the first or second aspect of the present invention cannot be achieved, that is, It proves that the present invention cannot be achieved by simple selection or combination. Such facts can also be well established by experiments such as nicotine and dioxin acute toxicity tests, as described in detail in the examples of the invention below, and such tests and their Without considering the results, it is not only possible to demonstrate the detoxification effect of the specific composition described above on nicotine and dioxin, but also the present invention cannot be completed.

Hereinafter, the present invention will be described in detail.
First, in the method for producing the composition according to the present invention, each herbal medicine according to the first or second aspect is washed, dried, chopped, and then added with about 10 to 30 times the weight of the herbal medicine. , Each crude drug extract is obtained by reflux extraction at 30-50 ° C. for about 30-50 hours, and each herb extract is obtained by concentrating them under reduced pressure until a predetermined solid content is obtained. Each extract obtained can be freeze-dried to obtain powder of each herbal extract, and then these powders can be mixed and produced in a certain amount according to the components. Here, a person having ordinary knowledge in this technical field understands that the reflux time can be appropriately adjusted according to the reflux temperature when refluxing for extracting the herbal medicine.

  Or before freeze-drying each herbal extract obtained by the above method into a powder, each extract is concentrated under reduced pressure at a predetermined concentration, or a certain amount is mixed and used without concentration. be able to. That is, the mixture composition of the herbal extracts according to the present invention is in any form of each crude drug extract stock solution, an extract obtained by concentrating the crude drug extract with a predetermined solid content, or a dry powder obtained by freeze-drying these extracts. , And can be mixed and used at a certain ratio.

  The composition according to the present invention preferably has about 1 to 20 black bean extracts on the basis of the state where each herbal extract is concentrated under reduced pressure with the same solid content or lyophilized into powder form. Parts by weight, about 5 to 30 parts by weight of the extract of cocoon, and about 5 to 30 parts by weight of the gold and silver flower extract can be mixed. When semi-summer is further contained, it may further include about 5 to 30 parts by weight of Keriko extract, about 1 to 20 parts by weight of licorice extract, and about 10 to 50 parts by weight of semi-summer extract.

  Those having ordinary knowledge in this technical field need to know whether each herbal extract is concentrated in the same solid content as described above, or is not lyophilized powder. It will be understood that the composition according to the present invention can be produced by adjusting the content of the extract to a ratio as described above according to the concentration.

  The composition according to the present invention is particularly desirable to be administered orally, and can be formulated in various forms for convenient administration. That is, a mixture of the extract extract can be taken in a disposable container or the like, or can be easily taken by formulating the mixed powder together with an appropriate carrier in the form of a capsule or a tablet. Can do.

  As described above, the composition according to the present invention can be used as an extract or a mixture of powders as it is or formulated together with a suitable carrier so that it can be taken in a certain amount every day. The composition can be added to foods and taken on a daily basis. That is, the composition may be added to drinks, jelly, candy, gum, snacks, confectionery, etc. to be ingested or mixed with water or milk to be produced in the form of a health supplement. .

  The composition according to the present invention exhibits a significant detoxifying effect of nicotine and / or dioxin in the body when taken at 350 mg or more, more preferably 400 mg or more per kg body weight per day based on the above mixed powder state. discovered. When taken at 350 mg or less, the detoxification effect was not so high. On the other hand, since each of the herbal medicines is a plant that is approved for consumption by the Pharmacopeia, or most of the composition is an extract of a plant that is often ingested and taken on a daily basis, If the dose is overdose, no side effects such as toxicity will occur.

Hereinafter, the manufacturing method of the said composition by this invention and its effect are demonstrated in detail based on an Example. These examples serve to illustrate the invention and do not limit the invention.

  Extraction of herbal extract and manufacture of extract powder

  Purchasing black beans, gold and silver blossoms, Koei Hou, Seiko, Hankao, and licorice in the Korean market, chopping each 500 g, adding 10 L of 40% ethanol for each herbal medicine, and reflux extraction at 40 ° C. for 48 hours . Each crude drug extract thus obtained was concentrated under reduced pressure at 40 ° C. until the solid content was 25%, and then freeze-dried to obtain a powder of each crude drug extract.

The powders of these herbal extracts were used according to the examples of the present invention by formulating the powder together with a carrier such as distilled water or in the form of a tablet. And the effect was compared for each herbal medicine or mixing and using the powder of 2 or more types of herbal extracts.

Nicotine acute toxicity test method

  After 22 male 3-week-old male ICR mice weighing about 12 ± 2 g (Samutako, located in Ulsan City, Gyeonggi-do, Korea) were divided into 11 for each of the control group and the treatment group, and adapted to the breeding environment for 1 week. Used for experiment.

