KR100793262B1 - HEALTH-SUPPLEMENTARY FOOD CONTAINING EXTRACT FROM Crotalaria sessiflora L., TO IMPROVE ANTICANCER ACTIVITY OR IMMUNITY FUNCTION - Google Patents

Abstract

The present invention relates to a bowel extract, a functional food or a composition for preventing and treating colon cancer containing the same.

In the present invention, the extract of the wild sprouts is separated into stems, leaves, roots and seeds, and then mixed with a halogenated hydrocarbon-based organic solvent and a C ₁ to C ₄ alcohol and left at room temperature for 3 to 5 days. It is prepared by separating and purifying by recrystallization method after repeated extraction, and the extract of the wild sprouts as an anti-cancer or immune function enhancing activity as well as antioxidant activity, containing as an active ingredient or a functional food or a composition for preventing and treating colorectal cancer It can be useful.

Sprouts Extract, Anticancer, Immune Function, Functional Food, Colon Cancer

Description

HEALTH-SUPPLEMENTARY FOOD CONTAINING EXTRACT FROM Crotalaria sessiflora L., TO IMPROVE ANTICANCER ACTIVITY OR IMMUNITY FUNCTION}

Figure 1 shows the total phenolic content of the extract for each part of the bowel of the present invention,

Figure 2 shows the results of the thin layer chromatography of the extract of Shrimp sprouts by the recrystallization method of the present invention,

Figure 3 is a result for the SOD-like activity of the extract of the wild sprouts isolated by the recrystallization method of the present invention,

4 is a result of the hydrogen peroxide scavenging ability of the extract of the wild sprouts isolated by the recrystallization method of the present invention,

Figure 5 shows the number of expression of ACF, a colorectal cancer pre-cancerous lesion after administration of colorectal carcinogens, to test the efficacy of colorectal cancer of the present invention.

The present invention relates to an active ingredient extract, a functional food containing the same or a composition for preventing and treating colorectal cancer, and more specifically, the active ingredient extract of the present invention, which is purified by recrystallization by extracting the active ingredient, By having anti-cancer or immune function enhancing activity, it relates to a functional food or a composition for preventing and treating colorectal cancer, which contains it as an active ingredient.

Sprout ( Crotalaria sessiflora L. ) is an annual herb, 20-70cm high, with long brown hairs except for the surface, and grows in wild weed rice and widely distributed throughout the country. However, wild sprouts have been known to be very poor in seed germination rate and to suppress the germination of other crops, which has made it difficult to resource them.

Recently, the low temperature treatment method for the dormant breakdown of wild sprout seeds, gibberellin (GA3), which is known to have a low temperature replacement effect, and the softening treatment of KNO ₃ and seedlings that control the moisture state of seeds, break through dormancy. A method for increasing germination has been developed, but a mass production method has not yet been established.

These wild herbs are legumes, called Guryongcho, wild lily, flame sensation, and ramyun lily, and contain alkaloids to treat various diseases from tumors, esophageal cancer, rectal cancer, lung cancer, stomach cancer and chronic bronchitis. Or as an antidote has been widely used in folk medicine.

However, only the effects on the bowels have been transmitted to the word of mouth, the active ingredient showing its efficacy, the study of the mechanism of action of the components have not been established yet.

Recently, it has been known to have anti-cancer effects due to the monocrotaline component contained in the wild sprouts, but it is only known to be toxic along with the anti-cancer effect of the monocrotaline component, and the research is only at an early stage.

Thus, the present inventors have tried to search for the active ingredient contained in the wild sprouts, by separating and purifying the extract after the extraction by the recrystallization method, to obtain an active extract of the wild herbs with excellent anti-cancer or immune function enhancement activity in addition to the antioxidant activity, The present invention has been completed by presenting a field of application containing the sprouts extract.

It is an object of the present invention to provide an extract of Sprout ( Crotalaria sessiflora L. ) having anticancer or immune function enhancing activity.

Another object of the present invention to provide a functional food containing the extract of the sprouts.

