JP2020132559A - Prophylactic/therapeutic agent for rhinitis - Google Patents

Abstract

To provide prophylactic/therapeutic agents for rhinitis containing ornithine, which has an established method for industrial mass production and has a low risk to the living body, as an active ingredient.SOLUTION: The above subject is solved by a prophylactic/therapeutic agent for rhinitis containing ornithine as an active ingredient. The concentration of ornithine is preferably 5.0 mass% or less.SELECTED DRAWING: Figure 1

Description

The present invention relates to a rhinitis preventive / therapeutic agent that can be used for the prevention or treatment of rhinitis.

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

Further, Patent Document 2 states that inhalation of the arginase inhibitor ABH in guinea pigs with allergic asthma improves the allergen-induced airway hyperresponsiveness (AHR).

WO2006 / 049286 Special Table 2010-510257

Patent Document 1 states that administration of amino acids such as valine and leucine to mice inducing symptoms of allergic asthma reduces the number of eosinophils. However, it is not stated whether these amino acids are effective against rhinitis. In addition, there is a description that administration of ornithine alone to the mice does not improve allergic symptoms.

Patent Document 2 also only verifies the amount of PC100 histamine when an arginase inhibitor is administered and when water is administered to mice that have induced symptoms of allergic asthma, and ornithine causes rhinitis. It is not stated whether or not it is effective against.

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

The above-mentioned problems will be solved by a rhinitis preventive / therapeutic agent containing ornithine as an active ingredient. In the rhinitis preventive / therapeutic agent, the concentration of ornithine is preferably 5.0% by mass or less.

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

It is a graph which shows the preventive effect of allergic rhinitis when the rhinitis preventive / therapeutic agent of Example 1 or Example 3 or the water of Comparative Example 1 is administered. The vertical axis shows the number of sneezes per 10 minutes. It is a graph which shows the preventive effect of allergic rhinitis when the rhinitis preventive / therapeutic agent of Example 1 or Example 3 or the water of Comparative Example 1 is administered. The vertical axis shows the number of nasal scratches per 10 minutes. The rhinitis model mouse to which the rhinitis preventive / therapeutic agent of Example 1 or Example 3 was administered, or the rhinitis model mouse to which water of Comparative Example 1 was administered on day 0, day 28, or day 35 of FIG. 1 or 2. It is a graph which shows the measurement result of the concentration of the egg white albumin-specific IgE antibody in blood. It is a graph which shows the therapeutic effect of allergic rhinitis when the rhinitis preventive / therapeutic agent of Example 2 or Example 3 or the water of Comparative Example 1 is administered. The vertical axis shows the number of sneezes per 10 minutes. It is a graph which shows the therapeutic effect of allergic rhinitis when the rhinitis preventive / therapeutic agent of Example 2 or Example 3 or the water of Comparative Example 1 is administered. The vertical axis shows the number of nasal scratches per 10 minutes. The rhinitis model mouse to which the rhinitis preventive / therapeutic agent of Example 2 or Example 3 was administered, or the rhinitis model mouse to which water of Comparative Example 1 was administered on day 0, day 28, or day 56 of FIG. 4 or FIG. It is a graph which shows the measurement result of the concentration of the egg white albumin-specific IgE antibody in blood.

Hereinafter, preferred embodiments for carrying out the present invention will be described.

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

The dosage form of the rhinitis preventive / therapeutic agent is not particularly limited, and for example, oral preparations such as tablets, capsules, granules, fine preparations, drops and syrups; injections; transdermal preparations such as lotions, ointments, aerosols and tapes. It can also be a nasal drop.

Rhinitis preventive / therapeutic agents shall include the form of processed foods. That is, the rhinitis preventive / therapeutic agent includes processed foods containing ornithine as an active ingredient. Examples of processed foods include dietary supplements. The processed food may be either a liquid such as a beverage or a solid such as a jelly or a tablet.

Examples of ornithine include L-ornithine, D-ornithine, and racemic ornithine. As these ornithines, commercially available ones can be used.

Additives other than ornithine may be added to the rhinitis preventive / therapeutic agent according to the dosage form. For example, a solvent, a thickener, dextrin, petrolatum, a preservative, or the like can be blended. Examples of the solvent include water. Examples of the thickener include locust bean gum, guar gum derivative, carrageenan, pectin, xanthan gum, gellan gum, dieutan gum, starch and the like. Examples of the preservative include paraoxybenzoic acid esters (paraben), benzalkonium chloride, chlorobutanol, cresol and the like. The additive preferably has the effect of realizing a desired dosage form, preventing putrefaction, and imparting viscosity, and also contains an amino acid other than ornithine in order to increase the content of ornithine. It is preferable not to do so.

