JP7244059B2 - Rhinitis prophylactic/therapeutic agent - Google Patents

Description

TECHNICAL FIELD The present invention relates to a prophylactic/therapeutic agent for rhinitis that can be used for prophylaxis or treatment of rhinitis.

It has been reported that specific amino acids such as valine, leucine, isoleucine, arginine, glycine, and serine are effective in suppressing symptoms of allergic asthma. For example, Patent Document 1 verifies the inhibitory effect on asthma using ovalbumin-induced asthma symptoms in mice. Specifically, amino acids such as valine and leucine are administered to mice in which asthma has been induced as described above, and the lungs of the mice are washed. Using the number of eosinophils contained in the physiological saline thus obtained as an index, the inhibitory effect on asthma has been verified. Patent Document 1 states that administration of amino acids such as valine and isoleucine reduces the number of eosinophils contained in lung lavage fluid.

In addition, Patent Document 2 states that allergen-induced airway hyperresponsiveness (AHR) is improved by inhaling arginase inhibitor ABH in guinea pigs with allergic asthma.

WO2006/049286 publication Special table 2010-510257 publication

Patent Document 1 states that administration of amino acids such as valine and leucine to mice in which symptoms of allergic asthma are induced reduces the number of eosinophils. However, it is not described whether these amino acids exert an effect on rhinitis. Also, there is a description that administration of ornithine alone to the mouse does not improve allergic symptoms.

Even in Patent Document 2, the amount of PC100 histamine was only verified in the case of administering an arginase inhibitor and the case of administering water to mice induced with symptoms of allergic asthma. It is not stated whether or not it is effective against

An object of the present invention is to provide a prophylactic/therapeutic agent for rhinitis containing ornithine as an active ingredient, which has a well-established method for industrial mass production and is less dangerous to the living body.

A prophylactic/therapeutic agent for rhinitis containing ornithine as an active ingredient solves the above problems. In the prophylactic/therapeutic agent for rhinitis, the concentration of ornithine is preferably 5.0% by mass or less.

INDUSTRIAL APPLICABILITY According to the present invention, a method for industrial mass production has been established, and it is possible to provide a prophylactic/therapeutic agent for rhinitis containing ornithine as an active ingredient, which is less dangerous to the living body.

1 is a graph showing the preventive effect of allergic rhinitis when the agent for preventing/treating rhinitis of Example 1 or Example 3 or the water of Comparative Example 1 is administered. The vertical axis indicates the number of sneezes per 10 minutes. 1 is a graph showing the preventive effect of allergic rhinitis when the agent for preventing/treating rhinitis of Example 1 or Example 3 or the water of Comparative Example 1 is administered. The vertical axis indicates the number of times of nose scratching per 10 minutes. Rhinitis model mice administered with the rhinitis preventive/therapeutic agent of Example 1 or Example 3 or rhinitis model mice administered with water of Comparative Example 1 on day 0, day 28, or day 35 of FIG. 1 or FIG. 1 is a graph showing the measurement results of ovalbumin-specific IgE antibody concentration in blood. 2 is a graph showing the therapeutic effect of allergic rhinitis in the case of administration of the rhinitis prophylactic/therapeutic agent of Example 2 or Example 3 or water of Comparative Example 1. FIG. The vertical axis indicates the number of sneezes per 10 minutes. 2 is a graph showing the therapeutic effect of allergic rhinitis in the case of administration of the rhinitis prophylactic/therapeutic agent of Example 2 or Example 3 or water of Comparative Example 1. FIG. The vertical axis indicates the number of times of nose scratching per 10 minutes. Rhinitis model mice administered with the rhinitis prophylactic/therapeutic agent of Example 2 or Example 3 or rhinitis model mice administered with water of Comparative Example 1 on day 0, day 28, or day 56 of FIG. 4 or FIG. 1 is a graph showing the measurement results of ovalbumin-specific IgE antibody concentration in blood.

Preferred embodiments for carrying out the present invention are described below.

The present invention is a prophylactic/therapeutic agent for rhinitis containing ornithine as an active ingredient.

