KR100857249B1 - Composition for increasing immune activity comprising hot water extract from capsosiphon fulvescens - Google Patents

Abstract

A composition comprising the extract of Capsosiphon fulvescens is provided to increase immune activity of complement system in human body by promoting production of IgA(immunoglobulin A) antibody, and inhibit growth of cancer cells without side effects and toxicity to human, so that the composition is useful to treat and prevent various diseases. A composition for increasing immune activity comprises the extract of Capsosiphon fulvescens with molecular weight of 300-500kDa which is prepared by extracting Capsosiphon fulvescens in hot water at 90-110 deg. C for 2-4 hours, precipitating the hot water extract of Capsosiphon fulvescens with alcohol, and ultrafiltering the precipitates, wherein the daily dosage of the composition is 1-1000 mg/kg.

Description

Composition for increasing immune activity comprising hot water extract from Capsosiphon fulvescens

Figure 1 shows the change in complementary activity and yield with respect to the extracting time of the extract of the hot water extracts of the perennial.

Figure 2 shows the change of complementary system activity and yield according to the extraction temperature of the falsified hot water extract according to the present invention.

Figure 3 shows the change in IgA in mouse feces to which the hot water extract was administered in accordance with the present invention.

Figure 4 is a graph showing the results of the solid rock growth inhibition experiment according to the present invention.

The present invention relates to a composition for increasing immune activity, which contains hot water extracts of Maesung, more specifically, the medicinal plants that promote complement system immune activity, intestinal immune activity, leukocyte increase in blood, and weight increase of immune-related organs contain hot water extract as an active ingredient. It relates to a composition for increasing immune activity and inhibiting cancer cell growth.

Industrialization for economic development is worsening the environment we live in, and changes in dietary habits change the homeostasis inside the human body, causing incurable diseases such as cancer, acquired immunodeficiency syndrome and adult diseases, which threaten human health and safety. Until now, various chemotherapy has been performed to solve these problems, but it is true that side effects thereof cannot be overlooked. Therefore, in recent years, studies are being actively conducted to treat or prevent various diseases through immunotherapy known to have relatively few side effects, that is, a method of enhancing immunity possessed by the human body. Some of the biochemical reactions that occur in vivo must occur in stages, which activate a first stage reaction and then a cascade reaction in which the second stage reaction is activated, which in turn activates the next stage reaction. . This continuous step reaction must be activated in sequence and not activated from the middle step without the previous step reaction. Biochemical reactions that take place step by step, combined with the function of antibodies, have a significant effect on host defense mechanisms.

Complement system (complement system) consists of about 20 kinds of serum circulating proteins and is a major defense mechanism of the body that performs the effective linkage of humoral immunity and removes non-infectious pathogens nonspecifically in the presence or absence of antibodies. One]. The activity of complement is the classical pathway that activates at the level of C1q by an immune complex comprising an antibody (IgM or IgG) and an antigen, and the minor pathway that directly activates C3 without the help of the antibody. It is activated by two pathways. So far known complement-activated polysaccharides are four pectic arabinogalactan isolated from the roots of Angelica acutiloba, two neutral arabinogalactan [2-4], and acidic heteroglycan [5] from the leaves of Artemisia princeps. Two acidic heteroglycans [6] from the stems of Coix lachryma-jobi var.ma-yuen, three acidic heteroglycans [7-8] from the leaves of Panax ginseng, and α- (of Pteridiym aquilinum). 1 → 2) -glucuronmannan back born [9], and many of these polysaccharides have been isolated from terrestrial higher plants.

On the other hand, marine life accounts for 80% of all living things on the planet, and the habitat environment is very different from that on land. Therefore, the present inventors searched for complement system active substances in vitro from 102 species of marine organisms, and among them, it is aimed at capsosiphon fulvescens , which is considered to be easy to industrialize due to its high activity and relatively rich resource. We reviewed the proper extraction and purification conditions of immune active substances, examined cytotoxicity against five human cancer cells, analyzed glycocomposition of purified immune active substances, confirmed intestinal immune activity in vivo, mouse sarcoma Cell growth inhibitory effect and acute toxicity test were performed.

