JP4102466B2 - Anti-cancer drug side effect reducing agent - Google Patents

Description

【００２７】
表１に示すように、５−フルオロウラシルまたはシクロフォスファミドを投与した実験群ｂ〜ｅでは、血球数の減少が見られた（骨髄毒性）。しかし、本発明抽出物を添加した実験群ｃとｅでは、血球数の減少が群ｂ、ｄよりも緩和されており、５−フルオロウラシルおよびシクロフォスファミド投与による副作用（骨髄毒性）の軽減作用が明確に認められる。
【００２８】
実験例３：抗癌剤誘発脱毛に対する軽減作用
実験動物に抗癌剤性脱毛を惹起するため、生後８日目、ＳＤラット（雌雄混合）にシトシンアラビノサイド２５ｍｇ／ｋｇを、８日間連続腹腔内投与した。また、本発明抽出物をシトシンアラビノサイドの投与と同期間、連続経口、腹腔、背部皮下および背部塗布投与し、脱毛予防作用を肉眼的観察により調べた。なお、経口、腹腔、背部皮下投与は、実施例１（３）の濃縮液を、固形含量５００ｍｇ／ｋｇで注入することにより、また背部塗布は、固形含量５０ｍｇ／ｍｌ溶液を１ｍｌ背部に塗布することにより行なった。また、脱毛度の評価基準は以下の通りである。
脱毛度Ａ：対照と同じレベルで、実質的に脱毛は認められない（正常）。
〃 Ｂ：体表面の半分以下の脱毛が認められる。
〃 Ｃ：体表面の半分以上の脱毛が認められるが全脱毛には至っていない。
〃 Ｄ：全脱毛。
【００２９】
【表２】

【００３０】
表２に示すように、本発明抽出物は、いずれの投与経路においてもシトシンアラビノサイドが誘発する脱毛を抑制した。
【００３１】
実験例４：抗癌剤誘発消化管障害および腎毒性に対する軽減作用
生後６週齢、雄性、フィッシャー系ラット５頭ずつからなる３つの群を用意し、対照群および２つの実験群とした。実験群のラットにはシスプラチン１ｍｇ／ｋｇを１２日間静脈内に連続投与し、うち一方の群については本発明の抽出物を添加しない飼料を与え、他方の群については実施例１（４）(b)の製剤を５％濃 度で混合した飼料を与え、それぞれ実験期間中自由摂食させた。投与最終日に採血を行ない、血液検査を実施した。消化管障害について血液検査した結果を表３に、腎毒性について血液検査した結果を表４に示す。
【００３２】
【表３】

