JP5219013B2 - Evaluation method and screening method for substances having activity to alleviate side effects of drugs, and side effect mitigating agents containing substances identified by these methods as active ingredients - Google Patents

Description

  The present invention relates to a method for evaluating and screening a substance having an activity to alleviate side effects of a drug on an organism having an acquired immune mechanism, using an organism having only an innate immune mechanism, and an edible oil having the activity as an active ingredient It relates to a side effect mitigating agent.

  In developed countries, cancer remains the leading cause of death. As a method for treating cancer, administration of chemotherapeutic agents such as anticancer agents is still performed in many cases. However, many anticancer drugs, such as mitomycin C, cause serious side effects such as weight loss, which may reduce the patient's Quality of Life, and the side effects may cause sufficient amounts to treat cancer. In some cases, it cannot be administered (Non-patent Document 1). Therefore, solving side effects such as anticancer agents has become an important issue. The administration method which suppresses a side effect by the administration of an antiemetic against the nausea caused by the administration of an anticancer agent has already been established (Non-patent Document 2). Attempts have also been made to establish a remedy for serious side effects such as weight loss, but few have been established.

  Until now, we focused on a specific substance, and examined the usefulness of the substance against side effects using an evaluation model of a mammalian anticancer agent. However, in practice, it is considered that a method of screening a large number of substances and obtaining a substance exhibiting an effect is useful. In conventional methods using mammals, it is impossible to conduct experiments using a large number of individuals for screening due to physical problems, costs, and ethical viewpoints (Non-Patent Documents 3 and 4). . Therefore, in the development of new drugs against side effects of drugs such as anticancer drugs, there is a need to search for experimental animals that can replace these mammals.

By the way, the present inventors have focused on silkworm larvae as experimental animals that can replace mammals, and have so far utilized humans such as silkworm larvae that have only an innate immune mechanism, or cultured cells thereof. It has been reported that an evaluation system has been developed for developing therapeutic agents against bacteria and viruses that infect living organisms having acquired immune mechanisms (Patent Documents 1 and 2, Non-Patent Documents 5 to 7).
However, it has never been known until now whether an organism having only these innate immunity mechanisms can be a model for evaluating side effects of a drug on an organism having an acquired immune mechanism.

WO2001 / 086287 WO2005 / 116269 N. Ishiki, H .; Onishi, andY. Macida, Evaluation of antitumor and toxic side effects of mitocin, C-stradiol conjugates, Int J Pharm 279 (2004) 81-93. R. Sharma, P .; Tobin, and S.M. J. et al. Clarke, Management of Chemotherapy-Induced Nausea, Vomiting, oral mucosis, and diarhoeea, Lancet Oncol 6 (2005) 93-102. V. Baumans, Use of animals in experimental research: an ethical dilemma? Gene Ther 11 Suppl 1 (2004) S64-66. F. B. Orlans, T .; L. Beauchamp, R.A. Dresser, D.M. B. Morton, and J.M. P. Gluck, The Human Use of Animals: Case Studies in Ethical Choice, ed. , Oxford University Press, New York and Oxford 1998. H. Hamamoto, K. et al. Kurokawa, C.I. Kaito, K .; Kamura, I .; Manitara Razanajatovo, H.M. Kusuhara, T .; Santa, and K.C. Sequimiz, Quantitative evaluation of the therapeutic effects of antibiotics using silkworms. C. Kaito, K .; Kurokawa, Y .; Matsumoto, Y. et al. Terao, S .; Kawabata, S .; Hamada, and K.K. Sekimizu, Silkworm pathological bacteria infection model for identification of novel virulence genes, Mol Microbiol 56 (2005) 934-944. C. Kaito, N .; Akimitsu, H .; Watanabe, and K.K. Sekimizu, Silkworm larvae as an animal model of bacterial pathogenetic to humans, Microb Pathog 32 (2002) 183-190.

  The present invention has been made in view of the above situation, and it is an object of the present invention to solve various problems in the past and achieve the following objects. That is, an object of the present invention is to provide a model of a side effect of a drug on an organism having an acquired immune mechanism using an organism having only an innate immune mechanism. Another object of the present invention is to provide a method for evaluating and screening a substance that alleviates a side effect of a drug on an organism having an acquired immune mechanism, efficiently and easily using the model. To do. Another object of the present invention is to provide a side effect mitigating agent for alleviating the side effect of a drug using a substance evaluated or screened by the above method.

