JP4690158B2 - Efficacy evaluation animal, method of onset of chronic obstructive pulmonary disease of the effect evaluation animal, and efficacy evaluation method using the animal - Google Patents

Description

The present invention relates to an animal for evaluating efficacy, which has no inflammation in the nasal cavity and pharynx and developed chronic obstructive pulmonary disease, a method for developing chronic obstructive pulmonary disease in the animal for evaluating efficacy, and a drug using the animal for evaluating efficacy It is related with the medicinal efficacy evaluation method.

Conventionally, as a method for evaluating the efficacy of anti-chronic obstructive pulmonary disease substances, tobacco smoke is aspirated into experimental animals as a cause of chronic obstructive pulmonary disease, and an animal for evaluating efficacy is experimentally developed for chronic obstructive pulmonary disease. It was prepared and the drug was evaluated (for example, see Patent Document 1). The medicinal efficacy evaluation animal disclosed in this document is prepared by inhaling mainstream smoke generated from Kentucky Reference Cigarette lRl in 6-week-old mice for 1 to 4 hours / day, 5 days / week for a total of 6 months, It took a long time from the start of the test to the evaluation of the drug, and had the disadvantage that an effective drug evaluation could not be performed.
In addition, in the method of inhaling tobacco smoke, since it passes through the oral cavity or nasal cavity, there is a possibility of causing an inflammatory reaction other than in the chronic obstructive pulmonary disease area, and the medicinal efficacy evaluation animal in which the affected area is selectively specified It was difficult to develop.

JP 2004-105173 A

  An object of the present invention is to provide a method for developing only a chronic obstructive pulmonary disease in an animal for evaluating drug efficacy, for example, having no inflammation in the nasal cavity and pharynx and developing only chronic obstructive pulmonary disease, and such an animal for evaluating drug efficacy, At the same time, it is intended to provide a method for evaluating selective drug efficacy against chronic obstructive pulmonary disease by administering a drug to such animal for drug efficacy evaluation.

The present invention is a method for developing chronic obstructive pulmonary disease in an animal for drug efficacy evaluation, wherein an aqueous solution obtained by dissolving tobacco smoke in water or physiological saline is directly administered to the lower respiratory tract of animals other than humans ,
In the direct administration, a bent sonde is inserted from the oral cavity, the tip is applied to the upper lower respiratory tract of the animal's pharynx, and the aqueous solution is administered directly from the other end of the sonde to the lower respiratory tract by spontaneous breathing of the animal. Provided is a method for developing chronic obstructive pulmonary disease in an animal for evaluating drug efficacy.
The present invention also provides an animal for evaluating drug efficacy, wherein the animal for evaluating chronic efficacy is caused to develop chronic obstructive pulmonary disease by the method for developing chronic obstructive pulmonary disease.
Furthermore, the present invention provides a drug efficacy evaluation method for administering a drug to the animal for drug efficacy evaluation and evaluating the action effect.

According to the present invention, it is possible to reliably provide an animal for drug efficacy evaluation that selectively develops only chronic obstructive pulmonary disease, and an evaluation method useful for the development of pharmaceuticals, particularly for screening for anti-chronic obstructive pulmonary disease substances It is possible to quickly provide a medicinal effect evaluation animal for performing the above.
The effects of the present invention are more specifically shown below.
(1) Although the conventional experimental animal model took 24 weeks or more until the symptoms of chronic obstructive pulmonary disease appeared, according to the present invention, it can be produced in a short period of 4 weeks or less.
(2) In the conventional experimental animal model, tobacco smoke was inhaled through the nasal cavity, but in the present invention, an aqueous solution in which tobacco smoke is dissolved in water or physiological saline is prepared and directly administered to the lower respiratory tract. As a result, the concentration of the components contained in the tobacco smoke in the aqueous solution can be changed. As a result, the degree of the pathological condition can be adjusted, and an animal for evaluation of medicinal effects with little variation can be produced. It becomes.
(3) In the conventional method in which cigarette smoke is inhaled, it passes through the oral cavity or nasal cavity, and therefore, an inflammatory reaction is caused outside the chronic obstructive pulmonary disease part. However, in the present invention, by administering an aqueous solution in which tobacco smoke is directly dissolved in the lower respiratory tract without going through the oral cavity or nasal cavity, it becomes possible to selectively identify the affected area and develop only chronic obstructive pulmonary disease. .
(4) Up until now, the creation of models required a device that generated tobacco smoke and an exposure chamber that allowed animals to inhale smoke. However, such a large-scale device was not required, and it was inexpensive and reliable. It is possible to produce an animal for evaluating drug efficacy.

