JP7246592B2 - Method for producing innate immunity activator - Google Patents

Description

TECHNICAL FIELD The present invention relates to an innate immunity activator containing neutral polysaccharides contained in broccoli as an active ingredient and a method for producing the same.

Innate immunity is a defense mechanism that rapidly eliminates foreign substances (bacteria, fungi, viruses, cancer cells) in vivo without relying on antibodies, and is a mechanism that is common to insects and mammals.
So far, the present inventors have discovered that silkworm muscle contraction occurs with the activation of innate immunity (Non-Patent Documents 1 and 2). Based on this result, the present inventor has established a method for identifying innate immunity activators in foods (Patent Document 1, Non-Patent Documents 3 and 4).

The present inventor has further attempted to extract a compound having an activity of activating natural immunity from broccoli extract using the above method (Patent Documents 2 and 3, Non-Patent Document 5).
However, the specific activity of the compounds obtained from broccoli extracts so far is not very high, and it is desired to search for compounds having higher specific activity.

WO2008/126905 JP 2010-030995 A WO2009/157409

Ishii K, Hamamoto H, Kamimura M. and Sekimizu K. (2008) Activation of the silkworm cytokine by bacterial and fungal cell wall components via a reactive oxygen species-mechanism, J. Biol. Chem., 283, 2185-2191. Ishii K, Hamamoto H, Kamimura M, Nakamura Y, Noda H, Imamura K, Mita K and Sekimizu K. (2010) Insect cytokine paralytic peptide (PP) induces cellular and humoral immune responses in the silkworm Bombyx mori, J. Biol. Chem., 285, 28635-28642. Dhital S, Hamamoto H, Urai M, Ishii K, Sekimizu K. (2011) Purification of innate immunostimulant from green tea using a silkworm muscle contraction assay. Drug Discov Ther. 5,18-25. Fujiyuki T, Hamamoto H, Ishii K, Urai M, Kataoka K. Takeda T, Shibata S, Sekimizu K. (2012) Evaluation of innate immune stimulating activity of polysaccharides using a silkworm (Bombyx mori) muscle contraction assay. Drug Discov Ther. 6, 88-93. Urai M, Kataoka K, Nishida S, Sekimizu K. (2017) Structural analysis of an innate immunostimulant from broccoli, Brassica oleracea var. italica. Drug Discov Ther. 11, 230-237.

The present invention has been made in view of the background art described above, and an object thereof is to provide a novel substance having a high innate immunity activating effect.

As a result of intensive studies to solve the above problems, the present inventors have found that neutral polysaccharides obtained from broccoli extracts are used in natural immunity using the silkworm muscle contraction activity measurement method established by the present inventors. As a result of measuring the activating action, the inventors have found that the specific activity is about 10 times or more higher than that of polysaccharides that have already been reported, and have completed the present invention.

That is, the present invention provides an agent for activating innate immunity, characterized by containing a neutral polysaccharide contained in broccoli as an active ingredient.

In addition, the present invention is a method for producing an innate immunity activator containing a neutral polysaccharide contained in broccoli as an active ingredient,
The present invention provides a method for producing an innate immunity activator, comprising at least a step of extracting broccoli with hot water and a step of recovering neutral polysaccharides.

The present invention also provides an innate immunity activator characterized in that it contains a neutral polysaccharide contained in broccoli as an active ingredient, and is obtained by the above-described method for producing an innate immunity activator. It provides.

The present invention also provides a method for producing food and drink for activating innate immunity, which is characterized by using the method for producing an innate immunity activator.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide an innate immunity activator having hitherto unknown high specific activity, that is, an extremely high innate immunity activating effect. In addition, it is possible to provide a food or drink having an extremely high innate immunity activating effect.

Moreover, the neutral polysaccharide of the present invention and the natural immunity activation agent of the present invention containing the neutral polysaccharide as an active ingredient are extremely safe and have no side effects. In addition, since it is easy to prepare various dosage forms and to be added to foods and drinks, it is possible to activate natural immunity extremely effectively as a safe natural immunity activator or functional food.

The present invention will be described below, but the present invention is not limited to the following specific embodiments, and can be arbitrarily modified within the scope of the technical idea.

The innate immunity activator of the present invention contains neutral polysaccharides contained in broccoli as an active ingredient.
Furthermore, the innate immunity activator of the present invention contains a neutral polysaccharide contained in broccoli as an active ingredient, and is obtained by the "method for producing an innate immunity activator" described later.