As a result of diligent studies to solve the above-mentioned problems, the present inventors have found that silkworms (arthropoda, submaxillary submaxillary) that have a fast generation change, can be easily reared in the laboratory, and have been genetically analyzed. Pay attention to the gates, insects and butterflies. Since silkworms have large larvae, C.I. Compared with small organisms such as elegans (which belongs to the order of the linear phylum, dinoid gland, Nematode elegans, and nematode), it is extremely easy to inject pathogens and drugs (Okada, E. et al. 1997. J. Seric. Sci. Jpn. 66: 116-122.). For this reason, a highly accurate experiment is attained by using a silkworm larva. In addition, the use of silkworm larvae is inexpensive, has no ethical problems, and is more useful than mammals.
Therefore, the present inventors tried to develop a model of a side effect of a drug using silkworm larvae. First, the present inventors examined whether silkworm larvae are useful as a side effect evaluation system for drugs. As a result, mitomycin C and bleomycin, which are known to exhibit side effects such as weight loss in mammals having acquired immune mechanisms, also inhibit the growth of silkworm larvae having only the innate immune mechanism. I found it. In addition, when screening for ingredients that act to alleviate the side effects from ingredients in the diet, it was found that soybean oil partially alleviates the side effects. Based on these findings, the silkworm larvae system was considered to be extremely effective for the evaluation and screening of substances that alleviate the side effects of drugs.

  The present invention is the world's first example of establishing an evaluation model of a side effect of a drug on an organism having an acquired immune mechanism using an organism having only the innate immunity mechanism. It is also the first example in the world that has been used to identify substances that actually reduce the side effects of drugs.

The present invention is based on the above knowledge, and means for solving the above problems are as follows. That is,
<1> A method for evaluating whether a test substance has an activity of alleviating side effects of a drug on an organism having an acquired immune mechanism,
(A) a step of administering the drug and the test substance to an organism having only an innate immune mechanism;
(B) detecting a side effect of the drug in an organism having only the innate immune mechanism, and
(C) A method comprising evaluating whether or not the test substance alleviates the side effect.
<2> A method for screening a substance having an activity to alleviate side effects of a drug on an organism having an acquired immune mechanism,
(A) a step of administering the drug and the test substance to an organism having only an innate immune mechanism;
(B) detecting a side effect of the drug in an organism having only the innate immune mechanism;
(C) a step of evaluating whether the test substance alleviates the side effect, and
(D) selecting a substance evaluated to alleviate the side effects.
<3> The method according to any one of <1> to <2>, wherein the drug is at least one of mitomycin C and bleomycin.
<4> The method according to any one of <1> to <3>, wherein the organism having only the innate immune mechanism is an organism belonging to insects.
<5> The method according to <4>, wherein the organism belonging to insects is a silkworm.
<6> A side effect mitigating agent for alleviating a side effect of a drug, characterized in that edible oil is an active ingredient.
<7> The edible oil is the side effect mitigating agent according to <6>, wherein the edible oil is derived from soybeans.

  ADVANTAGE OF THE INVENTION According to this invention, the conventional problems can be solved and the model of the side effect of the chemical | medical agent with respect to the organism which has an acquired immune mechanism using the organism which has only an innate immune mechanism can be provided. Further, according to the present invention, it is possible to provide a method for evaluating and screening a substance that alleviates the side effect of a drug on an organism having an acquired immune mechanism, efficiently and easily using the model. . In addition, according to the present invention, a side effect mitigating agent for alleviating a side effect of a drug using a substance evaluated or screened by the above method can be provided.

(Evaluation method, screening method)
The evaluation method of the present invention is a method for evaluating whether or not a test substance has an activity of alleviating side effects of a drug on an organism having an acquired immune mechanism, and includes the following steps (a) to (c): Including other steps as necessary.
Further, the screening method of the present invention is a method for screening a substance having an activity of alleviating side effects of a drug on an organism having an acquired immune mechanism, and includes the following steps (a) to (d), as necessary. And other steps.
The evaluation method and the screening method (hereinafter sometimes simply referred to as the “method”) include the evaluation or screening of a substance having an activity to alleviate the side effects of a drug on an organism having an acquired immune mechanism. It is characterized by using a living organism having only

<Process (a)>
In the method, first, a drug and a test substance for an organism having an acquired immune mechanism are administered to an organism having only an innate immune mechanism (step (a)).