In recent years, the increase in deaths from chronic obstructive pulmonary disease, which is thought to be caused mainly by smoking, has become a problem in Europe, the United States, and Japan, and the development of effective treatments and drugs is a very important medical issue. Therefore, development of a drug having a therapeutic effect has been desired, and an animal for evaluating drug efficacy for drug screening tests has been awaited.
According to the present invention, an animal for evaluating efficacy (hereinafter referred to as “chronic obstructive pulmonary disease model animal”) that has developed a chronic obstructive pulmonary disease desired in the market can be reliably provided.

Here, the chronic obstructive pulmonary disease referred to in the present invention will be briefly described below.
Chronic obstructive pulmonary disease is a pulmonary disease including chronic bronchitis and emphysema. Chronic obstructive pulmonary disease, although partially reversible, is generally characterized as progressive irreversible airway obstruction and is often associated with airway hyperresponsiveness. Chronic bronchitis is characterized by a chronic wet cough that lasts for more than 3 months in each year of two consecutive years. Emphysema is also an abnormal permanent swelling of the airspace distal to the terminal bronchioles with destructive changes in the alveolar walls and no obvious fibrosis. Destruction is defined as an irregular enlargement of the respiratory airspace, and the regular appearance of lung lobule and its components may be lost.

  As described above, chronic obstructive pulmonary disease is characterized by irreversible airway obstruction and has a different disease concept from asthma, which is a reversible airway obstruction disease. Furthermore, as a pharmacotherapy for bronchial asthma, inhaled steroids are the first in the Guideline for the Diagnostics of Management and Asthma (NHLBI, 2002), which is a global guideline for the treatment of asthma. Although it has been recommended as a single-choice drug and has proven to be of great utility, it has a similar global guideline, Global Initiative for Chronic Observative Lang Diseases (Global Initiative for Chronic Obstructive Lungs). disease) (GOLD; NHLBI / WHO, 1998) In, the effect of steroids is limited and not very recommended its use. Thus, the reactivity to drugs is different between chronic obstructive pulmonary disease and bronchial asthma.

  Risk factors for chronic obstructive pulmonary disease are harmful particulates due to smoking and air pollution, and the long-term exposure of these factors is thought to be caused by persistent chronic inflammatory conditions in the peripheral airways and alveoli. In other words, the Global Initiative for Chronic Obstructive Lange Disease (GOLD; NHLBI / WHO, 1998), which is the aforementioned global guideline, is a pulmonary disease in chronic obstructive pulmonary disease. It is clearly stated that inflammation is seen, and the important cause / progression factor is an imbalance between protease and protease inhibitor and inflammation due to oxidative stress.

  In the present invention, by selectively providing a chronic obstructive pulmonary disease model animal that has developed only chronic obstructive pulmonary disease that is different from the conventional chronic obstructive pulmonary disease model animal, the efficacy of the drug is accurately and temporally improved. Makes it possible to evaluate.

The chronic obstructive pulmonary disease model animal of the present invention can be administered directly to the lower respiratory tract of non-human animals with an aqueous solution in which tobacco smoke is dissolved in water or physiological saline (hereinafter referred to as “cigarette smoke solution”). Administration of smoke solution and endotoxin-containing aqueous solution (hereinafter, endotoxin-containing aqueous solution is simply referred to as “endotoxin-containing aqueous solution”) for direct administration to the lower respiratory tract of animals other than humans, or water or It is prepared by directly administering an aqueous solution in which tobacco smoke and endotoxin are dissolved in physiological saline (hereinafter referred to as “tobacco smoke / endotoxin-containing solution”) to the lower respiratory tract of animals other than humans. The “lower airway” here is defined as the airway, trachea, bronchi, bronchiole, alveoli, and the like. The term “directly administered” as used herein means that tobacco smoke liquid, endotoxin-containing aqueous solution, and tobacco smoke / endotoxin-containing aqueous solution are directly administered to the lower respiratory tract without touching the nasal cavity.

