ITTO990784A1 - PROCEDURE FOR THE STRENGTHENING OF ANTI-CANCER ACTIVITIES OF FARMACIGREZZI, COMPOSITION CONTAINING A RAW DRUG STRENGTHENING THE ACTIVITY - Google Patents

Description

DESCRIPTION

of the text of the patent for Industrial Invention

BACKGROUND OF THE INVENTION

The present invention relates to techniques such as a strengthening procedure, a procedure for evaluating and the like of the antitumor activity of crude drugs such as mushrooms containing polysaccharides having antitumor activity, particularly to techniques for increasing the manifestation of the antitumor effects in internal administration or the expected efficacy of antitumor activity in internal administration and the like.

Various types of mushrooms have so far been used in the field of Chinese medicine, dietary foods or the like. For example it is known that mushrooms such as GANODERMA (REISHI) {Fomes (SARUNOKOSHIKAKE)), PORIA (BUKURYO), maitake mushroom (MAITAKE, Grifola frondosu Dickson), and the like have antitumor activity such as carcinostatic activity, etc. Much research has been conducted relating to the active ingredients in mushrooms that exhibit anticancer activity, and it has generally been said that it is the polysaccharides contained in mushrooms that exhibit effective antitumor activity.

In fact, in the field of Chinese medicine, dried mushrooms are treated in infusion and used in hell as decoctions.

Various types of Chinese drugs have so far been used as natural medicines as they have fewer side effects than synthetic medicines. In the Japanese Pharmacopoeia, numerous Chinese drugs have been described as crude drugs.

On the other hand, with reference to the anticancer activities of such mushrooms, their infusions or extracts are administered to experimental animals such as mice, etc., together with food, comparing the mice of a control group to which they were not administered. Chinese drugs with those of a group to which such drugs have been administered to evaluate, for example, the condition of disappearance or transfer of the cancer, or the survival rate of mice or the like, in order to evaluate their effectiveness.

Furthermore, in these experimental animals, toxicity and efficacy are also examined, and after confirmation of sufficient safety for use in humans, these drugs are administered infernally and the efficacy of antitumor activity in humans is confirmed. through clinical tests.

SUMMARY OF THE INVENTION

Antitumor activities such as carcinostatic property, property of preventing cancer transfer, etc. of mushrooms, are already known as mentioned above, and their effectiveness is well known. They are used by infusing a dried mushroom product, ingesting the decoction or mixing an effective extracted ingredient with alcohol, etc. with other components such as vitamin preparations for preparing healing beverages or foods or medical agents.

With internal administration by ingestion of a decoction, such as the conventional one, the cases in which medical effects can be obtained and therapeutic effects are observed are unexpectedly few. With the agaricus mushroom {AGA-RIKUSUTAKE, which is also called "Agaricus") which has recently attracted attention for its remarkable medical effects, the same trend can be observed. Even in the case of the maitake mushroom (MAITAKE, Grifola frondosu Dickson), which has recently attracted attention, there are few examples to demonstrate its medical effects.

However, among the cases in which no medical effect is observed, there are cases in which an effective anticancer activity is clearly highlighted, albeit in an extremely limited number of examples.

The present inventor has therefore examined whether there is an effective method for obtaining the antitumor activity of the fungi in any case by studying the mechanism according to which differences in the manifestation of the antitumor effects of the fungi can occur.

Even when the efficacy of fungi having antitumor activity is confirmed in experimental animals, as mentioned above, the expected effects cannot be seen in almost all cases when it is actually administered internally to humans. To confirm the effect through internal administration in clinical experiments, it is necessary to continue the administration for a certain period of time by checking the administration system under constant conditions and with continuous observation. It would therefore take a considerable time to obtain a result by ascertaining whether or not the effects of the fungi are exhibited by internal administration to a patient.

This procedure is certainly effective but, for example, to examine a substance having an effective antitumor activity by selecting a certain number of natural products, an enormous amount of time is required, so it is necessary to develop a technique that allows to evaluate the antitumor activity. in a shorter period of time.

Furthermore, when a patient who is under clinical control suddenly has complications and an out-of-control treatment is urgently applied to the patient, confirmation of efficacy as far as the patient is concerned cannot be obtained. Hence, in a clinical trial over a long period of time, a large number of cases cannot be obtained to confirm efficacy so that it is not easy to perform the clinical trial over a long period of time while maintaining the efficacy of the test itself.

The present inventor has therefore deemed it necessary to develop a method that can highlight the manifestation of a certain degree of antitumor efficacy when a sample is administered to humans without carrying out such a clinical test.

Furthermore, in Chinese medicine, in addition to mushrooms, numerous substances with antitumor activity are known, and it is preferred to try to solve the aforementioned problems relating to mushrooms in order to extend them to all raw drugs.

An object of the present invention is to effectively develop the antitumor activity of crude drugs such as mushrooms and the like containing polysaccharides which have antitumor activity.

Another object of the present invention is to predict the anticancer activities of crude drugs such as mushrooms and the like when administered to humans, without carrying out clinical tests.

Another object of the present invention is to provide a composition containing crude drugs such as mushrooms and the like containing polysaccharides endowed with antitumor activity, so as to effectively manifest their antitumor activity.

The aforementioned objects and other objects and new features of the present invention are described in detail in the present description.

The present invention relates to a process for strengthening the antitumor activity which is applied to a crude drug containing polysaccharides having antitumor activity and comprises a step of roasting the aforementioned crude drug by means of far infrared rays and a fermentation step by adding microorganisms to strengthen the anticancer activity of the aforementioned raw drug with respect to the case in which roasting and fermentation are not carried out.

After the aforementioned fermentation step, the present invention comprises a step of preparation of an oily agent in which the aforementioned fermented raw drugs are contained, with an oily component of vegetable origin, such as for example toasted sesame or the like by means of far infrared rays. .

The present invention further encompasses the use of a mushroom that contains β-glucan.

The present invention also includes the use of at least one mushroom selected from the group consisting of the agaricus mushroom (AGARIKUSUTAKE, Agaricus blazer), the maitake mushroom (MAITAKE, Grifola frondosu), the shiitake mushroom {SHIITAKE, Cortinellus Shiitake), the matsutake mushroom ( MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKE, Flammulina velutipes).

When using the aforementioned agaricus mushroom (AGARIKUSUTAKE), the present invention comprises roasting the agaricus mushroom by far infrared rays.

The composition with enhanced antitumor activity containing the crude drug of the present invention comprises a crude drug to which the method of enhancing the antitumor activity of the crude drug corresponding to any of the aforementioned types has been applied.

The present invention is a method of evaluating the efficacy of the antitumor activity of the treatment to be applied to the crude drug containing polysaccharides having antitumor activity, which comprises the addition of the crude drug to which the aforementioned treatment is applied, to a system that forms peroxide. by ultraviolet irradiation of an unsaturated acid such as docosahexenoic acid and the evaluation of the treatment effect of strengthening the antitumor activity of the aforementioned crude anticancer drug which increases with increasing the ratio of the amount formed of the aforementioned lipid peroxide based on the increase of the concentration ratio of the aforementioned crude drug by treatment.

The present invention comprises mushrooms containing β-glucan for use as the aforementioned crude drug.

The present invention is a method of evaluating the efficacy of the crude drug for evaluating the efficacy of the antitumor effect presented by the crude drug containing polysaccharides having antitumor activity without performing clinical tests, which includes adding the crude drug to a system which form lipid peroxide by ultraviolet irradiation of an unsaturated fatty acid such as docosahexenoic acid and evaluation of the expected effect of antitumor activity by internal administration of the aforementioned crude drug which increases with increasing the ratio of the formed amount of the above lipid peroxide based on the high ratio of concentration of the aforementioned crude drug.

The present invention comprises the application of either of the aforementioned antitumor activity enhancement methods to the aforementioned crude drug.

The present inventor has conducted various studies on a substance that inhibits active oxygen in Chinese drugs and, by means of these studies, he has confirmed that active oxygen or lipid peroxides have significant effects on the human body and these substances lead to the breakdown of the tissues of the human body whereby they become the cause of various infectious diseases.