Still another object of the present invention is to provide a composition for preventing and treating colorectal cancer, which contains the extract of the wild sprouts.

In order to achieve the above object, the present invention separates the lobster ( Crotalaria sessiflora L. ) into stems, leaves, roots, and seeds, and then adds a halogenated hydrocarbon-based organic solvent and a mixed solvent of C ₁ to C ₄ alcohols at room temperature. It is left to stand for 5 days, and extracted two to three times, and then purified and separated by the recrystallization method to provide an extract.

It is preferable to use the mixed solvent which consists of a mixing ratio of 2: 1-9: 1 with respect to methylene chloride: ethanol.

The recrystallization method is carried out under any one solvent condition selected from the group consisting of methylene chloride / hexane, chloroform / hexane, methylene chloride / methanol and chloroform / methanol.

More preferably, the bowel extract having the above activity is to use a leaf extract or a stem extract.

The present invention provides a functional food containing the extract.

It is preferable to contain 30-100 mg / kg of sprouts from above.

The present invention also provides a composition for preventing and treating colorectal cancer, containing the extract of the bowel. At this time, it is preferable that the extract contains 30 to 100 mg / kg.

Hereinafter, the present invention will be described in detail.

The present invention is separated from the stem ( Crotalaria sessiflora L. ) into stems, leaves, roots, seeds and then mixed with a halogenated hydrocarbon-based organic solvent and C ₁ ~ C ₄ alcohol and left to stand at room temperature for 3 to 5 days, 2 After repeated extraction 3 times to separate and purified by the recrystallization method, to provide the prepared extracts.

Examples of the halogenated hydrocarbon-based organic solvents include methylene chloride, chloroform, trichlorethylene, trichloroethane, trichlortrifluoroethane and the like.

The alcohol used is preferably C ₁ to C ₄ alcohols, more preferably methanol or ethanol.

More preferably the mixed solvent of the present invention is carried out under any one solvent conditions selected from the group consisting of methylene chloride / hexane, chloroform / hexane, methylene chloride / methanol and chloroform / methanol, more preferably methylene chloride: ethanol Use a mixed solvent of. At this time, the preferred mixing ratio is 2: 1 to 9: 1, more preferably 3: 1.

After the extraction, the obtained extract can be separated and purified using methods commonly used by those skilled in the art, such as adsorbent (XAD-2) and column chromatography (silica and Diaion HP-). 20) A method can be used.

In particular, the present invention is characterized in that after the extraction, the obtained extract is separated and purified by recrystallization. More specifically, the extract of the present invention can be crystallized under soluble solvent conditions and poorly soluble solvent conditions to purify the extract. Preferred examples thereof include any one of solvent conditions selected from the group consisting of methylene chloride / hexane, chloroform / hexane, methylene chloride / methanol and chloroform / methanol, and more preferably methylene chloride is used as a soluble solvent, It is carried out using hexane as the solvent condition.

At this time, the anti-cancer or immune function enhancing activity of the present invention is contained in the leaf extract or stem extract of the extract of the wild sprouts, in particular, it is contained in a large amount of leaf extract ( FIG. 1 ).

The extract of the present invention ( crotalaria sessiflora L. ) has been known for its anticancer effect due to the monocrotaline compound represented by the following formula (1).

Thus, according to the present invention, as a result of separating and purifying an extract of a wild sprout by setting the monochromatin as an indicator, the extract of the present invention contains an active ingredient of another species in addition to the monocrotaline and the monochromatin. It was confirmed ( FIG. 2 ).

The extract of the present invention is higher in total phenolic content and electron donating ability than the monochromatin, which is the indicator, and has a higher SOD-like activity to remove the active oxygen ( FIG. 3 ), hydroxy radical scavenging ability and hydrogen peroxide scavenging. By showing high activity, it shows superior antioxidant activity than monocrotalin.