  Mice are kept in an environment set at a temperature of 22 ± 1 ° C, a humidity of 55 ± 5%, a ventilation rate of 12 times / day, and a light / dark cycle of 12 hours / day (lights on at 8 am, lights off at 8 pm). Each mouse was housed and the powdered feed for mice (Samuco, product number: Sam # 31) was ingested with water.

  At this time, the treated group of mice corresponds to 10% of the daily intake of about 4 g of feed, which is a mixture of the same amount of the powders of the six kinds of herbal extracts according to the present invention (hereinafter referred to as mixed powder A). 400 mg each was added to the feed and fed for 1 week from 4 weeks of age, and the control group fed only the same amount of powdered feed.

On the other hand, the nicotine lethal dose for the control group mice was determined in a preliminary experiment. For this purpose, 42 mg / kg of nicotine tartarate salt (Sigma No. N5260) in a pure nicotine amount was dissolved in 0.9% physiological saline, and this was then added to a 0.45 μL syringe filter ( Syringe filter) was used. After feeding the test feed (SamTako, product number: Sam # 31) for 1 week, nicotine is administered into the abdominal cavity of a 5-week-old mouse, and the time to start paralysis, the start and end time of seizure, and the time until death It was measured.

Confirmation method of nicotine detoxification effect in human body

Smoking allows many harmful substances to flow in directly or through tobacco smoke. In particular, nicotine among these harmful substances is used as a precursor for producing carcinogens such as NNK by metabolism in the body. According to a recent report, cotinine, the main metabolite of nicotine, is known to be a stable substance and at the same time suppress NNK activity and formation (Trushin, N., Hecht, SS Strereoselective metabolism of nicotine). tobacco-specific N-nitrosamines to 4-hydroxy-4- (3-pydidyl) butanoic acid in rats.Chem Res Toxicol.
12: 163-71 (1998)).

  In order to confirm the detoxification effect of nicotine in the human body, the aforementioned mixed powder A was produced as a tablet containing a daily intake of 1,000 mg. At this time, as described above, in order to compare the effect of each of the six kinds of herbal extracts according to the present invention with the effect of the mixture of two or more kinds of herbal extracts, each powder of one kind of herbal extract is used. And tablets containing a combination of two or more herbal extract powders. At this time, the total amount of the herbal extract powder contained in one tablet was set to 1,000 mg. The nicotine detoxification ability in the human body was measured by allowing the test subject to take the tablet thus produced by the method described later.

  The test subjects were 11 healthy men aged 20-30 with an average daily smoking amount of 1 box or more. The test subject took a tablet at the same time as urine collection, collected urine on the third day of administration, and at 10 am at weekly intervals, and quantified cotinine in the urine for about one month. The change in the amount of nicotine in the body before and after taking these tablets was confirmed.

Nicotine is very unstable compared to cotinine, which has a half-life of about 30 to 60 minutes and a half-life of 15 hours. Therefore, nicotine detoxification power is estimated by analyzing the amount of urinary cotinine that can reduce analysis errors. (Peach, H., Ellard, GA, Jenner, PJ and Morris, RW A simple inexpensive urine test of smoking. Thorax. 40: 351-357 (1985)). Confidence interval (CI) was calculated using Statistix and compared with 95% confidence limit (Statistix. Statistix for Windows. User's Manual, Analytical software. USA. (1996)) .

Cotinine assay

  Barlow et al. (Barlow, RD, Thompson, PAand Stone, RB Simultaneous determination of nicotine, cotinine and five additional nicotine metabolite in the urine of smokers using pre-column derivatisation and HPLC. BJ Chromatogr. 419: 375-380 (1987 )), The amount of cotinine in urine was measured.

That is, as described above, urine collected from each test subject was centrifuged at 450 × g for 10 minutes, and then 200 μL of the supernatant was put into a microtube, and 4M sodium acetate buffer (pH 4. 7) 100 μL, 1.5 M potassium cyanate 40 μL, 0.4 M chloramine-T 40 μL, 50 mM (v / v) 78 mM barbituric acid prepared with acetonitrile ) 200 μL was added in order and then mixed for 10 seconds. This mixture was reacted at room temperature for 15 minutes, and the reaction was stopped with 40 μL of 1M sodium metabisulphite, and then the absorbance was measured at 490 nm to quantify cotinine.