In the tissues of the human body, there is an enzyme called SOD (superoxide dismutase) to protect the tissues of the human body from this active oxygen and to eliminate the active oxygen. The amount of SOD decreases with age and the removal of active oxygen in the body can no longer be achieved. Its decrease is particularly noticeable beyond the age of 40, and the removal of the active oxygen present in the body cannot be sufficiently achieved as has been made clear in recent years, and it becomes the cause of various types of diseases in adults.

Also, when an excessive amount of active oxygen is formed in the body by chronic or acute stimulation or by chemicals, etc., the removal of active oxygen cannot be achieved sufficiently with the amount of SOD present in the body initially, so that the person is affected by various diseases.

Hence, for various diseases caused by insufficient elimination of active oxygen due to SOD deficiency, SOD administration treatment to resolve the SOD deficiency was considered. However, according to such treatment, effects can be obtained by administering the drug by injection, but not a noticeable effect is obtained in the case of a drug administered internally, so that it is currently believed that the effects have not been demonstrated.

With reference to the point where the effects as internal medicine are not highlighted, the present inventor has discovered that SOD is unstable in gastric juice and the molecular weight of SOD is 30,000 or more and therefore is not absorbed by the digestive organs as such. Furthermore, he found that there is an action limit such that SOD acts only on superoxide (02-) among the four existing types of active oxygen.

The present inventor has thoroughly investigated a substance that can remove excess active oxygen in the human body and can be administered internally, and as a result has found a composition which includes vegetable oils obtained from roasted plants which are added to a substance obtained by roasting under suitable conditions seeds or sprouts of plants and then proceeding to fermentation by addition of microorganisms, and this composition can be used as an agent for the elimination of active oxygen effective for internal administration (see Japanese patent No. 2,125,887).

In the aforementioned plant seeds or sprouts, there are low molecular weight active anti-oxygen substances such as flavonoids, polyphenols, tannin, tocopherol, vitamin B2, etc. The present inventor has found that such low molecular weight active anti-oxygen substances bind chemically with each other or with other components, and form a molecular compound or a complicated complex or a macromolecular compound by absorption or inclusion and when these are ingested in such conditions, an active oxygen suppression action expected from a low molecular weight active oxygen suppression substance cannot be obtained.

Therefore, when ingested in such a condition, many people cannot digest the substance by transforming it into low molecular weight active anti-oxygen substance by means of gastric juices, so that the active oxygen suppression action cannot be achieved. The present inventor emphasized this point in "Food Industry", vol. 35, n.

14, "Development and Improvement of DDS, SOD-like Function Foods from Natural Plants and Seeds and Their Pharmacological and Biochemical Consideration".

The present inventor has studied the way to effectively use the aforesaid active anti-oxygen substance contained in the seeds or sprouts of plants for the treatment or prevention of diseases, and the way to activate the aforesaid active anti-oxygen substances.

As a result, he found that by heating and roasting plant seeds or sprouts under moderate conditions, the active anti-oxygen substance contained in the plant seeds or their sprouts is partially released from the aforementioned complex forming an original low molecular weight substance or is partially activated by generating an active functional group due to the effect of chemical change. By carrying out these treatments, the active anti-oxygen action is significantly increased compared to that present before roasting.

On the other hand, many of the anticancer activity functions of fungi, such as carcinostatic property or tumor displacement inhibiting property, are generally based on polysaccharides such as β-glucan, etc., but such β-glucan is different from the aforementioned. substance with low molecular weight anti-active oxygen contained in the seeds or sprouts of plants, and does not fall within the scope of the aforementioned series of studies on active oxygen conducted by the present inventor.

However, the present inventor has examined the fact that the manifestation of the anticancer effects of mushrooms, administered as a decoction internally in the aforementioned way, is found on some people but such effects are not obtained for other people, it can be explained in a similar way due to the low molecular weight of the aforementioned active anti-oxygen substance.

Thus, while the action and mechanism of the antitumor activity of β-glucan has not been sufficiently elucidated, the fact that there are differences in the manifestation of the anticancer effects when the mushrooms are ingested indicates that there are patients in whom the active ingredient of fungi that manifest antitumor activity acts easily and patients in whom it acts with difficulty.

According to the way of internal administration, the ingested mushrooms are treated sooner or later by the gastric juices. Hence, the author considered that the fact that the effective ingredient exhibiting antitumor activity present in the aforementioned mushrooms acts effectively or not, is closely related to this treatment by gastric juices. Thus, the present inventor has believed that there are individual differences in gastric juice treatment and these individual differences cause the differences in the manifestation of the anticancer effects.

Therefore, the present inventor has constructed the following hypothesis between the ease of the manifestation of the antitumor action in internal use and the treatment of gastric juice in consideration of the clinical fact that there are individuals with gastric juices of different strength.

Thus, the effective ingredient that exhibits antitumor activity contained in mushrooms, which becomes the purpose of treatment with gastric juice, exists in the condition where the effects are hardly manifested and, as mentioned before, it is transformed to a condition in which the effects are easily manifested as a consequence of the action of gastric juices with differences from individual to individual. Therefore, if a person has, by its nature, a more energetic gastric juice, the ingredient having effects on the antitumor activity contained in the ingested mushrooms, assumes the state in which the action can be easily highlighted so that the anticancer effects are manifested. .

However, in a person who has a naturally weak gastric juice, the ingredient having anticancer effects is not transformed to the condition in which the action can be easily detected, so that the anticancer effects are not highlighted. Many people do not have strong gastric juices which can change the effective ingredient that exhibits antitumor activity of fungi in such a way that its action is easily manifested, and therefore it can be assumed that a number of examples that can confirm the efficacy of antitumor activity of fungi can hardly be observed by internal administration.

We can thus explain the fact that, in the case of a person who by nature has strong gastric juices, while the components of the fungi that have antitumor efficacy come in the form of a chain as, for example, in the case of polymerization, and exist in a stable condition, that is, in the inactive state in which they cannot be activated, these components are broken down by the gastric juice, altering their composition and transforming them into the activated state so that antitumor activity can be manifested in the body.

It can be assumed that many people's gastric juices are unable to break down the inactive chain into ingredients that may have anticancer efficacy. Hence, it can be explained that the effective ingredient for cancer is absorbed by the intestines in such a non-activated state that it cannot show its effect.

The present inventor has thus hypothesized that, if a treatment which can in advance activate the effective ingredient can be easily applied to the mushrooms, the aforementioned problems can be solved.

The present inventor has attempted various systems to activate the effective ingredients of such mushrooms in the inactive state, but has not easily succeeded in achieving an effective means. This is because sufficient knowledge of the state of effective ingredients cannot be obtained when they are in the non-activated state, as the current system is too complex to evaluate.

The present inventor has conducted various experiments, as mentioned above, and as a final experiment he has attempted to apply the aforementioned method as a means of activating active oxygen suppressing substances such as flavonoids, polyphenols, etc., although they have quite a composition different from that of β-glucan.

When such a procedure is to be adopted, the means refers to the activation of flavonoids and the purpose is quite different from the application to β-glucan, so the non-activated condition is considered to be quite different, and therefore differences in the method of activation of β-glucan which initially appear difficult.

However, the experimental results were quite different from the predictions of the present inventor and showed that the activation medium of the active oxygen suppressing substance can be effective for changing the structure of β-glucan even if it is different from that of substances that they suppress active oxygen such as flavonoids, etc. and can be activated easily.

Hence, the present inventor has found that mushrooms which are previously toasted by far infrared rays and then used in the experiment, exhibit an improved antitumor effect when administered internally compared to the case of unroasted mushrooms by far infrared irradiation. . Furthermore, it has been found that when infrared roasted mushrooms are fermented with the addition of microorganisms, particularly "Koji" (Aspergillus oryzae) or yeast, the anticancer effects are further enhanced.