【００３３】
【表４】

【００３４】
上記各表に示すように、シスプラチン投与により、一般血液状態の悪化が認められる。すなわち、グルコース、トリグリセライド、総蛋白質、アルブミンの減少、およびコリンエステラーゼの上昇から消化管障害の発症が示唆され、尿素体窒素およびクレアチニンの著増から腎障害の発症がそれぞれ示唆される。しかし、本発明の抽出物を接取したマウスでは血液状態の悪化が緩和されており、本発明抽出物は、シスプラチン投与により招来される上記に示すような各種の副作用を軽減する効果が明確に認められる。特に、表４の結果からは、シスプラチン投与による死亡例の主たる要因と考えられる、強度の腎障害が顕著に軽減されていることがわかる。
【００３５】
【発明の効果】
本発明のバシディオマイセテス（Basidiomycetes）培養抽出物は、抗癌剤投与による副作用を幅広くかつ顕著に抑制する。さらに、本発明抽出物の毒性は極めて低く、抗癌剤の副作用軽減剤としての有用性が高い。[0001]
BACKGROUND OF THE INVENTION
The present invention, anticancer agents to attenuate any adverse effects, particularly plant tissue raw material presence in Basi audio My ceteth of (Basidiomycetes, Basidiomycetes) regarding side effects reliever anticancer agents containing the components extracted bacteria from the liquid by cultivating belonging to .
[0002]
[Prior art]
In recent years, various types of anticancer agents have been developed. Anti-cancer agents have become the main treatment for many solid tumors as well as hematopoietic tumors. However, as is well known, most anticancer agents do not specifically act only on tumor cells, but also affect normal cells and induce side effects. Multi-drug combination therapy and short-term high-dose therapy have increased the effective rate of anticancer drugs and have been widely applied to clinical practice, but side effects due to increased dosage have become a more serious problem. There are many cases in which due to serious side effects, it is necessary to discontinue the medication and sufficient effects cannot be obtained.
[0003]
For example, anticancer agents currently used in clinical practice include alkylating agents such as nitrogen mustard and cyclophosphamide, antimetabolites such as 5-fluorouracil and cytosine arabinoside, antibiotics such as mitomycin and bleomycin, and others There are many types of anticancer drugs such as plant alkaloids, cisplatin, hormones, etc., and the side effects are almost bone marrow suppression, hair loss, vomiting, gastrointestinal disorders, hepatotoxicity, nephrotoxicity, cardiotoxicity, lung toxicity, stomatitis, skin disorders, neurotoxicity, etc. It extends to the whole body.
[0004]
Among them, bone marrow toxicity caused by anticancer drugs causes blood disorders. Anticancer drugs act on actively growing hematopoietic stem cells of the bone marrow, leukocytes / platelets, and erythroid immature cells to inhibit their division and maturation. The resulting bone marrow toxicity, such as leukocyte / thrombocytopenia and anemia, is the regulatory factor for most anticancer drugs.
[0005]
In addition, nephrotoxicity represented by cisplatin is a powerful side effect that sometimes threatens patients' lives. Cisplatin is an excellent anticancer agent useful for various types of cancer such as testicular tumors and ovarian cancer, but nephrotoxicity is a regulatory factor. In order to suppress nephrotoxicity, a large amount of circulatory fluid is administered at the same time as cisplatin administration to increase urine volume in the clinic, but a large amount of infusion is not an effective mitigation method because it places a burden on the circulatory system.
[0006]
In addition, hair loss, which is a side effect that appears strongly in relation to the dose and administration interval of an anticancer drug, is not a regulatory factor for anticancer drug administration, but is a serious problem that lowers the quality of life (QOL) of patients.
Conventionally, hair loss caused by an anticancer agent has been prevented by a scalp cooling method and a scalp tightening method, which are physical methods for reducing scalp blood flow. However, the scalp cooling method and the scalp tightening method temporarily reduce the blood flow of the head, so that the initial plasma half-life is long and the metabolite is not effective for drugs that exist in the blood for a long time. In addition, as an important problem, a case in which recurrence was observed in the scalp by the scalp cooling method has been reported.
Against this background, safe anti-cancer drug side effect reducing agents are currently being sought in order to improve the remission rate and QOL of patients by continuing administration of anti-cancer drugs.
[0007]
Various compounds have been studied as side effects reducing agents for anticancer agents. For example, JP-A-2-129914 describes that 4,4′-diaminodiphenyl sulfone, 4-aminodiphenyl ether or 4,4′-diaminodiphenyl ketone is effective for thrombocytopenia. . Japanese Examined Patent Publication No. 3-25407 describes that a neutropic compound has an effect of reducing hair loss. Japanese Patent Publication No. 5-2649 describes that fosfomycin is effective in alleviating nephrotoxicity. Although these compounds are effective against the side effects of specific anticancer agents, it is desired to provide side effect reducing agents effective against a wider range of side effects.
[0008]
[Problems to be solved by the invention]
An object of the present invention is to provide a new preparation which is non-toxic and can safely reduce a wide range of side effects caused by anticancer agents.
[0009]
[Means for Solving the Problems]
It is known that a culture extract of Basidiomycetes exhibits properties as a biological response modifier (BRM) (see JP-A-1-53701 and JP-A-8-259602). In view of the above problems, the present inventors have studied the further action of the culture extract of Basidiomycetes, and as a result, found the fact that various side effects induced by the anticancer agent are reduced, and have completed the present invention. It was.
[0010]
[Structure of the invention]
The present invention relates to the following side effects-reducing agents for anticancer agents.
1) A side effect reducing agent for an anticancer agent, comprising a component extracted from a mixture of a culture solution obtained by culturing a bacterium belonging to Basidiomycetes in the presence of a plant tissue raw material and a mycelium.
2) The side effect reducing agent of the anticancer agent according to 1 above, which reduces bone marrow suppression, hair loss, vomiting, gastrointestinal dysfunction, hepatotoxicity, nephrotoxicity, cardiotoxicity, pulmonary toxicity, stomatitis, skin disorder or neurotoxicity caused by an anticancer agent.
[0011]
The present invention is characterized by containing a component extracted from a mixture of a culture solution obtained by culturing a bacterium belonging to Basidiomycetes in the presence of a plant tissue raw material and a mycelium. Details of the bacterial species and culture conditions (these are the same as those described in JP-A-8-259602 above), extraction conditions, dosage forms, etc. are as follows.
[0012]
(1) Species Examples of bacteria belonging to Basidiomycetes used in the present invention include, for example, Lentinus edodes , Agaricus bisporus , and glyfora flow. Ndosa (Grifola frondosa, maitake), Foriota-nameko (Pholiota nameko, nameko), Puryurotasu-Osutoreatasu (Pleurotus ostreatus, oyster mushroom), Furamurina-Verachipesu (Flammulina velutipes, Flammulina), Gonoderuma-lucidum (Gonoderma lucidum, Ganoderma lucidum), Aurikararia・Auricularia auricula , Gonoderma applanalum , Coriolus lucidum , Grifola umbell, Grifola umbell ate, Ji Yoreimaitake), Schizophyllum-Commune (Schizphyllum commune, Schizophyllum commune), Vaulx-en-Vallier La Voruvaseae (Volvariella volvaceae, Fuku Rotake), and the like. These can be used alone or in combination of several kinds.
[0013]
(2) Culture conditions In the present invention, the above-mentioned bacteria are cultured in the presence of a plant tissue raw material.
The plant tissue raw material is not particularly limited as long as it is derived from plant tissue, and sawdust and the like can be used, but herbaceous plant-derived materials such as rice bran, bran, bagasse, corn rhizome, rice Rice bran, wheat straw, soybean meal and the like are preferable. These may be used alone or in combination. By using such raw materials, an active ingredient can be obtained efficiently. In these raw materials, a component that is soluble in hot water is particularly useful, and therefore, an extract extracted with hot water is preferably used.
[0014]