Accordingly, the present inventors have made a intensive study to overcome the problems of the prior art, as a result, the composition of the medicinal plants with immune activity using the hot water extract as an active ingredient, it is confirmed that can enhance the immune function in the human body, the present invention To complete.

Therefore, the main object of the present invention is to provide a composition for increasing immune activity, which contains hot water extract as an active ingredient, which is capable of increasing complement immune activity in vivo by searching for complement active substances in vitro from marine organisms. .

Another object of the present invention is to provide a composition for inhibiting cancer cell growth, wherein the periphery contains hot water extract as an active ingredient.

According to an aspect of the present invention, the present invention provides a composition for increasing immune activity, which contains the extract as an active ingredient.

In the composition of the present invention, the medicinal herb extract may be extracted with a hydrophilic organic solvent such as water or alcohol or a mixed solvent thereof, preferably hot water extracted. In addition, the hot water extraction is preferably hot water extraction for 2 to 4 hours at 90 ~ 110 ℃, preferably characterized in that hot water extraction for 3 hours at 100 ℃. In addition, the active substance of the present invention is stable to heat, and even if extracted with hot water, there is no big problem in maintaining the activity, and according to the current Korean Food Code, it is not possible to use solvents other than water and spirits in extracting food components.

In the composition of the present invention, the maesengyi extract generally shows higher activity as the molecular weight is higher, but it is preferable that the maeseng was purified by precipitation with alcohol and then purified by ultrafiltration in the range of 300 to 500 kDa molecular weight (Table 1). Reference). For example, in order to increase the immunological activity of the hot water extract, it can be fractionated and purified by sedimentation and ultrafiltration using a spirit that can be used as an extractant for food in the current food industry. In other words, every day, the hot water extract was concentrated in a vacuum to 1/20 volume and 4 times of alcohol was added thereto, and the mixture was allowed to stand at 4 ° C. for 24 hours, and then centrifuged to dissolve the precipitate in distilled water, followed by diafiltration for 24 hours. By using an extract having a molecular weight suitable for the purpose of the present invention. Ultrafiltration used in the present invention refers to a method of filtering colloidal particles or molecules that are difficult to be separated by ordinary filtration by applying pressure using dialysis using sulfate paper, bladder membrane, colloidion or the like.

In the composition of the present invention, the immune activity is preferably characterized by promoting complement system immune activity, intestinal immune activity, leukocyte increase in blood and weight increase of immune related organs.

In the present invention, some biochemical reactions occurring in vivo necessarily occur in stages, and these reactions are cascade reactions in which the second stage reaction is activated by activating the first stage reaction, which in turn activates the next stage reaction. ) Will happen. This continuous step reaction must be activated in sequence and not activated from the middle step without the previous step reaction. Biochemical reactions that occur step by step, combined with the function of antibodies, have a significant effect on host defense mechanisms, called the "complement system."

In the present invention, the gut-associated lymphocyte tissues composed of lymphocytes of the intestinal epithelium, lymphocytes of the lamina propria, Peyer's patch, and the like exist to form an effective bioprotective apparatus.

Leukocytes are the main constituents of the blood in the body and are composed of neutrophils, eosinophils, basophils, lymphocytes, monocytes, etc., and they act to protect bacteria that enter the body through phagocytosis. It is also involved in leukocytosis. In addition, it plays an important role as a primary effector cell by participating in an immune response that protects a living body from infection by the formation of an immune body. In addition, hepatic Kuffer cells, splenic macrophage of the spleen, and T-lymphocytes of the thymus are closely related to the immunity of the living body, and the increase in the weight of these organs suggests the increase of immune activity in the living body.

In the present invention, the intestinal immune activity is preferably a composition characterized by promoting IgA antibody production in the small intestine.