  The tobacco smoke liquid is produced, for example, by the method shown in the schematic diagram of FIG. That is, the filter suction port of the tobacco 1 is joined to one end of the introduction tube 4, and the other end of the introduction tube 4 is submerged in water in the container 3 or physiological saline 2. On the other hand, the suction pump 6 and the suction pipe 7 are joined, and the other end of the container 3 and the suction pipe 7 are joined so as not to touch the water surface. The suction pipe 7 is provided with a valve 5. After the cigarette 1 is ignited, the suction pump 6 is operated, and cigarette smoke is bubbled into the water while opening and closing the valve 5 as appropriate, thereby producing a tobacco smoke liquid.

  The concentration of the tobacco smoke in the tobacco smoke liquid and the tobacco smoke / endotoxin-containing aqueous solution may be a concentration sufficient to cause chronic obstructive pulmonary disease in the applied animal. For example, one or more cigarettes per 10 mL, preferably 5 It can be brought about by sucking more than a book of smoke. In addition, the amount of the tobacco smoke used is not limited as long as it is sufficient to cause chronic obstructive pulmonary disease in the applied animal, and preferably 20 μL or more, more preferably in one administration per animal. The range of 50 μL to 5 mL can be mentioned. The aqueous solution can contain water, physiological saline, and a water-soluble organic solvent that does not adversely affect animals other than humans. Here, examples of the organic solvent include organic solvents such as ethanol and dimethyl sulfoxide. The ratio of the organic solvent contained in these aqueous solutions varies depending on the animal to be used. A concentration of about 10% by weight can be mentioned.

Next, the endotoxin produced by the Gram-negative bacterium used in the present invention will be described.
The purpose of combining endotoxin produced by Gram-negative bacteria is thought to be the main cause of the development of chronic obstructive pulmonary disease, but secondary factors such as infection are said to exacerbate the symptoms. By adding this secondary factor, it is to create a disease model that is closer to the actual symptoms. The endotoxin produced by the gram-negative bacterium here refers to an endotoxin released from the cell wall of the gram-negative bacterium, and examples thereof include lipopolysaccharide. Such endotoxin is preferably administered in the form of a solution, for example, dissolved in physiological saline.

  The concentration of endotoxin produced by the gram-negative bacterium may be a concentration sufficient to cause chronic obstructive pulmonary disease in the applied animal. For example, it is preferable for water, physiological saline, or tobacco smoke. Can be 10 μg / mL or more, and more preferably in the range of 50 μg / mL to 2 g / mL. The endotoxin produced by the gram-negative bacterium may not be contained, but for the reasons described above, it is preferable that the endotoxin is contained in water, physiological saline or tobacco smoke.

  The optimal conditions for the dose and number of doses of tobacco smoke, endotoxin-containing aqueous solution, and tobacco smoke / endotoxin-containing aqueous solution vary depending on the type of experimental animal used. It is preferable to set conditions.

In addition, the number of administrations and the administration period of the tobacco smoke liquid, the endotoxin-containing aqueous solution, and the tobacco smoke / endotoxin-containing aqueous solution can be determined by the degree of onset of chronic obstructive pulmonary disease. The degree of onset is evaluated by the peak expiratory flow (mL / sec), and the number and period of decrease of 10% or more are set with respect to the physiological saline administration group as the control group. When the tobacco smoke liquid and the endotoxin-containing aqueous solution are administered in combination, the endotoxin-containing aqueous solution is preferably administered between the tobacco smoke liquid and the administration.
By administering endotoxin produced by gram-negative bacteria in addition to the aqueous solution, chronic obstructive pulmonary disease can be surely developed in a short period of time.

  The thus obtained animal model of chronic obstructive pulmonary disease has a characteristic that it can selectively cause inflammation at the target site of chronic obstructive pulmonary disease. For example, when the Evans Blue solution was administered to the lower respiratory tract, which is a feature of the present invention, and the colored part of the living body was confirmed, the colored part was not observed in the nasal part and nasal cavity, but was limited to the mouth and lower respiratory tract. The ratio was almost 100%.

