JP5167480B2 - Allergy prevention method or treatment method, food and drink, and oral medicine - Google Patents

Description

  The present invention relates to an allergy prevention method or treatment method, food and drink, and oral medicine. In particular, the present invention relates to a method for preventing or treating allergies, a food and drink, and an oral medicine that modify an immune response different from transdermal / intravenous administration by oral administration.

Tumor Growth Factor Beta (Transforming Growth Factor-β: TGF-β) is one of the biologically active molecules (cytokines) contained in a large amount in breast milk identified in the 1980s. Cell growth, survival, differentiation, etc. It is a multifunctional cytokine that regulates. (Non-Patent Document 1)
TGF-β has a gene sequence that is very well preserved in flies, frogs, mice, humans, etc., and that TGF-β receptors exist in almost all cells in the body. Mice lacking the gene (gene knockout mice) die in the embryonic period and are considered to be essential molecules for living organisms.

  According to past epidemiological studies, the amount of TGF-β ingested from breast milk is inversely proportional to the incidence of allergic diseases such as atopic dermatitis in infancy (Non-patent Document 2), and mothers are pregnant and breastfeeding. Ingestion of probiotics has been reported to reduce the incidence of allergic diseases in infants and the results are related to the amount of TGF-β in breast milk.

In addition, for the purpose of disease prevention / treatment, research on the transdermal / intravenous administration of TGF-β has been carried out in various animal disease mice. Some clinical trials have been conducted.
JP 7-258115 A "Transforming growth factor-beta (TGF-β) in human milk", Clin Exp Immunol Vol.94 (1): 220-224, 1993 Oct. "TGF-β in human milk is associated with wheeze in infancy", J Allergy Clin Immunol vol.112 (4): 723-728, 2003 Oct.

  However, the preventive / therapeutic effect of the disease by transdermal / intravenous administration of TGF-β has not been determined, and pharmaceuticals such as weak action on the disease and systemic side effects caused by the administration of TGF-β Many issues remain to be solved before it can be put into practical use.

Allergic diseases such as hay fever, bronchial asthma, atopic dermatitis, and allergic rhinitis are increasing year by year and become a social problem.
In recent years, food allergies have increased rapidly, especially in infants. Eczema and urticaria may develop due to ingestion of allergen-causing substances contained in food and drink, and in severe cases, death may occur due to anaphylactic shock. Each person is responsible for food allergies, but there are many familiar foods such as milk, eggs, wheat, rice, buckwheat, and seafood, and it is difficult to live a life that completely rejects allergens in modern society. It is.

The present inventor paid attention to a report (Non-patent Document 2) that the intake of TGF-β from breast milk is inversely proportional to the number of asthma episodes in infants, and prevention / treatment of food allergy by oral administration of TGF-β. The empirical research was advanced for the purpose.
Until now, it is generally considered that protein molecules taken orally are broken down in the gastrointestinal tract and do not exert their effects systemically, and TGF-β has not been sufficiently verified. As a result of examining the effect of oral TGF-β administration on the immune system using a model animal, the present inventors decompose TGF-β when a large amount of TGF-β is orally administered. Rather, it has been found that the immune response is modified in a manner specific to oral administration different from that of conventional transdermal / intravenous administration.

  The present invention provides an allergy prevention method or treatment method, food and drink, and oral medicine capable of preventing / treating allergic symptoms without trouble such as systemic side effects and immune deficiency by transdermal / intravenous administration of TGF-β The task is to do.

In the case of conventional percutaneous / intravenous administration, Th1 type immune response (cancer cells and pathogens) against cancer and infectious diseases as well as suppression of Th2 type immune response (production of IgE antibody etc.) related to allergic reaction Cell immunity such as a protective response to the immune system) is also suppressed, and the administration of TGF-β suppresses the immune response as a whole and may lead to an immunodeficiency state.

  This time, the inventor orally administered TGF-β to a food allergy model mouse and examined the effect on the systemic immune system. At the stage before verification, TGF-β, which is a protein molecule, was degraded in the gastrointestinal tract and was not expected to exert an immunosuppressive effect such as transdermal / intravenous administration. However, in practice, when TGF-β is administered orally, only the Th2-type immune response related to the allergic reaction is selectively suppressed, and the Th1-type immune response against cancer and infection is enhanced. It exhibited an immunosuppressive effect that was different from that of the cutaneous / intravenous administration. As a result, it was found that TGF-β administered orally while being a protein molecule is not degraded in the gastrointestinal tract, and that TGF-β modifies a specific immune response by oral administration. It came to be completed.

