JP2017032550A - Method of determining presence or absence of gastric mucosal atrophy - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a method of noninvasively determining the presence or absence of gastric mucosal atrophy.SOLUTION: A method of determining the presence or absence of gastric mucosal atrophy in mammals comprises steps of: (1) collecting breath exhaled from a subject mammal as a test sample at an arbitrary point in time within 60 minutes after orally administering a prescribed dose ofC-labeled carbonate compound, and measuring behavior ofCOexhaled with the breath; (2) comparing theCObehavior (measuredCObehavior) observed in the step (1) with correspondingCObehavior (referenceCObehavior) obtained from a mammal (control mammal) to be compared; and (3) determining the presence or absence of gastric mucosal atrophy in the subject mammal, based on difference between the measuredCObehavior and the referenceCObehavior.SELECTED DRAWING: None

Description

  The present invention relates to a method for determining the presence or absence of gastric mucosal atrophy.

  Chronic gastritis refers to cases where inflammation or the like has occurred in the gastric mucosa for a long period of time, and can be broadly classified into superficial gastritis and atrophic gastritis, for example. In superficial gastritis, inflammatory cells such as lymphocytes infiltrate into the surface of the gastric mucosa. In atrophic gastritis, the normal gland of the gastric mucosa shrinks and the gastric mucosa becomes thinner due to the progression of inflammation. Is recognized. Since it is known that the risk of developing gastric cancer increases as the mucosal atrophic change proceeds, it is very important in reducing gastric cancer risk to know the presence or absence of gastric mucosal atrophy (Non-patent Document 1).

  The main examination methods for gastric mucosal atrophy include endoscopy and pepsinogen method. Endoscopic examination is a very well-known examination method, but it is often painful both physically and mentally because a tube is inserted and the inside of the stomach is observed. The pepsinogen method is a method that can determine the presence or absence of gastric mucosal atrophy based on the ratio of pepsinogen I and II (PGI / PGII) present in blood, but this method requires blood collection (Non-patent Document 2). Thus, both of these methods are invasive.

  From the viewpoint of reducing the physical or mental pain of a patient, it is important to provide a method that can determine the presence or absence of gastric mucosal atrophy in a non-invasive manner.

Yasuhisa Kumakura et al., “High-risk group of gastric mucosal atrophy and gastric cancer in clinical examination of human dock”, Journal of Gastrointestinal Cancer Screening Society, Vol. 46, N0.6, pp.713-722, 2008 M. Kabuto1, Correlation between Atrophic Gastritis Prevalence and Gastric Cancer Mortality among Middle-aged Men in 5 Areas in Japan, J. Epidemiol, 1993, Vol. 3, No. 1, pp. 35-39.

  An object of this invention is to provide the method which can determine the presence or absence of gastric mucosa atrophy noninvasively.

