KR20220077979A - Composition containing Ucha-Shinki-hwan for preventing, improving or treating cancer associated cachexia - Google Patents

Abstract

The present invention relates to a composition for preventing, ameliorating or treating cancer cachexia or muscle loss containing oxalobacillus, and the present invention has shown an effect of improving weight loss in a mouse model induced with cancerous cachexia, IL6, CRP and Inhibits the expression of TNF-α, suppresses the increase in the expression of MURF1 and MAFbx in muscle tissue, and suppresses the decrease in the size of the muscle tissue, so it may be useful as a composition for the prevention, improvement or treatment of cancer cachexia or muscle loss can

Description

Composition containing Ucha-Shinki-hwan for preventing, improving or treating cancer associated cachexia

The present invention relates to a composition for preventing, improving, or treating cancerous cachexia or muscle loss, containing oxaxingihwan.

Cachexia refers to a complex syndrome that causes persistent muscle loss caused by various diseases such as cancer, heart failure, chronic obstructive pulmonary disease, chronic kidney disease, AIDS, and the like. Cancer cachexia is a complex metabolic disorder resulting from malignant tumors, with pathological loss of muscle and adipose tissue as the main symptom. is defined as decreasing. Cachexia can occur in a variety of medical conditions, but cancerous cachexia is closely related to terminal cancer. About 50% of all cancer patients, especially about 80% of terminal cancer patients, are affected by cancerous cachexia and suffer from decreased life and increased mortality due to loss of muscle and fat. Cancer cachexia is characterized by weight loss due to significant loss of muscle mass and adipose tissue, and has a different characteristic from muscle and fat loss through appetite suppression.

The diagnostic criteria for cancer cachexia include a reduction in weight loss of 5% or more, high levels of inflammatory marker expression in serum levels (IL6, CRP1, TNFr), decreased exercise capacity, and anorexia. Although the exact mechanism of cancer cachexia is not known, inflammatory cytokines such as IL-6, TNF-α, and C-reactive protein 1 (CRP1) are known to play a central role in muscle and fat loss.

Skeletal muscle weight loss and muscle wasting due to the inflammatory response activate the ubiquitin-proteasome proteolytic pathway and the autophagic/lysosomal proteolytic pathway, and muscle protein is degraded. In particular, increases in ubiquitin ligases such as MAFbx/Atrogin-1 (muscle atrophy F-Box) and MuRF1 (muscle RING finger-1) and autophagy-related genes such as Bnip3 It is directly involved in proteolysis.

Treatments for cancer cachexia include NSAIDs (non-steroidal anti-inflammatory drugs), β2-adrenergic agonists, corticosteroids, and ghrelin. There are α inhibitors.

A COX-2 inhibitor is a drug that reduces the loss of skeletal muscle by inhibiting COX-2, which is involved in the production of prostaglandins, which are inflammatory substances generated in large amounts in cancer tissues. As a COX-2 inhibitor, celecoxib is commercially available. However, side effects such as anemia, gastric ulcer, allergy, heart attack and stroke have been reported with celecoxib.

A TNF-α inhibitor is a drug that kills cancer cells by reducing the expression of TNF-α, which is often expressed in the bloodstream of patients with cancer cachexia. Thalidomide is commercially available as a TNF-α inhibitor. However, these drugs have little effect as a treatment for cancer cachexia, and have side effects such as depression, heart failure, shortness of breath, vomiting, rash, high blood pressure, and birth defects during pregnancy.

As such, most of the drugs being developed or marketed up to now have many side effects and lack of evidence for their therapeutic effect. Therefore, there is an urgent need to develop a treatment for cancer cachexia with a new mechanism.

On the other hand, Ucha-Shinki-hwan (牛車腎永丸) is a drug made by using chinensis, mussel, cornflower oil, sanyak, psyllium, taeksa, bokryeong, peony (Mokdanpi), and cinnamon (broiler chicken). It is easy to do, and the extremities are cold, and it is used for symptoms such as lower extremity pain, back pain, numbness, lack of vision in the elderly, itching, difficulty urinating, frequent urination, and edema with thirst due to reduced urine output or polyuria.

As a result of research to develop a therapeutic agent for cancer cachexia using a natural product, the present inventors have confirmed that there is a significant improvement in cancer cachexia in a mouse model in which Uchashin Ki-hwan induced cancer cachexia by injection of CT26 cells, and completed the present invention.

Korean Patent Publication No. 10-2019-0103985

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating cachexia or muscle loss.