That, allergy prevention or treatment method of the present invention, by orally administering 100 [mu] g / kg or more of tumor growth factor beta (TGF-beta) relative to the body weight of administration subject individual bodies, select the Th2 immune response It suppresses automatically.

According to this, since the Th2-type immune response is selectively suppressed, the onset of allergic symptoms can be prevented and allergic symptoms that have already developed can be alleviated or treated.
Furthermore, in the thing characterized by suppressing Th2-type immune response and maintaining or enhancing Th1-type immune response, in addition to allergy prevention or treatment, various diseases and infections including cancer and infectious diseases It can be used for prevention and treatment of symptoms.

  Here, since TGF-β is contained in a large amount in human breast milk and ingested and grows in infancy, it is considered that humans have a pathway for metabolizing TGF-β taken orally. Therefore, it is considered that oral administration can be safely performed within the range of intake of TGF-β that humans ingest from breast milk in early childhood.

Further, highly conserved beyond TGF-beta species, thereby to work since it can have contact between dissimilar may interact has been confirmed, the TGF-beta from heterologous orally Even when administered, it is considered that the same effects as those derived from humans can be obtained. However, the TGF-β used in the allergy suppressing agent and therapeutic agent, pharmaceutical composition, and food and drink of the present invention is preferably human-derived TGF-β in terms of safety.

  The allergy prevention method or treatment method of the present invention is characterized in that it is orally administered simultaneously with the intake of a causative substance of food allergy. According to this, it is possible to prevent the development of allergic symptoms such as eczema, urticaria and anaphylactic shock caused by ingestion of a causative substance of food allergy.

The food / beverage product of the present invention contains a tumor growth factor beta (TGF-β) in a predetermined amount to selectively suppress a Th2-type immune response, a food, a health food, and a functional food. , Nutritional supplements, foods for specified health use, and quasi drugs. According to this, TGF-β is ingested in a large amount orally by ingestion of food and drink, and the Th2-type immune response is selectively suppressed, so that allergic symptoms can be prevented or alleviated.
Furthermore, in foods and drinks characterized by suppressing the Th2-type immune response and maintaining or enhancing the Th1-type immune response, in addition to allergic prevention and alleviation effects, various foods including cancer and infectious diseases It can be effective in preventing diseases and maintaining health.

In addition, the food and drink of the present invention is characterized by adding tumor growth factor beta (TGF-β) in a predetermined amount to a food containing a causative agent of food allergy.
When a food or drink containing a food allergy causing substance is accidentally ingested by adding a predetermined amount of TGF-β to the food or drink containing the food allergy causing substance by a method such as mixing / mixing in advance, The onset of allergic symptoms can be prevented. In particular, it is effective for anaphylactic shock symptoms that are fatal allergic symptoms by ingestion.

  The predetermined amount is set in consideration of the type of food and drink, the daily intake, the age of the individual to be ingested, etc., and the total intake amount of TGF-β per day is the ingestion target. It is preferably set to be 100 μg / kg or more with respect to the weight of the individual. In particular, it is preferable in terms of safety that the total intake amount of TGF-β per day is set to 100 μg / kg to 200 μg / kg with respect to the body weight of the individual to be ingested.

  The food / beverage product of the present invention enhances Th1-type immune response by containing tumor growth factor beta (TGF-β) in a predetermined amount, food, health food, functional food, nutritional supplement It is selected from the group consisting of foods, foods for specified health use, and quasi drugs. According to this, since immunity is enhanced, effects such as prevention of various diseases such as cancer and infectious diseases, health maintenance, and health promotion can be exhibited.

An oral drug of the present invention comprises an allergy preventive agent, an allergy therapeutic agent, which selectively suppresses a Th2-type immune response by containing tumor growth factor beta (TGF-β) in a predetermined amount, It is selected from the group consisting of immunostimulants, antitumor agents, anti-infective agents, and antiviral agents. According to this, since a Th2-type immune response is suppressed by large doses of oral TGF-β, it can be used for the prevention and treatment of allergic symptoms.
Furthermore, in the oral medicine characterized by selectively suppressing the Th2-type immune response and maintaining or enhancing the Th1-type immune response, in addition to the effects of allergy prevention and alleviation, it includes cancer and infectious diseases. It can be used for prevention and treatment of symptoms of various diseases.