Dioxin acute toxicity test method

  Purchase 40 rats of 3 weeks old male rats (rat) TcnN (SD) BR (Sam Taco Co., Ulsan, Gyeonggi-do, Korea) weighing about 200 ± 20g. After 7 days of adaptation, it was used for experiments.

  The test animals are raised in an environment set at a temperature of 22 ± 1 ° C., a humidity of 55 ± 10%, a ventilation rate of 12 times / day, and a light / dark cycle of 12 hours / day (lights on at 8 am, lights off at 8 pm). One animal was raised per cage, and the rat chow was freely fed with water.

  Dioxin was used by using TCDD (AccuStandard Co., USA) and dissolving 1 mg of TCDD in 25 μL of DMSO and 225 μL of acetone, and then diluting it in 2.25 mL of corn oil. A mixed powder A produced by mixing the same amount of all the six kinds of herbal extract powders according to the present invention was orally administered after being dissolved in distilled water at a concentration of 400 mg / kg / day.

  As shown in FIG. 1, the first group (control group) contained physiological saline (0.4 mL), and the second group (carrier administration group) contained acetone, DMSO, and corn oil. The carrier alone, the third group (TCDD single administration group) was injected intraperitoneally with 0.4 mL of saline containing TCDD at a concentration of 5.0 μg per kg body weight of the rat. For the fourth group (premixed powder A pre-administered group), the powdered powder A was orally administered 7 days before the first intraperitoneal injection of TCDD, and for the fifth group (mixed powder A co-administered group), TCDD1 Simultaneously with the next peritoneal injection, the mixed powder A was orally administered, and the amount of these TCDD injections was set to 0.4 mL of physiological saline containing TCDD at a concentration of 5.0 μg per 1 kg body weight of the rat as in the third group. Saline and TCDD injections were administered twice at 1 week intervals.

  One week after the secondary peritoneal injection, the rats were anesthetized with ethyl ether and whole blood was collected from the heart, and then clinical values for blood, liver, pancreas, kidney, and heart function were analyzed. At this time, the least significant difference test (LSD) was performed at the p <0.05 level with nova (ANOVA) between each group using Statistix (Statistix. Statistix for Windows. User's Manual, Analytical software. USA. (1996)).

Comparison of the effects of herbal extracts by nicotine detoxification in the human body

  As described above, in order to compare the detoxification effect between one containing six kinds of herbal extracts according to the present invention and one containing a mixture of two or more kinds of herbal extracts, Tablets containing a total of 1,000 mg of powder or mixed powder of two or more extracts per tablet were produced. As described in the method for confirming the nicotine detoxification effect in the human body, each test subject took each of the manufactured tablets to measure and compare the nicotine detoxification effect in the human body.

  Among these, FIG. 2 shows, in particular, (1) tablets containing 1,000 mg each of black bean, cocoon and gold and silver flower extract powders; (2) black beans and cocoon, black beans and gold and silver flowers; (3) tablets containing about 333 mg each of three kinds of mixture powders of black beans, koei ei, gold and silver flowers, and 999 mg in total, and (4) ) Tablets containing 250 mg each of 4 kinds of herbal extract powders with a total of 1,000 mg mixed with black beans, koei kan, gold and silver flowers, and (5) black beans, koei kan, gold and silver flowers, semi-summer, It is the bar graph which compared and showed the effect at the time of taking the tablet which mixes all the powder of 6 types of herbal medicines of Keriko and licorice at the same quantity, and contains a total of 1,000 mg.

  The graph is composed of 11 bar groups each consisting of 6 bars. In each bar group, 6 bars are 11 test subjects who took the same tablet on the 0th day, 3rd day, 1st week, 2nd week, 3rd week, and 4th week from the day of taking each tablet. The average value of the cotinine concentration measured from urine was shown.

  Therefore, each bar group serves as an index for comparing the degree of nicotine degradation in urine according to the type of tablet. That is, in the graph, as described on the lower side of each bar group on the horizontal axis, the first bar group is a case where a tablet containing only the “black bean” extract is taken, and the second bar group Is the case of taking a tablet containing only the “Kong Koei” extract, the third group of bars is the case of taking a tablet containing only the “gold and silver flower” extract, and the fourth group of bars is “ The case of taking tablets containing a mixture of “black beans and 蒲公英” extract, the fifth group of bars is the case of taking tablets containing a mixture of “black beans and gold and silver flowers” extract, the sixth group of bars Is the case of taking a tablet containing a mixture of “Kong Koei and Golden Silver Flower” extract, and the seventh bar group is taking a tablet containing a mixture of three kinds of extracts “Black Bean, Xin Ying Kim, Gold and Silver Flower” The eighth group of sticks is a tablet containing a mixture of four kinds of extracts of “black beans, koei rin, gold and silver flowers and semi-summer”. The ninth bar group is the case of taking a tablet containing a mixture of four kinds of extracts of “black beans, Koei Sakaki, golden silver flowers, and Akemiko”, and the tenth bar group is “black beans” In the case of taking a tablet containing a mixture of four kinds of extracts, “Kimei, Jinyin, Golden Silver Flower and Licorice”, the eleventh bar group is six kinds of “Black Bean, Jin Yin, Golden Silver Flower, Semi-Summer, Koseko and Liquorice” The result of having measured the urinary cotinine density | concentration at the time of taking the tablet containing all the crude drug extracts of is shown.