The inventors of the present invention have made extensive studies in order to solve the above-mentioned problems. As a result, the presence or absence of gastric mucosa atrophy is determined depending on the behavior of ¹³ CO _{2 in} exhaled air after ¹³ C-labeled carbonic acid compound is orally administered. I found out that I can know. The present invention has been completed as a result of further studies based on this finding, and is as follows.
Item 1. A method for determining the presence or absence of gastric mucosal atrophy in a mammal,
(1) a predetermined dose of ¹³ breath test ^{mammal C-labeled} carbon compound is discharged at any time within 60 minutes after being orally administered as a test sample, the ¹³ CO ₂ emitted into the air the call Measuring the behavior,
(2) the step of ¹³ CO ₂ behavior (measured ¹³ CO ₂ behavior) obtained in (1), against a mammal (control mammal) ¹³ CO ₂ behavior corresponding to the obtained above for ( method having steps, and (3) determining the presence or absence of gastric mucosal atrophy mammalian subject based on the difference of the between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior is compared with the reference ¹³ CO ₂ behavior) .
Item 2. Item 2. The method according to Item 1, wherein the test sample is exhaled breath collected once at an arbitrary time point within 60 minutes after oral administration of the ¹³ C-labeled carbonate compound.
Item 3. The behavior of ¹³ CO ₂ ^{is, 13} C arbitrary determined from breath taken at Δ ¹³ C (‰) within 60 minutes after oral administration of the labeled carbon compound or Δ ¹³ C (‰) - area under the curve Item 3. The method according to Item 1 or 2, which is (AUC: Area under the curve).
Item 4. Item ^13. The behavior according to any one of Items 1 to 3, wherein the behavior of ¹³ CO ₂ is Δ ¹³ C (‰) obtained from exhaled breath collected at any time within 60 minutes after oral administration of the ¹³ C-labeled carbonic acid compound. Judgment method.
Item 5. The measurement ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior are derived from exhaled breath obtained by administering a ¹³ C-labeled carbonic acid compound to a fasting test mammal and a control mammal, respectively. The determination method according to any one of the above.
Item 6. Item 6. The determination method according to any one of Items 1 to 5, wherein the predetermined dose is 10 mg to 5 g.
Item 7. The ¹³ C-labeled carbonate compound is selected from the group consisting of an alkali metal carbonate, an alkaline earth metal carbonate, ammonium carbonate, an alkali metal bicarbonate, an alkaline earth metal bicarbonate, and an ammonium bicarbonate. Item 7. The determination method according to any one of Items 1 to 6, wherein the determination method is at least one carbonate compound.
Item 8. The ^{13 C-labeled} carbon compound is orally administered in step (1) is a solution of 200ml which was suspended ^{13 C-labeled} carbon compound in a liquid which does not adversely affect the oral administration possible and and ^{13 C-labeled} carbon compound, Item 8. The determination method according to any one of Items 1 to 7.
Item 9. Item 9. The determination method according to Item 8, wherein the liquid is water.
Item 10. Step (3) determines that the test mammal has no gastric mucosal atrophy when the measured ¹³ CO ₂ behavior is the same as or higher than a reference ¹³ CO ₂ behavior obtained from a control mammal without gastric mucosal atrophy. The determination method according to any one of Items 1 to 9, which is a step of determining that the test mammal has gastric mucosal atrophy when the measured ¹³ CO ₂ behavior is lower than the reference ¹³ CO ₂ behavior.
Item 11. Wherein the step (3), is measured ¹³ CO ₂ behavior, is higher than the preset cut-off value, determines that there is no gastric mucosal atrophy in mammalian subject, said measuring ¹³ CO ₂ behavior, the cut-off value Item 11. The determination method according to any one of Items 1 to 10, which is a step of determining that the test mammal has gastric mucosal atrophy when the test mammal is the same or lower.

  According to the present invention, the presence or absence of gastric mucosa atrophy can be determined non-invasively. Further, according to the present invention, it is possible to determine the presence or absence of gastric mucosa atrophy in a non-invasive and simple manner.

FIG. 1 shows the correlation between the determination result of the presence or absence of gastric mucosa atrophy by the method of the present invention and the determination result by the conventionally used pepsinogen method. FIG. 2 shows the ¹³ C (‰) value of exhaled breath 5 to 120 minutes after administration of the preparation. FIG. 3 shows the correlation between the determination result of the presence or absence of gastric mucosa atrophy by the method of the present invention and the determination result by the conventionally used pepsinogen method. FIG. 4 shows the correlation between the determination result of the presence or absence of gastric mucosa atrophy by the method of the present invention and the determination result by the conventionally used pepsinogen method. FIG. 5 shows the correlation between the determination result of the presence or absence of gastric mucosa atrophy by the method of the present invention and the determination result by the conventionally used pepsinogen method.

The present invention provides a method for determining the presence or absence of gastric mucosal atrophy in a mammal,
(1) a predetermined dose of ¹³ breath test ^{mammal C-labeled} carbon compound is discharged at any time within 60 minutes after being orally administered as a test sample, the ¹³ CO ₂ emitted into the air the call Measuring the behavior,
(2) wherein (1) the resultant of the ¹³ CO ₂ behavior (measured ¹³ CO ₂ behavior), the control to mammals (control mammal) ¹³ CO ₂ behavior corresponding to the obtained above for (reference ¹³ CO ₂ behavior), and (3) determining the presence or absence of gastric mucosal atrophy in the test mammal based on the difference between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior.

Hereinafter, the present invention will be described in more detail.
1. ¹³ C-labeled carbonic acid compound used for determination of gastric mucosal atrophy In the method of the present invention, exhalation of a test mammal discharged at any time within 60 minutes after the oral administration of ¹³ C-labeled carbonic acid compound is used as a test sample. To do.

^{The 13} C-labeled carbonic acid compound reacts with gastric acid in the stomach of the subject after oral administration, and in some cases is subsequently decomposed or metabolized to be excreted as ¹³ C-labeled carbon dioxide ( ¹³ CO ₂ ) in the breath. There is no particular limitation.