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

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

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

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

[Preparation of rhinitis model mouse]
A 6-week-old BALB / c female mouse was purchased from Japan Claire Co., Ltd. and bred in an animal breeding room for 1 week in order to adapt to the environment. The temperature of the animal breeding room was set to 24 ± 2 ° C., the humidity was controlled to 51 ± 13%, and the lights were turned on or off every 12 hours. The lighting time is from 7:00 to 19:00. Food and water were provided in an environment where they could be freely ingested. The solid feed CE-2 manufactured by Nippon Claire Co., Ltd. was used as the feed. The handling of mice was carried out in accordance with the "Notre Dame Seishin University Animal Experiment Guidelines".

[Method of whole body sensitization]
Sigma-Aldrich's egg white albumin (Grade V) 1 μg and LSL Co., Ltd. aluminum hydroxide gel (ALUM) 100 μg dissolved in 200 μl of physiological saline were added to mice on Day 0 and Day 5. Was administered intraperitoneally to cause systemic sensitization. One week after the purchase of the mouse was set as Day 0 (day 0).

[Method of local sensitization]
Then, from Day 14, 2 μl of ovalbumin solution was administered to both nasal cavities using a micropipette. In addition, in FIG. 1 and FIG. 2 and FIGS. 4 and 5, administration of ovalbumin to both nasal cavities is referred to as local sensitization. The egg white albumin solution was prepared by dissolving the above-mentioned egg white albumin in physiological saline so as to contain 100 mg of egg white albumin per ml. Hereinafter, this will be referred to as "100 mg / ml albumin physiological saline". In the experiment on the preventive effect shown in FIGS. 1 to 3 described later, 100 mg / ml albumin saline was administered every day from Day 14 to Day 35. In the experiment on the therapeutic effect shown in FIGS. 4 to 6 described later, 100 mg / ml albumin saline was administered daily from Day 14 to Day 28, and then on Day 56 three times a week. Until then, administration of 100 mg / ml albumin saline was repeated.

[Method of administration of preventive / therapeutic agents for rhinitis]
L-ornithine monohydrochloric acid (purity 98% by mass or more) purchased from Nacalai Tesque Co., Ltd. was dissolved in tap water to prepare a rhinitis preventive / therapeutic agent. At the time of preparation, 10 mg (Example 1), 30 mg (Example 2), or 100 mg (Example 3) of L-ornithine monohydrochloric acid was added to 100 ml of tap water, respectively. The concentration of ornithine in Example 1 is 0.0098% by mass, the concentration of ornithine in Example 2 is 0.0294% by mass, and the concentration of ornithine in Example 3 is 0.0979% by mass. Each concentration of rhinitis preventive / therapeutic agent or tap water (Comparative Example 1) was put into a water bottle and allowed to be freely ingested by the rhinitis model mice prepared above. In the experiments on the preventive effect shown in FIGS. 1 to 3, which will be described later, from Day 0 to Day 35, and in the experiments on the therapeutic effect shown in FIGS. 4 to 6, from Day 28 to Day 56. Each concentration of rhinitis preventive / therapeutic agent or tap water was administered. In FIGS. 1 and 2 and FIGS. 4 and 6, the administration period of the rhinitis preventive / therapeutic agent or water is referred to as ornithine administration.

[Experiment on preventive effect]
As described as Ornithine administration in FIGS. 1 and 2, from Day 0 to Day 35, the rhinitis model mice prepared above were subjected to the rhinitis preventive / therapeutic agent of Example 1 or Example 3, or Comparative Example 1. Water was freely ingested from the water absorption bottle.

Then, as described as Local sensitization in FIGS. 1 and 2, from Day 14 to day 35, 100 mg / ml albumin saline was administered daily to both nasal cavities of rhinitis model mice in the same manner as described above, and the local sensitization was performed. I made a sensitization.

In the experiment on the preventive effect, the rhinitis preventive / therapeutic agent was administered in advance from day 0 to day 14 before the local sensitization to the nasal cavity was performed. Therefore, it is possible to confirm the effect of the rhinitis preventive / therapeutic agent on preventing rhinitis. The preventive effects are summarized in the graphs of FIGS. 1 and 2. The preventive effect was evaluated by observing the following rhinitis symptoms. The data in FIGS. 1 and 2 are displayed with an average value ± 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. Statistical processing was performed by one-way ANOVA and multiple comparison by Tukey-Kramer method. The standard of significant difference was P <0.05. The meanings of error bars, asterisks, and statistical processing are the same for FIG. 3, which will be described later.

