KR100675269B1 - Pharmaceutical Composition for Alleviating Side Effects Resulting from Cancer Therapy Comprising Schisandra chinensis Extracts and Vitis coignetiae Extracts - Google Patents

Abstract

The present invention relates to a mitigating agent of anti-cancer treatment containing Schizandra chinensis and / or wild berry extract in order to alleviate the side effects associated with chemotherapy using natural substances, and the survival rate of normal cells by combining the mitigating agent of the present invention with chemotherapy Can increase the killing efficiency for cancer cells.

Schisandra chinensis, wild grapes, side effects of chemotherapy, alleviators

Description

Pharmaceutical composition for Alleviating Side Effects Resulting from Cancer Therapy Comprising Schisandra chinensis Extracts and Vitis coignetiae Extracts}

1 is a graph showing the weight change of mice receiving Schizandra chinensis and / or wild berry extract of the present invention with Dox.

Figure 2 is a graph showing the change in lipid peroxide formation in the heart tissue of mice receiving Schizandra chinensis and / or wild berry extract according to the present invention with Dox.

Figure 3a shows the change of bone marrow cell density in bone marrow tissue of mice receiving Schizandra chinensis or Wild grape extract according to the present invention [(A) -Dox, (B) -Dox + SF, (C) -Dox + VF].

Figure 3b shows the effect of cardiac toxicity in mice administered with Schizandra chinensis or Sansuma extract according to the present invention ((A) -Dox, (B) -Dox + SF, (C) -Dox + VF).

Figure 4a is a graph showing the results of treatment of Schizandra extract and Dox of the present invention in normal heart cell line and breast cancer cell line.

Figure 4b is a graph showing the results of treatment of wild berry extract and Dox of the present invention to normal heart cell line and breast cancer cell line.

5a and 5b are HPLC results of the Schizandra chinensis extract according to the present invention, and FIGS. 5c and 5d are graphs showing the HPLC results of the wild grape extract according to the present invention.

The present invention relates to a composition for alleviating side effects caused by anticancer treatment, and more particularly to a pharmaceutical composition for alleviating side effects caused by anticancer treatment, containing Schizandra chinensis and / or wild berry extract.

Cancer is defined as a disease in which certain cells grow indefinitely at any stage of the differentiation process to invade and destroy normal tissues. Many studies have been conducted on cancer, but the cause of the cancer is not known exactly. However, the development of cancer is known to be initiated by activation of cancer genes and loss of function of cancer suppressor genes after exposure to various chemical carcinogens, tumor-causing viruses or large amounts of radiation.

To date, treatment methods for cancer patients include surgery, chemotherapy, radiation therapy, hormone therapy and immunotherapy. Of these, hormonal therapy and immunotherapy are not widely used in clinical practice because the therapeutic effect and stability to the human body have not yet been established. Surgery is a treatment method for removing lesions (ie, cancer), but the initial cancer can be cured only by surgery. However, in the case of cancer after the middle stage, it is impossible to use the surgery itself.

On the other hand, chemotherapy, radiation therapy, or a combination of these treatments commonly produce free radicals to remove target cancer cells, but because of their low specificity, they also damage normal cells, such as hematopoietic and immune cells, that rapidly divide and proliferate. Vomiting, loss of appetite, oral mucositis, diarrhea, constipation, fever, infectious disease, leukopenia, thrombocytopenia, anemia, abdominal pain, kidney toxicity, hepatotoxicity, cardiac toxicity, peripheral nerve toxicity, central nerve toxicity, muscle pain, bone pain, etc. Causes side effects.

Therefore, many researchers are developing ways to minimize the side effects and maximize the usefulness of current chemotherapy and radiation therapy. Among these, food not only supplies the energy and nutrients that are essential for the normal recovery of damaged cells in the course of chemotherapy in cancer patients, but also that certain food ingredients play an important role in restoring or enhancing damaged bone marrow and immune function. Evidence is accumulating. In particular, the food ingredients include vitamins A, C, E, D ₃ and selenium, flavonoids from fruits and vegetables, fish oils, mushrooms, ginseng, and polymers from specialty crops that reduce the cytotoxicity induced by radiation and chemotherapy. It is believed that the usefulness of anticancer therapy can be increased.