The dosage form of the prophylactic/therapeutic agent for rhinitis is not particularly limited. or nasal drops and the like.

The prophylactic/therapeutic agents for rhinitis include processed foods. That is, the prophylactic/therapeutic agents for rhinitis include processed foods containing ornithine as an active ingredient. Processed foods include, for example, dietary supplements. Processed foods may be either liquids such as beverages or solids such as jelly or tablets.

Ornithine includes, for example, L-ornithine, D-ornithine, or racemic ornithine. These ornithine can use what is marketed.

Additives other than ornithine may be added to the prophylactic/therapeutic agent for rhinitis according to the dosage form. For example, solvents, thickeners, dextrin, petrolatum, preservatives, etc. can be added. Solvents include, for example, water. Thickeners include, for example, locust bean gum, guar gum derivatives, carrageenan, pectin, xanthan gum, gellan gum, diutan gum, starch and the like. Examples of preservatives include paraoxybenzoic acid esters (parabens), benzalkonium chloride, chlorobutanol, cresol, and the like. The additive preferably has the effect of realizing a desired dosage form, preventing putrefaction, and imparting viscosity. It is preferable not to .

The concentration of ornithine in the prophylactic/therapeutic agent for rhinitis or the processed food is preferably 5.0% by mass or less, more preferably 1.0% by mass or less, and 0.5% by mass or less. More preferably, it is 0.1% by mass or less. Although the lower limit of the concentration of ornithine is not particularly limited, it can be, for example, 0.0001% by mass or more. The concentration of ornithine can be adjusted, for example, by mixing the above additives with ornithine.

The method of use of the preventive/therapeutic agent for rhinitis is not particularly limited, as long as the method allows ornithine, which is an active ingredient, to be taken into the body. For example, it can be used by oral administration, transdermal administration, or injection.

Hereinafter, more specific description will be given with reference to Examples.

A mouse in which rhinitis symptoms were induced by the following method was prepared, and the rhinitis model mouse was orally ingested with a rhinitis prophylactic/therapeutic agent prepared by dissolving a predetermined concentration of ornithine in water by the following method, and rhinitis symptoms were observed. The effect on allergic rhinitis was observed separately for the preventive effect and the therapeutic effect.

[Preparation of Rhinitis Model Mouse]
Six-week-old BALB/c female mice were purchased from Clea Japan, Inc. and kept in an animal breeding room for one week to acclimatize them to the environment. The temperature in the animal breeding room was set at 24±2° C., the humidity was controlled at 51±13%, and the lighting was turned on or off at 12-hour intervals. Lighting hours are from 7:00 to 19:00. Food and water were provided freely. Solid feed CE-2 manufactured by CLEA Japan Co., Ltd. was used as the feed. Mice were handled in accordance with the "Notre Dame Seishin Women's University Animal Experiment Guidelines".

[Method of systemic sensitization]
A solution prepared by dissolving 1 μg of ovalbumin (Grade V) from Sigma-Aldrich and 100 μg of aluminum hydroxide gel (ALUM) from LSL Co., Ltd. in 200 μl of physiological saline was administered to mice on Day 0 and Day 5. systemic sensitization by intraperitoneal administration. One week after mouse purchase was defined as Day 0 (Day 0).

[Method of local sensitization]
Thereafter, from Day 14, 2 µl of the ovalbumin solution was administered to both nasal cavities using a micropipette. In FIGS. 1 and 2 and FIGS. 4 and 5, administration of ovalbumin to both nasal cavities is referred to as Local sensitization. An ovalbumin solution was prepared by dissolving the above ovalbumin in physiological saline to contain 100 mg of ovalbumin per ml. Henceforth, this is described as "100 mg/ml albumin physiological saline." In experiments on preventive effects, the results of which are shown in FIGS. 1 to 3, which will be described later, 100 mg/ml albumin in physiological saline was administered every day from Day 14 to Day 35. In addition, in the experiments on the therapeutic effects shown in Figures 4 to 6, which will be described later, 100 mg/ml albumin saline was administered daily from Day 14 to Day 28, and then on Day 56 at a frequency of three times a week. Administration of 100 mg/ml albumin saline was repeated until.