In the present invention, immunoglobulins (img) are a group of proteins that function as antibodies and play an important role in the humoral immunity of a living body. In particular, secreted IgA antibodies secreted from gut mucosal epithelial cells are produced in large quantities in the small intestine and play a central role in mucosal immune mechanisms through the inhibition, neutralization and removal of harmful substances.

According to another aspect of the present invention, the present invention provides a composition for inhibiting cancer cell growth, which contains hot water extract as an active ingredient.

In the composition of the present invention, the composition is characterized by inhibiting cancer cell growth due to increased immunity. In the embodiment of the present invention, it was shown that the administration of hot water extract inhibited the growth of solid cancer in vivo, but the effect of extending the lifespan when the extract was administered directly to the plural cancers was found to be insignificant. Supports the anticancer effect of increased immunity rather than direct cytotoxicity.

In the composition of the present invention, the cancer cells may be any cancer cells, such as solid cancer cells, hematologic cancer cells, which may inhibit the growth of cancer cells by increasing immunity, but are preferably solid cancer cells.

When the composition for increasing immune activity containing hot water extract of the perennial of the present invention is used as the pharmaceutical composition as described above, it may be prepared by a method known in the pharmaceutical field, a carrier per se or a pharmaceutically acceptable carrier, It may be mixed with excipients, diluents and the like to be prepared and used in the form of powders, granules, tablets, capsules, or injections. In addition, they may be administered orally or parenterally.

The dosage for administering the composition according to the present invention as an active ingredient may be appropriately selected according to the age, sex, weight, condition, and symptoms of the disease of the patient, preferably 1 to 1000 mg per day of adult can be administered. have.

Hereinafter, the present invention will be described in detail.

In the present invention, to examine the anti-cancer and immune activity of the hot water extract of the falsity, the proper extraction and purification conditions, sugar composition, cancer cell toxicity, sarcoma-180 growth inhibitory effect, leukocyte count change, complement system activity, intestinal immune activity, oral toxicity, etc. It was. Complementary active material of every life was extracted with 100 ℃ water for 3 hours, precipitated for 4 hours with 4 times alcohol, and then purified by dialysis and ultrafiltration to isolate and purify the active material with molecular weight of 300 ~ 500 kDa, and the sugar composition was xylose A) 19.01%, fucose 15.29%, mannose 4.23%, galactose 7.92%. In vitro, the toxicity of human active cancer cells to human cancer cells was mild at high concentrations (10 to 30 ㎍ / ml). Growth was inhibited by 40.13-59.42% compared to control. However, there was no statistically significant difference in life span extension among the experimental groups. As a result of the administration of the extracts to the abdominal cavity of the mice, purified leukocytes showed an increase in blood leukocyte count of 39%, 70% of 50 mg / kg group, and 83% of 100 mg / kg group compared to the control group. The weight of liver, spleen and thymus, which is related to immunity, was increased. As a result of oral administration of the extract to the mice and the amount of IgA antibody excreted in the feces, the control group was 2,092 ± 123.0 ㎍ / mg, but the control group was 2,454 ± 113.8 ~ 2,670 ± 133.1 ㎍ / mg. . As a result of measuring acute toxicity in mice, it was considered to be harmless to humans as it was not toxic until 2,000 mg / kg body weight was administered.

Hereinafter, the present invention will be described in more detail with reference to Examples. Since these examples are only for illustrating the present invention, the scope of the present invention is not to be construed as being limited by these examples.

Experimental Materials and Sample Preparation

1) Reagent

EDTA-gelatin veronal buffered saline (EDTA-GVB ⁺⁺ , pH 7.4), GVB ⁺⁺ (pH 7.4) and IgM hemolysin sensitized sheep erythrocytes (EA, 1 × 10 ⁹ cells) were purchased from Biotest Co., Japan, and NHS ( normal human serum) was prepared from blood collected with the voluntary consent of a healthy adult male (25 years old) who understood the purpose of the present invention. That is, blood obtained from venous blood of a normal person was left at 25 ° C. for 30 minutes, centrifuged at 2500 rpm for 10 minutes, and the supernatant was dispensed by 1 ml and used at -70 ° C. for storage.