  In addition, the oral pharmaceutical of the present invention contains a tumor growth factor beta (TGF-β) in a predetermined amount, thereby enhancing a Th1-type immune response, an immunostimulant, an antitumor agent, an antitumor agent, It is selected from the group consisting of infectious agents and antiviral agents. According to this, since immunity is enhanced by administration of TGF-β, it can be used for prevention and treatment of symptoms of various diseases including cancer and infectious diseases.

It said predetermined amount is intended to be set in consideration of the age, weight, symptoms and diseases prevention and treatment subject to be administered individual body, the dose per or once per day, administered subject individual It is preferable to set it to be 100 μg / kg or more with respect to the body weight. The daily dose or the dose per dose is preferably set to 100 μg / kg to 500 μg / kg relative to the body weight of the individual subject to administration, and safety is preferably set to 100 μg / kg to 200 μg / kg. Is particularly suitable.

According to the present invention, by administering a large amount of TGF-β orally, there is no trouble such as side effects or immunodeficiency observed when transdermally / intravenously administered TGF-β, thus preventing allergic diseases. Alternatively, treatment can be performed. Moreover, the onset of food allergy symptoms can be prevented by ingesting together with the allergen causing substance.
Furthermore, it can be used as an oral medicine used for prevention and treatment of allergic symptoms and various diseases, as well as foods and drinks having the effect of preventing various diseases including allergic symptoms, cancer and infectious diseases and maintaining health.

The present invention, by orally administering 100 [mu] g / kg or more of tumor growth factor beta (TGF-beta) relative to the body weight of administration subject individual body, characterized in that selectively inhibits Th2-type of immune response Allergy prevention method and treatment method. Furthermore, the present invention is an allergy prevention method and treatment method characterized by suppressing the Th2-type immune response and maintaining or enhancing the Th1-type immune response by oral administration of the TGF-β.

Here, the oral dose of 100 μ / kg or more of TGF-β indicates the daily intake of TGF-β, which is calculated based on experimental data of the inventors.
The dose of allergy prevention or useful TGF-beta as a therapeutic method is preferably set to 100μg / kg~500μg / kg body weight of the administration subject individual body. In particular, it is preferable to set the 150μg / kg~250μg / kg body weight of the administration subject individual body.
The amount of TGF-β contained in breast milk: 500 ng / ml to 1500 ng / ml, calculated from the weight of the breastfeeding infant, the amount of breastfeeding per day, etc. since the intake of TGF-beta on body weight is up to 200 [mu] g / kg to be, up to about a maximum 200 [mu] g / kg body weight of the administration subject individual body can be safely administered orally to humans It is estimated to be.

  The allergy prevention method or treatment method of the present invention is characterized in that it is orally administered simultaneously with the intake of a causative substance of food allergy, and the condition that it passes through a digestive system organ such as the gastrointestinal tract substantially simultaneously with the causative substance. If it can be satisfied, it may be administered in advance before ingestion of the causative substance, or may be administered after ingestion of the causative substance.

Moreover, it is considered that the subject of the allergy prevention method or treatment method of the present invention can be used not only for humans but also for mammals containing TGF-β in breast milk.
The present invention relates to a food, health food, functional food, nutritional supplement characterized by selectively suppressing a Th2-type immune response by containing tumor growth factor beta (TGF-β) in a predetermined amount. It is a food or drink selected from the group consisting of foods, foods for specified health use, and quasi drugs. Furthermore, the oral administration of the TGF-β suppresses a Th2-type immune response and maintains or enhances a Th1-type immune response. In addition, the present invention is a food or drink characterized by enhancing Th1-type immune response by oral administration of the TGF-β.