Looking at the prior art to alleviate the side effects of chemotherapy as described above are as follows.

Korean Patent Laid-Open Publication No. 2002-0015906 discloses a pharmaceutical composition comprising an extract of Leafy Mushroom (Grifola frondasa) capable of suppressing side effects of an anticancer agent. More specifically, the Leafy Mushroom Extract is currently widely used in clinical practice. It is disclosed to suppress renal toxicity, digestive toxicity, hematopoietic toxicity and the like caused by the administration of phosphorus cisplatin.

Korean Patent Laid-Open Publication No. 2004-0000757 discloses a herbal composition having an effect of inhibiting side effects of anticancer treatment, wherein the herbal composition has a dry weight ratio of 7-20 parts by weight of sucrose, 5-15 parts by weight, Angelica 5-15 parts, and Cheongung 5-15 It is composed of 1 part by weight, 5 to 15 parts by weight, 3 to 10 parts by weight of cornus oil, 1.5 to 5 parts by weight of bark skin, 1.5 to 5 parts by weight of wolfberry, 0.5 to 2.0 parts by weight of safflower and 1 to 3 parts by weight of licorice.

Korean Unexamined Patent Publication No. 2002-0040532 discloses an antioxidant drink having a side effect inhibitory effect according to an anticancer therapy prepared by using a low molecular weight red ginseng saponin and a glutate (GMT) having an active oxygen inhibitory effect and a method of preparing the same Doing. The antioxidant beverage can exert excellent free radical scavenging ability to suppress various side effects caused by free radicals.

In the case of conventional anticancer drug side effects inhibitors, most active ingredients are not clearly identified in the case of natural substances, and in the case of chemical substances, the side effects of the desired anticancer agents are reduced, but it is pointed out as a problem that causes another toxicity. For example, dexrazoxane, a derivative of ethylenediaminetetraacetic acid (EDTA), is used to prevent cardiac toxicity of anthracycline-based drugs including doxorubicin. (Levine BS et al. Cancer Treat Rep. 1980, vol. 64 (12), pp. 1211-1215; Von Hoff DD. Et al. Cancer Treat Rep. 1981, vol. 65 (3-4), pp. 249-252; Whittaker, P. et al. Environ Mol Mutagen. 2001, vol. 38 (4), pp.347-356; Levine, BS et al. Cancer Treat Rev. 1991, vol. 18 (4), pp .241-252).

The present inventors have not only alleviated various side effects caused by the anticancer drugs when administered Schisandra chinensis extract and / or wild grape extract with an anticancer agent used for anticancer therapy, but also increased the survival rate of the anticancer agent against cancer cells. It was confirmed to further promote the action and came to complete the present invention.

Accordingly, the present invention provides an agent for alleviating side effects of anticancer treatment containing Schizandra chinensis extract and / or Sansam extract.

In the present invention, as " Schisandra chinensis ", for example, North Schisandra chinensis, South Schisandra chinensis , Black Schisandra chinensis and the like can be used. Schisandra chinensis is known to strengthen the heart, lower blood pressure and boost immunity. In addition, it has been used to treat coughs and thirst due to strong lung function and antitussive and expectorant action.

In the present invention, " Vitis coignetiae " and the same genus belong to the genus ( Vitis amurensis ), the beetle ( Vitis amurensis for. Glabrescens ), the crow ( Vitis Thunbergii var. Sinuata ), Vine flexion ( Vitis flexuosa) ) May be used. Wild grapes are known to promote digestion, lower blood pressure and improve blood circulation.