2) Extraction of Sample

Capsosiphon fulvescens was collected in November in Wando, Jeonnam, transported to the laboratory at low temperature, washed with fresh water, dried in a hot air dryer (60 ℃), then crushed and 20 times distilled water in a powder that passed 0.84 mm standard. Proper extraction temperature and time were investigated by adding, and the extraction yield was expressed as a percentage of the weight of the sample used for extraction.

Example 1. Measurement of Immune Activity

Complement system activity was measured by Mayer's method [10] as a method for measuring the immune activity of hot water extracts. That is, the sample (50 ㎕) in serum (50 ㎕) and gelatin veronal buffer (GVB ⁺⁺⁾ was added to 50 ㎕ In 37 ℃ 30 minutes of reaction and then the amount of GVB ⁺⁺ sensitized red blood cells was added to 750 ㎕ 250 ㎕ After reacting at 37 ° C. for 1 hour, PBS (2.5 ml) was added and centrifuged to measure the absorbance (412 nm) of the supernatant. Complement activity was inhibited by total complement hemolysis (TCH50) of the control group. (inhibition total complement hemolysis, ITCH50). The method is a method of measuring the activity of the complement system by measuring the hemolysis degree by adding a red blood cell sensitized with hemolysin after reacting a certain amount of the active substance with complement, and has been widely used in the European Pharmacopoeia or the Korean biological standards and test methods. .

1) Extraction of Immune Active Substances

Considering that the complement system active substance, which is a kind of immune active substance, is easily extracted from hot water, and the complement system activating polysaccharide-related products currently distributed are health functional foods, a suitable method for extracting hot water from Korea Food Corporation, Optimal conditions were reviewed. In other words, after cleaning the debris with deionized water, desalting, drying with a hot air dryer, pulverizing, adding 20 times water, the yield and complement activity were observed according to extraction time and temperature. First, the extraction temperature is set at 100 ° C. and the change in complementary activity and yield according to extraction time is shown in FIG. 1. Yield yields of complementary activators of maesengi increased with time up to 4 hours of extraction, but decreased after 3 hours of extraction. Therefore, it was found that the proper extraction condition of the active material of the complement system was 3 hours, and the material extracted after 3 hours and showing the increase of yield had less effect on the activity of the complement system. And the extraction time is 3 hours, the change in the complementary system activity and yield for each extraction temperature is shown in FIG. Yield of maize was increased as the extraction temperature increased to 110 ℃, but the activity was the highest at 100 ℃ with 90.8%. Therefore, it is most suitable to extract the active material of the prosthetic complement system for 3 hours at 100 ℃.

2) Purification of Immune Active Substances

In order to increase the activity of immune active substances, sedimentation and alcoholic ultrafiltration were performed using alcohol (alcohol), which can be used as an extractant for food, in the current Food Code. In other words, every day vacuum concentrated hot water extract to 1/20 volume, then 4 times of alcohol was added to remove the weakly active low molecular substance, and then allowed to stand at 4 ℃ for 24 hours and then centrifuged to dissolve the precipitate in distilled water. The dialysis was performed by ultrafiltration and fractionated by molecular weight. Complementary system activity and yield by molecular weight are shown in Table 1. As a result of fractionation by distillation and ultrafiltration of the complement system active substance, the activity of the molecular weight 300 ~ 500 kDa fraction was 97.6%, which was 6.8% higher than 90.8% before purification and the yield was 23.1%. Therefore, it is recommended to take fractions of 300-500 kDa molecular weight using ultrafiltration membranes after alcohol precipitation and dialysis. Molecular weights of plant-derived system-activated polysaccharides reported so far are neutral polysaccharide BR-5-I (18 kDa) in Shiho, acid polysaccharide CAP-IIIb (120 kDa) in red pepper, and PA-IIa, which is an acid complex polysaccharide isolated from ferns. -1 (1 kDa), PA-IIa-2 (450 kDa), HPA-2-IVa (500 kDa), and Yamada et al. Showed that the higher the molecular weight, the higher the activity [12-16].