Another object of the present invention is to provide a health functional food composition and health food composition for preventing or improving cachexia or muscle loss.

Another object of the present invention is to provide an anticancer adjuvant composition.

In order to achieve the above object,

The present invention provides a pharmaceutical composition for the prevention or treatment of cachexia or muscle loss, including oxalomycetes.

In addition, the present invention provides a health functional food composition and health food composition for the prevention or improvement of cachexia or muscle loss, including oxalobium.

In one embodiment of the present invention, the cachexia may be caused by cancer.

Furthermore, the present invention provides an anticancer adjuvant composition comprising the pharmaceutical composition.

The present invention showed a significant improvement in weight loss in a mouse model in which cancer cachexia was induced by injection of CT26, a colorectal cancer cell line. The expression of α was suppressed, and the expression of MURF1 and MAFbx, which are muscle proteolytic factors, was suppressed in muscle tissue caused by cancerous cachexia, and had an effect of inhibiting the size reduction of muscle tissue, thus preventing cancer cachexia or muscle loss; It may be useful as a composition for improvement or treatment.

1 shows the inhibitory effect on weight loss due to cancerous cachexia of woochashingihwan.
Figure 2 shows the inhibitory effect on the expression of inflammatory cytokines (IL-6, CRP, and TNF-α) in the serum by the cancerous cachexia of woochashingihwan.
Figure 3 shows the inhibitory effect on the reduction in muscle tissue (tibialis anterior; TA) weight due to the cancerous cachexia of the right shin Ki-hwan.
4 shows the results of confirming the histological changes and MURF1 changes in the muscle tissue (TA) of the cancerous cachexia model caused by oxalic cystitis through hematoxylin & eosin (H&E) staining (top) and immunohistochemical staining (bottom). it has been shown
5 is a quantification of the muscle tissue size (left) and MURF1 expression level (right) in the muscle tissue (TA) of a cancerous cachexia model caused by right hypochondrium through hematoxylin & eosin (H&E) staining and immunohistochemical staining. the results are shown.
FIG. 6 shows the results of confirming the expression of MURf1 and MAFbx in muscle tissue (TA) according to the treatment of oxalobacilli by western blot and quantification thereof.

Hereinafter, the present invention will be described in detail.

Pharmaceutical composition for preventing or treating cachexia or muscle loss

The present invention provides a pharmaceutical composition for the prevention or treatment of cachexia or muscle loss, including oxalomycetes.

As used herein, the term 'cachexia' refers to a high degree of general weakness that can be seen in the late stages of cancer, tuberculosis, diabetes, acquired immunodeficiency syndrome, AIDS, etc., and gastric cancer, esophageal cancer, pancreatic cancer It is frequently seen in patients with gastrointestinal cancer such as colon cancer and lung cancer. It shows symptoms such as decreased appetite, decreased body weight and stamina due to muscle and fat reduction, anemia, lethargy, and indigestion. indicates a declining state. When cachexia develops, it shows a low response to chemotherapy or radiation therapy, which reduces the patient's quality of life, shortens life expectancy, and causes death due to weight loss in 10 to 20% of all cancer patients. do.

In the pharmaceutical composition according to the present invention, the cachexia may be caused by cancer. In one embodiment of the present invention, cachexia induced by the cancer is expressed as 'cancer cachexia'.

As used herein, the term 'muscle loss' refers to a decrease in muscle in the body, for example, a decrease in muscle tissue caused by not using the muscle, a decrease in muscle tissue due to a disease of the muscle itself It may be a decrease or a decrease in muscle tissue due to damage to the nerves controlling the muscle, and such muscle loss may be caused by, for example, cachexia or muscular atrophy (muscular atrophy).

In the pharmaceutical composition according to the present invention, the muscle loss may be caused by cancer or cancerous cachexia.

Cancer as defined herein is leukemia, lymphoma, myeloma, myelodysplastic syndrome, breast cancer, head and neck cancer, esophageal cancer, stomach cancer, colorectal cancer (= colon cancer), rectal cancer, anal cancer, hepatocellular liver cancer, bile duct cancer, gallbladder cancer, pancreatic cancer, lung cancer (non-small cell) Lung cancer, small cell lung cancer), thymus cancer, kidney cancer, bladder cancer, prostate cancer, testicular cancer, ovarian cancer, cervical cancer, sarcoma, gastrointestinal stromal tumor (GIST, GIST), cancer of unknown primary site, mesothelioma, melanoma, It may include neuroendocrine tumors, skin cancers, blood cancers, and the like.