[Observation of rhinitis symptoms]
In the observation of rhinitis symptoms, first, the rhinitis model mice were left in an observation cage (21 cm × 31 cm × 13 cm) for 10 minutes to acclimatize. Then, as described above, immediately after nasal instillation of 2 μl of 100 mg / ml albumin physiological saline into both nasal cavities, the patient was returned to the observation cage and the number of sneezing and the number of nasal scratching movements were measured for 10 minutes. Observation of rhinitis symptoms was performed once a week from Day 0 to the end of the experiment.

As shown in FIGS. 1 and 2, in the group to which the rhinitis preventive / therapeutic agents of Examples 1 and 3 were administered, the exacerbation of allergic rhinitis induced by ovalbumin was suppressed. From this, it can be seen that the rhinitis preventive / therapeutic agents of Examples 1 and 3 exert a preventive effect on allergic rhinitis.

[Measurement of egg white albumin-specific IgE antibody]
A rhinitis model mouse to which the rhinitis preventive / therapeutic agent of Example 1 was administered, a rhinitis model mouse to which the rhinitis preventive / therapeutic agent of Example 3 was administered, and a rhinitis model mouse to which water was administered of Comparative Example 1 in the above experiment on the preventive effect. The concentration of IgE antibody that specifically reacts with ovalbumin was measured for each of the above. For measurement, on Day 0, Day 28, and Day 35, blood was collected from the tail vein of the mouse using a heparin-treated hematocrit capillary, and the collected blood was centrifuged using a hematocrit centrifuge (KUBOTA 3200). (12,000 rpm, 5 minutes) and the plasma was cryopreserved at −30 ° C. Then, 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, it can be seen that the higher the concentration of ornithine contained in the rhinitis preventive / therapeutic agent, the stronger the increase in the concentration of egg white albumin-specific IgE antibody in the blood is suppressed.

[Experiment on therapeutic effect]
As described as Ornithine administration in FIGS. 4 and 5, from Day 28 to Day 56, the rhinitis model mice prepared above were subjected to the rhinitis preventive / therapeutic agents of Examples 2 and 3 or Comparative Example 1. Water was freely ingested from the water absorption bottle.

In experiments on the therapeutic effect, rhinitis preventive / therapeutic agents were administered from day 28 to day 56 after local sensitization to the nasal cavity. Local sensitization to the nasal cavity with 100 mg / ml albumin saline is performed from day 14 to day 56 before administration of the rhinitis preventive / therapeutic agent. Since the administration of the rhinitis preventive / therapeutic agent is started after the local sensitization is performed, the effect of the rhinitis treatment / preventive agent on the rhinitis can be confirmed. The therapeutic effects are summarized in the graphs of FIGS. 4 and 5. The therapeutic effect was evaluated by the same method as in the above-mentioned "observation of rhinitis symptoms". The data of FIGS. 4 and 5 are displayed with an average value ± 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. Statistical processing was performed by one-way ANOVA and multiple comparison by Tukey-Kramer method. The standard of significant difference was P <0.05.

As shown in FIGS. 4 and 5, the group to which the rhinitis preventive / therapeutic agent of Example 2 and Example 3 was administered had a rhinitis preventive / therapeutic agent as compared with the group to which the water of Comparative Example 1 was administered. The symptoms of allergic rhinitis induced by ovalbumin have been suppressed since the start of administration. From this, it can be seen that the rhinitis preventive / therapeutic agents of Examples 2 and 3 exert a therapeutic effect on allergic rhinitis.

[Measurement of egg white albumin-specific IgE antibody]
A rhinitis model mouse to which the rhinitis preventive / therapeutic agent of Example 2 was administered, a rhinitis model mouse to which the rhinitis preventive / therapeutic agent of Example 3 was administered, and a rhinitis model mouse to which water of Comparative Example 1 was administered in the above experiment on the therapeutic effect. The concentration of IgE antibody that specifically reacts with ovalbumin was measured for each of the above. The method for measuring IgE antibody was the same as the method of "Measurement of egg white albumin-specific IgE antibody" described above. The results are shown in FIG. The data in FIG. 6 was displayed as an average value ± standard error (n = 7). “*” In FIG. 6 indicates P <0.05, which is compared with Comparative Example 1. Statistical processing was performed by one-way ANOVA and multiple comparison by Tukey-Kramer method. The standard of 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 increase in the concentration of egg white albumin-specific IgE antibody in the blood even when allergic rhinitis has already developed. It can be seen that it is greatly suppressed.

The rhinitis preventive / therapeutic agents 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 if the content is small, the dose can be reduced, and the effect of reducing the possibility of side effects is expected. In addition, the manufacturing cost can be reduced accordingly.

Claims (2)

A rhinitis preventive / therapeutic agent containing ornithine as an active ingredient. The rhinitis preventive / therapeutic agent according to claim 1, wherein the concentration of ornithine is 5.0% by mass or less.

Images