As mushrooms containing polysaccharides which have anticancer activity, one can use for example the agaricus mushroom (AGARIKUSUTAKE) which has recently attracted particular attention for its medical effects. Furthermore, mushrooms that have so far been considered to have powerful anticancer activity such as Fomes (SARUNOKOSHIKAKE), PORIA (BUKU-RYO), etc. can be used without problems.

Incidentally, with reference to the agaricus mushroom (AGARIKUSUTAKE), it has been found that, when it is toasted with infrared rays and fermented under various conditions after harvesting and drying, a sufficient effect is not obtained. Hence, it is preferable to subject to the treatment in undried conditions, i.e. in the raw state, i.e. shortly after harvesting.

In addition, the agaricus mushroom (AGARIKUSUTAKE) is easily spoiled and, if left for a long time before infrared roasting treatment, it should not be left in the raw state for long. Therefore, in this case it is preferable to keep it in the refrigerator at a low temperature and carry out the toasting with infrared rays within three days of harvesting. Furthermore, after the aforementioned infrared roasting and fermentation treatment, it has been found that the anticancer effects are more enhanced when the resulting product is subjected to an oily treatment. For example, an oily component obtained from plants such as sesame, etc., subjected to infrared roasting, is added to the mushrooms covering the resulting material after fermentation treatment with the oily component.

As mentioned earlier, it could not initially have been foreseen that the means for transforming the active anti-oxygen substance contained in plant seeds and sprouts into a low molecular weight compound could also be effectively employed for the manifestation of antitumor activity. of mushrooms containing polysaccharides having antitumor activity which have a rather different structure, for example, from the active anti-oxygen substance. However, it can be clarified that significant effects can be obtained on the material on which the experiments leading to the present invention were performed.

The present inventor has also found that the antitumor activity that deserves more attention can be manifested by carrying out the aforementioned treatment even when the fungi have not attracted specific attention from the point of view of the antitumor activity. For example, it has been found that infrared roasting of the maitake mushroom (MAI-TAKE, Grifola frondosu Dickson), echoes. and following fermentation treatment with microorganisms, an antitumor effect comparable or superior to that of the agaricus mushroom (AGARIKUSU-TAKE) can be obtained.

The anticancer effect of fungi such as the maitake mushroom (MAITAKE, Grifola frondosu Dickson) was already known, but was generally not thought to have an effect comparable to that of the agaricus mushroom (AGARIKUSUTAKE) which has recently attracted attention. By applying the aforementioned treatments of the present invention to mushrooms, the antitumor effects of mushrooms such as the specifically possessed maitake mushroom (MAITAKE, Grifola frondosu Dickson) are sufficiently manifested and an antitumor activity comparable to that of the agaricus mushroom (AGARIKUSUTAKE) is highlighted.

Therefore, the potential antitumor activity possessed by fungi cannot currently be sufficiently highlighted and the efficacy cannot be clearly confirmed. However, by applying the treatment of the present invention to fungi, their anticancer activities are clearly shown and the antitumor activity can be manifested to a level that could not even be predicted.

In each of the mushrooms, β-glucan is more or less present and the aforementioned treatment is considered a procedure capable of strongly promoting its action and manifestation. Therefore, in addition to the maitake mushroom (MAITAKE, Grifola frondosu Dickson), the treatment of the present invention can be adopted for mushrooms that have normally been used as raw food products, such as the shiitake mushroom (SHIITAKE, Lentinus edodes), the matsutake mushroom (MATSUTAKE, Tricholoma matsutake), the shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes), the enokitake mushroom (ENOKITAKE, Flammulina velutipes), etc. As a result, it has been found that the process can strongly develop the potential antitumor activity present in the fungi themselves.

When the above process of the present invention is adopted to enhance the antitumor activity of fungi, the β-glucan of fungi that can be found in a simple and inexpensive way can be activated without the use of rare and expensive fungi, and even when the amount of β-glucan is lower than that of rare mushrooms, a significant antitumor efficacy can be developed compared to the case in which rare mushrooms are used with the conventional method. This leads to a reduction in the cost to the patient.

Furthermore, the composition containing fungi to which said antitumor activity strengthening procedure is applied, significantly increases its antitumor activity compared to the conventional composition in which the fungi to which antitumor activity is attributed are dried and pulverized and simply mixed.

Furthermore, during the series of the aforementioned mushroom researches, mushrooms are added by progressively increasing their concentration to the system in which lipid peroxide is formed by ultraviolet irradiation of an unsaturated fatty acid such as docosahexenoic acid, etc., and the quantities of Formed lipid peroxides are determined with the respective fungi at various concentrations. As a result it can be seen that the greater the increase in the amount of lipid peroxide formed by the fungi, the greater the antitumor effect obtained with internal administration.

Then, various types of fungi are added to the system which forms the aforementioned lipid peroxide with the same concentration, and the amounts of the respective lipid peroxides formed are measured. When the amount of lipid peroxide formed is greater, the anticancer effects can occur more potently with internal administration.

If this procedure is adopted, it is possible to evaluate the anticancer effects presented by the mushrooms without having to resort to internal administration. Even when a clinical test is not performed on a patient to experience internal administration, the method can be used to evaluate the efficacy or antitumor effect by internal administration.

Furthermore, for example, various types of treatment are applied to the mushrooms, and the amounts of lipid peroxide formed are compared by adding the mushrooms to which the respective treatment is practiced, thus evaluating the effectiveness of the treatments themselves.

Furthermore, the above explanation is made independently of the fungi, and the process can be applied to a crude drug containing polysaccharides having antitumor activity, other than fungi. The application is effective on at least one crude drug containing β-glucan.

The process can be applied to crude drugs other than fungi, such as natural crude drugs including those derived from plants, crude drugs derived from animals and crude drugs derived from minerals. Also, many of the Chinese medicines use naturally grown raw drugs, but there can be raw drugs collected in a crop under suitable conditions or they can be grown by artificial culture such as culture water, etc.

Also, as crude drugs, crude drugs other than those described, for example, in the Japanese Pharmacopoeia, etc., can be used if they exhibit effective antitumor activity when applying the present invention, and such products may be natural or cultured products.

DESCRIPTION OF THE FAVORITE REALIZATIONS

In what follows, embodiments of the present invention will be described in detail, with reference to examples.

As regards the enhancement of the anticancer activity of crude drugs, a composition containing crude drugs with enhanced anticancer activity, a method of evaluating the anticancer efficacy of the treatment of crude drugs and a method of evaluating the anticancer efficacy of crude drugs, will be explained with the use of fungi which are well known as crude drugs containing β-glucan which is a polysaccharide having antitumor activity.

(REALIZATION 1)

In embodiment 1 of the present invention, a process for strengthening the antitumor activity of crude drugs and a composition containing crude drugs strengthened as antitumor activity is described.

In the procedure for strengthening the anticancer activity of raw drugs, mushrooms containing β-glucan are first toasted by means of far infrared rays, then microorganisms such as "Koji", etc. are added. and fermentation is carried out in predetermined conditions of humidity and temperature. Once the fermentation phase is complete, the resulting product is suitably transformed into a fine powder and this is covered with an oily component obtained by sesame squeezing which has been toasted by far infrared to prepare an oily agent.

Furthermore, a product obtained by subsequently subjecting mushrooms containing β-glucan to the aforementioned infrared roasting step, to the fermentation step by means of microorganisms and to the step of preparation of the oily agent, becomes a composition showing an effective antitumor activity by internal administration, containing fungi to which the procedure of strengthening the antitumor activity has been applied.

In what follows, the present invention will be explained in more detail with reference to examples.

(EXAMPLES)

As mushrooms to be used in the present example, β-glucan containing mushrooms are employed. In particular, the anticancer effects are strongly manifested when using mushrooms which contain a lot of β-glucan and have general anticancer effects, which is preferred.