The pharmaceutical composition according to the present invention may be to improve one or more symptoms selected from the group consisting of decreased appetite, weight loss, increased fatigue, increased inflammation, muscle loss, fat loss, and hematopoietic toxicity, preferably body weight It may be to improve one or more symptoms selected from the group consisting of reduction, muscle loss, and fat loss.

In addition, the pharmaceutical composition according to the present invention may be to reduce the expression of inflammatory cytokines in the hypothalamus, blood or bone marrow. In this case, the inflammatory cytokine may be one or more selected from the group consisting of CRP, TNF-α, IL-1β, IL-6, and IL-17.

In addition, the pharmaceutical composition according to the present invention may inhibit the expression of one or more genes selected from the group consisting of MAFbx, MuRF1 and Bnip3 in muscle.

According to one embodiment of the present invention, it showed an effect of improving weight loss in a mouse model of cancer cachexia induced by the injection of CT26, a colorectal cancer cell line, in the vicinity of the thigh, and IL6 acting as a major indicator of cancer cachexia in the serum. , showed the effect of inhibiting the expression of CRP and TNF-α, and it was confirmed that it showed the effect of reducing the expression of MURF1 and MAFbx, which are muscle proteolytic factors in muscle tissue, and inhibiting the size reduction of muscle tissue (Example) 1 to 6).

In the present invention, the 'woochashingihwan' is Rehmanniae Radix, hyssop (Achyranthis Radix), Corniflora (Corni Fructus), Sanyak (Dioscoreae Rhizoma), Psyllium (Plantaginis Semen), Taxa (Alismatis Rhizoma), Bokryeong (Poria Sclerotium) ), peony (Moutan Radicis Cortex) and cinnamon (Cinnamomi Cortex) may refer to a mixed extract extracted from six medicinal substances, and the mixed extract may be a mixture of further addition of Aconiti Lateralis Radix Preparata. Alternatively, a commercially available oxacingihwan may be used.

In the present invention, the rehmanniae (Rehmanniae Radix) may be a root of Rehmannia glutinosa Liboschitz ex Steudel belonging to the Scrophulariaceae, and may include Saengjihwang or Sukjihwang. The hyssop (Achyranthis Radix) may be a root of Achyranthes japonica Nakai or hyssop ( Achyranthes bidentata Blume) belonging to the family Amaranthaceae. The cornus oil (Corni Fructus) may be a ripe fruit of Cornus officinalis Siebold et Zuccarini belonging to the Cornaceae family, from which seeds have been removed. The acid medicine (Dioscoreae Rhizoma) is a rhizome from which the bark of Dioscorea batatas Decaisne or Dioscorea japonica Thunberg belonging to the family Dioscoreaceae has been removed, and may be dried as it is or steamed. The psyllium (Plantaginis Semen) may be a ripe seed of Plantago asiatica Linne or Plantago depressa Willdenow belonging to the Plantaginaceae. The taxa (Alismatis Rhizoma) may be a tuber of a plant of the Alismataceae family, which is a tuber of Alisma orientale Juzepzuk, from which fine roots and bark are removed. The bokryeong (Poria Sclerotium) may be a sclerotia of bokryeong ( Poria cocos Wolf) belonging to the family Polyporaceae. The peony skin (牡丹皮, Moutan Radicis Cortex) may be the root bark of the peony family plant Mokdan (Paeonia suffruticosa Andrews), also called mokdanpi. The cinnamon (Cinnamomi Cortex) is also called broiler (肉桂), and may be used as the stem bark of the Lauraceae plant, Cinnamomum cassia Presl, as it is or by slightly removing the bark. The father and son (附子, Aconiti Lateralis Radix Preparata) are made by processing the root of the cucumber belonging to the Ranunculaceae (烏頭, Aconitum carmichaeli Debeaux); It may be aspirational (砲附子), preferably, it may be a number (修治)bujamal (末). The above numerical values are powdered to reduce the toxicity of the father and son, and for example, may refer to the wealthy father, Dangpo wealthy, Gyeongpo wealthy, Yeom seed rich man, Daegupo rich man, etc., but is not limited thereto. All rich people who have gone through processing to reduce the toxicity of the rich can use it.