-Organisms with only innate immune mechanism-
The “innate immunity mechanism” means an immune biological defense mechanism (innate immunity mechanism) that does not depend on an acquired immunity (acquired immunity) mechanism. Vertebrate animals have an acquired immune mechanism that protects the living body using molecules that specifically recognize invaders such as antibodies against the invasion of pathogens, but invertebrates and plants have such an acquired immune mechanism. I don't have it. That is, the “organism having only the innate immunity mechanism” is, in other words, invertebrates and plants having no acquired immunity mechanism.

  Therefore, the “organism having only the innate immunity mechanism” is not particularly limited and can be appropriately selected from invertebrates and plants according to the purpose. Among them, organisms belonging to insects are preferable. Can be mentioned. The “insects” refers to a class of arthropods from the Greater Submaxillary, which is composed of four subclasses of caterpillars, flying beetles, sterile insects, and moth insects. The “organism belonging to insects” is not particularly limited and may be appropriately selected depending on the intended purpose. However, larvae are preferred for convenience of handling. There is no restriction | limiting in particular as said larva, It can select suitably according to the objective, For example, the larva of the Lepidoptera (including moth and butterfly), Coleoptera (including beetle), etc. are mentioned. In addition, it is preferable that the said larva is a large larva from a viewpoint of the ease of administration of the said chemical | medical agent and the said test substance. The “large larva” refers to a larva having a body length of 1 cm or more. As the larvae, for example, silkworm (larvae) larvae are preferably mentioned.

  The organism having only the innate immunity mechanism other than the organism belonging to the insects is not particularly limited and can be appropriately selected according to the purpose. For example, arthropods other than insects such as spiders and scorpions , Mollusks such as slugs and snails, annelids such as earthworms, terrestrial animals such as starfish and sea urchins, linear animals such as leopards and roundworms, coelenterates such as hydra, sea anemones and jellyfish, rice, radish, etc. And other plants. These organisms can also be used as organisms having only the innate immune mechanism.

-Drugs for organisms with acquired immune mechanisms-
The “drug against organism having acquired immune mechanism” is not particularly limited and can be appropriately selected according to the purpose. For example, fish such as carp, birds such as chicken, mice, rats, rabbits, dogs, Examples include drugs for mammals such as cats, monkeys, and humans.

  The “drug” is not particularly limited as long as it exhibits side effects on an organism having an acquired immune mechanism such as a human, and can be appropriately selected according to the purpose. For example, an antibiotic anticancer drug (doxorubicin) , Pirarubicin, daunorubicin, mitomycin C, actinomycin D, bleomycin, etc., antimetabolites (methotrexate, fluorouracil, etc.), alkaloid anticancer agents (etoposide, irinotecan, etc.), alkylating agents (cyclophosphamide, etc.), others (cisplatin) Etc.). It is also possible to use drugs showing cytotoxicity such as chemotherapeutic agents for bacterial / fungal infections.

-Test substance-
The “test substance” is not particularly limited, and a desired substance for which the mitigating action against the side effect of the drug can be evaluated can be used. For example, a cell extract, a cell culture supernatant, a fermented microorganism product, a marine product Examples include biological extracts, plant extracts, purified proteins, crude purified proteins, peptides, non-peptidic compounds, synthetic low-molecular compounds, natural compounds, and the like.

-Administration-
The method for administering the drug or the test substance to an organism having only the innate immune mechanism is not particularly limited and can be appropriately selected according to the purpose. For example, intraperitoneal administration, into the blood Injection, addition to feed (food), injection into the intestine, and the like.
Further, the dose of the drug or the test substance to the organism having only the innate immune mechanism is not particularly limited. For example, the type of the target organism, the drug, the test substance, It can be appropriately selected depending on the degree of side effects. Generally, the dose per body weight in humans can be administered in terms of silkworm, and those skilled in the art can select an appropriate dose.