The method for producing an innate immunity activator of the present invention is a method for producing an innate immunity activator containing a neutral polysaccharide contained in broccoli as an active ingredient,
It is characterized by including at least a step of extracting broccoli with hot water and a step of recovering neutral polysaccharides.

The "active ingredient of the natural immunity activator" contained in broccoli obtained by the production method of the present invention is obtained by, for example, extracting broccoli with hot water and extracting the ethanol precipitated fraction with DEAE cellulose, as shown in the examples below. The innate immunity activating component is a neutral polysaccharide by subjecting it to chromatography and obtaining it as a column non-adsorbed fraction.

As described above, the innate immunity activator of the present invention contains a neutral polysaccharide contained in broccoli as an active ingredient, and is obtained by the "method for producing an innate immunity activator" described later.
The neutral polysaccharide, which is an active ingredient of the innate immunity activator, may be artificially synthesized, for example, and may be produced using a production method other than the "method for producing an innate immunity activator" described later. , it may be possible to obtain a similar neutral polysaccharide.
However, "neutral polysaccharides specified by the production method described later" are distinguished from "other neutral polysaccharides as substances", and simply the chemical structure, composition, each content concentration, impurity concentration of the neutral polysaccharide , physical property parameters, etc.

Therefore, identifying all the substances contained in the above neutral polysaccharides and specifying their concentrations takes a huge amount of time and effort, and is impractical. "Neutral polysaccharides" can only be identified by the method of production.

In the hot water extraction, any tissue of broccoli may be used, and flower buds, flower buds, flower stalks, etc. of broccoli may be used. Also, multiple tissues of broccoli may be used, or the whole broccoli may be used without dividing into individual tissues.
It is preferable to extract from flower buds and/or flower buds in that an excellent innate immune activation effect can be obtained.

The hot water extraction method is not particularly limited. For example, a hot water extract can be obtained by adding a solvent such as water to an autoclave device and heating the device.
There are no particular restrictions on the solvent when using an autoclave, but water is preferred as shown in the Examples.

As for the hot water extraction conditions, the temperature of the hot water extraction is preferably 80° C. or higher and 160° C. or lower, more preferably 90° C. or higher and 150° C. or lower, in that a natural immunity activating component with high specific activity can be obtained, and 100° C. More than 140° C. or less is particularly preferable.
The hot water extraction time is preferably 1 minute or more and 60 minutes or less, more preferably 5 minutes or more and 50 minutes or less, and particularly preferably 10 minutes or more and 40 minutes or less.

Further, in the production method of the present invention, it is preferable to crush, shred or grate the broccoli before the hot water extraction, since it is possible to obtain a natural immunity activating component with a high specific activity.
The means for pulverizing, shredding or grating is not particularly limited as long as the plant tissue is destroyed or the surface area is increased so that the natural immunity activating component can be easily extracted, and known means are used. can be used. The device for pulverizing, shredding, or grating is not particularly limited, but includes a pulverizer such as a homogenizer.

The above-mentioned "process of pulverizing, shredding or grating broccoli" is preferably carried out at 10°C or less in order to obtain natural immunity activating components with high specific activity. For example, when performing "the step of grinding, chopping or grating broccoli" using a grinder such as a homogenizer, it can be performed on ice.

It is also possible to add other processes as a pre-process and/or a post-process of the above "process of pulverizing, chopping or grating broccoli". "Other processes" include disruption of cell tissue, freezing, drying, heating, preservation, chemical treatment, enzyme treatment, and the like. These are performed to facilitate the elution of components other than the innate immunity activating components, or facilitate the elution of the innate immunity activating components. In the "other step", it is preferred that at least the innate immunity activating component is not substantially eluted or taken out of the system.

The method for recovering the neutral polysaccharides from the hot water extract of broccoli is not particularly limited, and known means can be used. For example, as described in the Examples, after recovering saccharides from a broccoli hot water extract by ethanol precipitation, the saccharides can be separated by chromatography to recover the target neutral polysaccharides.
The method for producing the innate immunity activator of the present invention preferably includes a step of ethanol precipitation after the hot water extraction.