In the present example, PORIA (BUKURYO), agaricus mushroom (AGARIKUSUTAKE), POLYPORUS (CHOREIMAITAKE, Polyporus umbellatus FRIES), maitake mushroom (MAITAKE, Grifola frondosu Dickson), matsutake mushroom (MATSUTAKE, Tricholoma matsutake) are used as mushrooms. (SHIITAKE, Cortinellus Shiitake). As regards the agaricus mushroom (AGARIKUSUTAKE), the present inventor has experimentally found that it has a more powerful antitumor activity when used in its raw state than when used after drying, so it is stored in a refrigerator at a low temperature (4 ° C) after harvesting and used within 3 days of harvesting.

With reference to other fungi, no major differences are observed when they are used in the raw state and in the dried state, as hitherto provided as Chinese medicine, applying the procedure of strengthening the antitumor activity of the present invention so that in the present example dry materials of PORIA (BUKURYO), POLYPORUS (CHOREIMAITAKE, Polyporus umbellatus FRIES), maitake mushroom (MAITAKE, Grifola frondosu Dickson), matsutake mushroom {MATSUTAKE, Tricholoma matsutake) and shiitake mushroom (SHIITAKE, Cortìnellus Shiitake)

Incidentally, in the present embodiment, two types of maitake mushrooms are employed. Those produced in Japan are mainly grown in Niigata prefecture which is a snowy country and are formally called snow maitake mushrooms (Yukiguni maitake) or black maitake mushrooms (Kuromaitake), and in the present description they are referred to as maitake (black) mushroom.

On the other hand, the maitake mushroom (MAITAKE, Grifola frondosu Dickson) from China is mentioned as POLYPORUS (CHOREIMAITAKE, Polyporus umbellatus FRIES), in this description to distinguish it from the aforementioned maitake (black) mushroom as appropriate. In particular, in POLYPORUS (CHO-REIMAITAKE, Polyporus umbellatus FRIES), the root is mainly used.

The first step of the antitumor activity enhancement process of the present example is a far infrared roasting step in which the various aforementioned species of mushrooms are roasted by far infrared heating. The infrared rays to be used in this roasting step are preferably infrared rays having a wavelength from about 4 to 14 μm. Mushrooms are roasted using a device such as a plate, oven, etc., in which a ceramic that emits infrared radiation at the wavelength in the aforementioned field is applied to the internal surface of an equipment such as a plate, oven, etc. ., mixing ceramic powder that radiates far infrared, with stone, sand, gravel, etc. of metal oxides.

To carry out this roasting, you can use mushrooms other than the agaricus mushroom (AGARIKUSUTAKE), which are the ones that were normally dried in normal Chinese medicines. With reference to the agaricus mushroom (AGARIKUSUTAKE), this is used in its raw state as mentioned above.

The aforementioned mushrooms are roasted in the aforementioned device, for example, in an earthenware container prepared from substances that emit far infrared radiation such as granite, ceramic, Tenshouseki stone, etc. and emit infrared radiation with a wavelength between 4 and 14 μm, without burning them, for a time from 30 to 90 minutes by shaking them gradually and continuously.

The roasting process is not limited to the above process, and any process can be adopted as long as it promotes the release of the β-glucan of the mushrooms. For example, as a source of far infrared to be used for roasting, in addition to the aforementioned devices, others can be used as long as they include a product capable of emitting infrared rays with a wavelength between 4 and 14 μm, such as platinum wave fiber electromagnetic, etc.

After this infrared roasting phase, the fermentation phase is carried out. After the application of the infrared roasting treatment in the aforementioned way, fermentation is carried out using bacteria such as Koji "or yeast, in an environment containing humidity and at a temperature from 30 to 36 ° C for a time from 48 to 72 hours. Fermentation can be done in a fermenter to decrease the fermentation time.

As a microorganism to be used as fermentation it is also possible to use a microorganism other than "Koji".

Furthermore, to carry out the fermentation, in addition to the microorganism "Koji", it is possible to use a substance with fermentative power such as ripe papaya, Pompeii juice, fig bark, grape skin, young bamboo bark, etc. Furthermore, digestive enzymes such as diastase, pancreatin, etc., proteolytic enzymes derived from microorganisms such as protease, pepsin, trypsin, etc., lytic enzymes of polysaccharides such as hemicellulase or a precursor substance which forms the aforementioned digestive enzymes or proteolytic enzymes, etc. .

However, according to the experiments performed by the present inventor, the most preferred results can be obtained by carrying out the fermentation with the use of microorganisms and among these the "Koji" is the one which provides the strengthening of the greatest antitumor activity. Once this fermentation phase is completed, the starting products are pulverized.

For spraying, commercial sprayers can be used, but a machine that causes high temperatures during spraying should preferably be avoided. Pulverization must be done with a system that does not generate high temperatures, such as a stone mill.

The products of the starting mushrooms to which the infrared toasting phase are applied in the fermentation phase, are then subjected to a treatment phase with an oily agent. This procedure for forming the oily agent is for the purpose of enclosing the aforementioned fine powder of the starting product after fermentation into an oily component obtained from sesame which has been roasted with far infrared irradiation.

As an oily component to be used for the preparation of the oily agent, an oil (hereafter referred to as sesame paste oil) collected from toasted sesame can be used. Sesame paste oil is an oil obtained by subjecting raw sesame to infrared irradiation at a temperature not exceeding 100 ° C and slowly for an extended period, then grinding and compressing the sesame after roasting. In this sesame paste oil, there remain fine solid products that are formed when the sesame is ground so that it takes on a pasty appearance.

To this sesame paste oil the aforementioned pulverized mushrooms are added after fermentation to enclose the mushrooms in a fine powder with an oily component of the sesame. By enclosing the mushrooms, their permeation strength in the cells of the diseased part is improved compared to the case in which such an oily paste shell is not used. This step of preparing the oily agent is an important step in providing effective targeting in view of the drug delivery system (DDS) which has attracted attention in recent years.

Incidentally, the step of preparing the oily agent is an important step as stated, and providing such step is specifically preferred, but if there is any difficulty in procuring such an oily agent, the step of its preparation can be omitted. The reason for this statement lies in the fact that, as indicated below, it was found through tests that if the infrared roasting phase and the fermentation phase are currently performed, the effectiveness of strengthening the antitumor activity becomes significantly great. even when the step of preparing the oily agent is not carried out, as mentioned below.

Also, for preparing an oily agent, more preferred results can be obtained when using a mixed oil in which the aforementioned sesame oily paste and an oil collected from crude sesame are mixed at a suitable ratio.

By mixing the oil obtained from raw sesame, the oil droplets can be made smaller, thus improving the ability to penetrate the target cells significantly compared to the case where the mushrooms are enclosed with sesame paste oil having high viscosity and large droplets.

The oil collected from raw sesame is an oil obtained by grinding raw sesame as such and squeezing the resulting material, then separating the solid residue, yielding a normal sesame oil of the commercially available type.

The mixing ratio of sesame paste oil with the aforementioned sesame oil obtained from raw sesame can vary depending on the amount of the starting fine powder that must be added by suitably passing from 1 to 3 parts by weight of sesame oil for one part of sesame paste oil. Further, the mixing ratio of the oil with the starting fine powder can be, for example, about 4 to 5 parts by weight of starting fine powder per 1 part by weight of the oil.

Fungi in which the anticancer activities have been enhanced by the above process are added to a system which forms lipid peroxide by irradiation with ultraviolet rays of docosahexenoic acid, controlling the amount of lipid peroxide formed. Furthermore, their anticancer effects by internal administration are also evaluated through clinical tests.

Furthermore, to compare the strengthening of the antitumor activity of the aforementioned procedure, the amount of lipid peroxide formed above and the efficacy in clinical tests are checked with reference to the case in which only the infrared roasting to the aforementioned mushrooms is used, to the case in which only the fermentation of the polysaccharide with lytic enzyme is applied, in the case in which only the "Koji" fermentation is applied, in the case in which the infrared roasting and the 'Koji "fermentation is applied and in the untreated case in which no treatment is performed.

To effect the above fermentation with the polysaccharide lytic enzyme, various polysaccharide lytic enzymes can be used but, in the present example, hemicellulase is used which is a representative case among such enzymes.