  As can be seen from the graph, when taking tablets containing all 6 kinds of herbal extracts (11th bar group), it is the third day after taking compared to before taking (day 0). The urinary cotinine concentration was the highest. Next, in the case of taking a tablet further containing one kind of herbal extract in addition to “Black Bean, Koei Hou, Gold and Silver Flower” (8th, 9th, 10th bar group) The cotinine concentration on one day appeared higher than before taking (day 0). In the case of tablets (seventh group of rods) containing three kinds of extracts of “Black Bean, Koei Sasa, and Gold and Silver Flowers”, the cotinine concentration on the third day after taking the tablet is compared with the day of taking the tablet. Such an aspect appears as a tablet containing only one herbal extract (first to third bars) or a tablet containing only two herbal extracts (fourth to sixth bars). ) Is markedly different from the case of taking).

  Tablets containing only one other herbal extract not shown in FIG. 2, or two or three or more herbal extracts that essentially do not contain “Kurome, Koei, and Gold and Silver Flowers” Although these were also tested, they showed a lower detoxification effect than those shown in the drawings, and the illustration was omitted.

  From these results, the composition containing all the six kinds of herbal extracts according to the present invention has the best detoxification effect of nicotine in the human body, and at least three kinds of herbal medicines containing at least “black beans, sung kouei and gold and silver flowers”. It has been discovered that a mixture of extracts has a significant nicotine detoxifying effect in human body compared to a single or a mixture of two or more other herbal extracts.

  Therefore, in the following examples, using a composition containing all six herbal extracts that have been proved to be most effective in the present invention, nicotine acute toxicity test, nicotine detoxification effect in human body and Dioxin detoxification effect was tested.

Nicotine acute toxicity test results

  The mouse acute toxicity test for the composition according to the present invention is a treatment in which a mixed powder A containing the same amount of all six herbal extracts according to the present invention at a dose of 400 mg / day per week is administered together with a feed. The group and the control group administered only the same amount of powdered feed were used as subjects.

The nicotine lethal dose obtained in the preliminary experiment, 42 mg / kg, was adjusted according to body weight and injected intraperitoneally. Nicotine, which is water-soluble, is toxic enough to be absorbed into the body immediately when administered intraperitoneally, and in response to nerve cell membrane receptors, it increases blood pressure, induces convulsions of skeletal muscle fibers, and seizures It is known to cause such oral paralysis (Craig, CR and Stizel, RE Modern pharmacology 4 th ed. Little BROWN. USA. p171-173 (1994)).

(Table 1) Protective effect of the composition according to the present invention on mice injected with lethal dose of nicotine (42 mg / kg) ¹⁾

As can be seen from Table 1 above, the onset time of paralysis initially occurring due to acute nicotine toxicity is 55 seconds on average in the control group, and 2 minutes and 20 seconds in the treatment group administered with the mixed powder A according to the present invention. It has been found that the intake of the herbal extract according to the invention delays the onset time of paralysis more than twice.

  In terms of seizure onset and completion time, the control group showed seizure symptoms from 1 minute 26 seconds to 3 minutes 39 seconds for 2 minutes 13 seconds, whereas the mixed powder administration group according to the present invention exhibited 3 minutes 13 seconds to 4 minutes. Seizures were exhibited for 1 minute and 10 seconds up to 23 minutes. The time required from the start of paralysis to the start of seizure was 31 seconds for the control group, and 53 seconds for the mixed powder treatment group of the herbal extract according to the present invention, which was longer than the control group.

  In the case of the control group, 9 out of 11 animals died after the seizure had stopped and were in coma for an average of 4 minutes and 6 seconds, and 7 minutes and 45 seconds after the injection of a lethal dose of nicotine. Two animals were confirmed to die for the first time and two recovered. In contrast, in the mixed powder A administration group according to the present invention, 3 out of 11 animals died for the first time after the seizure stopped, but the remaining 8 recovered.