<Step (b)>
Next, in the method, the side effect of the drug in an organism having only the innate immune mechanism is detected (step (b)).
The side effect of the drug to be detected is not particularly limited and can be appropriately selected depending on the purpose. For example, suppression of weight gain, weight loss, killing of organisms having only the innate immunity mechanism, amount of food to eat Decreased (anorexia), increased susceptibility to bacterial infection (decreased immunity), decreased blood cell count (leukopenia), emesis, diarrhea, decreased exercise (depression), abnormal muscle contraction / abnormality Relaxation (convulsion, paralysis, etc.), periodic abnormalities of back vasoconstriction (abnormal heart rate) (in parentheses are examples of corresponding symptoms in an organism having an acquired immune mechanism).
Moreover, there is no restriction | limiting in particular as the detection method of the said side effect, According to the kind of side effect to detect, it can select suitably from well-known detection methods, for example.

<Step (c)>
Next, in the method, it is evaluated whether or not the test substance alleviates the side effect (step (c)).
As an evaluation method of whether to reduce the side effects, there is no particular limitation, can be appropriately selected according to the purpose, for example, compared with the case where the test substance is not administered (control), When the test substance is administered, when the degree of side effects of the drug in the organism having only the innate immune mechanism is reduced (relieved), the test substance is a drug against the organism having the acquired immune mechanism. It can be evaluated that it has the activity to alleviate the side effects of.
As mentioned above, the said evaluation method can be performed by the said process (a)-process (c). By the evaluation method, it can be evaluated efficiently and easily whether or not the test substance has an activity to alleviate the side effect of the drug on the organism having the acquired immune mechanism.

<Step (d)>
In the screening method, a substance evaluated to alleviate the side effects in the step (c) is further selected (step (d)).
Perform evaluations in the steps (a) to (c) using various test substances, and then select a substance that alleviates the side effects from various test substances in this step (d). Thus, it is possible to efficiently and easily screen for a substance having an activity to alleviate the side effect of a drug on an organism having the acquired immune mechanism.

In addition, the substance identified as a substance having an activity to alleviate the side effect of the drug on the organism having the acquired immune mechanism by the above method is, for example, chemical synthesis or purification depending on its structure or origin. Depending on the technique, it can be generated appropriately. The substance can be administered directly or mixed with a pharmaceutically acceptable carrier, for example, to a living body for the purpose of alleviating side effects.
In addition, the present inventors have actually found that edible oils have the activity of alleviating the side effects of drugs on organisms having the acquired immune mechanism by using the above method (see Example 2 below). . Therefore, the edible oil is useful, for example, as a side effect mitigating agent of the present invention described later.

(Side-effect reducing agent)
The side effect mitigating agent of the present invention can alleviate the side effects of the drug, contains edible oil as an active ingredient, and further contains other ingredients as necessary.

-Edible oil-
There is no restriction | limiting in particular as the said edible oil, According to the objective, it can select suitably, The plant origin, the animal origin, or what was synthesize | combined artificially may be sufficient. Among these, soybean oil is a preferred example of the edible oil.
There is no restriction | limiting in particular as content of the said edible oil in the said side effect mitigating agent, According to the objective, it can select suitably.

-Other ingredients-
Moreover, there is no restriction | limiting in particular as said other component, According to the objective, it can select suitably, For example, a pharmacologically acceptable carrier etc. are mentioned. The pharmaceutically acceptable carrier is not particularly limited and may be appropriately selected depending on the intended purpose. For example, when the side effect reducing agent is used in various dosage forms, depending on the dosage form. It can be selected appropriately.
There is no restriction | limiting in particular as content of the said other component in the said side effect reducing agent, According to the objective, it can select suitably.

-Dosage form-
There is no restriction | limiting in particular as said dosage form, For example, it can select suitably according to a desired administration method, For example, oral solid preparations (a tablet, a coated tablet, a granule, a powder, a capsule etc.), an oral liquid agent (Internal liquids, syrups, elixirs, etc.), injections (solutions, suspensions, solids for erection, etc.), inhaled powders and the like.