According to an embodiment of the present invention, the mixed extract may be prepared by extracting a mixture of the above medicines, or prepared by preparing an extract of each of the medicinal substances and then mixing the extracts. Preferably, 0.5~6:1~4:1~4:1~4:1~4:1~4:1 of the rehmannia, hyssop, cornflower oil, sanyak, psyllium, texas, bokryeong, peony skin and cinnamon It may be a mixed extract prepared by mixing in a weight ratio of ~4:1 to 4:1, and may be prepared by further mixing the rich material in an amount of 1 to 3 times the weight based on the weight of cinnamon.

In the present invention, 5:3:3:3:3:3:3:3: Rehmannia glutinosa, hyssop, cornflower oil, sanyak, psyllium, taeksa, bokryeong, peony skin (mokdan skin), cinnamon (broiler chicken), and Suibujamal are mixed with 5:3:3:3:3:3:3:3: The mixed extract extracted by mixing in a weight ratio of 1:1 was concentrated and dried, and then granulated was named 'Ucha-Shinki-hwan (牛車腎气丸)', and in one embodiment of the present invention, granulated As an herbal supplement in the form of herbal medicine, a granulated preparation containing 60% by weight or more of the granulated mixed extract per packet was purchased from Tsumura (Tsumura, Japan) and used.

The "extract" refers to a preparation concentrated by squeezing the extraction target with an appropriate leachate and evaporating the leachate, but is not limited thereto, but is not limited thereto, It may be a dried product obtained, a crude product or a purified product thereof. The woochasingi ring can be prepared using general extraction methods, separation and purification methods known in the art. The extraction method is not limited thereto, but preferably, methods such as hot water extraction, hot water extraction, cold extraction, reflux cooling extraction, or ultrasonic extraction may be used.

The method may further include fractionating the extract with water, C1 to C4 alcohol, or a combination thereof. The extract may be fractionated with water, ethanol, methanol, butanol, ethyl acetate, hexane, or a combination thereof.

The method may further include performing concentration, pulverization, granulation, centrifugation, filtration, adsorption, or chromatography of the extract.

For example, the woochasingihwan of the present invention is a mixture of Rehmannia, Wooseul, Cornus officinalis, Sanyak, Chajeon, Taxa, Bokryeong, Peony Peel (Mokdanpi), Cinnamon (broiler chicken) and Suibujamal, C1 to C4 alcohol, Obtaining an extract by immersing in ethyl acetate, or a mixed solvent thereof; and pulverizing or granulating the extract; may be prepared by a method comprising, but is not limited thereto. In this case, the C1 to C4 alcohol may be ethanol, methanol, butanol, or a combination thereof.

The pharmaceutical composition according to the present invention may be formulated for oral administration, intramuscular administration, intravenous administration, intraperitoneal administration, subcutaneous administration, intradermal administration, or topical administration.

Uchashingihwan of the present invention can be administered in various oral and parenteral formulations during clinical administration, and when formulated, commonly used fillers, extenders, binders, wetting agents, disintegrants, diluents such as surfactants or excipients are used. is manufactured by

Solid preparations for oral administration include tablets, patients, powders, granules, capsules, troches, and the like, and these solid preparations include at least one or more excipients, for example, starch, calcium carbonate, It is prepared by mixing sucrose, lactose, or gelatin. In addition to simple excipients, lubricants such as magnesium stearate talc are also used. Liquid formulations for oral administration include suspensions, solutions, emulsions, or syrups. In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. can

Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspension solutions, emulsions, lyophilized formulations, suppositories, and the like. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerol, gelatin, and the like can be used.

The pharmaceutical composition may further include a carrier, excipient or diluent. Carriers, excipients, or diluents include, for example, lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil.

In addition, the pharmaceutical composition may further include a known active ingredient having an activity to prevent or treat cancer cachexia or muscle loss.

The effective dose for the human body of the present invention may vary depending on the patient's age, weight, sex, dosage form, health status and disease level, and is generally about 0.001 to 100 mg/kg/day, preferably is 0.01-35 mg/kg/day. Based on an adult patient weighing 70 kg, it is generally 0.07 to 7000 mg/day, preferably 0.7 to 2500 mg/day, and once a day at regular time intervals according to the judgment of a doctor or pharmacist It may be administered in several divided doses.

Health functional food and health food composition for preventing or improving cachexia or muscle loss

In addition, the present invention provides a health functional food composition for preventing or improving cachexia or muscle loss, including oxalomycetes.