Various carbon sources or nitrogen sources may be added to the medium in addition to the plant tissue raw material. Examples of the carbon source include glucose, shochu, maltose, saccharose, sucrose, brown sugar, molasses, molasses, malt extract and the like. Examples of nitrogen sources include meat extract, peptone, gluten meal, soy flour, dry yeast, yeast extract, ammonium sulfate, ammonium tartrate, urea and the like. In addition, if necessary, add inorganic salts such as sodium salt, magnesium salt, manganese salt, iron salt, calcium salt, phosphate, and vitamins such as inositol, vitamin B1 hydrochloride, L-asparagine, biotin, etc. May be. The culture may be carried out in accordance with the usual culture of mesophilic bacteria, and is carried out at a pH of 2 to 6, 10 to 45 ° C, preferably 15 to 30 ° C. The duration of culturing is usually 4 to 20 days, preferably about 6 to 12 days, although it depends on the amount of bacteria and the form of plant tissue raw material.
[0015]
(3) The active ingredient of the side effect reducing agent of the extracted anticancer agent can be obtained by extracting from the mixture of the culture solution and mycelium obtained as described above. For extraction, enzymes such as cellulase, amylase, protease, pectinase and chitinase are added to the above mixture, and the reaction is carried out for 2 to 20 hours under optimum temperature conditions to disrupt the cells. Inactivation is carried out by removing mycelium residues from the treated product by centrifugation or the like. The liquid part obtained contains the active ingredient of the present invention. The specific structure and the like of the active ingredient is unknown, but the main component is a polysaccharide having a substituent, in addition to substances derived from plant tissue materials, substances derived from mycelium, fungal metabolites, etc. These are considered to act synergistically.
[0016]
(4) Dosage Form and Formulation The side effect reducing agent of the present invention is a pharmacologically and pharmaceutically acceptable manufacturing aid that is subjected to a treatment such as drying as necessary to the mixture obtained as described above. It is manufactured according to a conventional method by adding. Examples of production aids include sucrose, starch, mannitol, sorbitol, lactose, glucose, cellulose, calcium phosphate, calcium carbonate and other excipients as well as conventional binders, disintegrants, lubricants, preservatives, Stabilizers, dispersants, diluents, fragrances, sweeteners and the like can be mentioned. Both oral administration and parenteral administration are possible.
Oral administration agents can be solid preparations such as powders, granules, capsules, microcapsules, tablets and troches, or liquid preparations such as drinks, syrups and elixirs. In addition, examples of parenteral administration agents include injections, ointments, suppositories, and the like.
The dose of the side effect reducing agent of the anticancer agent of the present invention may be increased or decreased depending on the age, weight and symptoms, dosage form, etc. of each patient to be treated, but generally 0.01 to 10 g, preferably 0.5 to It is administered 1 to 4 times a day with 5 g as a daily dose.
[0017]
(5) Acute toxicity The side effect reducing agent of the present invention has very low toxicity and can be judged to be sufficiently safe for use as a medicine.
[0018]
(6) Side effect reducing effect The side effect reducing agent of the present invention is effective against various anticancer agents. Specifically, alkylating agents such as cyclophosphamide, antimetabolites (nuclear base derivatives) such as 5-fluorouracil and cytosine arabinoside, antibiotics such as mitomycin and bleomycin, other plant alkaloids, platinum such as cisplatin Reduce side effects caused by compounds and hormones. In particular, it is excellent in reducing side effects of alkylating agents, antimetabolites (nuclear base derivatives), and platinum compounds. Side effects that are alleviated are widespread and include bone marrow suppression, hair loss, vomiting, gastrointestinal disorders, hepatotoxicity, nephrotoxicity, cardiotoxicity, lung toxicity, stomatitis, skin disorders and neurotoxicity.
[0019]
【Example】
EXAMPLES Hereinafter, although an Example and a test example demonstrate this invention further in detail, the scope of the present invention is not limited by these examples. The numerical values in the tables shown in the examples, except for Table 2, were converted to 100% as the value of the control test and expressed as percentages (%).
[0020]
Example 1:
(1) Using a glass petri dish with a culture diameter of 90 mm, inoculate bacteria from Lentinus edodes cultured in a solid medium (maltose 1%, peptone 0.2%, ammonium tartrate 0.2%, agar 1.5%). 8 liters (Add 800 ml of water to 150 g of rice bran and heat at 120 ° C. for 15 minutes, then add 10 g of maltose, 2.5 g of peptone and 2.0 g of ammonium tartrate to the filtered extract and add an appropriate amount of water. PH 4.0) The inoculated 10 liter culture bottle was inoculated and cultured at 20 ° C. for 7 days with aeration. Subsequently, 8 liters of this culture solution was inoculated into a culture tank containing 300 liters of a liquid medium having the same composition, and cultured with gentle agitation at 23 ° C. for 9 days under aeration.
[0021]
(2) Extraction The mixture of the culture solution and mycelium obtained by the above culture is heated to 90 ° C., 8 g of amylase is added and reacted for 3 hours, then cooled to 60 ° C., and 15 g of cellulase and 15 g of protease are added. The reaction was carried out at 55 ° C. for 10 hours. The enzyme was inactivated by heating at 120 ° C. for 20 minutes. The mycelium residue was removed from the cultured product by centrifugation to obtain an extract. The concentration of solid content in the extract was 2.5% by weight.
[0022]
(3) Concentration The extract obtained in the extraction step is concentrated 10 times under reduced pressure to obtain a concentrate.
[0023]
(4) Formulation
(a) Formulation Example A: The extract of Example 1 (2) is aseptically dispensed into vials to obtain a drink.
(b) Formulation Example B: 4.8 kg of cyclodextrin and 19.2 kg of branched dextrin are added to 330 liters of the concentrated liquid of Example 1 (3), freeze-dried by a conventional method, and pulverized to obtain 80 kg of powder.
(c) Formulation Example C: Microcapsules are prepared by a conventional method at a blending ratio of 100 kg of the powder obtained in (b) above and 40 kg of hardened oil to obtain 130 kg of microcapsules.
[0024]
Experimental Example 1: Acute toxicity test To confirm the safety of the extract of the present invention, a toxicity test was conducted by single oral administration. The animals used were male 4-week-old male and female 5-week-old female SD rats in the same number of males and females. The preparation of Example 1 (4) (b) was administered once at a maximum dose of 12500 mg / kg, and the presence or absence of lethality within one week was observed. As a result, neither male nor female death was observed in the maximum dose group. That is, it was shown that the extract of the present invention has high safety.
[0025]
Experimental Example 2: Reducing effect on anticancer drug-induced myelotoxicity Groups ae consisting of 5 male mice and 5 ddY mice were prepared, and the following drugs were administered to each group.
(a) Group a (control): No drug administration
(b) Group b: Cyclophosphamide 100 mg / kg was administered intraperitoneally once a day for 14 consecutive days.
(c) Group c: Cyclophosphamide 100 mg / kg was administered intraperitoneally once a day for 14 consecutive days. In addition, the preparation of Example 1 (4) (b) was mixed with feed at a concentration of 5% and allowed to eat freely during the same period.
(d) d group: 5-fluorouracil 50 mg / kg was intraperitoneally administered once a day for 14 consecutive days.
(e) e group: 50 mg / kg of 5-fluorouracil was intraperitoneally administered once a day for 14 days. In addition, the preparation of Example 1 (4) (b) was mixed with feed at a concentration of 5% and allowed to freely eat during the same period.
The a, b and d groups were fed freely with the same feed as the c and e groups except that the extract of the present invention was not added. Blood was collected on the 14th day after administration, and the number of red blood cells and white blood cells were counted. The results are shown in Table 1.
[0026]
[Table 1]