  Thus, mice fed the mixed powder of the herbal extract according to the present invention, in the acute toxicity experiment injected with a lethal dose of nicotine, the occurrence of paralysis and the delay in seizure start time, shortening the seizure duration compared to the control group, It was also confirmed that the survival rate from nicotine lethal dose injection increased from 18% to 73%.

  These results show that iridoid glucosides and polyphenolic compounds contained in other herbal extracts such as gold and silver flowers in the herbal medicine are decomposed into nicotine into cotinine. It is considered that the paralysis symptom induced by convulsions, seizures, etc. of skeletal muscle fibers is alleviated and mortality due to nicotine is greatly reduced.

Nicotine detoxification test in human body

  In order to test the efficacy of nicotine degradation for the composition according to the present invention (mixed powder A), a calibration curve was prepared using standard cotinine, and a linear equation such as the following Equation 1 was obtained.

(Equation 1)
y = 0.0009x + 1.344 (R ² = 0.992). . . . (1)

  In the above formula 1, x is the absorbance of the specimen, y is the amount of cotinine due to the absorbance, and R means that the closer the square value is to 1, the higher the accuracy.

  Cotinine was quantified from the urine of the test subject, and the propensity of cotinin to be excreted in the urine is shown in FIG. In addition, FIG. 3 isolate | separates and shows only the 11th bar | burr group among the bar graphs which are the data experimented in order to compare the effect between crude drug extracts in Example 1. FIG.

  As shown in FIG. 3, the average value of 197.4 ± 4.9 uM was obtained on the day 0 sample taken before the subject took tablets made of the mixed powder of 6 kinds of herbal extracts according to the present invention. A high cotinine excretion pattern was shown, and on the third day after ingestion, the excretion amount increased rapidly to 302.9 ± 50.4 uM. On the 7th day, the amount of cotinine excretion decreases sharply to 27.6 ± 8.5uM, rises slightly on the 14th day, and shows a stable discharge tendency to 10uM or less after 21 days. I understood.

  According to a report by Kyung et al. (Kyung, YJ and Lee, DH Enhanced conversion to cotinine from nicotine by green tea extract. Korean J. Environmental Agriculture. 19: 147-153 (2000)), nicotine with green tea extract in vitro. As a result of promoting the degradation of, the production rate of cotinine was remarkably increased, and it was confirmed that the green tea extract might promote the degradation of nicotine. In addition, in a nicotine degradation clinical experiment for green tea and mulberry leaves, ginkgo, cinnamon, licorice, apple and lemon extract mixed preparation, the extract was mixed with the urine collected three times at 30-minute intervals from the control group drinking only water. In the case of the administration group in which the preparation was drunk, the average amount of cotinine excretion was further eliminated by 20 uM, and the extract preparation acted as an accelerator for decomposing nicotine into cotinine in the body, or nicotine was converted to nitrosamines. It is said that the amount converted to cotinine is relatively increased (Kim, JH, Lee, DH, Cho, HJ and Jeong, JM Effects of phyto-extract mixture on the nicotine decomposition. J. Fd Hyg. Safety. 17: 8-14 (2002)).

  As a result of this experiment, the degradation of blood nicotine maintained above a certain level by smoking was promoted for one week after taking the mixed powder tablets of the six kinds of herbal extracts according to the present invention, and the amount of cotinine excretion in urine increased. After one week, the urinary cotinine content decreased to a level of 5% or less compared to before taking the mixed powder tablet of the herbal extract due to a decrease in the accumulated blood nicotine concentration, and powerful nicotine excretion And the detoxification power could be confirmed.

Dioxin (TCDD) acute toxicity test

  (1) Effect of the composition according to the invention on clinical and chemical indices of rat blood exposed to TCDD

  The results of testing the effect of feeding the composition according to the invention (mixed powder A) on the clinical and chemical index of rat blood induced acute toxicity with TCDD are shown in Table 2.

Table 2 Effect of the composition according to the invention on the clinical chemistry index of blood of male rats exposed to TCDD

  As can be seen from Table 2, the white blood cell count decreased by 46% in the third group administered with TCDD compared to the first group administered with saline alone, and the fourth and fifth groups. There was also a decrease in the group (p <0.05). In the case of red blood cell count, there was no significant difference in all groups administered TCDD compared with the first group, but a slight decrease phenomenon appeared, and TCDD administration induced TCDD acute toxicity. (Hwang, SY, Kim, SK, Kim SH, Kwak, YSand Jeong, YJ Effect of korean red ginseng on clinical chemical parameters in male guinea pigs exposed acutely to 2, 3,7,8-Tetrachlorodibenzo-p-dioxin. J. Korean Soc. Food Sci, Nutr. 28: 1349-1354 (1999)).