Moreover, in the method for producing an innate immunity activator of the present invention, the step of recovering the neutral polysaccharide is preferably a step of recovering a fraction not adsorbed to the anion exchanger.
Further, it is preferred that the anion exchanger is a DEAE cellulose carrier.

As shown in Non-Patent Document 5, etc., the present inventors have so far recovered saccharides from a broccoli hot water extract, and have performed natural immunity against column-adsorbed fractions (acidic polysaccharides) obtained by subjecting them to DEAE celluloseography. It is reported to have an activating effect.
On the other hand, the innate immunity activating component found in the examples of the present application is a neutral polysaccharide, and has a different structure from the component described in Non-Patent Document 5. In addition, the specific activity of the component described in Non-Patent Document 5 is 130 units / mg, and the neutral polysaccharide found in the examples of the present application is about 10 times higher, and the component described in Non-Patent Document 5 etc. are clearly different.

In addition, as shown in Patent Documents 2 and 3, the present inventors have so far washed broccoli heated by microwave irradiation with running water at 0 to 40 ° C. to remove components other than natural immunity activation components. A method of eluting and recovering (extracting) the innate immunity activating component is described.
However, the innate immunity activating component found in the examples of the present application is subjected to DEAE cellulose chromatography using an anion exchanger as a carrier, and only the unadsorbed component is recovered. Clearly different from the listed ingredients. Moreover, the specific activity is about 10 times higher than that of the components described in Patent Documents 2 and 3, and from this point, it is clearly different from the components described in Patent Documents 2 and 3.

The content of the neutral polysaccharide, which is an active ingredient in the innate immunity activator produced by the production method of the present invention, relative to the whole innate immunity activator is not particularly limited, and can be appropriately selected according to the purpose. can be done. When the whole innate immunity activator is 100 parts by mass, the total amount of polysaccharides is preferably 0.01 to 100 parts by mass, more preferably 0.1 to 99 parts by mass, particularly preferably 1 The content is up to 95 parts by mass, more preferably 10 to 90 parts by mass.

The innate immunity activator of the present invention can contain "other ingredients" in addition to the above polysaccharides as active ingredients.
The "other ingredients" are not particularly limited, and can be appropriately selected depending on the intended purpose as long as they do not impair the effects of the present invention. Examples thereof include pharmaceutically acceptable carriers.
Such a carrier is not particularly limited, and can be appropriately selected according to, for example, the dosage form described later. Also, the content of the "other component" in the natural immunity activator is not particularly limited, and can be appropriately selected according to the purpose.

The dosage form of the agent for activating natural immunity of the present invention is not particularly limited, and can be appropriately selected, for example, according to the desired administration method as described below.
Specifically, for example, oral solid formulations (tablets, coated tablets, granules, powders, hard capsules, soft capsules, etc.), oral liquid formulations (oral liquid formulations, syrups, elixirs, etc.), injection formulations (solvents, suspensions, etc.) agents, etc.), ointments, patches, gels, creams, powders for external use, sprays, inhalation dusts, and the like.

As the innate immunity activator, for example, the active ingredient, excipients, and if necessary additives such as binders, disintegrants, lubricants, coloring agents, flavoring agents, and the like, It can be manufactured by a conventional method.

Examples of the excipient include lactose, white sugar, sodium chloride, glucose, starch, calcium carbonate, kaolin, microcrystalline cellulose, silicic acid and the like.
Examples of the binder include water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, hydroxypropyl cellulose, hydroxypropyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, polyvinylpyrrolidone, and the like. be done.
Examples of the disintegrant include dry starch, sodium alginate, agar powder, sodium hydrogen carbonate, calcium carbonate, sodium lauryl sulfate, monoglyceride stearate, lactose and the like.
Examples of the lubricant include refined talc, stearate, borax, polyethylene glycol and the like.
Examples of the coloring agent include titanium oxide and iron oxide.
Examples of the flavoring/flavoring agent include sucrose, orange peel, citric acid, and tartaric acid.

The above-mentioned oral liquid can be produced, for example, by adding additives such as flavoring agents, buffering agents, stabilizers, edible (processed) oils, animal and vegetable oils to the above-mentioned active ingredients, and by a conventional method.

Examples of the flavoring/flavoring agent include sucrose, orange peel, citric acid, and tartaric acid. Examples of the buffering agent include sodium citrate and the like. Examples of the stabilizer include tragacanth, gum arabic, gelatin and the like.