As the oral solid preparation, for example, an additive, an additive such as a binder, a disintegrant, a lubricant, a colorant, a corrigent / flavoring agent and the like are added to the active ingredient, if necessary, and manufactured by a conventional method. can do.
Examples of the excipient include lactose, sucrose, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, and silicic acid. Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxypropyl starch, methylcellulose, ethylcellulose, shellac, calcium phosphate, polyvinylpyrrolidone and the like. It is done. Examples of the disintegrant include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, stearic acid monoglyceride, and lactose. Examples of the lubricant include purified talc, stearate, borax, and polyethylene glycol. Examples of the colorant include titanium oxide and iron oxide. Examples of the flavoring / flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like.

The oral solution can be produced by a conventional method, for example, by adding additives such as a flavoring / flavoring agent, a buffering agent, and a stabilizer to the active ingredient.
Examples of the flavoring / flavoring agent include sucrose, orange peel, citric acid, tartaric acid and the like. Examples of the buffer include sodium citrate. Examples of the stabilizer include tragacanth, gum arabic, and gelatin.

As the injection, for example, a pH adjuster, a buffer, a stabilizer, a tonicity agent, a local anesthetic, and the like are added to the active ingredient, and injection for subcutaneous, intramuscular and intravenous injections is performed by a conventional method. Agent can be produced.
Examples of the pH adjusting agent and the buffering agent include sodium citrate, sodium acetate, sodium phosphate and the like. Examples of the stabilizer include sodium pyrosulfite, EDTA, thioglycolic acid, thiolactic acid, and the like. Examples of the isotonic agent include sodium chloride and glucose. Examples of the local anesthetic include procaine hydrochloride and lidocaine hydrochloride.

-Drug-
Further, the drug that is a target of side effect alleviation of the side effect alleviating agent is not particularly limited and can be appropriately selected according to the purpose. For example, as exemplified in the items of the evaluation method and screening method, Examples thereof include drugs that exhibit side effects on organisms having acquired immune mechanisms such as humans.

-Administration-
The side effect reducing agent can be administered orally or parenterally, for example, depending on the dosage form.
The organism to be administered with the side effect alleviating agent is not particularly limited and can be appropriately selected according to the purpose. For example, it may be an organism having an acquired immune mechanism such as a human or a silkworm, etc. It may be an organism having only an innate immune mechanism.
The dose of the side effect alleviating agent is not particularly limited. For example, when administered to humans, a doctor or the like considers the type of disease, the age, weight, symptoms, genetic background, etc. of the patient. Thus, it can be appropriately selected.

[effect]
The present inventors can evaluate a side effect of a drug on an organism having an acquired immune mechanism using an organism having only an innate immune mechanism, for example, silkworm larvae, and further, a substance having an activity to alleviate the side effect For the first time in the world. Candidate substances that relieve side effects of drugs more efficiently and more easily by using organisms that have only an innate immune mechanism, such as silkworm larvae, compared to the case of using mammals as in the past Can be isolated and identified. In addition, an organism having only the innate immunity mechanism is lower in cost and easier to handle than a mammal. Therefore, it is possible to reduce the experimental cost and the experimental space, that is, it is possible to develop a side effect mitigating agent that alleviates the side effect of the drug at low cost and simply.

Examples of the present invention will be described below, but the present invention is not limited to these examples.
The silkworm larvae used in this example were purchased from Ehime Sanyu (Ehime, Japan) from silkworm larvae eggs [Hu · Yo x Tukuba · Ne], hatched at 27 ° C. and grown. Silkworm larvae were fed with Silkmate 2S (Nosan Corporation, Yokohama, Japan) as a bait unless otherwise noted, and were reared until the 4th sleep. Mitomycin C was purchased from Kyowa Hakko Kogyo Co., Ltd., and bleomycin was purchased from Sigma.

(Example 1: Growth suppression by administration of anticancer agent in silkworm larvae)
Anticancer agents such as mitomycin C and bleomycin are known to exhibit side effects such as weight loss in mammals. Therefore, in order to clarify whether side effects due to anticancer agents can be examined in silkworm larvae, silkworm larvae were injected with either mitomycin C or bleomycin to examine whether growth was affected. Specifically, various doses of mitomycin C or bleomycin were administered to 5th day of silkworm larvae, food was added, they were raised at 27 ° C., and their body weights were individually measured over time. Ten animals were used per group.