In addition, the present invention is the result of the forced swimming load test conducted to verify the immune boosting effect by the physical strength or endurance of the experimental animal using the extract of the wild sprouts, immunity by significantly reducing the dead time in proportion to the administration concentration of the extract Functional enhancement effect can be confirmed ( Table 3 ). In addition, as a result of the carbon removal ability test using the extract of the wild sprouts, the phagocytic activity was increased by dose-dependently confirmed the immune function enhancing activity ( Table 5 ).

Accordingly, the present invention provides a functional food containing the extract of Crottaaria sessiflora L ..

Since the extract is excellent in antioxidant activity, it can provide a functional food containing it, and by containing the extract, it can be used to preserve foods or to maintain the freshness and quality of food for a long time.

Examples of the functional food containing the active ingredient extract of the present invention include various foods, fish, meat and processed foods thereof (e.g. ham, sausage cornbeans), beverages (including alcoholic beverages), breads and noodles (E.g. udon, soba noodles, ramen noodles, spagate, macaroni, etc.), fruit juices, various drinks, chocolate, snacks, confectionery, pizza, ramen, other noodles, gums, ice cream, frozen foods and various seasonings (e.g. miso) , Soy sauce, source, etc.), vitamin complexes and other health supplements may be applied, but is not limited thereto.

At this time, in order to optimize the anticancer or immune function enhancing activity, the extract of the wild sprouts is to contain 30 to 100 mg / kg, more preferably 30 to 60 mg / kg. At this time, if the extract is less than 30 mg / kg, there is a problem in the induction of the effect, when it exceeds 100 mg / kg, especially when it exceeds 120 mg / kg, the concentration-dependent tendency to anti-cancer and the immune function It is out of the preferred dose range because it shows a decrease in immobility time and a decrease in phagocytosis which is an indicator of enhanced activity.

In addition to the antioxidant activity of the present invention, the extract shows an inhibitory effect on the proliferation of liver cancer cells, breast cancer cells and colon cancer cells, and more preferably has inhibitory activity on the generation of Aberrant Crypt Foci, a colon cancer precancerous lesion ( 4 and Table 1 ).

Thus, the present invention provides a composition for preventing and treating colorectal cancer, which contains the extract of the wild herbs as an active ingredient.

The composition containing the extract of the present invention as an active ingredient can be administered in an effective amount through various routes of administration. The use together contains a pharmaceutically acceptable carrier. Examples of suitable carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, ziitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose And microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil. The pharmaceutical composition may further include fillers, anti-coagulants, lubricants, wetting agents, fragrances, emulsifiers, preservatives and the like. The pharmaceutical compositions of the invention may be formulated using methods well known in the art to provide rapid, sustained or delayed release of the active ingredient after administration to a mammal. The formulations may be in the form of tablets, pills, powders, sachets, elixirs, suspensions, emulsions, solutions, syrups, aerosols, soft or hard gelatin capsules, sterile injectable solutions, sterile powders and the like.

The pharmaceutical compositions of the invention can be administered via several routes including oral, transdermal, subcutaneous, intravenous or intramuscular.

Typical daily dosages of the extracts of the present invention are in the range of 1 to 1,000 mg / kg body weight, preferably 10 to 100 mg / kg body weight, and may be administered once or in several doses. However, it is to be understood that the actual dosage of the active ingredient should be determined in light of several relevant factors such as the route of administration, the age, sex and weight of the patient, and the severity of the disease, and therefore the dosage should in any way be regarded as the present invention. It does not limit the scope of.

Although the present invention has been described in detail only with respect to the described embodiments, it will be apparent to those skilled in the art that various changes and modifications can be made within the technical scope of the present invention, and such modifications and modifications belong to the appended claims.

< Example  1> Bow  Preparation of extract for each part

After separating the sprouts into stems, leaves, roots, and seeds, methylene chloride (CH ₂ Cl ₂ ): ethanol (EtOH) was added in a mixing ratio of 3: 1 and left at room temperature (25 ° C.) for about 3 days. After repeated extractions were filtered and concentrated under reduced pressure. Extracts were separated by XAD-2 and column chromatography (silica and Diaion HP-20) to obtain the extracts of each part of the sprouts.