In the present invention, the extract can be prepared using the raw materials of Schizandra chinensis and / or wild grapefruit (ie, fruit, root, leaf, stem or mixture thereof), but it is preferable to use fruit. Specifically, Schizandra chinensis and / or wild grape extract of the present invention is added to 70 to 90% ethanol corresponding to 2.5 to 15 times, preferably 5 to 10 times the weight of Schizandra chinensis and / or wild grape raw material, at 20 to 60 ℃ at normal pressure , Preferably at 30 to 50 ° C., after extraction for 1 to 10 hours, preferably 1 to 6 hours, and then the extract thus obtained is filtered, and the filtered liquid is extracted with excess ethanol using, for example, a vacuum distillation apparatus. Can be obtained by evaporation. If reflux is accompanied during the extraction process, it is suitable to extract for 2 to 3 hours. Through this extraction process it is possible to improve the extraction efficiency of the anthocyanin and phenol component in Schizandra chinensis and / or wild grape. On the other hand, Schisandra chinensis and / or wild grape extract thus obtained may be, for example, lyophilized to be prepared in a solid phase.

On the other hand, anti-cancer treatments such as chemotherapy and radiotherapy, which are commonly used in clinical practice, have various side effects. The term "chemotherapy" refers to a method of treating cancer using an anticancer agent, and the anticancer agent includes, but is not limited to, cisplatin, carboplatin, procarbazine, and meclo. Mechlorethamine, cyclophosphamide, ifosfamide, melphalan, chlorambucil, bisulfan, nitrosourea, diactinomycin (dactinomycin), daunorubicin, doxorubicin, bleomycin, bleomycin, plicomycin, mitomycin, etoposide, tamoxifen, taxol (taxol), transplatinum, 5-fluorouracil, vincristin, vinblastin, methotrexate and the like are used. In the present invention, "radiation therapy" generically refers to treatment with radiation such as X-ray irradiation and γ-ray irradiation.

The "side effects of anticancer treatment" which can be alleviated with Schizandra chinensis and / or wild berry extract according to the present invention vary depending on the cancer treatment or the anticancer agent used, but in general vomiting, decreased appetite, oral mucositis, diarrhea, constipation, fever, infection Includes leukopenia, thrombocytopenia, anemia, abdominal pain, kidney toxicity, liver toxicity, cardiac toxicity, peripheral neurotoxicity, central nervous system toxicity, muscle pain, bone pain.

Schizandra chinensis and / or wild berry extract according to the present invention can alleviate at least one or more of the side effects of the anticancer treatment described above, in particular myelosuppression (ie, leukopenia, thrombocytopenia, anemia), cardiotoxicity, The term "mitigating" in the specification means alleviation, reduction or elimination of any one of the aforementioned side effects.

One aspect of the present invention is to mitigate the side effects caused by doxorubicin using the schizandra extract and / or wild berry extract of the present invention. Doxorubicin is an anthracycline antibiotic isolated from Streptomyces peucetius and is a chemotherapeutic agent with potent anticancer activity against a wide range of hematological and solid cancers. In particular, doxorubicin is effective against breast cancer, esophageal carcinomas osteosarcoma, Kaposi's sarcoma, soft-tissue sarcomas, Hodgkin's and non-Hodgkin's lymphoma. . However, cumulative doses of doxorubicin cause cardiac toxicity, such as congestive heart failure, as well as conventional side effects caused by other chemotherapy agents such as inhibition of hematopoiesis, vomiting, hair loss, and the like. Schizandra chinensis and / or wild grape extract according to the present invention has been found to have a particularly relieving effect on cardiac toxicity, the extract of the present invention is preferably used with doxorubicin.

Although the Schizandra chinensis and / or wild grape extract according to the present invention may be used as such, it is preferably provided as a pharmaceutical composition (ie, emollient) in a form incorporated into a pharmaceutically acceptable carrier.

The amount of Schizandra chinensis extract per weight of the emollient according to the invention is 1 to 99% by weight, preferably 5 to 80% by weight, more preferably 10 to 50% by weight. In addition, the wild berry extract is 1 to 99% by weight, preferably 5 to 50% by weight, more preferably 10 to 30% by weight, based on the total weight of the emollient.