  As a result, it was found that mitomycin C inhibited the growth of 5th instar silkworm larvae by administration of 0.1 mg and bleomycin administration of 10 μg (FIGS. 1 and 2). Therefore, it was shown that the presence or absence of side effects of the anticancer drug on the body weight of mammals can be examined using silkworm larvae.

Example 2: Alleviation of growth inhibition of silkworm larvae caused by mitomycin C by addition of soybean oil to the diet
Next, we examined whether there is a factor that acts to suppress the weight loss of silkworm larvae caused by anticancer drugs. Specifically, first, silkworm larva artificial feeds were prepared according to Table 1 by mixing powders / liquids and heating and melting them. Subsequently, the food excluding the respective components, or the diet added 5 times, was given to silkworm larvae (day 5) on which mitomycin C was injected into the blood, and weight gain was individually measured over time. Ten animals were used per group.

  As a result, it was found that soybean oil has an effect of partially alleviating the side effects of anticancer agents (FIG. 3). Thus, silkworm larvae have been shown to be useful as a system for evaluating and searching for substances that act to alleviate the side effects of anticancer drugs.

  From this example, the present inventors have found that administration of an anticancer agent to silkworm larvae causes the side effect of weight loss in silkworm larvae as in mammals (Example 1). Moreover, it discovered that the side effect of this anticancer agent was relieve | moderated by soybean oil (Example 2). These findings found by the present inventors mean that an organism having only an innate immune mechanism such as a silkworm larva can serve as a model of a side effect of a drug on an organism having an acquired immune mechanism such as a human. Therefore, according to this example, it is possible to evaluate the side effect of a drug on an organism having an acquired immune mechanism using an organism having only an innate immune mechanism, and to evaluate and search for a substance that acts to alleviate the side effect. Was shown to be possible.

  Since the evaluation method of the present invention and the screening method of the present invention can efficiently and easily evaluate and search for substances that act to alleviate side effects of drugs, for example, to reduce side effects such as anticancer drugs Therefore, it is considered useful for the development of side effect mitigating agents. In addition, the side effect reducing agent of the present invention is considered useful for alleviating side effects such as weight loss caused by anticancer agents such as mitomycin C.

FIG. 1 is a graph showing the growth inhibitory effect of mitomycin C on silkworm larvae. To 5th silkworm larvae, 0.1 mg of phosphate buffer (PBS) or mitomycin C was administered, food was given, and body weight was measured over time. Ten silkworm larvae were used for each group, and the average value was determined. FIG. 2 is a graph showing the growth inhibitory effect of bleomycin on silkworm larvae. To 5th-year silkworm larvae, 0.9% NaCl or bleomycin 1, 10, 100 μg was administered, food was given, and body weight was measured over time. Ten silkworm larvae were used for each group, and the average value was determined. FIG. 3 is a diagram showing the side effect mitigation effect of mitomycin C in soybean oil. Artificial feed to which normal amount of soybean oil was added and artificial feed to which 5-fold amount was added were given to mitomycin C 0.1 mg administration group and non-administration group to 5-year-old silkworm larvae, and weight gain was measured over time. Ten silkworm larvae were used for each group, and the average value was determined.

Claims (3)

A method for evaluating whether a test substance has an activity of alleviating side effects of a drug on an organism having an acquired immune mechanism,
(A) a step of administering the drug and the test substance to a silkworm that is an organism having only an innate immune mechanism;
(B) detecting a suppression of weight gain or a decrease in weight, which is a side effect of the drug in a silkworm that is an organism having only the innate immune mechanism; and
(C) The method of evaluating whether the said to-be-tested substance suppresses the suppression of the body weight increase which is the said side effect , or a reduction | decrease in body weight, The method characterized by the above-mentioned. A method for screening a substance having an activity to alleviate side effects of a drug on an organism having an acquired immune mechanism,
(A) a step of administering the drug and the test substance to a silkworm that is an organism having only an innate immune mechanism;
(B) a step of detecting suppression of weight gain or weight loss that is a side effect of the drug in a silkworm that is an organism having only the innate immune mechanism;
(C) a step of evaluating whether or not the test substance suppresses weight gain suppression or weight loss , which are the side effects; and
(D) including a step of selecting a substance that is evaluated to suppress the increase in body weight or the decrease in body weight, which are the side effects.   The method according to claim 1, wherein the drug is at least one of mitomycin C and bleomycin.