Yield of each extract obtained above was stem (4.47%)> leaf (3.95%)> seed (2.90%)> root (2.67%).

< Experimental Example  1> Bow  In vitro of the extract for each site ( in vitro Of antioxidant effect in

1. Quantitative Phenolic Compounds

Na ₂ CO ₃ was added and mixed with each fraction, and the mixture was left at room temperature and absorbance was measured at 750 nm.

Figure 1 shows the total phenolic content of the extract of each part of the bowel of the present invention, the total phenolic content was the highest in the leaves, the lowest content in the stem. On the other hand, phenol content was higher than tocopherol and crothalin, except for sesamol, among conventional antioxidants.

2. Determination of Superoxide dismutase (SOD) -like activity

After adding the buffer solution and pyrogallol to the extract for each part of the bowels for 10 minutes, the absorbance was measured at 420 nm.

As a result of the measurement, the leaf extract activity was the highest, and the seed portion was the lowest.

< Example  2> Bow  Separation of Functional Substances in Extracts

Methyl chloride (CH ₂ Cl ₂ ): ethanol (EtOH) is added to the leaves of the active shoots with excellent yield and activity in a mixing ratio of 3: 1, and left at room temperature (25 ° C.) for about 3 days before extraction. The herb leaf extract was prepared, and the leaf of the active ingredient was dissolved in methylene chloride, which is the most soluble solvent, and left for one day. The precipitated material was collected, rinsed twice with the least insoluble hexane, and then dissolved again with water to remove sugar. It produced by the recrystallization method to obtain the sample.

As a result of TLC analysis of the sample, monocrotaline, which is an anticancer substance contained in conventional bowel sprouts, was used as an indicator, and the bowel extract of the present invention contained an active substance of another species in addition to monocrotaline. It was confirmed (FIG. 2).

Experimental Example 2 In Vitro of Leaf Extract of Wild Sprouts in vitro Of antioxidant effect in

1. Quantitative Phenolic Compounds

Na ₂ CO ₃ was added to the extracts of the actives, mixed, left to stand at room temperature, and absorbance was measured at 750 nm.

As a result, in the case of the extract of the bowel sprouts obtained by the recrystallization method, the total phenol content is higher than that of the monochromalin, which is an indicator of the present invention, and thus anticancer activity stronger than that of the monochromatin can be predicted.

2. Electronic donation ability

DPPH reagent was added to the extracts of the actives, mixed, and reacted at room temperature for 30 minutes, and then absorbance was measured at 517 nm.

As a result of the electron donating ability of the extract of the bowel sprouts obtained by the recrystallization method, it was also higher than the monochromalin which is an indicator of the present invention.

3. Determination of Superoxide dismutase (SOD) -like activity

After adding a buffer solution and pyrogallol to the bowel extract for 10 minutes, absorbance was measured at 420 nm.

As shown in Figure 3 , the extract of the bowel sprouts obtained by the recrystallization method shows a higher SOD-like activity than the monochromatin which is an indicator of the present invention, it is possible to predict the improved anticancer activity than monochromatin.

4. Determination of hydroxy radical scavenging ability

After the reaction for 2 hours by mixing FeSO ₄ / EDTA solution, 2-deoxyribose, phosphorylation buffer solution, hydrogen peroxide to the bowel extract, trichloroacetic acid solution and thiobarbituric acid solution was added, 15 minutes After heating, it was rapidly cooled to measure absorbance at 532 nm.

As a result of measuring the hydroxy radical scavenging ability of the extract of the wild lobster, it was observed that the hydroxy radical scavenging activity was higher than that of the monochromalin which is an indicator of the present invention.

5. Determination of hydrogen peroxide scavenging activity

Oxidation lipid production was measured by the 2-thiobarbituric acid reactive-substance (TBARS) method using a certain amount of egg yolk lecithin as a substrate.