[Method of administration of prophylactic/therapeutic agent for rhinitis]
L-ornithine monohydrochloride (purity of 98% by mass or more) purchased from Nacalai Tesque Co., Ltd. was dissolved in tap water to prepare a rhinitis preventive/therapeutic agent. For the preparation, 10 mg (Example 1), 30 mg (Example 2), or 100 mg (Example 3) of L-ornithine monohydrochloride was added to 100 ml of tap water. The concentration of ornithine in Example 1 is 0.0098% by weight, the concentration of ornithine in Example 2 is 0.0294% by weight, and the concentration of ornithine in Example 3 is 0.0979% by weight. Each concentration of the rhinitis preventive/therapeutic agent or tap water (Comparative Example 1) was placed in a water supply bottle, and the rhinitis model mice prepared above were allowed to ingest freely. In addition, in the experiments on the preventive effect whose results are shown in FIGS. 1 to 3 described later, from Day 0 to Day 35, in the experiments on the therapeutic effect whose results are shown in FIGS. 4 to 6, from Day 28 to Day 56 Each concentration of prophylactic/therapeutic agent for rhinitis or tap water was administered. In FIGS. 1 and 2 and FIGS. 4 and 6, the administration period of the prophylactic/therapeutic agent for rhinitis or water is referred to as Ornithine administration.

[Experiment on preventive effect]
From Day 0 to Day 35, as described as Ornithine administration in FIGS. of water ad libitum from the water absorption bottle.

1 and 2, from Day 14 to Day 35, 100 mg/ml albumin physiological saline was administered to both nasal cavities of rhinitis model mice in the same manner as described above. I made a impression.

In experiments on the preventive effect, a rhinitis preventive/therapeutic agent was pre-administered from day 0 to day 14 before local sensitization of the nasal cavity. Therefore, it is possible to confirm the effect of the rhinitis preventive/therapeutic agent for preventing rhinitis. The preventive effect is summarized in the graphs of FIGS. 1 and 2. The preventive effect was evaluated by observing the symptoms of rhinitis described below. In addition, the data in FIGS. 1 and 2 are expressed as mean±standard error (n=7). "*" in FIGS. 1 and 2 indicates P<0.05, and "**" indicates P<0.01, both of which are compared with Comparative Example 1. FIG. Statistical processing was performed by one-way analysis of variance and multiple comparison by the Tukey-Kramer method. The criterion for significant difference was P<0.05. The meaning of error bars, the meaning of asterisks, and statistical processing are the same for FIG. 3, which will be described later.

[Observation of symptoms of rhinitis]
For observation of symptoms of rhinitis, first, rhinitis model mice were left in observation cages (21 cm×31 cm×13 cm) for 10 minutes to acclimatize. Then, as described above, 2 μl of 100 mg/ml albumin physiological saline solution was administered to both nostrils by nasal drops, immediately after which the animals were returned to the observation cage, and the number of sneezing and nose picking movements was counted for 10 minutes. Rhinitis symptoms were observed once a week from Day 0 to the end of the experiment.

As shown in FIGS. 1 and 2, in the groups to which the prophylactic/therapeutic agent for rhinitis of Examples 1 and 3 was administered, exacerbation of ovalbumin-induced allergic rhinitis symptoms was suppressed. From this, it can be seen that the rhinitis prophylactic/therapeutic agents of Examples 1 and 3 exert a prophylactic effect on allergic rhinitis.

[Measurement of ovalbumin-specific IgE antibody]
Rhinitis model mice administered with the rhinitis preventive/therapeutic agent of Example 1, rhinitis model mice administered with the rhinitis preventive/therapeutic agent of Example 3, rhinitis model mice administered with water of Comparative Example 1 , the concentration of IgE antibody that specifically reacts with ovalbumin was measured. For the measurement, on Day 0, Day 28, and Day 35, blood was collected from the tail vein of the mouse using a heparinized hematocrit capillary tube, and the collected blood was centrifuged using a hematocrit centrifuge (KUBOTA 3200). (12,000 rpm, 5 minutes) and the plasma was stored frozen at -30°C. After that, an EIA kit (Cayman Chemical) was used to measure the concentration of IgE antibody that specifically reacts with ovalbumin. The results are shown in FIG.