The term "health functional food" used in the present invention refers to food manufactured and processed in the form of tablets, capsules, pills, liquids, powders and granules, etc. using raw materials or ingredients useful for the human body. Here, 'functionality' refers to obtaining useful effects for health purposes, such as regulating nutrients or physiological effects on the structure and function of the human body. The health functional food of the present invention can be manufactured by a method commonly used in the ordinary technical field, and at the time of the preparation, it can be prepared by adding raw materials and components commonly added in the conventional technical field. In addition, the dosage form of the health functional food may also be manufactured without limitation as long as it is a dosage form recognized as a health functional food. The health functional food composition of the present invention can be prepared in various types of dosage forms, and unlike general drugs, it has the advantage of not having side effects that may occur during long-term administration of drugs using food as a raw material, and has excellent portability, The health functional food of the present invention can be ingested as an adjuvant for enhancing the effect of a cancer cachexia therapeutic agent or an anticancer agent.

Furthermore, the present invention provides a health functional food composition for preventing or improving cachexia or muscle loss, including oxalomycetes.

The health functional food composition or health food composition according to the present invention may be added to health functional food or health food such as food or beverage for the purpose of preventing or improving cancer cachexia or muscle loss.

There is no particular limitation on the type of the food. Examples of foods that can be added to the present invention include drinks, meat, sausage, bread, biscuits, rice cakes, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gums, dairy products including ice cream , various soups, beverages, alcoholic beverages and vitamin complexes, dairy products, and dairy products, and includes both health food and health functional food in the ordinary sense.

The health functional food and health food composition according to the present invention may be added to food as it is or used together with other food or food ingredients, and may be appropriately used according to a conventional method. The mixing amount of the woochashingihwan according to the present invention may be suitably determined according to the purpose of its use (for prevention or improvement). In general, the amount of woochashingi ring in health food and health functional food may be added in an amount of 0.1 to 90 parts by weight of the total food weight. However, in the case of long-term intake for health maintenance or health control, the amount may be less than the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

The health food and health functional food composition of the present invention is not particularly limited in other ingredients except for containing the oxalic acid ring according to the present invention as an essential ingredient in the indicated ratio, and various flavoring agents or natural carbohydrates, such as a conventional beverage, are added. It can contain as an ingredient. Examples of the above-mentioned natural carbohydrates include monosaccharides such as glucose, fructose and the like; disaccharides such as maltose, sucrose and the like; and polysaccharides such as conventional sugars such as dextrin and cyclodextrin, and sugar alcohols such as xylitol, sorbitol, and erythritol. As flavoring agents other than those described above, natural flavoring agents (taumatin, stevia extract (eg, rebaudioside A, glycyrrhizin, etc.) and synthetic flavoring agents (saccharin, aspartame, etc.) can be advantageously used. The proportion of the natural carbohydrate is generally about 1 to 20 g, preferably about 5 to 12 g per 100 g of the dietary supplement of the present invention.

In addition to the above, the health food and health functional food composition containing Uchashinkihwan according to the present invention are various nutrients, vitamins, minerals (electrolytes), flavoring agents such as synthetic flavoring agents and natural flavoring agents, coloring agents and thickening agents (cheese, chocolate) etc.), pectic acid and its salts, alginic acid and its salts, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin, alcohols, carbonation agents used in carbonated beverages, and the like. In addition, the health food and health functional food composition of the present invention may contain natural fruit juice, fruit juice, and fruit for the production of a vegetable drink.

These components may be used independently or in combination. The proportion of these additives is not limited thereto, but is generally selected from 0.1 to about 20 parts by weight per 100 parts by weight of the health food and health functional food composition containing the active substance of the present invention.

Anti-cancer adjuvant composition

In addition, the present invention provides an anticancer adjuvant composition comprising a pharmaceutical composition for the prevention or treatment of cachexia or muscle loss comprising the woochashingihwan.

The term "anticancer adjuvant" used in the present invention may be used to increase the anticancer therapeutic effect, suppress or improve the side effects of anticancer agents, and may be administered to a patient in combination with an anticancer agent.

The anticancer adjuvant composition of the present invention exhibits an effect of suppressing or improving cachexia symptoms such as muscle loss, weight loss, fat reduction, hematopoiesis, and decreased appetite caused by cancer, thereby increasing the anticancer effect of the anticancer agent. .

Existing anticancer agents as defined herein include cyclophosphamide, methotrexate, 5-fluorouracil, doxorubicin, mustine, vincristine, procarba Procarbazine, prednisolone, bleomycin, vinblastine, dacarbazine, etoposide, cisplatin, epirubicin, cisplatin ( cisplatin), capecitabine, oxaliplatin, and the like.