As a result, the extract of the bowel sprouts obtained by the recrystallization method showed higher hydrogen peroxide scavenging activity than the monochromalin, which is an indicator of the present invention, and thus anticancer activity can be predicted to be higher than that of monocrotalin.

Experimental Example 3 Colorectal Cancer Efficacy Evaluation Experiment in vivo Experiment)

56 four-week-old SD male rats (Orient Co., Ltd.) were used to measure colorectal cancer precancerous lesions (ACF), and the rats were housed in three polycarbonate cages. During the experiment, the breeding environment was maintained at a temperature of 22 ± 2 ℃, a relative humidity of 55 ± 5%, artificial illumination (12 hours on, 12 hours off) and roughness 150 ~ 300 lux. Purified water filtered using a filter was freely fed.

As a colorectal carcinogen, 1,2-dimethylhydrazine (hereinafter referred to as 'DMH', manufactured by Sigma Aldrich Co., Ltd.) was dissolved in sterile physiological saline at a dose of 20 mg / kg per 1 kg of body weight, and once per week, intraperitoneally. Three weeks in total, three doses were administered.

The test substance administration group was dissolved in cottonseed oil at the dosage of 30, 60, and 120 mg / kg of Sprouts extract, orally administered once daily for 8 weeks from 1 week after the last administration of 1,2-dimethylhydrazine, a colorectal carcinogen. It was. The positive control group 1 was intraperitoneally administered bleomycin at a dose of 250 ㎍ / kg once a week from 3 weeks after the administration of 1,2-dimethylhydrazine to the end of the test. Tallinn (monocrotaline, hereinafter referred to as 'MCT') was administered once daily at the dose of 0.5 mg / kg from 3 weeks after 1,2-dimethylhydrazine administration to the end of the test.

Aberrant Crypt Foci (hereinafter referred to as 'ACF') and Aberrant Crypt (hereinafter referred to as 'AC') of the colon were measured as follows.

ACF and AC of the large intestinal mucosa of rats were observed after staining with methylene blue according to the method of Bird et al. Ten weeks after the start of the experiment, the animals were sacrificed and the colon tissues were mixed with physiological saline and 10% neutral buffered formalin in a 1: 1 ratio. Unfolded and fixed in 10% neutral buffered formalin. The immobilized tissue was divided into three portions and stained with 0.2% methylene blue solution for 1 minute. Stained tissues were observed by light microscopy (40x and 100x) to determine the number of ACF and AC in the colon. The results are shown in Table 1 below.

Results of the measurement of the number of ACF and AC, precancerous lesions of colorectal cancer Classification Dose (mg / kg) Number of total ACF ∑≤ 3AC ∑≥ 4AC Total ACF / colon DMH administration group 20 94.3 ± 14.31 32.7 ± 4.18 143.3 ± 12.57 Live Sprout Extract Administration Group 30 54.3 ± 7.97 16.3 ± 6.12 70.7 ± 12.88 60 33.0 ± 9.85 11.3 ± 2.73 47.7 ± 15.38 120 72.3 ± 21.07 19.3 ± 9.82 91.7 ± 30.34 Positive Control 1 0.25 29.7 ± 6.17 6.3 ± 1.86 36.0 ± 8.02 Positive Control 2 0.5 25.8 ± 9.20 6.8 ± 1.80 32.5 ± 7.42

No ACF was found in SD rats of the normal control group, and ACF, a colorectal cancer precancerous lesion, was induced in the DMH alone group ( FIG. 5 ).

On the other hand, the administration of the 30%, 60, 120 mg / kg of the extract of the sprouts showed a decrease in the number of ACF, and most preferably, the effect of reducing the ACF number in the 60 mg / kg administration group. In addition, the reducing effect of the extract of the sprouts showed a similar result to the case of the positive control group 1 (Bleomycin administration group) and the positive control group 2 (MCT administration group, which is an indicator of the active sprout extract).

Experimental Example 4 Stability Test of Wild Sprout Extract

To characterize the stability test of the extract, the extract was administered to rats for 8 weeks to observe the effect on the internal organs.