Next, a method for directly administering tobacco smoke, tobacco smoke and endotoxin-containing aqueous solution or tobacco smoke / endotoxin-containing aqueous solution to the lower respiratory tract of animals other than humans will be described.
For administration, a metal sonde can be used, but a commonly used stainless steel sonde can be used, and a preferred metal sonde such as size and shape can be selected depending on the type of animal used in the experiment. . Usually, it is preferable to use the tube tip processed so as to ensure direct administration to the lower respiratory tract. As a processing method, for example, there is a method of bending the tip of a stainless steel sonde so that it can be easily administered directly to the lower respiratory tract. The degree and shape of the curve vary depending on the type and shape of the animal used, but in general, the curve is formed so that the angle formed by the portion corresponding to the curved tip and the portion corresponding to the base of the sonde is 90 to 150 °. It is preferable to make it. In direct administration, a curved sonde is inserted from the oral cavity, its tip is applied to the upper lower airway of the pharynx of the animal, and the aqueous solution is administered directly from the other end of the sonde to the lower airway by spontaneous breathing of the animal.

  As the experimental animal used in the present invention, an animal having a lower respiratory tract and performing pulmonary respiration can be used. For example, rodents such as guinea pigs, mice, rats, and hamsters, heavy teeth such as rabbits, goats, Mammals such as sheep, pigs, etc., horses, donkeys, etc., dogs, cats, etc., primates such as monkeys, chimpanzees, etc. Guinea pigs, mice, rats, hamsters, rabbits, monkeys, chimpanzees, dogs, goats, sheep, pigs, etc. are preferred, and considering the advantages of experimental operation due to shape, etc., and the cost of experimental animals, guinea pigs, mice, rats, Rodents such as hamsters are particularly preferred.

As a drug efficacy evaluation method, a drug is administered to the chronic obstructive pulmonary disease model animal, and its action effect is evaluated. What is necessary is just to perform evaluation with a change of a respiratory function for an effect. Evaluation of chronic obstructive pulmonary disease in the animal model of chronic obstructive pulmonary disease of the present invention is performed by evaluating changes in respiratory function normally used as an evaluation parameter of the disease, that is, specific airway resistance, peak expiratory flow, tidal volume It is performed with the number of minute breaths and the minute ventilation, and these can be measured by a known method. For example, specific airway resistance is described by Pennock BE, et al, Journal of Applied Physiology (1979), 46, No. 2, p. In accordance with a method described in 399-406 (hereinafter referred to as “Penock et al. Method”), a total respiratory function measurement system (for example, PULMOS-I; M.I.P.S, etc.) via a flow sensor. Can be measured. In addition, the measurement of the peak expiratory flow, the tidal volume, the number of minute breaths, and the minute ventilation is, for example, PULMOS-I; I. P. S can be used for measurement.
In chronic obstructive pulmonary disease, neutrophils infiltrate into the cell membrane among inflammatory cells, so check the degree of inflammation by measuring the number of neutrophil cells. Is also possible.

  In order to describe the contents of the present invention more specifically, the following examples are shown. However, the scope of the present invention is not limited to these examples.

Example 1
<Creation of chronic obstructive pulmonary disease model animal>
The production of a chronic obstructive pulmonary disease model animal when the experimental animal is a guinea pig and evaluation results of a drug using the same were shown.
1. Preparation of tobacco smoke liquid Tobacco smoke liquid was prepared by the apparatus according to the schematic diagram of FIG.
1) Number of cigarettes used: 40 2) Amount of physiological saline: 40 mL
3) Pump intake volume and intake time: Using a suction pump (intake volume: 6.5 mL / min.), A cigarette smoke is sucked into physiological saline with a suction time of about 5 minutes per cigarette as a guide. And the desired tobacco smoke liquid was prepared.

2. The preparation of the solution with Li Po polysaccharide as endotoxin produced by preparing gram negative bacteria lipopolysaccharide solution were carried out as follows. That is, 50 mg of lipopolysaccharide was dissolved in 100 mL of physiological saline to prepare a 500 μg / mL lipopolysaccharide solution.

3. Development of disease animals And 2. Disease animals were developed using the tobacco smoke liquid and lipopolysaccharide liquid prepared in (1) as follows.
1) Animals used: 6 Hartley male guinea pigs (purchased at 4 weeks of age and preliminarily raised for 1 week before the test)
2) Dosage form and administration period: Tobacco smoke solution 200 μL / animal / dose was administered once a day for 4 consecutive days, and lipopolysaccharide solution 500 μL / animal / day was administered on the fifth day. With this as one cycle, the same operation was repeated on the 6th and 11th days, and 200 μL of tobacco smoke / animal / dose was administered on the 16th to 19th days.
3) Administration method: In direct administration, a bent sonde is inserted from the oral cavity, the tip is applied to the upper lower respiratory tract of the animal's pharynx, and the animal's spontaneous breathing causes tobacco smoke and lipopolysaccharide solution from the other end of the sonde. Was administered directly into the lower respiratory tract.