[0027]
As shown in Table 1, in the experimental groups b to e administered with 5-fluorouracil or cyclophosphamide, a decrease in blood cell count was observed (bone marrow toxicity). However, in the experimental groups c and e to which the extract of the present invention was added, the decrease in the blood cell count was mitigated as compared with the groups b and d, and the side effect (bone marrow toxicity) alleviated by administration of 5-fluorouracil and cyclophosphamide. Is clearly recognized.
[0028]
Experimental Example 3: Reducing effect on anticancer drug-induced hair loss In order to induce anticancer drug-induced hair loss in experimental animals, cytosine arabinoside 25 mg / kg was administered intraperitoneally for 8 days to SD rats (mixed male and female) on the 8th day after birth. Further, during the same period as the administration of cytosine arabinoside, the extract of the present invention was administered continuously orally, abdominally, subcutaneously on the back and applied on the back, and the hair loss preventing action was examined by visual observation. For oral, abdominal and subcutaneous dorsal administration, the concentrated solution of Example 1 (3) is injected at a solid content of 500 mg / kg, and for back application, a solid content 50 mg / ml solution is applied to the back of 1 ml. Was done. Moreover, the evaluation criteria of the degree of hair loss are as follows.
Degree of hair loss A: At the same level as the control, virtually no hair loss is observed (normal).
〃 B: Hair loss of less than half of the body surface is observed.
〃 C: Hair loss of more than half of the body surface is observed, but total hair loss has not been reached.
Ｄ D: Total hair loss.
[0029]
[Table 2]