The internal organs were examined for damage to the liver, spleen, kidney, lung, heart and thymus.

Results of Stability Test of Green Sprout Extract division Dose (mg / kg) Organ weight (g) liver spleen kidney lungs Thymus Heart Control 0 10.75 ± 1.41 0.59 ± 0.12 1.33 ± 0.13 1.40 ± 0.08 0.44 ± 0.01 1.58 ± 0.09 DMH administration group 20 12.26 ± 2.62 0.83 ± 0.23 1.40 ± 0.18 1.57 ± 0.20 0.45 ± 0.08 1.67 ± 0.46 Live Sprout Extract Administration Group 30 11.41 ± 0.58 0.67 ± 0.06 1.37 ± 0.14 1.38 ± 0.06 0.61 ± 0.17 1.54 ± 0.15 60 12.89 ± 1.51 0.73 ± 0.14 1.47 ± 0.08 1.58 ± 0.09 0.61 ± 0.14 1.65 ± 0.15 120 12.03 ± 1.07 0.68 ± 0.16 1.39 ± 0.15 1.53 ± 0.18 0.55 ± 0.15 1.50 ± 0.15 Positive Control 1 0.25 11.90 ± 0.07 0.62 ± 0.02 1.33 ± 0.12 1.44 ± 0.05 0.45 ± 0.05 1.72 ± 0.16 Positive control 2 0.5 12.74 ± 1.40 0.87 ± 0.04 * 1.42 ± 0.28 1.45 ± 0.18 0.39 ± 0.06 1.80 ± 0.30

As shown in Table 2 , no specific features or lesions were found in the gross findings at the time of necropsy, and significant changes were obtained when the weight of major organs was measured to compare and analyze the absolute organ weights and relative organ weights with respect to body weight. Was not induced.

Therefore, it was confirmed that the long-term administration of the sprout extract does not have a significant effect.

Experimental Example 5 Observation on Flourishing Sprouts of Immune Function

Forced swimming test was performed as follows to verify the immune-promoting effect by enhancing the stamina or endurance of the test animal using the extract of the wild sprouts.

Five male ICR-based mice were set as a control group and a butterfly extract administration group as one group. The control group was orally administered 0.5% CMC used as an excipient. The administration group of the extracts of the wild herbs were suspended in 0.5% CMC at doses of 30 mg / kg, 60 mg / kg and 120 mg / kg, respectively, and orally administered to ICR mice at a dose of 10 ml / kg for 2 weeks.

On the day of the swimming load test, the mice were forced into a 300 x 420 x 500 mm polycarbonate cage filled with water at a temperature of about 23 to 25 ° C and observed for 10 minutes.

It was determined that the mice were immotile when their heads stood out above the water, stationary and floating upwards. A dead time of at least 2 minutes of 10 minutes of observation was recorded. When the immobility time is reduced compared to the control group, it can be determined that there is an immune enhancing effect. The results are shown in Table 3 below.

Dead time (sec) measurement result division Dead time (sec) Control 365.2 ± 24.49 Active Sprout Extract group, 30 mg / kg 267.0 ± 74.76 Live Sprout Extract administration group, 60 mg / kg 179.8 ± 67.12 Livestock extract administration group, 120 mg / kg 176.2 ± 42.96

As shown in Table 3 above, the experiment was conducted in a forced swimming test by placing the mouse in a water tank, and the immobilization time was remarkably reduced in proportion to the administration concentrations of 30 mg / kg and 60 mg / kg of the sprout extract, thereby improving immune function. It was confirmed. However, in the administration group of 120 mg / kg administered with the high concentration of the sprout extract, the tendency of decreasing the dead time is small, suggesting the dosage range of the extract of the sprout extract against the increase of immunity.

Experimental Example 6 Observation of Immune Function Enhancement of Sprouts Extract-Carbon Removal Ability Test

1. Optical Density (OD) Measurement

Five male 5 week-old ICR mice were administered orally for 2 weeks in one group, which was set as a control, 30 mg / kg, 60 mg / kg, and 120 mg / kg administration group.