Carriers used in the emollients of the present invention include carriers, adjuvants and vehicles commonly used in the pharmaceutical art and are collectively referred to as “pharmaceutically acceptable carriers”. Pharmaceutically acceptable carriers that may be used in the laxatives of the present invention include, but are not limited to, ion exchange, alumina, aluminum stearate, lecithin, serum proteins (eg, human serum albumin), buffer materials (eg, various Phosphates, glycine, sorbic acid, potassium sorbate, partial glyceride mixtures of saturated vegetable fatty acids), water, salts or electrolytes (e.g. protamine sulfate, disodium hydrogen phosphate, hydrogen carbonate, sodium chloride and zinc salts), colloidal silica , Magnesium trisilicate, polyvinylpyrrolidone, cellulose-based substrates, polyethylene glycols, sodium carboxymethylcellulose, polyarylates, waxes, polyethylene-polyoxypropylene-blocking polymers, polyethylene glycols and wool, and the like.

Emollients of the present invention can be administered via any general route as long as the desired tissue can be reached. Therefore, the emollients of the present invention may be administered topically, orally, parenterally, intraocularly, transdermally, rectally, intestine, etc., and may be formulated into solutions, suspensions, tablets, pills, capsules, sustained release preparations, and the like. As used herein, the term “parenteral” includes subcutaneous, intranasal, intravenous, intraperitoneal, intramuscular, intraarticular, synovial, intrasternal, intracardiac, intradural, intralesional, and intracranial injection or infusion techniques. .

In one embodiment, when the emollients of the present invention are formulated as oral solids, diluents (e.g., lactose, dextrose, sucrose, cellulose, corn starch or potato starch), ointments, together with Schizandra chinensis and / or wild berry extracts (E.g. silica, talc, stearic acid, magnesium or calcium stearate and / or polyethylene glycol), binders (e.g. starch, arabic gum, gelatin methylcellulose, carboxymethylcellulose or polyvinyl pyrrolidone), disintegrants ( Starch, alginic acid, alginate or sodium starch glycolate), formal mixtures, dyes, sweeteners, wetting agents (e.g. lecithin, polysorbates, laurylsulfate) and pharmacologically inerts generally used in pharmaceuticals May contain substances. These agents can be prepared by known methods, for example by means of mixing, granulating, tableting, dragging or film-coating processes.

In another embodiment, the emollients of the present invention may include inert diluents commonly used (e.g., purified water, ethanol) when formulated into liquid compositions for oral administration such as solutions, emulsions, suspensions, syrups and elixirs. have. If desired, the liquid composition may further comprise auxiliaries such as wetting and suspending agents, sweetening agents, flavoring aromas and preservatives.

In another embodiment, the emollients of the present invention may be prepared in an aqueous solution for parenteral administration. Preferably, suitable buffer solutions such as Hanks' solution, Ringer's solution, or physically buffered saline can be used. Aqueous injection suspensions can be added with a substrate that can increase the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol or dextran. In addition, suspensions of Schizandra chinensis and / or wild berry extracts are suitable oily injection suspensions, and suitable lipophilic solvents or carriers include fatty acids such as sesame oil or synthetic fatty acid esters such as ethyl oleate, triglycerides or liposomes. . Polycationic amino polymers may also be used as carriers. Optionally, the suspension may use suitable stabilizers or agents to increase the solubility of the compound and to prepare high concentration solutions.

Preferred emollients of the present invention may be in the form of sterile injectable preparations as sterile injectable aqueous or oily suspensions. Such suspensions can be formulated according to techniques known in the art using suitable dispersing or wetting agents (eg Tween 80) and suspending agents. Sterile injectable preparations may also be sterile injectable solutions or suspensions (eg solutions in 1,3-butanediol) in nontoxic parenterally acceptable diluents or solvents. Vehicles and solvents that may be used include mannitol, water, Ringer's solution, and isotonic sodium chloride solution. In addition, sterile, nonvolatile oils are conventionally employed as a solvent or suspending medium. For this purpose, any non-volatile oil can be used including synthetic mono or diglycerides. Fatty acids such as oleic acid and its glyceride derivatives can be used in the preparation of injections as well as pharmaceutically acceptable natural oils (for example olive oil or castor oil), in particular their polyoxyethylated ones.