Incidentally, in the case in which only toasting is adopted, in the far infrared of the mushrooms, in the case in which only fermentation with the lithic enzyme of the polysaccharide is adopted, in the case in which only "Koj" fermentation is adopted and in the case in which both roasting and "Koji" fermentation are adopted, at the moment in which the respective treatments are carried out, the material is pulverized into a fine powder with the above-mentioned procedure, and used in the subsequent experiments.

The degree of pulverization of the respective samples is such as to have a variable average particle size, so as to exclude the effect of the pulverized particle size alone. In the case of untreated product, the agaricus mushroom (AGARIKUSUTAKE) is used in finely chopped form in the raw state, and the other mushrooms indicated above, which are used to prepare fine powders after drying.

The measurement of the aforementioned amount of lipid peroxide formed is performed as indicated below. A known quantity of docosahexenoic acid is placed in various tubes, and to these are added different quantities of the above mentioned mushrooms previously weighed and then mixed in the tubes. Furthermore, the contents of the respective tubes are irradiated with ultraviolet rays, and the amount of lipid peroxide formed is determined.

Docosahexenoic acid is used by previously preparing a diluted solution in which the starting liquid is diluted with ethanol 10 times and the diluted solution is further diluted so as to obtain a final concentration diluted 200 times.

Samples of mushrooms to which the above treatment is applied (including untreated ones) are added to a predetermined quantity of ethanol in order to obtain a mushroom concentration of 60 mg / ml and the ethanol is left at room temperature for a period of time. about 2 to 6 weeks to perform the ethanol extraction. To use the sample in the test, the extract is used by diluting to the final concentration of 0.6 mg / ml.

By mixing 0.05 ml of the obtained dilute docosahexenoic acid solution prepared in the aforementioned manner, 0.1 ml of a mushroom sample and 0.85 ml of ethanol in a test tube are brought to 1 ml. This tube is irradiated with ultraviolet rays for 3 hours to form a lipid peroxide. The lipid peroxide formed is measured by the TBA reaction (thiobarbituric acid reaction).

By heating the lipid peroxide formed under acidic conditions, the released malonodialdehyde (MDA) reacts with thiobarbituric acid (TBA) and the maximum absorption is determined to indirectly obtain the amount of lipid peroxide formed.

To carry out the TBA reaction, 0.2 ml of a 7% solution of sodium dodecyl sulfate, 2 ml of 0.1 N hydrochloric acid, 0.3 ml of phosphotungstic acid and 1 ml of the reagent prepared by mixing 0.67% of TBA and acetic acid with a ratio of 1: 1, are added to the respective tubes irradiated with ultraviolet rays, and the reaction with TBA is carried out by keeping the tubes at 95 ° C for 12 minutes after addition.

The mixture is then cooled to room temperature, and the reaction layer with TBA is extracted with n-butyl alcohol, then applying the fluorospectrophotometric method to this layer to measure the aforementioned quantity of MDA. To perform the measurement, light with a wavelength of 515 nm and an excitation light and fluorometry at a wavelength of 535 nm are used. The fluorospectrophotometer used is the F2000 type manufactured by Hitachi Ltd., using a 400 V photomultiplier, and the measurement is performed by passing the band through irradiation of excited light at 20 nm by measuring the fluorescence at 20 nm.

With reference to the effects of docosahexenoic acid on the fungi to which the respective aforementioned treatments are applied by ultraviolet irradiation in the lipid peroxide formation system, they are examined to evaluate the MDA content (which is equal to the amount of MDA detected by absorbance) measured in the above manner. The results are reported in Table 1.

On the other hand, among the aforementioned mushrooms, PORIA (BUKURYO), agaricus mushroom (AGARIKUSU-TAKE), maitake mushroom (MAITAKE, of the black maitake type (Kuromaitake) (snow maitake mushroom (Yukiguni maitake)) and POLYPORUS ( CHOREIMAITAKE, Polyporus umbellatus FRIES), clinical tests are carried out on breast cancer, stomach cancer and lung cancer in the respective cases of untreated fungi, after fermentation treatment with lytic polysaccharide enzyme, "Koji" fermentation treatment, treatment of far infrared roasting plus "Koji" fermentation treatment, far infrared roasting treatment plus "Koji" fermentation plus oily agent preparation treatment The results are shown in Tables 2, 3 and 4.

Table 2 shows the results of treatments on breast cancer patients with fungi to whom the various types of treatment above have been applied. Tables 3 and 4 show the effects of treatment on stomach cancer and lung cancer patients with the various types of fungi to which the same treatments have been applied as in Table 2. Incidentally, in Tables 2 and 4, they are also reported the actual ratios calculated by the following formula.

Effectiveness ratio = (Number of effective cases Number of ineffective cases) x 100 / (Number of effective cases Number of ineffective cases Number of ineffective cases)

Table 1 Effects of various fungi on MDA production

The respective mushrooms are added in quantities of 6 mg / ml

Table 2 Effects of treatment with various fungi on breast cancer patients

The number in brackets indicates the total number of patients examined.

Effectiveness ratio =

The same formula is used for the calculation in Tables 3 and 4.

Table 3 Effects of treatment with various fungi on stomach cancer patients

The number in brackets indicates the total number of patients examined.

Table 4 Effects of treatment with various fungi on

The number in brackets indicates the total number of patients examined.

From the above Table 1, it can be understood that the value of MDA which is an index of the quantity

of lipid peroxide formed becomes greater, with the same type of mushroom, when roasting far infrared compared to the untreated case, when fermentation with the lytic enzyme of the polysaccharide is performed compared to the case in which it is performed, the far infrared roasting only, when performing 'Koji "fermentation compared to fermentation with polysaccharide lytic enzyme, when performing both far infrared roasting and" Koji "fermentation compared to fermentation only 'Koji "and when performing the three treatments of far infrared roasting," Koji "fermentation and preparation of the oily agent compared to the case in which far infrared roasting and" Koji "fermentation are performed.

This tendency is common regardless of the type of mushroom used in the tests, except for the case of PORIA (BUKURYO) between far infrared roasting and fermentation with lytic enzyme of the polysaccharide, and the case of the matsutake mushroom (MATSU-TAKE, Tricholoma matsutake), between fermentation with the lytic enzyme of the polysaccharide and fermentation "Koji".

Basically the same MDA value can be obtained (to be precise, the value of one of the processing methods is about 1% higher than the other method) when performing far infrared roasting and when fermenting with a lytic enzyme of the polysaccharide on the PORIA (BU-KURYO), and when performed with the fermentation of the lytic enzyme of the polysaccharide and the fermentation "Koji" on the matsutake mushroom (MATSUTAKE, Tricholoma matsutake).

On the other hand, from Tables 2 to 4 which represent the clinical test results of the administration of PORIA (BUKURYO), agaricus mushroom (AGA-RIKUSUTAKE), maitake mushroom (MAITAKE, black maitake mushroom (Kuromaitake) (snow maitake mushroom ( Yukiguni maitake)) and POLYPORUS (CHOREIMAITAKE, Polyporus umbellatus FRIES), who have undergone the respective aforementioned treatments, in patients with breast cancer, stomach cancer and lung cancer it can be understood that the efficacy ratios in the respective cases of breast cancer, stomach cancer and lung cancer patients becomes higher, for the same type of mushroom, when applying the lytic enzyme fermentation of the polysaccharide compared to the untreated case, when applying the "Koji" fermentation compared to to the case where the lytic enzyme fermentation of the polysaccharide is applied, when both the far infrared roasting and the "Koji" fermentation are applied compared to the case in c ui only "Koji" fermentation is applied, and when both far infrared roasting and "Koji" fermentation and the preparation of the oily agent are applied compared to the case in which far infrared roasting and fermentation are adopted "Koji".