Injectable ink was diluted three times with India sterile saline solution for injection, and Na ₂ CO ₃ solution was prepared by adding 100 ml of distilled water in 0.1g. One Indian ink prepared by diluting the mouse vein was injected by calculating the amount of 0.1 ml per 10 g of body weight.

Immediately after completion of the Indian ink injection, the time was measured. At 2 and 10 minutes after injecting the ink, 40 μl of blood was taken from the intravenous gland of the eye, respectively, and placed in 4 ml of Na ₂ CO ₃ solution. The optical density was measured at 600 nm wavelength of the spectrophotometer, and the Na ₂ CO ₃ solution was It was blank. After sacrifice, the liver and spleen were removed, and blood on the surface of the organ was removed by filter paper, and the organ weight was measured. Phagocytosis index indicates the rat's carbon-removing ability. When the experimental group's eating index is significantly higher than that of the control group, the test result was positive.

At this time, the absorbance measurement results, the results of measuring the optical density (OD) is shown in Table 4 below.

Optical density measurement results Time (min) Dose (mg / kg) Control 30 60 120 2 0.0932 ± 0.00183 0.0864 ± 0.00384 0.0955 ± 0.00542 0.0862 ± 0.00552 10 0.0970 ± 0.00527 0.0854 ± 0.00548 0.0992 ± 0.00486 0.0902 ± 0.00503

As shown in Table 4, no difference was observed in the OD measurement results of the control group and the Sprout extract administration group by the phagocytic activity of 2 and 10 minutes after Indian ink administration.

2. Statistical Analysis of Compensated Leukocyte Index

Using the OD values of the control group and each test substance administration group, the white blood cell index (Phagocytic index) K was calculated by using Equations 1 and 2, and the corrected phagocytic index was corrected in consideration of the weight of the animal and the weight of the spleen and liver. ) "a" was calculated. The significance of the control group compared to the value of the Sprout extract administration group is shown in Table 5 below.

Corrected white blood cell index a results division cock Control 2.8513 ± 0.1653  30 mg / kg 3.1472 ± 0.4329  60 mg / kg 3.7680 ± 0.3095 120 mg / kg 3.0658 ± 0.3602

As shown in Table 5, in the case of males, the dose of up to 60 mg / kg of the sprout extract was increased in comparison with the control group. However, in the case of 120 mg / kg, since the phagocytosis was decreased, it is considered to be a side effect due to an excessive dose, and the dose is preferably less than 120 mg / kg in relation to the phagocytosis and the dose.

As a result of the above, the present invention

First, the present invention provides a method for the separation and purification of the active ingredient of the active ingredient in optimum conditions,

Second, by checking the anticancer or immune function-promoting activity of the extract of the Sprouts in vitro and in vivo system, to provide a novel anti-cancer material having antioxidant and anticancer activity,

Third, in vivo experiments have shown that it is suitable as a functional food or composition for preventing and treating colorectal cancer containing the extract, thereby providing a novel use of the extract.

Although the present invention has been described in detail only with respect to the embodiments described, it will be apparent to those skilled in the art that various modifications and variations are possible within the technical scope of the present invention, and such variations and modifications belong to the appended claims. will be.

Claims (8)

delete delete delete delete After separating the Sprouts of Crotalaria sessiflora L. into stems, leaves, roots and seeds, methylene chloride: ethanol was added to the mixture and the mixture was added at a ratio of 2: 1 to 9: 1 and left to stand at room temperature for 3 to 5 days. After repeated extraction 2 to 3 times, the extract of the bowel sprouts prepared by separating and purifying by recrystallization of the solvent conditions of methylene chloride / hexane, the anticancer or immunity characterized in that it contains a leaf extract excellent in anticancer or immune function enhancement activity Health supplements effective for function enhancement activity. The health supplement foods effective for the anticancer or immune function enhancing activity according to claim 5, wherein the extract of the Sprouts sprout contains 30 to 100 mg / kg. delete delete

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