For the above-mentioned injections, for example, pH adjusters, buffers, stabilizers, tonicity agents, local anesthetics, etc. are added to the above-mentioned active ingredients, and subcutaneous, intramuscular, and intravenous injections are carried out according to conventional methods. etc. can be manufactured.
Examples of the above pH adjuster and buffer 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 tonicity agent include sodium chloride and glucose. Examples of the local anesthetic include procaine hydrochloride and lidocaine hydrochloride.

The above ointment can be produced, for example, by blending the above active ingredient with known bases, stabilizers, wetting agents, preservatives and the like, and mixing the mixture in a conventional manner.
Examples of the base include liquid paraffin, white petrolatum, bleached beeswax, octyldodecyl alcohol, and paraffin. Examples of the preservative include methyl parahydroxybenzoate, ethyl parahydroxybenzoate, and propyl parahydroxybenzoate.

The patch can be produced, for example, by applying a cream, gel, paste or the like as the ointment to a known support by a conventional method. Examples of the support include woven fabrics and nonwoven fabrics made of cotton, staple fiber, and chemical fibers, films of soft vinyl chloride, polyethylene, polypropylene, polyurethane, foam sheets, and the like.

The innate immunity activator of the present invention is, for example, an individual in need of activation of the innate immune system (e.g., an individual in need of health maintenance or recovery from fatigue; an individual in need of prevention or treatment of cancer or lifestyle-related diseases. individuals; individuals infected with bacteria, fungi, viruses, etc.; etc.).

Animals to be administered with the agent for activating innate immunity of the present invention are not particularly limited, but for example, humans; experimental animals such as mice and rats; monkeys; horses; pets such as dogs;

In addition, the method for administering the natural immunity activator is not particularly limited, and can be appropriately selected according to, for example, the dosage form described above. Injection into the inside, etc. are mentioned.

In addition, the dosage of the above-mentioned innate immunity activator is not particularly limited, and can be appropriately selected according to the age, body weight, degree of desired effect, etc. of the individual to be administered. The daily dosage of is preferably 1 mg to 30 g, more preferably 10 mg to 10 g, particularly preferably 100 mg to 3 g, as the total amount of the above neutral polysaccharide as an active ingredient.
Also, the timing of administration of the natural immunity activator is not particularly limited and can be appropriately selected according to the purpose. For example, it may be administered prophylactically or therapeutically. .

Moreover, the manufacturing method of the natural-immunity activation food-drinks of this invention is characterized by using the manufacturing method of the said natural-immunity activator.
That is, the food and drink for activating natural immunity contain the neutral polysaccharide or the activating agent for natural immunity.

The content of the neutral polysaccharide or the natural immunity activator in the food and drink for activating natural immunity is not particularly limited, and can be appropriately selected according to the purpose and the aspect (type) of the food and drink. , When the total amount of food and drink is 100 parts by mass, the total amount of the innate immunity activator is preferably 0.001 to 100 parts by mass, more preferably 0.01 to 100 parts by mass, especially The content is preferably 0.1 to 100 parts by mass.

Also, one of the neutral polysaccharides and the innate immunity activator may be used alone, or two or more of them may be used in combination. When two or more substances are used in combination, the content ratio of each substance in the food or drink is not particularly limited, and can be appropriately selected according to the purpose.

The food and drink for activating natural immunity can further contain "other ingredients" in addition to the neutral polysaccharide or the agent for activating natural immunity of the present invention.
The above-mentioned "other ingredients" in the natural immunity activation food and drink having such natural immunity activation action are not particularly limited, and can be appropriately selected according to the purpose within a range that does not impair the effects of the present invention. , for example, various food raw materials. In addition, the content of "other components" is not particularly limited and can be appropriately selected according to the purpose.

The type of food and drink for activating natural immunity is not particularly limited, and can be appropriately selected according to the purpose. Examples include confectionery such as jelly, candy, chocolate, and biscuits; Dairy products such as fermented milk, yogurt, and ice cream; Processed vegetable and fruit products such as vegetable drinks, fruit drinks, and jams; Liquid foods such as soup; Processed grain products such as bread and noodles; seasoning; and the like.
The method for producing these foods and beverages is not particularly limited, and for example, they can be produced as appropriate according to the usual methods for producing various kinds of foods and beverages.