For example, from Table 2, in PORIA (BUKU-RYO), it can be seen that the efficacy ratio increases when applying far infrared roasting plus "Koji" fermentation plus preparation of the oily agent than in the non-case. treated, from 7.5% to 20.0%, i.e. about three times. With the agaricus mushroom (AGARIKUSUTAKE), the effectiveness ratio increases from 7.5% to 35%, that is, approximately 4.9 times. With the maitake mushroom (MAITAKE), the efficacy ratio strongly increases, passing from an anticancer effect of 10.0% to 57.5%, i.e. 5.75 times. This trend is the same in Tables 3 and 4.

From the above results, it can be seen that the treatment method in succession with the far infrared roasting step, the "Koji" fermentation step, and the oily agent preparation step, can be effectively adopted as a method for increasing or manifesting properties of antitumor activity for fungi, that is, as a method of strengthening antitumor activity.

Table 5 Amount of free β-glucan in the respective types of untreated and treated mushrooms by far infrared roasting and "Koji" fermentation

On the other hand, the amount of free β-glucan as an effective ingredient that exhibits antitumor activity is measured in the case of the treatment to reinforce the manifestation of the aforementioned antitumor activity and in the case of the untreated product. The mushrooms used are agaricus mushroom (AGARIKUSU-TAKE) and maitake mushroom (MAITAKE, black maitake mushroom (Kuromaitake) which shows a high efficacy ratio in Tables 2 to 4. Incidentally, the analyzes are carried out at the Food Analysis Center in Suita , Osaka, Japan The results are shown in Table 5.

From the results of Table 5, it will be understood that the amounts of free β-glucan increase when the far infrared roasting treatment is performed plus the "Koji" fermentation treatment on agaricus mushroom (AGARIKUSUTAKE) and on. maitake mushroom (MAITAKE). With reference to the maitake mushroom (MAI-TAKE), as well as with the black maitake mushroom (Kuromaitake), the same effects can also be obtained with POLYPORUS {CHOREIMAITARE, Polyporus umbellatus FRIES). On the other hand, in the results of the clinical tests of Tables 2 to 4, it can be seen that the anticancer effects are higher in the case of application of the far infrared roasting treatment plus the "Koji" fermentation treatment compared to the case of the non-product. treaty.

Therefore, by examining the results of Tables 2 to 4 and those of Table 5 in combination, it can be considered that among the β-glucans effective as antitumor activity in fungi, particularly when the amount of free β-glucan is large, the antitumor effects are manifest in a remarkable way.

The present inventor has continued the aforementioned series of studies by advancing the aforementioned hypothesis, that in untreated fungi the β-glucans that have antitumor action are in a non-decomposable state by means of a link between them forming a chain, which is the so-called position not activated, and when the mushrooms are administered internally the chain cannot be cleaved by the gastric juice of a normal person so the antitumor activity would not be presented as expected. However, as mentioned above, it can be found that the increase in the quantity of free β-glucan strongly influences the manifestation of the antitumor action and the results that can be obtained according to the hypothesis of the present inventors.

Furthermore, the composition produced by the far infrared roasting step, the fermentation step and the oily agent preparation step which constitute the method of strengthening the antitumor activity, as previously mentioned is, as seen in Table 5 , etc., a composition in which the amount of free β-glucan is increased and effectively exhibits antitumor activity by internal administration. Compared to the composition containing fungi (untreated fungi) to which the above method of strengthening the antitumor activity is not applied, its antitumor activity is consistently strengthened.

Furthermore, from Table 1, it can be seen that the order of magnitude of the MDA value in mushrooms changes according to the types of treatment, including untreated ones. In the order indicated in Table 1, with reference to the three types of PORIA (BUKU-RYO), Matsutake mushroom (MATSUTAKE) and shiitake mushroom (SHIITAKE), these present a possibility of changing the efficacy of the antitumor activity in the three procedures treatment from the untreated type, "Koji" fermentation and far infrared roasting plus' Koji "fermentation.

For example, it can be seen that in the case of the untreated type, the antitumor activity becomes high in the order of shiitake mushroom (SHIITAKE), matsutake mushroom (MATSUTAKE) and PORIA (BUKURYO), while if infrared roasting is applied far and the "Koji" fermentation the antitumor activity becomes high in the order of PORIA (BUKURYO), Matsutake mushroom (MATSUTAKE) and shiitake mushroom (SHIITAKE). It will be understood that, by applying the above method of strengthening the antitumor activity according to the present invention, it is possible to increase the evaluated order for untreated fungi which has been estimated lower as antitumor activity.

In the field of Chinese medicine that uses a natural raw drug, there are few species that cannot ensure a stable yield even when it is a fungus with high antitumor activity. However, when adopting the aforementioned method of the present invention, instead of such few species, it is possible to use mushrooms which can ensure sufficient yield or can be cultivated. This can stabilize the supply and lower the price of the product so that many people can effectively enjoy the anticancer effects of mushrooms.

Incidentally, in the aforementioned explanation as the best embodiment, only the treatment in the three steps of far infrared roasting, fermentation treatment and preparation of the oily agent is explained. However, if only far infrared roasting is applied, or only fermentation with lytic enzyme of the polysaccharide such as hemicellulose, etc., or only fermentation treatment by microorganisms such as "Koji", etc., or only roasting at all ' far infrared and fermentation treatment, there is an increase in the manifestation of anticancer effects compared to the untreated case.

Thus, even when there are reasons why the preparation of the oily agent cannot be carried out due to the condition of the equipment, the treatments of far infrared roasting and microorganism fermentation can be carried out in combination and the anticancer activity can be achieved. considerably developed compared to the untreated case.

Also, in the treatment where the above-mentioned mushrooms are applied the three kinds of treatment of far infrared roasting, fermentation treatment and oily agent preparation, or two kinds of treatment are applied to mushrooms such as far infrared roasting and the fermentation treatment, in addition to the fermentation treatment with the use of microorganisms such as "Koji" or yeast bacteria, a fermentation treatment with a lytic enzyme of the polysaccharide can be used.

In the preparation phase of the oily agent in the aforementioned method of strengthening the antitumor activity, a sesame paste oil obtained from toasted sesame is used, but it is also possible to use vegetable oils other than sesame obtained by toasting soybeans, corn, safflower , primrose, rice husk, rapeseed, etc., in far infrared, before pressing.

• Furthermore, in the composition containing the fungi to which the aforementioned procedure for strengthening the antitumor activity is applied, in addition to the products obtained by treating fungi in the aforementioned way, other components can be added that do not bring negative side effects and show antitumor activity, such as vitamin agents, etc.

As a composition having such a constitution, a preparation developed by the present inventor having an active oxygen suppression effect can be formulated. The crude drug treated to enhance antitumor activity, containing a compound having such a constitution, may in fact be prepared in the form of an agent which can be easily administered internally by encapsulating in a gelatin capsule, etc.

Also, using a suitable excipient, binder, etc., a tablet can be prepared, or it can be in the form of granules or pills without any difficulty. Furthermore, although the present invention is a method of strengthening the antitumor activity by internal administration, the possibility of preparing a product for injection using mushrooms to which the method of strengthening the antitumor activity of the present invention has been applied can be kept in mind, providing the effect of increasing the manifestation of the antitumor activity with respect to the case in which the method of strengthening the antitumor activity of the present invention is not applied.

Incidentally, in the foregoing, the case in which an agaricus mushroom (AGARIKUSUTAKE) is used has been explained, but the invention is not necessarily limited only to mushrooms of the aforementioned type, and any mushroom can be used as long as it contains β-glucan. For example, shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes), enokitake mushroom (ENOKITAKE, Flammulina velutipes), etc. can be used.

Furthermore, a crude drug containing a polysaccharide having an antitumor activity other than β-glucan can be used. Furthermore, the present invention can be used as a process for enhancing the manifestation of the antitumor effect with a polysaccharide having antitumor activity such as β-glucan in a natural or cultivated product derived from plants which also include fungi.

(Realization 2)

In the present embodiment, the method of evaluating the efficacy of the antitumor activity for a crude drug and a method of evaluating the antitumor efficacy of the treated crude drug are explained.