TABLE 1 Complement Activity and Yield by Molecular Weight of Hot Water Extracts

Example 2 Analysis of Sugar Composition of Immune Active Substances

In order to determine the sugar composition of the complementary active substance extracted from maeseng, 20 times of distilled water was added to (SP-0), heated at 100 ° C. for 3 hours, and centrifuged to obtain supernatant (SP-1). The extract was concentrated in vacuo to a volume of 1/20, and then 4 times the amount of alcohol was added thereto, followed by standing at 4 ° C. for 24 hours, followed by centrifugation to dissolve the precipitate in distilled water, followed by dialysis for 24 hours to vacuum freeze drying (SP-2). Each was provided as a sample for glucose analysis. In the sugar composition analysis, 0.2 mg of the lyophilized sample was pretreated according to Chaplin's method [11] and analyzed by gas chromatography (GC) (Table 2). Quantification was performed by TMS of internal standard (myo- inositol) and standard reagents (D-xylose, D-fucose, D-mannose, D-glucose, D-galactose) in the same way as the sample to calculate the relative area ratio of each concentration. .

TABLE 2 Analysis of Gas Chromatography for Sugar Composition

Table 3 shows the results of analyzing the sugar composition of raw materials (SP-0), hot water extract polysaccharide (SP-1) and alcohol purified polysaccharide (SP-2) using GC. In other words, the sugar composition analyzed by GC was 6.32% xylose, 4.77% fucose, 2.14% mannose, 5.27% glucose, and 0.77% galactose. The hydrothermal extract polysaccharides were xylose 19.97%, fucose 16.34%, mannose 3.67%, glucose 3.86%, and alcohol refined polysaccharides were xylose 19.01%, fucose ( fucose) 15.29%, mannose 4.23%, galactose 7.92%. Complementary system-activated polysaccharides reported so far are mostly acidic complex polysaccharides, varying with no apparent commonality in sugar composition, and are known to have no direct effect on activity [12-14, 17,18].

TABLE 3 Sugar Composition (%) of Perennial Raw Material (SP-0), Hot Water Extracted Polysaccharide (SP-1) and Alcohol Purified Polysaccharide (SP-2) Using GC

Example 3. Intestinal Immunity Activity Measurement

32 mice (Balb / c) were divided into the control group (PBS administration) and the falconus extract (SP-2) administration group (20, 50, 100 mg / kg body weight) and administered to the oral cavity for 7 days, respectively, followed by the production of IgA antibodies in the small intestine. The amount of IgA antibody in feces was measured to determine the effect on the response. Take 2 ~ 4 eggs of fresh fecal matter and measure the weight, add protease inhibitor solution to 0.1 g / ml, and leave at 4 ℃ for 1 hour, then stir strongly by vortex mixer to make suspension and centrifuge. The solution was taken and the absorbance was measured at 450 nm to quantify the IgA antibody. At this time, mouse IgA was used as a standard material.

Intestinal immune activity was measured to examine the feasibility of functional food use of extracts from P. falciparum. In other words, the amount of IgA antibody in feces was measured in order to determine the effect on the IgA antibody production response of the small intestine after administering the extract (SP-2) to the mouse. Compared to ± 123.0 μg / ml, the perennial extract administration group was 2,454 ± 113.8 ~ 2,670 ± 133.1 μg / ml, indicating that the perennial extract promotes IgA antibody production in the small intestine of mice (FIG. 3). The digestive tract is an important immune organ that not only digests and absorbs nutrients, but also excludes harmful substances such as viruses and pathogens. The gut contains lymphoid tissues composed of lymphocytes of the intestinal epithelium, lymphocytes of the lamina propria, and Peyer's patch to form an effective biological defense mechanism. Immunoglobulin (Ig) is a group of proteins that functions as antibodies and plays an important role in the humoral immunity of living organisms. In particular, secreted IgA antibodies secreted from gut mucosal epithelial cells are produced in large quantities in the small intestine and play a central role in mucosal immunity through inhibition, neutralization and removal of harmful substances [19].