The anti-cancer adjuvant composition may additionally include one or more active ingredients exhibiting the same or similar function in addition to including oxaxingi-hwan as an active ingredient. The anticancer adjuvant can be administered orally or parenterally during clinical administration, and when administered parenterally, intraperitoneal injection, intrarectal injection, subcutaneous injection, intravenous injection, intramuscular injection, intrauterine dural injection, intracerebrovascular injection, or thoracic It can be administered by internal injection, and can be used in the form of general pharmaceutical preparations.

The anticancer adjuvant may be used alone or in combination with methods using surgery, radiation therapy, hormone therapy, chemotherapy, and biological response modifiers. The daily dose of the anticancer adjuvant is about 0.0001 to 1000 mg/kg, preferably 1 to 100 mg/kg, and it is preferable to divide and administer it once to several times a day, but the patient's weight, age, sex, health condition, The range varies depending on the diet, administration time, administration method, excretion rate, and the severity of the disease. The anticancer adjuvant of the present invention may be administered in various parenteral formulations during actual clinical administration. When formulated, a diluent or excipient such as a commonly used filler, extender, binder, wetting agent, disintegrant, surfactant, etc. is prepared Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Non-aqueous solvents and suspensions may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base of the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

Hereinafter, the present invention will be described in more detail with reference to the following examples. However, the following examples are merely illustrative of the present invention, and the content of the present invention is not limited by the following examples.

<Example 1> Preparation of cancerous cachexia-induced mouse model

In order to induce cancerous cachexia, BALB/c mice were divided into 3 groups (7 mice in each group), and among them, group 2 contained 5×10 ⁵ cells of CT26 colorectal cancer cell line, mixed with PBS and matrigel 1:1, and a total volume of 200 μl was injected subcutaneously in the vicinity of the right thigh to induce cancerous cachexia (Cachexia "+"). The normal control group was not injected with anything (Cachexia "-").

After 1 week of induction of cancerous cachexia (A) Normal control group injected with PBS for an additional 4 weeks, (B) Disease control group injected with PBS for an additional 4 weeks (C) Urinary tract Shingi-hwan (USH, 0.5g/kg/day) for an additional 4 weeks (D) Experimental group injected with celecoxib (celecoxib, 0.05 g/kg/day) for an additional 4 weeks, and (E) Uchashin Ki-hwan (USH, 0.5 g/kg/day) and celecox for an additional 4 weeks The experiment was conducted with a different treatment in the experimental group injected with sieve (celecoxib, 0.05 g/kg/day). At this time, the oxalopod was purchased from Tsumura (Tsumura, Japan; catalog number: 005301-002) and used.

<Example 2> Effect of improving weight loss due to cancerous cachexia of Shin Ki-hwan Woo

In order to confirm the effect of induction of cancerous cachexia and improvement of oxalgia, the weights of mice were measured after treatment for a total of 5 weeks in the same manner as in Example 1 above. At this time, in order to correct for the difference in body weight depending on the cancer tissue, the cancer free body weight was compared from the total body weight of the mice minus the weight of the cancer tissue. In addition, a body weight ratio was calculated to represent the weight loss rate.

The results of the experiment on the effect of woochashinkihwan on the inhibition of weight loss due to cancerous cachexia are shown in FIG. 1 .

As a result, it was confirmed that in the disease control group injected with the CT 26 colorectal cancer cell line, body weight excluding the cancer tissue ( FIG. 1A , cancer free body weight) was significantly reduced compared to the normal control group, confirming that cancerous cachexia was induced. It was confirmed that the improvement effect on weight loss caused by the cancerous cachexia was shown. In addition, the weight loss rate was significantly improved in the experimental group treated with oxalomycetes compared to the disease control group (FIG. 1B). As it showed an improvement effect on both the weight of the body weight and the ratio values according to the change in weight loss, it was confirmed that woochashinkihwan significantly inhibited the weight loss induced by cancerous cachexia compared to the disease control group (CC).

<Example 3> Effect of improving expression on cancer cachexia index factor

Serum inflammatory cytokines interleukin-6 (IL-6), tumor necrosis factor alpha (TNFα), C-reactive protein 1 (C-reactive protein 1, The level of CRP1) was measured.