The previously prepared liquid composition is usually sterilized by incorporating a bactericide or spinning by filtration through a bacterial capture filter or the like. The sterilized composition can be solidified by obtaining a solid composition, for example by lyophilization, which, in use, is dissolved in sterile water or sterile diluent.

In general, the effective dosage of a palliative agent according to the invention is from 0.1 μg to about 500 mg per kg of body weight per day, preferably from about 1 μg to about 250 mg per kg of body weight and most preferably from about 2 μg to about 100 mg per kg of body weight. . As will be apparent to those skilled in the art, the specific amount of emollient to be administered to a patient will depend on a number of factors, including the patient's symptoms, the depth of side effects, and the like.

The palliative agent according to the present invention may not only alleviate the side effects caused by the anticancer treatment already treated, but may also prevent and alleviate the side effects caused by the anticancer treatment in parallel with the anticancer treatment.

Hereinafter, the examples are only for illustrating the present invention in more detail, and the scope of the present invention is not limited by these examples in accordance with the gist of the present invention, those skilled in the art. Will be self-evident.

<Example 1>

Preparation of Schizandra chinensis Extract

A. Preparation of Animal Experimental Samples

After lyophilization of Schisandra chinensis and Vitis coignetiae fruits, 80% ethanol corresponding to 10 times the weight of the fruit was added at normal pressure and 30 ° C., and extracted under reflux for 3 hours, followed by filtration. The filtered solution was used as a sample of the animal experiment of Example 2 by evaporating the excess ethanol in a vacuum distillation apparatus and freeze-dried the remaining filtrate.

The Schisandra chinensis extract thus obtained was called "SF", and the wild grape berries extract was called "VF".

B. Preparation of Samples for Cell Culture Experiments

The animal test samples were semi-purified by solid phase extraction using a Sep-Pak C18 cartridge (Waters, USA). First, the C18 cartridge was activated with acidic methanol and acidified water, and the animal test sample (1 g) was dissolved in distilled water (10 mL) and injected into the cartridge. In order to remove impurities and concentrate the active ingredient, it was washed with acidic water and the active ingredient was dissolved from the C18 cartridge using acidic ethanol. The ethanol sample solution thus obtained was distilled under reduced pressure, and then dissolved in acidic water and UV / Vis spectrophotometer HP8453, Hewlett-Packard Co., Ltd. using extinction coefficient of standard material (cyanidin 3-glucoside). USA) was used as a sample of the cell culture experiment of Example 3 after correcting the concentration of the semi-purified extract by colorimetric determination at 520nm.

<Example 2>

Confirmation of Side Effects Relief from Anticancer Treatment of Schizandra chinensis Extract by Animal Experiments

A. Animal Breeding

Four-week-old male C57BL / 6 mice were purchased from a central laboratory animal and allowed to acclimate to the animal breeding environment for one week before being used for the experiment. After adaptation, weights were divided into five groups of 10-15 rats using the weight-dependent ovary method. Each experimental animal was raised in a cage, and the diet provided a solid blend and the diet and water were free to eat. The five groups classified above were divided into one control group and four test groups, and all four test groups contained doxorubicin (Dox) at 10 mg / kg B.W. Intraperitoneal injection at concentration. At this time, the three experimental groups of the experimental group, respectively, SF extract alone (SF administration group), VF extract alone (VF administration group), and a mixture of SF and VF extract ("SV" mixed administration group) a total of 10 days before the dox intraperitoneal injection It was administered orally once a day at a concentration of 500 mg / kg BW for days.

B. Treatment and Tissue Collection of Laboratory Animals

On the fifth day after the intraperitoneal injection, the blood was collected through orbital blood collection, and after laparotomy, the ascites was observed, and the heart and spleen were removed and weighed. The heart and femur were also soaked in 10% neutral formalin fixative for 2-3 days for tissue observation.