Example 4 Determination of Leukocyte Count and Immune-Related Organ Weights

Purified Maeseng Yiyi extract (SP-2) was administered to the abdominal cavity of the experimental animal (Female ICR mouse) continuously for 10 days at different concentrations (0, 20, 50, 100 mg per kg of body weight) for 1 day from the last day of sample administration. On the 2nd, 4th, and 7th day, the blood was collected and the leukocyte count was measured by a hemacytometer. On the 8th day, the animals were lethal by the cervical dislocation method and weighed. Were extracted and each weight was expressed as a percentage of the weight.

1) Effect on blood white blood cell count

The leukocytes of the control group administered with the same amount of deionized water were measured on day 1, day 2, day 4, and day 7 after the extract was administered to the abdominal cavity continuously for 10 days. Compared to the number 7,654 ± 728 cells / mm ³ , the periphery showed an increase of 39% of the extract, 70% of the 50 mg / kg group, and 83% of the 100 mg / kg group. In the case of stopping the administration of the extract, the leukocyte count decreased, and in the 100 mg / kg administration group, the leukocyte count was 13,972 ± 475 cells / mm ^{3 at} day 1 and then decreased to 12,860 ± 619 cells / mm ³ after 7 days. (Table 4).

TABLE 4 Effect of Perennial Hot Water Extract on the Leukocyte Count in ICR Mice

Leukocytes are the main constituents of the blood in the body and are composed of neutrophils, eosinophils, basophils, lymphocytes, monocytes, etc., and they act as a defense against bacteria entering the body by phagocytosis. It is also involved in leukocytosis. In addition, it plays an important role as a primary effector cell by participating in an immune response that protects a living body from infection by the formation of an immune body [20]. Therefore, the increase in the leukocyte count in the blood due to the administration of the extracts of Maeseng may be related to the increase of immune function in vivo.

2) change in weight of immune-related organs

After extracting the maeseng this extract into the abdominal cavity of the mouse for 10 days, the weight of the organs related to immunity such as liver, spleen, thymus, etc. was measured and shown in Table 5 as the weight to weight ratio. All organs such as liver, spleen, and thymus showed an increase in weight by the administration of the extract of Maeseng, and statistical significance was found at the dose of 50 mg / kg or more. Kuffer cells in the liver, splenic macrophage in the spleen, and T-lymphocytes in the thymus are closely related to the immunity of the living body, and the increase in the weight of these organs suggests the increase of immune activity in the living body.

TABLE 5 Effect of Hot Water Extract on Immune-Related Organ Weights of ICR mice

Example 5 Toxicity Testing of Cancer Cells In Vitro

Cytotoxicity of Maeseng Yiyi extract (SP-2) was measured by MTT assay (Methylthiazoletetrazolium assay) on five human cancer cells (lung cancer, ovarian cancer, pigment cell cancer, central nervous system cancer, colon cancer).

Anti-cancer activity by complement system-activated polysaccharides has been reported to be mostly due to enhanced function of the host immune system, rather than direct cytotoxicity. However, cytotoxicity of some seaweed components has been reported. It was confirmed in vitro (Table 6). However, the cytotoxicity of the extracts of Maesai erectus against human cancer cells was insignificant at high concentrations (10-30 ㎍ / ml). Although most cytotoxic substances are known to be toxic by penetrating or entrapping into cells as low-molecular substances, complementary active polysaccharides are inferior in direct cytotoxicity because they are high molecular materials of 300 kDa or more.

TABLE 6 Cytotoxicity of Human Cancer Cells from Hot Water Extracts of Progeny (Net growth as% of control)

Example 6 Inhibition of Solid Cancer Growth in Vivo

After transplanting the tumor cells (Sarcoma-180) to the left inguinal part of the mouse, the medicinal herb extract (SP-2) was administered to the abdominal cavity continuously for 24 days after 24 hours, and the weight of the solid cancer was measured on the 32th day. Tumor growth inhibition rate was measured accordingly.