After a total of 5 weeks of treatment in the same manner as in Example 1, blood was collected from mice, and serum was separated using a centrifuge at 4000 RPM, 4°C, and 20 minutes, and then IL-6, CRP1, and TNFα were Serum was added to the coated plate and reacted, followed by reaction with a secondary antibody, and the difference in color according to the expression level was analyzed using an ELISER analyzer (VERSAmax microplate reader, Molecular Devices, CA, USA).

Fig. 2 shows the results of an experiment on the effect of woochashinkihwan on changes in inflammatory cytokines that occur in the process of cancerous cachexia.

As a result, it was confirmed that Woochashinkihwan showed the effect of reducing the increase in the expression of inflammatory cytokines induced by cancer cachexia for all of IL-6, TNFα and CRP1 in the serum. In particular, compared to celecosib, which is used as a treatment for cancer cachexia, a higher rate of decrease was observed in oxalobium.

<Example 4> Effect of inhibiting muscle tissue reduction due to cancerous cachexia of Shin Ki-hwan Woo

In order to confirm the effect of induction of cancerous cachexia and improvement of oxalgia, the weight of muscle used as an important index in cancerous cachexia was measured. After a total of 5 weeks of treatment in the same manner as in Example 1, the weight of the tibialis anterior (TA) muscle tissue of the mice was measured.

3 shows the results of testing the effect of woochashingihwan on the inhibition of muscle tissue reduction.

As a result, the right shin Ki-hwan significantly inhibited the decrease in the tibialis anterior muscle (TA), a muscle tissue, due to cancerous cachexia.

<Example 5> Improving effect on histological changes in muscle tissue of a cancerous cachexia model of Shin Ki-hwan

In order to confirm the effect of woochashinki-hwan on the histological changes occurring during the onset of cancer cachexia, hematoxylin & eosin staining (H&E staining) and immunohistochemistry staining were performed.

In H&E staining, a colorless tissue is first made as a sample and then dyed for observation with an optical microscope. Since the dye, which is an acidic or basic compound, forms an electrostatic or salt linkage with the ionic group of the tissue, Stain using hematoxylin and eosin. Hematoxylin meets with acids such as the nucleus and other acidic structures of cells (the RNA-rich structure and cartilage matrix in the cytoplasm) and dyes them blue, while eosin meets with basics and turns other cytoplasmic components and collagen into red. dye it The staining sequence was stained with hematoxylin, washed with alcohol, then stained with eosin, and finally treated with xylene. The results of confirming the histological changes in the anterior tibialis anterior muscle tissue in which the cancerous cachexia was induced through H&E staining and the quantification results are shown in FIGS. 4 and 5 .

Next, for immunohistochemical staining, after a total of 5 weeks of treatment in the same manner as in Example 1, the muscle tissue (TA) samples of mice were formalin fixed and then paraffin tissue was sectioned to a thickness of 5 μm. was manufactured. Five slides per mouse were prepared for analysis. The specimen sections were parraffin-free with xylene and hydrated with a series of alcohols. After treatment with 0.1% trypsin working solution (composed of 0.4ml trypsin, 0.01g calcium and 0.01g chloride, 7ml D.W.) for 15 minutes, the sections were blocked using fetal bovine serum (FBS). . For immunohistochemical (IHC) analysis, sections were incubated o/n (over night) at 4 °C with a 1:50 dilution of primary antibody of MURF1, followed by DAB (3,3′-Diaminobenzidine) staining at room temperature. was carried out for 10 minutes. After that, the sections were dehydrated and mounted in the usual way. Five slides were randomly selected and analyzed for each group. Slides were examined using an Olympus IX71 Research Inverted Phase microscope (Olympus Co., Tokyo, Japan), and density was measured with ImageJ 1.47v software (National Institute of Health, Bethesda, MD, USA). The results of confirming changes in MURF1, a factor that decomposes muscle protein, through immunohistochemical staining, and the results of quantifying the expression level of MURF1 are shown in FIGS. 4 and 5 .

As a result, it was confirmed that the size of the root canal in the muscle tissue was decreased in the cancerous cachexia-induced disease control group, and the pathological decrease in which these muscles were lost was inhibited when administered with oxalic acid, thereby increasing the size of the muscle and root canal area. (Fig. 4 top and Fig. 5 left graph). In addition, as a result of confirming the change in MURF1, a muscle degrading enzyme, it was significantly increased when cancerous cachexia was induced, but it was confirmed that the increase in MuRF1 expression was significantly decreased when oxalic acid was treated (lower graph of FIG. 4 and right graph of FIG. 5) .