As shown in FIG. 1, the average body weight of the Dox-administered group was 20.8 g, which was significantly reduced compared to 23.4 g of the non-Dox-administered group. In the Dox group, the weights of the spleen and heart were slightly reduced, but there was no significant difference compared to the organ weight per body weight. Examination of the effect of the extract showed that administration of SF and / or VF slightly improved the weight loss due to Dox administration. The body weight gradually increased after the start of the experiment, and then showed a sharp decrease in body weight after administration of Dox. The decrease in body weight decreased with the administration of SF and / or VF. In Dox-only group, body weight decreased 2.1g after administration, but weight loss decreased significantly by 1.2g in SF group and 1.1g in SV group.

In addition, in addition to weight loss, ascites was observed in most of the subjects in the Dox-administered group at the time of dissection. In the administration group, SF and SV were found to be significantly reduced to improve the ascites formation. However, administration of VF alone did not reduce ascites formation (Table 1).

Comparison of the Effect of Extracts on Multiple Formation revenge Dox (%) Dox + SF (%) Dox + VF (%) Dox + SV (%) ＞ 1ml 8/15 (53) 2/10 (20) 4/9 (44) 2/10 (20) <1 ml 7/15 (47) 8/10 (80) 5/9 (56) 8/10 (80)

From the above results, side effects such as weight loss and ascites caused by Dox administration were somewhat improved by SF alone or SF and VF administration, but VF alone showed no significant improvement in weight loss or ascites. .

C. Blood Cell Analysis

In addition, the blood collected in A of Example 2 was measured for blood indicators such as platelet count, erythrocyte count, leukocyte count, lymphocyte, monocyte, granulocyte, eosinophil, basophils in blood. Measurement of blood indicators was performed using the ADIVA 120 Hematology System (Bayer, USA).

Dox administration induced the toxicity of the hematopoietic system that occurs when general anticancer drugs were administered. As a result of the changes in blood cells, Dox administration significantly decreased the levels of erythrocytes, leukocytes, hemoglobin, etc., and the WBC differential analysis showed that the neutrophil ratio increased and the lymphocyte ratio decreased. . Investigating the effects of SF and / or VF, SF alone or a combination of SF and VF did not increase white blood cell counts or hemoglobin levels but significantly increased red blood cell counts and was increased by Dox treatment. Leukocyte differentiation analysis, including neutrophil ratio and decreased lymphocyte ratio, was confirmed to be close to the control. The administration of VF was significantly increased in erythrocytes compared to the Dox group, but did not significantly affect the leukocyte count, hemoglobin, and leukocyte differential counts (Table 2).

Effect of Extracts According to the Invention on Blood Cells in Blood Control Experimental group Dox Dox + SF Dox + VF Dox + SV Erythrocytes (× 10 ⁶ / μl) 11.0 ± 0.36 9.7 ± 0.24 10.2 ± 0.20 ^** 10.1 ± 0.36 ^* 10.3 ± 0.18 ^** Leukocytes (× 10 ³ / μl) 5.4 ± 0.72 4.6 ± 1.20 4.7 ± 0.66 4.9 ± 1.19 4.9 ± 1.03 Hemoglobin (g / L) 17.1 ± 0.5 15.2 ± 0.5 15.5 ± 0.2 15.1 ± 0.6 15.7 ± 0.6 Leukocyte Differential Analysis Neutral sphere (%) 11.5 ± 4.9 23.4 ± 8.3 12.3 ± 3.1 ^** 23.6 ± 10.4 12.0 ± 5.1 ^** Lymphocyte (%) 86.5 ± 4.8 75.0 ± 8.4 86.0 ± 3.0 ^** 74.0 ± 1.05 86.3 ± 4.8 ^** Monocytes (%) 1.9 ± 0.9 1.6 ± 1.2 1.8 ± 0.9 2.4 ± 1.1 1.9 ± 1.0