As a compound that appears as ¹³ C-labeled carbon dioxide gas in the expired air immediately after reacting with gastric acid in the stomach, ¹³ C-labeled carbonate ion ( ¹³ CO ₃ ^-2 ) or ¹³ C-labeled bicarbonate ion ( ¹³ C-labeled carbonic acid compounds that generate H ¹³ CO ₃ ⁻¹ ) can be widely exemplified. As such a ¹³ C-labeled carbonate compound, for example, an alkali metal carbonate such as sodium carbonate or potassium carbonate, an alkaline earth metal carbonate such as calcium carbonate, magnesium carbonate or barium carbonate, or ammonium carbonate is narrowly defined. In addition to the above carbonate compounds, alkali metal salts of hydrogen carbonate such as potassium hydrogen carbonate and sodium hydrogen carbonate, alkaline earth metal salts of hydrogen carbonate, hydrogen carbonate compounds such as ammonium hydrogen carbonate and the like can be mentioned. Preferred examples of the ¹³ C-labeled carbonate compound include calcium carbonate, magnesium carbonate, sodium carbonate, potassium carbonate, sodium bicarbonate, and potassium bicarbonate. These may be used alone or in combination of two or more.

The labeling method with an isotope ( ¹³ C) is not particularly limited, and a commonly used method is widely adopted. In addition, as the ¹³ C-labeled carbonic acid compound labeled with the isotope, known and commercially available compounds are widely used (Sasaki, “5.1 Application of stable isotopes to clinical diagnosis”: Chemistry. Pp.149-163 (1975) Nanedo; Sugawara, RADIOISOTOPES, 41, 45-48 (1992), etc.).

In the method of the present invention, the ¹³ C-labeled carbonic acid compound may be orally administered as the ¹³ C-labeled carbonic acid compound itself (single substance), and is contained in a composition containing the ¹³ C-labeled carbonic acid compound and other components. The ¹³ C-labeled carbonic acid compound is not limited as long as it can be orally administered to the test mammal. As the other ingredients, pharmaceutically acceptable ingredients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid and the like; simple syrup, glucose solution, Binders such as starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone; dry starch, sodium alginate, agar powder, laminaran powder, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid Disintegrants such as monoglyceride, starch and lactose; Absorption accelerators such as quaternary ammonium base and sodium lauryl sulfate; Moisturizers such as glycerin and starch; Lubricants such as purified talc, stearate, boric acid powder and polyethylene glycol Agent Other additives (e.g. flavoring, flavoring agents, stabilizers, etc.) and the like. These may be used alone or in combination of two or more.

^{The form of the 13} C-labeled carbonic acid compound and the composition may be solid, semi-solid, or liquid, and can be prepared in various forms such as powder, granules, tablets, pills, and liquids. However, in the present invention, it is preferable that the ¹³ C-labeled carbonic acid compound dissolves quickly in the stomach, and from this point of view, the form of the capsule encapsulated in the capsule substrate is not very preferable. Similarly, from the viewpoint of immediate solubility, the granules, tablets, pills, and the like are preferably not coated with a pH-dependent soluble film or a sugar coating that is difficult to dissolve.

The predetermined dose of ¹³ C-labeled carbonic acid compound is not limited as long as the effects of the present invention can be obtained, but it is usually 10 mg to 5 g, preferably 10 mg to 4 g, more preferably 20 mg to 2 g per individual (1 body). It can be selected appropriately. The dosage can be appropriately adjusted according to the type of the test mammal and the body weight of the individual. In the range satisfying the predetermined dose, the ¹³ C-labeled carbonic acid compound and the composition are usually in the range of 10 mg to 20 g, preferably 10 mg to 10 g per unit dose, from the viewpoint of ease of taking. it can. Administration may be selected as appropriate based on the expiration collection time described later, but it is performed within 60 minutes, preferably within 30 minutes, more preferably within 15 minutes, and the number of administrations is usually 1-2. Times, preferably once.

2. Method for Determining Presence or Absence of Mammalian Gastric Mucosal Atrophy As described above, the present invention relates to a method for judging the presence or absence of mammalian gastric mucosal atrophy and includes the steps (1) to (3).

The present invention is performed non-invasively using exhaled breath discharged from a mammal to which ¹³ C-labeled carbonate compound is orally administered as a test sample.

  Mammals targeted by the present invention are particularly limited as long as they have a respiratory system and digestive system (especially from the viewpoint of gastric mucosal atrophy) that function similarly to the human respiratory system and digestive system. Instead, humans, monkeys, dogs, cats, rabbits, guinea pigs, rats, mice and the like are exemplified. The mammal is preferably a human, and when an experimental animal is used, a dog, rabbit, guinea pig, rat, and mouse are preferable because of easy availability and handling.