  In the case of the hemoglobin concentration, there was no significant difference in the TCDD single administration group (third group) compared with the first group, but it decreased, and the fourth group and the simultaneous administration of the mixed powder A according to the present invention were reduced. In the administered fifth group, the amount of decrease in hemoglobin is partially alleviated.

  The number of platelets decreased with the administration of TCDD, but the result of the administration of the mixed powder A according to the present invention showed a significant difference and recovered close to the steady value (p <0.05). This is due to the administration of red ginseng extract in TCDD acute toxicity experiments (Hwang, SY, Kim, SK, Kim SH, Kwak, YS and Jeong, YJ Effect of korean red ginseng on clinical chemical parameters in male guinea pigs exposed acutely to 2 , 3,7,8-Tetrachlorodibenzo-p-dioxin. J. Korean Soc. Food Sci, Nutr. 28: 1349-1354 (1999)) TCDD administration was confirmed to be a platelet and leukocyte production disorder substance.

  As can be seen from the second group, the hematocrit associated with anemia decreased the hematocrit significantly with the administration of DMSO and acetone carrier, and was significantly different with all TCDD administration groups. Diminished. In addition, the average red blood cell volume (MCV) is not significantly different between the treatment groups, and in the case of the average red blood cell hemoglobin amount (MCH), it partially increases in the group treated with carrier or TCDD, and the average red blood cell hemoglobin concentration (MCHC) is , The remaining treatment group increased significantly with respect to the first group, and it was confirmed that the administration of the carrier and TCDD also affected the occurrence of anemia, and the administration of the mixed powder A according to the present invention showed no significant difference. Although not, it has been shown to partially block MCHC elevation.

  In addition, Neutrophil Segment (%), Lymphocyte (%), and Monocyte (%) are immune to acute chronic inflammation, leukemia, allergic diseases, hematopoietic cell disorders, etc. The second group in which the neutrophil (%) was decreased by TCDD administration but the simultaneous administration of the mixed powder A according to the present invention was significantly different from the carrier treatment group. It was confirmed that the immunity was restored to the standard. It was confirmed that lymphocytes (%) increased significantly in all administration groups of TCDD. Mononuclear cells (%) decreased in the third group which was the TCDD administration group, but recovered to the same level as the first group in the fourth group and the fifth group to which the mixed powder A according to the present invention was administered. I found out that

  (2) Effect of composition according to the present invention on intrahepatic clinical and chemical indices of rats exposed to TCDD

  The effect of the composition according to the invention on the intrahepatic clinical and chemical indices of male rats exposed to TCDD is shown in Table 3.

Table 3 Effect of the composition according to the invention on the intrahepatic clinical chemistry index of male rats exposed to TCDD

As can be seen from Table 3, the blood protein content was not significantly different in all the test groups, and the albumin content was significantly decreased (p <0.05) by TCDD administration. The administration of powder A recovered to a value similar to that in the first group as the control group. In addition, GOT (glutamate oxaloacetate transferase) activity was significantly increased when TCDD was administered as compared to the first group, but was found to be significantly suppressed by administration of mixed powder A according to the present invention. (Glutamate pyruvate transferase) activity was slightly increased by TCDD administration, but it was confirmed that the activity was reduced by administration of mixed powder A according to the present invention. Alkaline phosphatase also increased significantly with TCDD administration, but recovered to the first group level with prior administration and simultaneous administration of the mixed powder according to the present invention.

  In the hepatotoxicity test of mice against TCDD, in the case of the TCDD single administration group, the appearance was normal and the body weight did not show a significant difference between the treatment groups for 2 weeks after the TCDD administration, but at the time of necropsy, the liver was discolored due to jaundice Became thickened and many punctate moribunds were observed. However, in the case of the mixed powder A administration group according to the present invention, the hue was faded as compared with the first group, but it was observed that there were almost no punctate moribund lesions. (Zhang, HS, Nam, SH and Kang, JK Protective Effects of Bear Bile against Hepatotoxicity induced by 2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) in Mice. Kor. J. Pharmacogn. 32: 121-127 (2001)).