D. Analysis of Lipid Peroxidation in Cardiac Tissue

Some of the cardiac tissues obtained in A of Example 2 were immediately frozen with liquid nitrogen and stored at −70 ° C. until the experiment. After the heart tissue was added to phosphate buffer solution of 20-fold by taking a duplicate 0.5ml was homogenized in a glass homogenizer to homo mixed well so that here end _{1mM FeSO 4, 0.01mM BHT, 1mM} L- ascorbic acid from 37 to 30 ℃ Incubated for a minute. After standing, 10% TCA solution was added, centrifuged at 12,000 g for 10 minutes, the supernatant was taken, 1% TBA reagent was added, and left to develop in a 95 ° C. water bath for 10 minutes, followed by cooling at room temperature for 30 minutes. The supernatant was collected by centrifugation and colorimetrically determined at 532 nm. The malonaldehyde standard curve was used to obtain a curve by reacting 1,1,3,3, -tetramethoxypropane with a standard solution.

Dox is known to cause various ROS, and in particular, it is known to cause various heart diseases by directly increasing oxidative stress of cardiac tissues, causing DNA and lipid peroxidation, apoptosis, and the like. As shown in FIG. 2, lipid peroxidation was increased in cardiac tissue of the Dox-administered group in this example. SF alone or a combination of SF and VF significantly reduced the concentration of cardiac lipid peroxides (indicated by MDA) that resulted from Dox administration. VF also decreased the amount of lipid peroxides, but showed no significance. Therefore, SF alone or a combination of SF and VF is expected to have a protective effect against cardiotoxicity caused by Dox.

E. Cardiac tissue and bone marrow blood cell observation

The cardiac tissue and femur fixed in accordance with A of Example 2 were demineralized in the case of the femur, and then each tissue was prepared into 4 μm-thick paraffin tissue sections. Tissue sections were stained with hematoxylin-Eosin and observed under an optical microscope.

As shown in (A) of FIG. 3A, the number of blood cells in bone marrow was decreased in the Dox-administered group, and Dox in the animals administered with SF or VF as shown in (B) and (C) of FIG. 3A. It was observed that the density of bone marrow cells was significantly increased compared to the dosing group.

Dose-dependent cardiomyopathy is known as a side effect of Dox. As shown in (A) of FIG. 3B, condensed and fragmented nuclei were found in some of the heart tissue sections of the Dox-administered animals, and morphological changes such as disorganization were observed. However, as shown in (B) and (C) of FIG. 3B, in the SF and VF-administered groups, the cardiac toxicity exhibited by Dox was alleviated.

<Example 3>

Cell culture experiment

H9C2 (rat, cardiomyocytes) and MCF-7 (human, breast adenocarcinoma) cell lines were distributed from ATCC (Rockville, MD) and used in the experiment. MCF-7 cell line was cultured in DMEM medium, H9C2 cell line was present at 37 ° C., 5% CO ₂ in modified DMEM medium supplemented with 10% FBS, streptomycin (100 μg / ml) and penicillin (100 U / ml) Incubated in a CO ₂ incubator. H9C2 cell lines were passaged at 70% confluency to prevent loss of differentiation potential. The cells that reached the confluency were treated with trypsin-EDTA to detach the cells from the culture dish to obtain a single cell suspension, and to divide the cells at a concentration of 1-1.5 × 10 ⁴ cells / sphere on 96-spheres and to 37 ° C. CO ₂ The incubator was incubated for 24 hours. Dox was added thereto, followed by incubation for another 24 hours, and then changed to a fresh medium. The semi-purified SF and semi-purified VF samples obtained in Example B were added at a concentration of 0-100 µg / ml and incubated for 24 hours. Cell viability was then measured via SRB and / or MTT assays.

As shown in Figures 4a and 4b SF or VF resulted in an increase in cell viability of up to about 30% compared to the group administered only Dox in normal cardiac cell lines. In addition, MCF-7 cells, cancer cell lines, showed about 20-30% increase in cytotoxicity at the highest concentration of each sample without affecting the cytotoxicity caused by Dox. Therefore, the results of the cell culture experiments are expected that the two samples may enhance Dox's effect on cancer cells as a supplement to Dox and protect against the toxicity of normal heart cells.