4). Evaluation of the disease On the 20th day, specific airway resistance, peak expiratory flow, tidal volume, number of minute breaths, and minute volume were measured on the animals developed in (1).
The results are shown in Table 1 (average value of 6 animals).

According to this example, the affected animals have increased specific airway resistance, decreased peak expiratory flow, decreased tidal volume, decreased number of minute expirations, decreased minute ventilation, chronic It was confirmed that obstructive pulmonary disease had developed.
Therefore, from the results of Example 1, according to the present invention, an experimental chronic obstructive pulmonary disease model animal having a strong reaction in a short period of time without causing inflammation in other parts such as the oral cavity and the nasal cavity is obtained. It became clear that it can be manufactured without using a large-scale apparatus such as the above.

<Lung organ and lung tissue findings>
Regarding the chronic obstructive pulmonary disease of the above-mentioned animals, a photograph of lung organs and a micrograph of lung tissue in such guinea pigs were presented.
That is, FIG. 2 is an external view of a lung of a guinea pig that has not developed by administration of only physiological saline, and FIG. 3 is an external view of the lung of a guinea pig that has developed according to this example. In the guinea pig in which this example developed, pulmonary hyperinflation, which is a characteristic of chronic obstructive pulmonary disease, was confirmed.
4 is a photomicrograph of pulmonary tissue of a guinea pig that has not developed by administration of only physiological saline, and FIG. 5 is a photomicrograph of the pulmonary tissue of a guinea pig that developed in this example. Compared with normal guinea pigs, the guinea pigs were confirmed to have a cavity of the alveolar wall that is characteristic of chronic obstructive pulmonary disease.
From the above photographs of lung organs and micrographs of lung tissue, the symptoms of chronic obstructive pulmonary disease were confirmed, and it was revealed that the guinea pig of this example developed the disease.

<Measurement of neutrophil count in alveolar lavage fluid>
In chronic obstructive pulmonary disease, it is known that among inflammatory cells, neutrophils infiltrate the cell mucosa and cause inflammation. Therefore, the neutrophil count was measured for the purpose of confirming that the chronic obstructive pulmonary disease reaction occurred in the experimental model of the present invention by measuring the neutrophil count.
5. Leukocyte collection method After measuring the respiratory function of the affected guinea pigs, the animals were exsanguinated, and the blood vessels were perfused with physiological saline from the pulmonary artery, and physiological saline (fluid 5 mL × 2 / An animal was injected to perform alveolar lavage (BAL), and BAL fluid (BALF) was collected. BALF was centrifuged with a refrigerated centrifuge (4 ° C., 400 × g, 10 minutes), hemolyzed, centrifuged again under the same conditions, and the resulting pellet was suspended in physiological saline.
6). 4. Measurement of total white blood cell count 100 μL of Turku's solution was added to 25 μL of BALF obtained in the above and stained, and the total white blood cell count was counted using a Birkelturk hemocytometer (Elma sales).
7). 5. Cell smear preparation method and neutrophil measurement Based on the total white blood cell count, adjust the BALF leukocyte concentration to 3 × 10 ⁵ cells / mL with physiological saline, collect 25 μL from it, precipitate the cells on a slide glass, and prepare a Diff-Quick stained sample. Neutrophils were counted under microscopic examination.
The results (average value in 6 animals) are shown in Table 2.

In the present Example, it became clear that the neutrophil count increased in the tobacco smoke liquid and lipopolysaccharide administration group compared to the non-stimulated group, and symptoms of chronic obstructive pulmonary disease developed.

(Example 2) Confirmation of effect of known drug theophylline Confirmation of effect using theophylline known as a therapeutic agent for chronic obstructive pulmonary disease for the purpose of clarifying its usefulness as a method for evaluating the drug of the present invention. Went.
In this example, tobacco smoke liquid preparation, lipopolysaccharide preparation, diseased animal onset, and disease evaluation were performed in the same manner as in Example 1. Theophylline was orally administered (10 mg / kg) one hour before administration of tobacco smoke and LPS solution.
The results are shown in Table 3 (average value of 6 animals).

  According to the present example, the theophylline was confirmed to have an effect of improving respiratory function in the evaluation method according to the present invention, and it was revealed that the evaluation method is useful as a method for evaluating chronic obstructive pulmonary disease.