  In addition, as a result of administration of red ginseng to guinea pigs that induced TCDD acute toxicity, it was confirmed that there was no change in albumin and total protein content, which was different from this experiment, and GOT, GPT, alkaline phosphatase increased with TCDD administration, Administration of red ginseng is known to recover close to the control group and reduce liver toxicity, and it was confirmed that the mixture of herbal extracts according to the present invention also significantly reduces liver toxicity (Hwang, SY, Kim, SK, Kim SH, Kwak, YS and Jeong, YJ Effect of korean red ginseng on clinical chemical parameters in male guinea pigs exposed acutely to 2,3,7,8-Tetrachlorodibenzo-p-dioxin.J. Korean Soc. Food Sci, Nutr. 28: 1349-1354 (1999)).

  (3) Effect of the composition according to the invention on the clinical and chemical index of rat pancreas exposed to TCDD

  The effect of the composition according to the invention on the clinical and chemical index of male rat pancreas exposed to TCDD is shown in Table 4.

Table 4 Effect of the composition according to the invention on the clinical chemical index of male rat pancreas exposed to TCDD

In all animals, administration of dioxin, or TCDD, is known to increase the total blood cholesterol and neutral fat content (Brewster, DW and Matsumura, F. 2,3,7,8-Tetrachlorodibenzo-p -dioxin reduces lipoprotein lipase activity in the adipose tissue of the guineapig. Biochem. Biophys. Res. Commun. 122: 810-817 (1989)).

  Table 4 is an analysis of the effect of TCDD on the pancreas. The total cholesterol content is significantly increased in all TCDD administration groups, and the administration of the mixed powder A according to the present invention shows the total cholesterol value by TCDD as the first value. Although it did not recover to the group level, it was confirmed that it recovered about 70% with a significant difference (p <0.05). In the case of blood neutral lipid, it was confirmed that there was a significant difference between the carrier and TCDD administration, and it was confirmed that the administration of the mixed powder A according to the present invention reduced the neutral lipid increased by TCDD by about 50%. It was done.

  HDL cholesterol (high-density lipoprotein cholesterol) increased with TCDD alone with a significant difference compared to the first group. However, the first group of the mixed powder A according to the present invention was pretreated and coadministered. Fell to the level. This trend was found to be similar to the study on the effect of red ginseng on guinea pigs that induced TCDD acute toxicity (Hwang, SY, Kim, SK, Kim SH, Kwak, YS and Jeong, YJ Effect of korean red ginseng on clinical chemical parameters in male guinea pigs exposed acutely to 2,3,7,8-Tetrachlorodibenzo-p-dioxin. J. Korean Soc. Food Sci, Nutr. 28: 1349-1354 (1999)).

  Other lipases and amylases were confirmed to have little difference between treatment groups.

  (4) Effect of the composition according to the invention on the clinical and chemical index of rat kidneys exposed to TCDD.

The effect of the composition according to the invention on the clinical and chemical indices of male rat kidneys exposed to TCDD is shown in Table 5.

Table 5 Effect of the composition according to the invention on the clinical chemistry index of male rat kidneys exposed to TCDD

  As can be seen from Table 5 above, the kidney-related hematological index by TCDD administration shows that blood urea nitrogen has almost no difference in all treatment groups, and in the case of creatinine, only TCDD In the third group to which shampoo was administered, 0.53 ± 0.23 increased significantly compared to the first group, but the fourth and fifth groups which were the mixed powder A administration group according to the present invention In this case, the same numerical value as in the carrier-administered group (second group) was shown, and it was confirmed that the increase due to TCDD was suppressed. The concentration of uric acid (Uric acid) was not significantly different between the treatment groups, but increased in the carrier and TCDD administration groups, and showed a tendency to decrease with the administration of the mixed powder A according to the present invention.

  And although the content of calcium (Ca) and phosphoric acid (P) is not changed between treatment groups, the iron (Fe) content in blood decreased with TCDD administration with a significant difference. It was found that pre-administration (fourth group) and simultaneous administration (fifth group) completely recovered above normal levels.

The kidneys are responsible for waste excretion, maintaining homeostasis by regulating body fluids, and hormone-related endocrine functions.The kidneys have a high blood flow and are exposed to toxic substances contained in the blood in the process of filtering waste products. During high and temporary storage, the concentration of toxic substances occurs and becomes the target of toxic substances (Hwang, SY, Yang, JB, Chang, CS, Lee, YC and Lee, HY Protective effect of cornu parvum extract on toxicity induced by 2,3,7,8-tetrachlrodibenzo-p-dioxin in rat. Korea J Oreintal Physiology & Pathology. 16: 647-679 (2002)). From such a result, it is determined that such a result appears due to damage caused by TCDD deposited in the adipose tissue because the kidney is surrounded by adipose tissue.

  (5) Effect of the composition according to the invention on the clinical and chemical indices of rat hearts exposed to TCDD.