[0030]
As shown in Table 2, the extract of the present invention suppressed hair loss induced by cytosine arabinoside in any administration route.
[0031]
Experimental Example 4: Mitigating Action on Anticancer Agent-Induced Gastrointestinal Disorder and Renal Toxicity Three groups of 6 weeks old, male, 5 Fischer rats were prepared, and used as a control group and two experimental groups. Rats in the experimental group were continuously administered intravenously with 1 mg / kg of cisplatin for 12 days, and one group was given a feed to which the extract of the present invention was not added, and Example 1 (4) for the other group. A feed prepared by mixing the formulation of b) at a concentration of 5% was given and allowed to eat freely during the experiment. Blood was collected on the last day of administration and a blood test was performed. The results of blood tests for gastrointestinal disorders are shown in Table 3, and the results of blood tests for nephrotoxicity are shown in Table 4.
[0032]
[Table 3]

[0033]
[Table 4]

[0034]
As shown in the above tables, general blood conditions are worsened by cisplatin administration. That is, a decrease in glucose, triglyceride, total protein, albumin, and an increase in cholinesterase indicate the onset of gastrointestinal tract, and a marked increase in urea body nitrogen and creatinine suggests the onset of renal damage. However, in the mice treated with the extract of the present invention, the deterioration of the blood state was alleviated, and the extract of the present invention clearly has the effect of reducing various side effects as shown above caused by cisplatin administration. Is recognized. In particular, it can be seen from the results in Table 4 that severe renal damage, which is considered to be the main cause of death due to cisplatin administration, is significantly reduced.
[0035]
【The invention's effect】
The culture extract of Basidiomycetes of the present invention broadly and remarkably suppresses side effects caused by administration of anticancer agents. Furthermore, the toxicity of the extract of the present invention is extremely low, and its usefulness as an anticancer drug side effect reducing agent is high.

Claims (1)

Bone marrow suppression, hair loss, gastrointestinal tract disorder, characterized by containing a component extracted from a mixture of a culture solution of mycelia and Lentinus edodes ( Lentinus edodes ) in the presence of plant tissue raw materials Anti- cancer side effect reducing agent selected from nephrotoxicity .