In addition, the food and drink for activating natural immunity may be manufactured as oral solid preparations such as tablets, granules and capsules, or oral liquid preparations such as oral liquid preparations and syrups. The method for producing the above solid oral preparations and oral liquid preparations is not particularly limited, and can be appropriately selected according to the intended purpose. can.

The above foods and drinks are considered to be particularly useful as functional foods, health foods, and the like for the purpose of activating the innate immune system.

In the method for producing the food and drink for activating natural immunity of the present invention, steps well known to those skilled in the art can be performed. A person skilled in the art can appropriately combine the steps of mixing the neutral polysaccharide of the present invention with other components, the molding step, the sterilization step, the fermentation step, the baking step, the drying step, the cooling step, the granulation step, the packaging step, and the like. , it is possible to make the desired food and drink.

<Action/Principle>
As already described in Patent Documents 2 and 3, "broccoli heated by microwave irradiation is washed with running water of 0 to 40 ° C. and then components are extracted. In the method, components other than natural immunity activation components are It is possible that there was a lot mixed in.
The innate immunity activating components found in the examples of the present application are "neutral polysaccharides" by excluding "acidic polysaccharides that are adsorbed components" by DEAE cellulose chromatography, so Patent Document 2 and It is considered that the specific activity of innate immunity activation is about 10 times or more higher than that of the component described in 3.

EXAMPLES The present invention will be described in more detail below based on examples and study examples, but the present invention is not limited to the specific scope of the following examples and the like. In the following, references to "%" mean "% by weight" when it relates to weight.

Example 1
<Preparation of neutral polysaccharides (Immunocoli) from broccoli extract>
On ice, 15 g of flower buds of broccoli were cut with scissors, added with 30 mL of purified water, and homogenized using a Polytron homogenizer. The sample was autoclaved (121° C., 20 minutes), then centrifuged (8,000 rpm, 10 minutes), and the supernatant was used as a hot water extract.
Next, ethanol precipitation was performed and the precipitate was collected. The resulting precipitate was dissolved in 10 mM Tris-HCl buffer (pH 7.9) and subjected to DEAE cellulose column chromatography to collect an unadsorbed fraction.
The non-adsorbed fraction was suggested to be neutral polysaccharides because it was not adsorbed by DEAE cellulose column chromatography. Hereinafter, the neutral polysaccharide may be referred to as "Immunocoli".
Next, the conditions for extracting immunocoli were examined in Examination Examples 1 to 4.

In the following study examples, the specific activity was measured by silkworm muscle contraction using the method described in Hamamoto H. et al., J. Biol. Chem. Jan. 25; 283(4):2185-91 (2008). Activity was calculated.
Specifically, 0.05 mL of the sample was intrabloodally administered to the decapitated muscle specimen of the 5th instar silkworm, and the degree of muscle contraction of the silkworm larvae was evaluated by contraction value (C value). The body length was measured when the C value reached the maximum, and "'Body length of silkworm before sample injection (cm)' - 'Body length of silkworm after sample injection (cm)'" was changed to "Body length of silkworm before sample injection (cm)". It is a value divided by body length (cm).
In addition, the above sample was serially diluted to 1/1, 1/4, 1/16, 1/64, and 1/256 with 0.9% physiological saline, and the C value was measured. The slow muscle contraction activity at 0.15 was defined as 1 unit, and the specific activity (the number of units per unit mass) was determined.

Study example 1
<Study of conditions for dissolving the ethanol precipitate in the solvent>
After autoclave treatment (hot water extraction) of broccoli, the temperature and time for dissolving the precipitate obtained by ethanol precipitation in a buffer solution (Tris-HCl (pH 7.9)) were investigated. Table 1 shows the results of the study.

As shown in Table 1, it was found that the specific activity was reduced by heating when the ethanol precipitate was dissolved in the buffer.
In the study examples below, the ethanol precipitate was dissolved in the buffer solution at room temperature.

Study example 2
<Study of conditions for homogenization>
Next, the temperature at which broccoli is homogenized using a Polytron homogenizer was investigated. Table 2 shows the examination results.
In addition, the effect of drying and/or autoclaving the ethanol precipitate before subjecting it to DEAE cellulose chromatography on the specific activity was also investigated. In Table 2, "+" indicates that the treatment was performed, and "-" indicates that the treatment was not performed.

As shown in Table 2, it was found that a high specific activity can be obtained when the homogenization treatment is performed on ice and the ethanol precipitated fraction is not dried and autoclaved.