  The effect of the composition according to the invention on the clinical and chemical indices of the heart of male rats exposed to TCDD is shown in Table 6.

Table 6 Effect of the composition according to the invention on the clinical chemical index of the heart of male rats exposed to TCDD

As can be seen from Table 6, in the blood chemistry index related to the heart, in the case of lactate dehydrogenase, which is a numerical value that discriminates the degree of fatigue of liver disease, myocardium, skeletal muscle, etc., TCDD is mostly administered Although it had no effect, the value was lower than that of the control group (first group) at the time of pre-administration (fourth group) and simultaneous administration (fifth group) of the mixed powder A according to the present invention, and muscle fatigue It is judged that it had a positive effect on the decrease in the degree of stamina and the increase in endurance.

  Creatine phosphokinase (CPK) is an enzyme that regulates so that sufficient ATP level can be maintained during muscle contraction by phosphorylating ADP formed from depletion of ATP. The TCDD administration significantly decreased 67% compared to the normal group, and the pre-administration (the fourth group) and the co-administration (the fifth group) of the mixed powder A according to the present invention had almost no effect. It was confirmed. In addition, CK-MB, which is the value of CPK in the muscle-brain, is decreased with a significant difference by the administration of TCDD as in CPK, but the mixed powder A according to the present invention is reduced. Pre-administration (fourth group) and simultaneous administration (fifth group) were confirmed to have little effect. The propensity of CPK and CK-MB is to prevent TCDD, an environmental hormone that acts as a biological disruptor, from disrupting the activity of CPK and supplying sufficient energy to the myocardium and other muscles. It is thought to increase muscle fatigue and decrease motility.

From these results, the organs in which TCDD is mainly metabolized are different, and the toxicity that appears in each organ differs depending on the organ. Therefore, it is considered that the toxicity reducing action of the composition according to the present invention appears differently between tissues. It is done. It has been reported that the metabolic processes of toxic chemicals and drugs are mostly carried out in the liver, and reactions that occur in the kidney, intestine and skin play an important role in drug detoxification and carcinogenic processes of carcinogenic precursors. Vecchini, F., Mace, K., Magdalou, J., Mahe, Y., Bernard BA and Shroot, B. Constitutive and inducible expression of drug metabolizing enzymes in cultured human keratinocytes. Dermatol. 132: 14-21 (1995)).
Thus, in view of changes in the clinical chemistry index of blood, TCDD administration exhibits a more markedly toxic effect on blood and pancreas than on liver, kidney and heart, and the six herbal extracts according to the present invention. Toxicity-reducing action by ceramide was remarkably expressed in the liver and pancreas, and it was confirmed that the change in the clinical index by TCDD administration was restored to a steady value. We were able to confirm that it was a substance.

  In the embodiments described above, only the effect of nicotine and dioxin on reducing the in vivo toxicity of the composition of the present invention has been mainly described, but it is related to other in-vivo metabolites or other physical diseases related to the toxic substance. Nevertheless, those skilled in the art will appreciate that the compositions of the present invention can have significant effects. Such an effect belongs to the scope of the present invention.

FIG. 3 shows a protocol for administration of mixed powder A and 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) according to the present invention for a dioxin acute toxicity test. 2 is a graph showing the nicotine detoxification effect in the human body after administration of a tablet containing each herbal extract alone or a mixture according to the present invention. It is a graph which shows the cotinine density | concentration in urine after taking the tablet manufactured with the mixed powder A by one Example of this invention.

Claims (5)

A composition comprising an extract of black beans, koei kan, gold and silver flowers, semi-summer, seiko and licorice for decomposing nicotine and dioxins in the body. Each of the extracts is 1 to 20 parts by weight of a black bean extract, 5 to 30 parts by weight of a rice cake, 5 to 30 parts by weight, based on those concentrated to have the same solid content. gold and silver flower extract parts, last seed-sowing day extract 10-50 parts by weight, the composition according to claim 1, characterized by comprising a licorice extract determined Akiko extract and 1-20 parts by weight of 5 to 30 parts by weight object. The composition according to claim 1 or 2 , wherein the composition is intended for oral administration. The composition according to claim 1 or 2 , wherein each herbal extract is in a powder state obtained by concentrating and drying the extract. Each of the herbal medicines according to claim 1 or claim 2 is washed and dried, then chopped, and each herbal medicine is refluxed and extracted with 10 to 30 times the weight of each herbal medicine, and each extracted herbal medicine extract is concentrated. 3. The method for producing a composition according to claim 1, wherein the composition is freeze-dried or mixed without being freeze-dried.

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