Specific foods and drinks include artificial milk for infants and ready-made foods, dairy products such as yogurt and milk drinks, confectionery, noodles and grains, instant foods, supplements, and mammal food (pet food). It is done.
In addition, foods and beverages containing food allergies include beverages such as milk and yogurt, confectioneries such as chocolate, biscuits and pudding, okonomiyaki flour, flour and wheat flour, soba and udon In addition to noodles, eggs and seafood can be considered.
Similar to the allergy prevention method or treatment method described above, since the maximum daily intake of TGF-β of the infant is 200 μg / kg, the total daily intake is based on the body weight of the subject individual. It is estimated that up to 200 μg / kg can be safely ingested as food and drink.

  The present invention relates to a prophylactic agent for allergy which selectively suppresses a Th2-type immune response by containing tumor growth factor beta (TGF-β) in a range of 100 μg / day to 100 mg / day. An oral medicine characterized by being selected from the group consisting of an allergy therapeutic agent, an immunostimulant, an antitumor agent, an anti-infective agent, and an antiviral agent. Furthermore, the oral pharmaceutical preparation is characterized by suppressing Th2-type immune response and maintaining or enhancing Th1-type immune response by oral administration of TGF-β. In addition, the present invention is an oral medicine characterized by enhancing Th1-type immune response by oral administration of the TGF-β.

Examples of oral pharmaceutical forms include tablets, granules, powders, capsules, suspensions and the like.
Similar to the above-mentioned allergy prevention method or treatment method, since the maximum intake amount of TGF-β per day for an infant is 200 μg / kg, the dose per day or per dose is the individual to be administered. It is estimated that up to 200 μg / kg of the body weight can be safely orally administered as a pharmaceutical product.

This also previous experiments, the dose or per per day TGF-beta to prophylactic or therapeutic effects are confirmed, the 100μg / kg~500μg / kg body weight of the administration subject individual body It is about the scope and is expected to be applied to oral medicines as preventive and therapeutic agents for each disease. However, as described above, the amount is estimated to safe as the dose for humans, because it is up to 200 [mu] g / kg body weight of the administration subject individual body, 200 [mu] g / kg to body weight of administration subject individual body For oral administration of 500 μg / kg or 500 μg / kg or more of TGF-β, it is considered necessary to sufficiently clinically examine side effects and the like.

  Embodiments of the present invention will be described below based on examples, but the present invention is not limited to these examples.

* Example 1: Prevention of food allergy symptoms by large doses of TGF-β
<Inhibition of IgE antibody production>
Using a transgenic mouse (DO11.10 mice) into which an ovalbumin (OVA) -specific T cell receptor was introduced, the effect of preventing allergic symptoms by oral administration of TGF-β was examined.
In the transgenic mice (DO11.10 mice), OVA-specific IgE (immunoglobulin E) antibody is produced in the blood when OVA is repeatedly orally administered, and anaphylactic shock can be induced by intravenous injection of OVA. It is a food allergy model mouse. (Shida et al., J Allergy Clin Immunol vol.105: 788, 2000)
For the food allergy model mouse (DO11.10, age 6-8 weeks, body weight 20-30 g), OVA: 100 mg (Sigma Chemical Inc.) + Vehicle, OVA: 100 mg + TGF-β: 5 μg (R & D Inc.) was repeated every other day for a total of 6 times. Two days after the last feeding (9 days later), a total of three sera were collected from each model mouse and a model mouse not fed with OVA as a control, and the amount of IgE antibody produced was determined by ELISA (enzyme linked immunosorbent). assay method).

  As shown in FIG. 1 (a), IgE antibody was remarkably elevated in model mice fed with OVA + vehicle, but IgE antibody was detected from the serum of model mice fed TGF-β simultaneously with OVA. (ND: non ditectable). From this, it is considered that the production of IgE antibody was suppressed by TGF-β administered orally simultaneously with OVA causing allergy. Moreover, since this result is the same even after 9 days have passed since the last feeding, it is considered that suppression of IgE antibody production by TGF-β is not transient.

  FIG. 1 (b) shows the results of measuring the production of OVA-specific IgE antibody in serum using the passive cutaneous anaphylaxis (PCA method). It can be seen that the production of OVA-specific IgE antibody is suppressed by TGF-β, similarly to the result of the ELISA method.

<Inhibition of production of immune response-related molecules>
FIG. 2 shows five immune response-related molecules (OVA-specific IgG antibody and cytokine) related to immune responses other than IgE antibodies: (a) IgG1 (immunoglobulin G1), (b) IgG2a (immunoglobulin Ga), ( c) IL-4 (interleukin-4), (d) IFN-γ (interferon-γ), (e) IL-12 (interleukin-12) serum concentration and measurement in mouse spleen cells The results are shown.