Examination example 3
<Examination of extraction solvent used for hot water extraction>
Next, the solvent used for hot water extraction was investigated. Table 3 shows the examination results.

As shown in Table 3, the samples extracted with Tris-HCl (pH 7.9) or sodium phosphate (Na-phosphate, pH 6.0) buffer had lower specific activities than those extracted with water. Decreased.
Therefore, it was found that it is better to use water as a solvent when the target neutral polysaccharide is extracted with hot water.

Study example 4
<Effect of 37°C treatment after homogenization>
After broccoli was homogenized on ice, it was examined whether the activity of the DEAE cellulose-unadsorbed fraction was affected when treated at 37° C. for 1 hour. Table 4 shows the results.

As shown in Table 4, when the homogenized product (homogenate) was treated at 37°C, the specific activity decreased to about 1/10 of that of the untreated product. It is conceivable that the innate immunity activating component was degraded by the degrading enzyme present in the homogenized product.

Study example 5
<Examination of temperature and extraction time during hot water extraction>
After homogenizing broccoli on ice, the temperature and time of hot water extraction were investigated. Table 5 shows the examination results.

As shown in Table 5, when the extraction temperatures were compared, the specific activity was higher at 121°C. Moreover, when the extraction time was compared, it was found that the specific activity increased when the extraction was performed for 20 minutes or longer.

Study example 6
<Measurement of specific activity per dry weight of DEAE cellulose non-adsorbed fraction>
Regarding the immunocoli obtained in Example 1, the specific activity per dry weight was calculated for the sample whose specific activity per 1 mg of sugar was around 3,000 units/mg (sugar). Table 6 shows the results.

<Summary of Examples>
Table 7 shows the specific activity of the innate immune activity of each polysaccharide published in Non-Patent Document 4. These specific activities were measured by the same method as described in the above examination example.

As shown in Table 7, it has been reported that polysaccharides such as β-glucan have innate immune activity according to the silkworm muscle contraction activity assay constructed by the present inventors. However, curdlan, which has the highest specific activity among them, has a specific activity of 100 units/mg. Yeast β-glucan, which is widely used as a health food, is 20 to 33 units/mg. Therefore, the specific activity of immunocoli, which is a neutral polysaccharide obtained by the production method of the present invention, is significantly higher than the activities of polysaccharides already reported.

INDUSTRIAL APPLICABILITY According to the present invention, a neutral polysaccharide having a high effect of activating innate immune function can be obtained, so that it can be widely used in various foods and drinks, tablets, granules, chewable tablets and the like. In addition, the neutral polysaccharide obtained by the production method of the present invention can be used as a functional food that has an action to activate the natural immune function in vivo, a food for specified health use, food for sick people and food for nursing care that have a problem of decreased immune function. It can be particularly suitably used as a material. Furthermore, it can be widely used as a raw material for pharmaceuticals.

Claims (8)

A method for producing an innate immunity activator containing a neutral polysaccharide contained in broccoli as an active ingredient,
At least, a step of hot water extraction of broccoli flower buds or flower buds , and a step of recovering neutral polysaccharides,
A method for producing an innate immunity activator, wherein the step of recovering the neutral polysaccharide is a step of recovering a fraction not adsorbed to an anion exchanger. The method for producing an innate immunity activator according to claim 1, wherein the anion exchanger is a DEAE cellulose carrier. The method for producing an innate immunity activator according to claim 1 or 2, comprising a step of ethanol precipitation after the hot water extraction. The method for producing an innate immunity activator according to any one of claims 1 to 3, comprising a step of pulverizing, shredding or grating the broccoli before the hot water extraction. The method for producing an innate immunity activator according to claim 4, wherein the step of pulverizing, shredding or grating is performed at 10°C or lower. The natural immune activity according to any one of claims 1 to 5, wherein the hot water extraction temperature is 80 ° C. or higher and 160 ° C. or lower, and the hot water extraction time is 1 minute or longer and 60 minutes or shorter. manufacturing method of the agent. It contains a neutral polysaccharide contained in broccoli as an active ingredient, and is obtained by the method for producing an innate immunity activator according to any one of claims 1 to 6. Innate immune activator. 7. A method for producing a natural immunity-activating food or drink, characterized by using the method for producing a natural immunity-activating agent according to any one of claims 1 to 6.