<Example 6> Improvement effect on changes in the expression of proteolysis-related factors in muscle caused by cancerous cachexia of Shin Ki-hwan Woo

In order to confirm the expression changes of MAFbx and MURF1, which are key factors for intramuscular proteolysis in the anterior tibialis anterior muscle induced with cancerous cachexia, western blot analysis was performed.

After a total of 5 weeks of treatment in the same manner as in Example 1, the muscle tissue (TA) of mice was mixed with RIPA buffer [50 mM Tris-HCl (pH7.5), 0.1% SDS (sodium dodecyl sulphate), 0.1% Triton X). -100, 1% Nonidet P-40, 0.5% sodium deoxycholate, 150 mM NaCl, 1 mM phenylmethylsulphonyl fluoride], followed by 8% SDS-polyacrylamide gel electrophoresis and transfer to a nitrocellulose membrane. After blocking with 5% skim milk, various primary antibodies and secondary antibodies to which horseradish peroxidase (Jackson Lab.) were attached were reacted, and the target protein was identified using ECL solution (Amersham Bioscience, Piscataway, NJ). The results of confirming the expression changes of the target proteins MAFbx and MURF1 through Western blot and the quantification of the relative protein expression levels are shown in FIG. 6 .

As a result, the protein expression of MAFbx and MURF1 was increased in the anterior tibialis anterior muscle tissue of mice induced with cancerous cachexia, and the expression of both factors was significantly decreased when the oxiasis was administered. Therefore, it was confirmed that the present invention can improve cancer cachexia by inhibiting protein degradation in muscle tissue induced by cancer.

So far, with respect to the present invention, the preferred embodiments have been looked at. Those of ordinary skill in the art to which the present invention pertains will understand that the present invention can be implemented in a modified form without departing from the essential characteristics of the present invention. Therefore, the disclosed embodiments are to be considered in an illustrative rather than a restrictive sense. The scope of the present invention is indicated in the claims rather than the foregoing description, and all differences within the scope equivalent thereto should be construed as being included in the present invention.

Claims (14)

A pharmaceutical composition for preventing or treating cachexia (cachexia) or muscle loss, including woochashingihwan. According to claim 1,
The cachexia is characterized in that caused by cancer, a pharmaceutical composition. According to claim 1,
The composition is characterized in that it improves one or more symptoms selected from the group consisting of decreased appetite, weight loss, increased fatigue, increased inflammation, muscle loss, fat loss, and hematopoietic toxicity, a pharmaceutical composition. According to claim 1,
The composition is characterized in that it reduces the expression of inflammatory cytokines in the hypothalamus, blood or bone marrow, a pharmaceutical composition. 5. The method of claim 4,
The inflammatory cytokine is one or more selected from the group consisting of CRP, TNF-α, IL-1β, IL-6, and IL-17, a pharmaceutical composition. According to claim 1,
The pharmaceutical composition is characterized in that it inhibits the expression of one or more genes selected from the group consisting of MAFbx, MuRF1 and Bnip3 in the muscle, the pharmaceutical composition. According to claim 1,
The woochasingihwan is Rehmanniae Radix, hyssop (Achyranthis Radix), Cornifolia (Corni Fructus), Sanyak (Dioscoreae Rhizoma), Chajeon (Plantaginis Semen), Taxa (Alismatis Rhizoma), Bokryeong (Poria Sclerotium), Peony (Poria Sclerotium), Peony (Peony Peony) Moutan Radicis Cortex) and cinnamon (Cinnamomi Cortex), characterized in that it comprises a pharmaceutical composition. A health functional food composition for preventing or improving cachexia (cachexia) or muscle loss, comprising woochashingihwan. 9. The method of claim 8,
The cachexia is a health functional food composition, characterized in that caused by cancer. 9. The method of claim 8,
The health functional food is a food in the form of tablets, capsules, pills or liquid, health functional food composition. A health food composition for preventing or improving cachexia or muscle loss, comprising woochashingihwan. 12. The method of claim 11,
The cachexia is characterized in that caused by cancer, health food composition. 12. The method of claim 11,
The health food is selected from various drinks, meat, sausages, bread, candy, snacks, noodles, ice cream, dairy products, soups, ionic drinks, beverages, alcoholic beverages, gum, tea and vitamin complexes, health food composition . An anticancer adjuvant composition comprising the pharmaceutical composition of claim 1.

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