  The amount of OVA-specific IgG1 and IgG2a antibodies in the serum of model mice was evaluated by optical density (OD unit) by ELISA. As for IgG1 antibodies that are considered to be involved in Th2-type immune responses (allergic reactions etc.) As shown in FIG. 2 (a), production is suppressed similarly to IgE by co-administration of TGF-β, but it is considered to be involved in Th1-type immune responses (such as protective immunity against cancer and infectious diseases). As for the IgG2a antibody, as shown in FIG. 2 (b), the result that the production amount was increased by a factor of 2 or more where the production was not suppressed was obtained.

In addition, as shown in FIG. 2 (c), the production amount of IL-4 specifically producing IgE antibody in mouse spleen cells by the administration of TGF-β was suppressed in the same manner as in the case of IgG1 antibody. Regarding the production of interferon γ (IFN-γ), which is considered to be involved in the action, immunity enhancing action, and antitumor action, and IL-12, which is supposed to activate NK cells and cellular immunity, in mouse spleen cells (d) As shown in (e), the result of enhancement similar to the IgG2a antibody was obtained.
From the above results, a large dose of TGF-β selectively suppresses the production of bioactive molecules involved in the Th2-type immune response and enhances the production of bioactive molecules involved in the Th1-type immune response. It became clear.

<Influence by anti-TGF-β antibody>
For food allergy model mice (DO11.10, age 6-8 weeks, weight 20-30 g) OVA: 100 mg + excipient feeding, 2. (OVA: 100 mg + TGF-β: feeding 5 μg) + subcutaneous injection of IgG antibody (control antibody); (OVA: 100 mg + TGF-β: feeding of 5 μg) + subcutaneous injection of anti-TGF-β antibody (αTGF-β Ab) was repeated every other day for a total of 6 times for feeding and injection. Two days after the last feeding (9 days later), a total of 3 sera from each model mouse was collected, and the production amount of IgE antibody was measured by ELISA and PCA. As a result, the production amount of IgG2a antibody was determined by ELISA method. The measurement results are shown in FIG.

As shown in FIGS. 3 (a), (b) and (c), anti-TGF-β antibody (αTGF-β Ab) administered by subcutaneous injection suppresses IgE antibody production and IgG2 antibody by oral TGF-β administration. It can be seen that any enhancement of the production of is inhibited. This confirms that Th2-type immune response involving IgE antibody is suppressed and T1-type immune response involving IgG2 antibody is enhanced by TGF-β which is orally administered and absorbed / metabolized from the gastrointestinal tract. We were able to.
From the above results, orally administered TGF-β is not degraded in the gastrointestinal tract and is metabolized by a route different from that of transdermal / intravenous TGF-β to produce an immune response. It is considered to be modified.

<Inhibition of anaphylactic shock>
OVA: 100 mg + vehicle, OVA: 100 mg + TGF-β: 5 μg, TGF-β: 5 μg every other day for allergic disease model mice (DO11.10, age 6-8 weeks, body weight 20-30 g) A total of 6 feedings were made. Table 1 shows the results of observing whether or not anaphylactic shock was caused by intravenously injecting OVA two days after the last feeding.

For the conditions of OVA: 100 mg + vehicle, OVA: 100 mg + TGF-β: 5 μg, an experiment was performed on 6 model mice each, and control (no feeding) and TGF-β: 4 μg for the conditions of 5 μg each. Experiments were conducted on model mice.
Most mice that received OVA developed fatal anaphylactic shock at “++” to “++++” levels, whereas model mice that received TGF-β orally received “−” to “+” It became clear that it was suppressed to a state that caused almost no shock.
This revealed that anaphylactic shock, which is a severe allergic symptom, can be suppressed by administering a large amount of TGF-β orally at the same time as the allergic causative agent.

<Comparison with transdermal / intravenous administration>
As a comparison experiment with conventional transcutaneous / intravenous administration, OVA: 100 mg feeding + TGF-β for allergic disease model mice (DO11.10, age 6-8 weeks, body weight 20-30 g) : 5 μg subcutaneous injection every other day for a total of 6 feedings and injections. Two days after the last feeding, the production amount of serum IgE antibody was measured by the PCA method. The production amount of serum IgG2a antibody was measured by the ELISA method, and the results are shown in FIG.