(Example 3) Production of a chronic obstructive pulmonary disease model animal A chronic obstructive pulmonary disease model animal was prepared in the same manner as in Example 1 using dog, sheep, and monkey experimental animals. As confirmation of the onset, the number of neutrophils was measured by the same method as in Example 1.
The results (average value in 6 animals) are shown in Table 4.

  From this example, in each experimental animal, the onset of chronic obstructive pulmonary disease was clarified in the tobacco smoke liquid and lipopolysaccharide administration groups.

  The animal model of chronic obstructive pulmonary disease of the present invention is useful because it can be used for screening of compounds in the development of pharmaceuticals, particularly in the development of anti-chronic obstructive pulmonary disease therapeutic agents.

It is a schematic diagram which shows the method of producing the tobacco smoke liquid used by this invention. BRIEF DESCRIPTION OF THE DRAWINGS It is an external view of the lungs of a guinea pig which does not develop by administering only the physiological saline used as the comparison object in Example 1. BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is an external view of an affected guinea pig lung in Example 1. It is a microscope picture of the lung tissue of the lung of a guinea pig which does not develop by administering only the physiological saline used as a comparison object in Example 1. 2 is a photomicrograph of lung tissue of a guinea pig lung that developed in Example 1. FIG.

Explanation of symbols

1 Tobacco 2 Water or physiological saline 3 Container 4 Introduction pipe 5 Valve 6 Suction pump 7 Suction pipe

Claims (18)

A method for developing chronic obstructive pulmonary disease in an animal for drug efficacy evaluation, wherein an aqueous solution obtained by dissolving cigarette smoke in water or physiological saline is directly administered to the lower respiratory tract of animals other than humans ,
In the direct administration, a bent sonde is inserted from the oral cavity, the tip is applied to the upper lower respiratory tract of the animal's pharynx, and the aqueous solution is administered directly from the other end of the sonde to the lower respiratory tract by spontaneous breathing of the animal. A method for developing chronic obstructive pulmonary disease in an animal for evaluating drug efficacy. The method for onset of chronic obstructive pulmonary disease in an animal for drug efficacy evaluation according to claim 1 , wherein the aqueous solution contains endotoxin produced by a gram-negative bacterium. The method for developing chronic obstructive pulmonary disease in an animal for drug efficacy evaluation according to claim 2 , wherein the endotoxin produced by the gram-negative bacterium is lipopolysaccharide. The method for developing chronic obstructive pulmonary disease in an animal for drug efficacy evaluation according to any one of claims 1 to 3 , wherein the aqueous solution contains a water-soluble organic solvent. An animal for evaluation of drug efficacy, wherein chronic obstructive pulmonary disease is developed by the method for developing chronic obstructive pulmonary disease of the animal for drug efficacy evaluation according to any one of claims 1 to 4 . The animal for drug efficacy evaluation according to claim 5 , wherein the animal is a rodent. The animal for drug efficacy evaluation according to claim 5 , wherein the animal is a carnivorous. The animal for drug efficacy evaluation according to claim 5 , wherein the animal is a heavy tooth. The animal for drug efficacy evaluation according to claim 5 , wherein the animal is an artiodactyla. The animal for drug efficacy evaluation according to claim 5 , wherein the animal is a primate. The animal for evaluating drug efficacy according to claim 6 , wherein the rodent animal is a guinea pig, a mouse, a rat, or a hamster. The animal for medicinal efficacy evaluation according to claim 7 , wherein the animal that is carnivorous is a dog or a cat. The animal for evaluating drug efficacy according to claim 8 , wherein the animal having a heavy tooth is a rabbit. The animal for evaluating medicinal effects according to claim 9 , wherein the animal that is an artiodactyla is a goat, a sheep, or a pig. The animal for evaluating medicinal effects according to claim 10 , wherein the primate animal is a monkey or chimpanzee. A drug efficacy evaluation method, comprising administering a drug to the drug efficacy evaluation animal according to any one of claims 6 to 15 and evaluating the action and effect thereof. The drug effect evaluation method according to claim 16 , wherein the action effect is a change in respiratory function. The method for evaluating efficacy according to claim 17 , wherein the change in respiratory function is specific airway resistance, peak expiratory flow, tidal volume, number of minute breaths, and minute ventilation.

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