<Example 4>

Analysis of Active Ingredients Contained in Extracts

SF and VF extracts are known to have high content of anthocyanins with antioxidant effects. Therefore, anthocyanins in both samples were analyzed using HPLC (Jasco, Japan).

Acidic methanol is added to the freeze-dried SF or VF sample, shaken and extracted, filtered through a filter paper, and concentrated by evaporating excess methanol in a vacuum distillation apparatus. This was semi-purified using solid phase extraction (SPE) via a Sep-Pak C18 cartridge (Waters, USA) and used for HPLC analysis. For aglycone analysis, it was used after acid decomposition using 2N HCl.

Specific HPLC analysis conditions for anthocyanin analysis in SF and VF extracts are shown in Table 3 below.

HPLC analysis conditions HPLC conditions  detection UV 520 nm  column Allsphere ODS-2 5u, 4.6mm × 150mm  Flow 0.7 ml / min  Gradient condition A; 4% phosphoric acid, B; ACN  Aglycon 88 (0) -88 (5) -85 (30) -80 (50) -88 (55) A%  Glycoside 90 (0) -87 (35) -90 (40) -90 (50) A%

Analysis results of SF and VF according to the present invention is shown in Figures 5a to 5d and Table 4. 5A and 5B are anthocyanin glycosides and aglycone analysis results of SF, respectively, and FIGS. 5C and 5D are anthocyanin glycosides and aglycone analysis results of VF, respectively. SF extract was composed mostly of cyanine and its glycosides among the various anthocyanins, but its content was not high. VF extracts contained various anthocyanins, of which delpinin, petunidine, and malvidin were the highest.

Component Analysis Table for SF and VF sample % Of total peak area  Anthocyanins (aglycones)  Anthocyanins (glycosides) SF  Delphinidine (Dp) 9.0  Dp-3-glucose 16.5  Cyanidin (Cy) 69.7  Cy-3-glucose 14.7  Petunidine (Pt) -  Pt-3-glucose -  Pellagonidine (Pg) -  Pl-3-glucose -  Pheonidine (Pn) -  Pn-3-glucose -  Malvidin (Mv) -  Mv-3-glucose - VF  Delphinidin 30.8  Dp-3-glucose 21.7  Cyanidin 15.4  Cy-3-glucose 5.6  Petunidine 19.7  Pt-3-glucose 13.5  Pelagonidine -  Pl-3-glucose -  Peonidine 8.0  Pn-3-glucose 3.8  Malvidin 22.1  Mv-3-glucose 11.4

The palliative agents according to the present invention not only alleviate the various side effects caused by anticancer treatment but also increase cancer cell killing ability by anticancer agent or radioactivity while increasing the survival rate of normal cells.

Claims (6)

A side effect alleviator of anticancer treatment containing Schisandra chinensis extract or Wild grape extract or both. The emollient according to claim 1, wherein the Schizandra extract is 1 to 99% by weight based on the total weight of the emollient. The emollient according to claim 1, wherein the wild berry extract is 1 to 99% by weight based on the total weight of the emollient. According to claim 1, the Schizandra chinensis extract was added to 70 to 90% ethanol corresponding to 2.5 to 15 times the weight of Schizandra chinensis extract and extracted for 1 to 10 hours at a temperature of 20 to 60 ℃ under normal pressure and then filtered the extract thus obtained. And obtained by evaporating ethanol from the filtrate. The extract of claim 1, wherein the extract is extracted by adding 70 to 90% ethanol corresponding to 2.5 to 15 times the weight of the raw material of the wild berry extract at a temperature of 20 to 60 ° C. under normal pressure for 1 to 10 hours, and then filtering the extract thus obtained. And obtained by evaporating ethanol from the filtrate. The method of claim 1, wherein the side effects of the chemotherapy include nausea, loss of appetite, oral mucositis, diarrhea, constipation, fever, infection, leukopenia, thrombocytopenia, anemia, abdominal pain, kidney toxicity, liver toxicity, cardiac toxicity, peripheral nerves Palliative agent, characterized in that selected from the group consisting of toxicity, central nervous system toxicity, muscle pain, bone pain.

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