To the test system in which the lipid peroxide is formed by ultraviolet irradiation explained in the previous embodiment 1, a crude drug is added to evaluate the antitumor activity, such as fungi, and the amount of lipid peroxide formed is checked. When mushrooms are added by increasing the amount, a mushroom which has a tendency to greatly increase the amount formed of lipid peroxide exhibits potent anticancer activity when administered.

Hence, when the increased ratio of the formed amount of lipid peroxide is large relative to the increased concentration ratio of the employed mushroom, the mushroom can be considered to have a powerful antitumor effect when administered internally. Thus, various types of fungi are introduced into the aforementioned experimental system, and when comparing the ratio of increase in the amount of lipid peroxide formed to the ratio of the concentration of the fungus employed, the order of efficacy of the antitumor effect per administration can be determined. internal of various types of mushrooms.

In the following, the two aforementioned evaluation methods of embodiment 2 are explained based on the above examples. When the MDA values with respect to the respective treatment method of Table 1 previously mentioned are compared with each other, it will be understood in which case any other treatment such as infrared roasting, etc. applied, will lead to a higher MDA value than in the case of no treatment.

When performing the far infrared roasting treatment or the "Koji" fermentation, the MDA value is higher in the mushrooms to which the "Koji" fermentation has been applied. When applying "Koji" fermentation and far infrared roasting in combination, the MDA value is higher than when applying one of the treatments between far infrared roasting and "Koji" fermentation alone. Also, the MDA value when applying the three steps of far infrared roasting, "Koji" fermentation and oily agent preparation will be greater than when applying two of the treatments such as far infrared roasting and fermentation. "Koji".

On the other hand, from the aforementioned Tables 2, 3 and 4 in which clinical tests related to breast cancer, stomach cancer and lung cancer were performed in the cases of untreated product, fermentation treatment with lytic enzyme of the polysaccharide and "Koji" fermentation treatment, far infrared roasting plus "Koji" fermentation treatment and far infrared roasting plus "Koji" fermentation treatment plus oily agent preparation treatment, on PORIA (BUKURYO), agaricus mushroom (AGARIKUSUTAKE), maitake mushroom (MAITAKE, black maitake mushroom (Kuromaitake), snow maitake mushroom (Yukiguni maitake)) and POLYPORUS (CHOREIMAITAKE, Polyporus umbellatus FRIES) on the aforementioned mushrooms, it is known that, in the same mushroom, the clinical effect becomes high when the treatment that results in the highest MDA value reported in Table 1 is applied.

From these results, it is possible to evaluate the importance of the clinical effect, i.e. the efficacy of the antitumor effect, using the MDA value as an index, and it can be said that the treatment that makes the MDA value high effectively indicates an effect. antitumor.

Hence, when the MDA value is large, which shows an amount of lipid peroxide formed by ultraviolet irradiation of docosahexenoic acid, the treatment can be considered highly effective as an antitumor activity of fungi.

Furthermore, when the results of Table 1 and those of Tables 2 to 4 are compared, with the same method of treatment of fungi, the order of the MDA content agrees with the order of the efficacy ratio of the clinical effects. Thus, if a different fungus is added to the system that forms lipid peroxide by ultraviolet irradiation of the aforementioned docosahexenoic acid, to which the same treatment is applied in the same concentration, and the amount formed of lipid peroxide has an MDA value (quantity) in the same way of the aforementioned embodiment, it can be said that the fungus forming the high amount of MDA is effective as an anticancer.

It is therefore possible to expect a high antitumor activity, at the time of internal administration, from the various types of fungi which, upon treatment with ultraviolet irradiation, show a high value of MDA as a quantity of lipid peroxide formed by docosahexenoic acid. Thus, without carrying out clinical tests, it is possible to evaluate the efficacy of the antitumor activity of the respective types of fungi at the time of administration.

Incidentally, in the previous explanation, the case applied to mushrooms such as the agaricus mushroom (AGARIKUSUTAKE), etc., was explained, but the type of mushroom to which it can be applied is not limited only to the aforementioned types. The present invention can be

because they contain β-glucan

Furthermore, the present invention can be applied to mushrooms containing polysaccharides endowed with antitumor activity, other than β-glucan. Furthermore, the present invention can be applied to broaden the field of crude drugs, such as natural or cultured products derived from plants other than fungi and containing polysaccharides having antitumor activity, such as β-glucan.

In the above explanation, docosahexenoic acid was used, but a fatty acid other than docosahexenoic acid can also be used which is, for example, an unsaturated fatty acid in which a certain amount of lipid peroxide is formed by ultraviolet irradiation by adding of the aforementioned mushrooms. In addition, such saturated fatty acid derived from living bodies in which a quantity of lipid peroxide is formed by irradiation with ultraviolet rays by adding the aforementioned fungi, can be used in the field where the aforementioned evaluation method can be applied, even when it is not derived from a living body, for example a synthetic fatty acid.

By adding mushrooms containing a carcinostatic polysaccharide to the system that forms lipid peroxide by ultraviolet irradiation of docosahexenoic acid, as previously explained, at various concentrations, and by comparing the amounts of lipid peroxides formed with the respective concentrations, an administration dose can be easily assessed. media adapted to the stage of progress of the malignant tumor such as cancer without carrying out clinical tests on the respective fungus to be used.

Hence, the relationship between the mushroom concentration and the MDA value in the respective mushroom types is linked in the aforementioned manner and the MDA value and its efficacy in the respective progress stage in cancer are confirmed in advance by the clinical test in a certain type of mushroom.

By operating in the above way, if the concentration of said mushroom is calculated in such a way as to obtain an MDA value confirmed as effective in the respective stage of advancement of the tumor such as cancer, etc., an average internal administration dose can be easily evaluated from the concentration of the fungus .

Furthermore, in the present invention, the antitumor activity of a crude drug can be compared with a common index such as the MDA value, so that the antitumor activity of the various types of crude drug compared by carrying out clinical tests can be obtained by comparison of the one. with the other without carrying out clinical tests, that is, without the risk of harming a patient.

The meanings that the present inventor has found for the value of MDA as a parameter capable of constituting a common measure for the respective types of crude drugs and allowing a mutual evaluation of the antitumor activity, finds a significantly large comparison between the value of MDA and the antitumor activity itself, to the point that it is possible to carry out an evaluation of numerous crude drugs in a simple and easy way.

According to the method of strengthening the antitumor activity of crude drugs of the present invention, the efficacy of the antitumor activity of crude drugs such as fungi in case of internal administration, can be enhanced with respect to the case in which this method is not applied.

Hence, the method of enhancing the antitumor activity of crude drugs of the present invention, even in crude drugs such as mushrooms, etc., which are not considered effective when the method of the present invention is not applied, can be applied to an antitumor agent with greater effectiveness in internal administration.

The enhanced antitumor activity of the crude drug composition containing crude drugs such as fungi to which the antitumor activity enhancing method of the present invention has been applied, has high antitumor efficacy with internal administration compared to the crude drug containing composition to which this method has been applied. has not been applied.

According to the method of evaluating the antitumor efficacy of the treatment with the crude drugs according to the present invention, the efficacy of the antitumor activity of the various types of treatment applied to the crude drugs can be evaluated in a simple and rapid way.

According to the method of evaluating the antitumor efficacy of crude drugs according to the present invention, the efficacy of crude drugs to internal administration can be predicted without carrying out clinical tests. Therefore, compared to the conventional method in which results can be obtained by carrying out experiments on animals or long clinical tests, a research on a substance effective as an anticancer by selecting a large number of natural products can be carried out in a simple way compared to the conventional one.