As shown in FIG. 3, the higher the concentration of ornithine contained in the prophylactic/therapeutic agent for rhinitis, the stronger the suppression of the increase in the concentration of ovalbumin-specific IgE antibodies in the blood.

[Experiment on therapeutic effect]
From Day 28 to Day 56, as described as Ornithine administration in FIGS. of water ad libitum from the water absorption bottle.

In the experiment on therapeutic effects, a prophylactic/therapeutic agent for rhinitis was administered from day 28 to day 56 after local sensitization of the nasal cavity. Local sensitization of the nasal cavity with 100 mg/ml albumin physiological saline was performed from day 14 to day 56 before administration of the prophylactic/therapeutic agent for rhinitis. Since the administration of the rhinitis preventive/therapeutic agent is started after the local sensitization, the effect of the rhinitis therapeutic/preventive agent for treating rhinitis can be confirmed. The therapeutic effect is summarized in the graphs of FIGS. 4 and 5. FIG. The therapeutic effect was evaluated by the same method as in the above "observation of symptoms of rhinitis". The data in FIGS. 4 and 5 are expressed as mean±standard error (n=14). "*" in FIGS. 4 and 5 indicates P<0.05, and "**" indicates P<0.01, both of which are compared with Comparative Example 1. FIG. Statistical processing was performed by one-way analysis of variance and multiple comparison by the Tukey-Kramer method. The criterion for significant difference was P<0.05.

As shown in FIGS. 4 and 5, in the groups administered with the prophylactic/therapeutic agent for rhinitis of Examples 2 and 3, compared with the group administered with water of Comparative Example 1, the effect of the agent for prophylactic/therapeutic rhinitis was higher. Symptoms of allergic rhinitis induced by ovalbumin have been suppressed since the start of administration. From this, it can be seen that the rhinitis prophylactic/therapeutic agents of Examples 2 and 3 exert therapeutic effects on allergic rhinitis.

[Measurement of ovalbumin-specific IgE antibody]
Rhinitis model mice administered with the rhinitis preventive/therapeutic agent of Example 2, Rhinitis model mice administered with the rhinitis preventive/therapeutic agent of Example 3, Rhinitis model mice administered with water of Comparative Example 1 , the concentration of IgE antibody that specifically reacts with ovalbumin was measured. The method for measuring the IgE antibody was the same as the method for "measurement of ovalbumin-specific IgE antibody" above. The results are shown in FIG. The data in FIG. 6 are expressed as mean±standard error (n=7). “*” in FIG. 6 indicates P<0.05 and is compared with Comparative Example 1. Statistical processing was performed by one-way analysis of variance and multiple comparison by the Tukey-Kramer method. The criterion for significant difference was P<0.05.

As shown in FIG. 6, the higher the concentration of ornithine contained in the rhinitis preventive/therapeutic agent, the higher the concentration of ovalbumin-specific IgE antibody in the blood, even when allergic rhinitis has already developed. It can be seen that it is greatly suppressed.

The prophylactic/therapeutic agents for rhinitis of Examples 1 to 3 contain only a very small amount of ornithine as an active ingredient. Although the content of ornithine is small, it exerts a preventive or therapeutic effect on allergic rhinitis. Since such an effect is exhibited even when the content is small, it is possible to reduce the dosage, and the effect of reducing the possibility of side effects is expected. Moreover, along with this, it is possible to reduce the manufacturing cost.

Claims (2)

A prophylactic/therapeutic agent for rhinitis containing ornithine as an active ingredient,
A prophylactic/therapeutic agent for rhinitis (excluding ornithine and valine) for preventing or treating rhinitis by suppressing an increase in blood IgE antibody concentration . The preventive/therapeutic agent for rhinitis according to claim 1, wherein the concentration of ornithine is 5.0% by mass or less.

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