  The production of serum IgE antibody is suppressed by the administration of TGF-β in FIG. 5 (a) as in the case of oral administration, but in the case of oral administration, as shown in FIG. 4 (b). The production of the enhanced serum IgG2a antibody is also suppressed. That is, in the case of transdermal TGF-β administration, protective immunity (Th1-type immune response) against infectious diseases and cancer is suppressed together with allergic reaction (Th2-type immune response). This can lead to an immune deficiency.

On the other hand, oral administration of TGF-β can suppress only the allergic reaction (Th2-type immune response) without reducing the immune response necessary for life support (suppressing the Th1-type immune response). it can. Thus, allergic diseases can be prevented or treated without problems such as side effects and immune deficiencies observed when administering TGF-β transcutaneously / intravenously.
In addition, the oral administration of TGF-β enhances the Th1-type immune response involved in cellular immunity, and is therefore effective in the prevention and treatment of various diseases including allergic symptoms, cancer and infectious diseases. it is conceivable that.

* Example 2: Treatment of allergic symptoms <Treatment of atopic dermatitis>
Model mice (DO11.10, age 6-8 weeks, body weight 20-30g) that actually develop atopic dermatitis, TGF-β: 5μg / mouse / mouse every other day for a total of 3 Oral administration was performed multiple times. FIG. 5 shows clinical severity levels (Clinical Skin Severity) before and after oral administration of TGF-β one week after administration, and IgE levels in serum.

As shown in FIG. 5 (a), the model mice to which TGF-β was orally administered alleviated the inflammatory state of atopic dermatitis compared to before administration, as shown in FIG. 5 (b). It was revealed that the IgE level also decreased. In addition, symptoms such as side effects caused by oral administration of TGF-β were not observed in the model mice.
Thus, allergic symptoms that have already developed allergies can be treated and alleviated by administering a large amount of TGF-β orally.

2 is a graph showing suppression of production of serum IgE antibody by oral TGF-β in Example 1. It is a graph which shows the influence which oral TGF-beta has on the production of bioactive molecules (a) IgG1 antibody, (b) IgG2a antibody, (c) IL-4, (d) IL-12, (e) IFN-γ . It is a graph which shows the influence which the anti- TGF- (beta) antibody administered intravenously has on the effect | action to an immune system by oral TGF- (beta). It is a graph which shows production suppression of (a) serum IgE antibody and (b) serum IgG2 antibody by intravenously administered TGF-β. It is a graph which shows the healing effect of (a) allergic symptom (dermatitis) and (b) production suppression of IgE antibody by oral TGF-β of Example 2.

Claims (12)

An oral allergy preventive agent containing TGF-β,
An allergy prevention agent for preventing allergies caused by food allergy sources taken at the same time. An oral allergy therapeutic agent containing TGF-β,
An allergy remedy for treating allergies caused by food allergy sources taken at the same time. An oral antiallergic agent containing TGF-β,
An oral antiallergic agent for allergies caused by food allergy sources taken at the same time. An oral medicine that is an allergy preventive agent containing TGF-β,
To prevent allergies caused by food allergy sources taken at the same time
An oral medicine that is an allergy preventive agent for selectively suppressing a Th2-type immune response . An oral medicine that is an allergy therapeutic agent containing TGF-β,
To treat allergies caused by food allergy sources taken at the same time
An oral drug which is an allergy therapeutic agent for selectively suppressing a Th2-type immune response . An oral drug that is an oral antiallergic agent containing TGF-β,
For allergies caused by food allergy sources taken at the same time,
An oral drug that is an oral antiallergic agent for selectively suppressing a Th2-type immune response . The allergy preventive agent according to claim 1, which is used for non-human mammals. 3. The allergy therapeutic agent according to claim 2, which is used for a non-human mammal. 4. The antiallergic agent according to claim 3, which is used for a non-human mammal. 5. The oral medicine which is an allergy preventing agent according to claim 4, which is used for a non-human mammal. 6. The oral medicine which is an allergy therapeutic agent according to claim 5, which is used for a non-human mammal. The oral medicine which is an antiallergic agent according to claim 6, which is used for a non-human mammal.