Claims (29)

CLAIMS 1. Process for strengthening the antitumor activity applied to a crude drug containing polysaccharides having antitumor activity, which includes a roasting step, to toast the raw drug by means of far infrared rays and a fermentation step by adding microorganisms to strengthen the antitumor activity of the raw drug compared to the case in which roasting and fermentation are not carried out. 2. A process for enhancing the antitumor activity of the crude drug according to claim 1, wherein a mushroom containing β-glucan is used as the crude drug. 3. A process for strengthening the antitumor activity of a crude drug according to claim 2, in which at least one mushroom selected from the group consisting of agaricus mushroom (AGARIKUSUTAKE), maitake mushroom (MAITAKE, Grifola frondosu), shiitake mushroom ( SHIITAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKE, Flammulina velutipes). 4. A process of enhancing the antitumor activity of a crude drug according to claim 3, in which the agaricus mushroom (AGA-RIKUSUTAKE) is used, then the crude agaricus mushroom is toasted by far infrared rays. 5. A process for strengthening the antitumor activity of a crude drug according to claim 1, wherein after the fermentation step, an oily agent preparation step is carried out, in which the fermented crude drug is enclosed in an oily component obtained from plants such as toasted sesame by means of far infrared rays, thus strengthening the antitumor activity of the raw drug compared to the case in which these stages of roasting, fermentation and preparation of the oily agent are not carried out. 6. A process for enhancing the antitumor activity of a crude drug according to claim 5, wherein a mushroom containing β-glucan is used as the crude drug. 7. A process for strengthening the antitumor activity of a crude drug according to claim 6, in which at least one mushroom selected from the group consisting of agaricus mushroom (AGARIKUSUTAKE), maitake mushroom (MAITAKE, Grifola frondosu), shiitake mushroom ( SHIITAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKE, Flaramulina velutipes). 8. A method of enhancing the antitumor activity of a crude drug according to claim 7, wherein the agaricus mushroom (AGA-RIKUSUTAKE) is used, then the crude agaricus mushroom is toasted by far infrared irradiation. 9. Composition containing a crude drug enhanced as antitumor activity, which comprises a crude drug to which the antitumor activity enhancement procedure has been applied comprising a roasting step, roasting the crude drug by far infrared irradiation and a step of fermentation by adding a microorganism, whereby the antitumor activity of said crude drug is strengthened with respect to the case in which roasting and fermentation are not carried out. 10. A composition containing a crude drug enhanced as an antitumor activity, according to claim 9, wherein a mushroom containing β-glucan is employed as said crude drug. 11. Composition containing a crude drug enhanced as an antitumor activity according to claim 10, wherein at least one mushroom selected from the group consisting of agaricus mushroom (AGARIKUSUTAKE), maitake mushroom (MAITAKE, Grifola frondosu), shiitake mushroom (SHIITAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKE, Flammulina velutipes). A composition containing a crude drug enhanced as anticancer activity according to claim 11, wherein the agaricus mushroom (AGARIKUSUTAKE) is used, then the raw agaricus mushroom is toasted by far infrared irradiation. A composition containing a crude drug enhanced as an antitumor activity according to claim 9, in which, after the fermentation step, an oily agent preparation step is carried out in which the crude drug is enclosed in an oily component obtained from plants such as toasted sesame by far infrared irradiation so as to strengthen the antitumor activity of the raw drug with respect to the case in which the toasting, fermentation and preparation phases of the oily agent are not carried out. 14. A composition containing a crude drug enhanced as an antitumor activity, according to claim 13, wherein, as said crude drug, a mushroom containing β-glucan is employed. 15. Composition containing a crude drug enhanced as antitumor activity, according to claim 14, in which, as a mushroom, at least one mushroom selected from the group consisting of agaricus mushroom (AGARIKUSUTAKE), maitake mushroom (MAITAKE, Grifola frondosu), shiitake mushroom is used (SHIITAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKE, Flammulina velutipes). A composition containing a crude drug enhanced as an antitumor activity according to claim 15, in which the agaricus mushroom (AGARIKUSUTAKE) is employed, then the raw agaricus mushroom is toasted by far infrared irradiation. 17. Process for evaluating the efficacy of the antitumor activity of a treatment to be applied to a crude drug containing polysaccharides having antitumor activity, which comprises the addition of the crude drug to which the treatment is applied to a system that forms lipid peroxide by means of irradiation with ultraviolet rays of an unsaturated fatty acid such as decosahexenoic acid and evaluation of the treatment effect for strengthening the antitumor activity of the crude drug which increases with the increase in the ratio of the amount of lipid peroxide formed by the treatment of the crude drug, with increasing the ratio of the concentration of the crude drug. 18. Process for evaluating the efficacy of the antitumor activity of a treatment to be applied to a crude drug according to claim 17, wherein a mushroom containing β-glucan is used as said crude drug. 19. Process for evaluating the efficacy of the antitumor activity of a treatment to be applied to a crude drug according to claim 18, in which, as a mushroom, at least one mushroom selected from the group consisting of agaricus mushroom (AGARIKUSUTAKE), mushroom maitake (MAITAKE, Grifola frondosu), shiitake mushroom (SHIITAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, - Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom, Flammutipulina (ENOKITAKE). 20. Process for evaluating the efficacy of the antitumor activity of a treatment to be applied to a raw drug according to claim 19, in which the agaricus mushroom (AGARIKUSU-TAKE) is used, then the raw agaricus mushroom is toasted by irradiation to the far infrared. 21. Crude drug efficacy assessment process to evaluate the efficacy of the antitumor effect presented by the crude drug containing polysaccharides having anticancer activities without conducting clinical trials, which includes adding the crude drug to a system that forms a lipid peroxide by irradiation with ultraviolet rays of an unsaturated fatty acid such as docosahexenoic acid, evaluating the expected effect of antitumor activity of the crude drug by its internal administration, said effect being greater with the increase in the ratio of the amount formed of lipid peroxide with respect to the ratio the concentration of the crude drug. A method for evaluating the efficacy of the antitumor activity of a crude drug according to claim 21, wherein said crude drug to which the antitumor activity enhancement process is applied comprising a roasting step for roasting the crude drug by means of far infrared irradiation and a fermentation step by adding microorganism, whereby the antitumor activity of said crude drug is increased with respect to the case in which roasting and fermentation are not carried out. 23. A method of evaluating the efficacy of the antitumor activity of a crude drug according to claim 22, wherein a mushroom containing β-glucan is used as the crude drug. 24. Process for evaluating the efficacy of the antitumor activity of a crude drug according to claim 23, in which, as a mushroom, at least one mushroom selected from the group consisting of the agaricus mushroom (AGARIKUSUTAKE), the maìtake mushroom (MAITAKE, Grifola) is used frondosu), shiitake mushroom (SHII-TAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKEes, Flammulina velutipipa). A method for evaluating the efficacy of the antitumor activity of a crude drug according to claim 24, in which the agaricus mushroom (AGARIKUSUTAKE) is used, then the crude agaricus mushroom is toasted by far infrared irradiation. 26. A process for evaluating the efficacy of the antitumor activity of a crude drug according to claim 21, wherein said crude drug to which the antitumor activity strengthening process is applied comprising a step of preparation of an oily agent, in which the fermented raw drug is enclosed with an oily component obtained from plants such as toasted sesame by far infrared irradiation after the fermentation phase, so that the strengthening of the antitumor activity of the raw drug is greater than in the case of not roasting, fermentation and preparation of the oily agent. 27. A method for evaluating the efficacy of the antitumor activity of a crude drug according to claim 26, wherein a mushroom containing β-glucan is used as the crude drug. 28. Process for evaluating the efficacy of the antitumor activity of a crude drug according to claim 27, in which, as a mushroom, at least one mushroom selected from the group consisting of the agaricus mushroom (AGARIKUSUTAKE), the maitake mushroom (MAITAKE, Grifola) is used frondosu), shiitake mushroom (SHII-TAKE, Cortinellus Shiitake), matsutake mushroom (MATSUTAKE, Tricholoma matsutake), shimejitake mushroom (SHIMEJITAKE, Lyophyllum decastes) and enokitake mushroom (ENOKITAKEes, Flammulina velutipipa). 29. A process for evaluating the efficacy of the antitumor activity of a crude drug according to claim 28, in which the agaricus mushroom (AGARIKUSUTAKE) is used, then the raw agaricus mushroom is toasted by far infrared irradiation,