As a result of administering the extract of Maeseng to the abdominal cavity of mice, it was confirmed that the number of leukocytes in blood and the weight of immune-related organs were increased. This increase in immunity has been reported to inhibit the growth of cancer cells in the body. To examine this, after transplanting sarcoma-180 to the left inguinal part of the mouse, the extracts were administered to the abdominal cavity and the tumor growth inhibition rate was measured. 7 is shown. The growth of mouse solid cancer was inhibited by 40.13 ~ 59.42% compared to the control group by the administration of the extract of the maeseng, the growth inhibition rate tended to increase slightly in proportion to the dosage of the extracts (Fig. 4). Cho et al. Reported that tumor inhibition rate of glycoproteins extracted from hearing was not significantly different between 49.16% at 50 mg / kg concentration and 53.30% at 100 mg / kg concentration [21].

TABLE 7 Anticancer Activity of Perennial Hot Water Extracts in ICR Mice with Sarcoma-180 Cells

Example 7. Life Extension Experiment

After implanting the tumor cells (Sarcoma-180) into the abdominal cavity of the mouse, the sample (SP-2) was administered to the abdominal cavity for 24 days to 10 days and observed for survival up to 35 days, and the average lifespan was calculated from the following equation. The life extension rate was obtained.

Experimental results of life extension effects of the extract of Maeseng on sarcoma-180 cell ascites cancer in mice are shown in Table 8. In this experiment, the survival time of the mice ranged from 16 to 22 days, and life expectancy of the control group was 5.51 ~ 9.48%, but there was no statistical difference between the groups at p <0.05. It has been shown that the administration of the extract of Maesengyi inhibits the growth of solid cancer in vivo, but the effect of extending the lifespan is insignificant when the extract of Maeseng is directly administered to the ascites cancer. It is assumed to be due to the increase.

TABLE 8 Effects of Progeny Hydrothermal Extract on Life Extension of ICR Mice

Example 8. Acute Toxicity Test

Oral administration of Perennial Tooth Extract (SP-2) at 100, 500, 1,000 and 2,000 mg / kg concentrations in mice and death for 14 days, weight change and general condition (hair, convulsions, diarrhea, tears, walking, etc.) Was observed.

Maesengi is a seaweed that has been traditionally used in the south coast of Korea. It is not considered to be a problem for human safety due to toxicity. However, it is likely that certain components are concentrated in the process of extracting complementary active substances from Maesui. Therefore, oral toxicity was measured for the extracts of the falciparum in mice (Table 9). As a result, all test groups died for 14 days and no abnormal condition was found, and the weight gain was 0.48 ~ 0.49 g / day, which is similar to the control 0.48 g / day administered with the same amount of distilled water. There is no risk to humans due to toxicity.

TABLE 9 Acute Toxicity of Perennial Hot Water Extracts in ICR Mice

Statistical analysis of animal test results

The experimental results were expressed as mean ± standard error, and the significance between each group was tested by Duncan's multiple range test at p <0.05 level using SAS Enterprise Guide (Version 9.13).

As described above, according to the present invention, every single hot water extract promotes the production of IgA antibodies to increase intestinal immune activity, increases leukocyte count, and increases the weight of immune-related organs. In addition, every single hot water extract is not harmful to humans due to toxicity, and has the effect of inhibiting the growth of cancer cells may be used in diseases in which the therapeutic and preventive effects are expected due to an increase in immunity such as cancer.

[references]

Claims (8)

Complementary immunosuppressive activity comprising the step of extracting hot water at 90-110 ° C. for 2-4 hours, the step of precipitating the hot water extract to alcohol and the step of ultrafiltration to purify the precipitate to 300-500 kDa Method of producing a extract of the falsified herb. A composition for increasing immune activity, which comprises an extract of Mae jae as an active ingredient, having a molecular weight of 300-500 kDa prepared by the method of claim 1. delete delete delete delete delete delete

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