3. In the step (1) of measuring the behavior of ¹³ CO ₂ excreted in exhaled breath, the test mammal is excreted at an arbitrary time within 60 minutes after the prescribed dose of ¹³ C-labeled carbonic acid compound is orally administered. This is a step of measuring the behavior of ¹³ CO ₂ discharged into the exhalation using the exhaled breath as a test sample.

The method for oral administration of the ¹³ C-labeled carbonic acid compound to the test mammal is not particularly limited.

For example, the ^{13 C-labeled} carbon compounds or may be directly orally administering the composition, a liquid which does not adversely affect the water or buffer solution is orally administered such and ^{13 C-labeled} carbon compound, ^{13 C-labeled} carbon compound Or you may administer orally after suspending this composition. The liquid is preferably water (for example, a temperature of about 0 to 60 ° C.).

More preferably, the ¹³ C-labeled carbonic acid compound or the composition is orally administered after being suspended in the liquid. In addition, when suspended in a liquid in this way, the ¹³ C-labeled carbonate compound or the composition is not limited as long as it can be suspended, but from the viewpoint of easy oral administration, 10 mg to 5 g of the ¹³ C-labeled carbonate compound is added. It is exemplified that it is suspended and orally administered as a 5 to 1000 ml solution, more preferably a 50 to 500 ml solution, still more preferably 50 to 500 ml. From the viewpoint that it can be more easily orally administered, 200 ml is particularly preferred. Illustrated.

Moreover, it is preferable to administer on an empty stomach from the viewpoint of being hardly affected by meals. In this case, it is more preferable to orally administer the ¹³ C-labeled carbonate compound after at least 4 hours have elapsed since the start of fasting, and more preferably after 8 hours have elapsed.

However, in order to measure the behavior of ¹³ CO ₂ excreted in exhaled breath while suppressing variation in measured values among individuals, it is preferable to measure the behavior of ¹³ CO ₂ after stimulating the stomach. In this case, test foods and drinks (for example, water, caffeinated beverages, alcoholic beverages, consomme soup such as solid or liquid foods such as Calorie Mate (registered trademark)) or gastric stimulants (for example, Histamine hydrochloride, betazole hydrochloride, gastrin, insulin, etc.) is injected to stimulate the stomach, and the ¹³ C-labeled carbonate compound is preferably administered orally within 1 hour.

Further, the present invention is not limited to this, but from the viewpoint of further reducing individual differences and determining more accurately, it is preferable to orally administer the ¹³ C-labeled carbonate compound while avoiding a certain period of time after waking up and intense exercise. Illustrated.

When the ¹³ C-labeled carbonic acid compound orally administered to the test mammal enters the stomach, it reacts with gastric acid to produce ¹³ C-labeled carbon dioxide ( ¹³ CO ₂ ), which is sequentially discharged into the exhaled breath. ^The reaction formula for producing ¹³ C-labeled carbon dioxide gas ( ¹³ CO ₂ ) is shown below, taking as an example the case of using ¹³ C-labeled calcium carbonate (Ca ¹³ CO ₃ ) as the ¹³ C-labeled carbonate compound.

As "the behavior of ¹³ CO _2" as measured by the present invention can be exemplified by the following (a) ~ (d).
(A) the amount of ¹³ CO ₂ excreted into the exhaled breath at any time within 60 minutes after the prescribed dose of ¹³ C-labeled carbonate compound is orally administered;
(B) After ^{the 13} C-labeled carbon dioxide compound of a given dose is orally administered, the ratio of ¹³ CO ₂ amount to the ¹² CO ₂ amount discharged during expiration at any time within 60 minutes ⁽¹³ _CO ^2/12 CO ₂ concentration ratio: δ ¹³ C value);
(C) After a predetermined dose of ¹³ C-labeled carbonic acid compound is orally administered, the amount of ¹³ CO ₂ relative to the amount of ¹² CO ₂ contained in exhaled breath at any time within 60 minutes (at the time of exhalation collection (t)) “The ratio” (hereinafter referred to as “δ ¹³ C _t ”) and “the ratio of the ¹³ CO ₂ amount to the ¹² CO ₂ amount included in the exhaled breath before the oral administration of the ¹³ C-labeled carbonic acid compound” (hereinafter referred to as “ the difference between [delta] ¹³ that C ₀ ^{") [Δ 13 C (‰)} = δ 13 C t -δ 13 C 0] ( hereinafter, this" Δ ¹³ C _{(‰) t} "or" Δ ¹³ C (‰) ");
(D) “Δ ¹³ C (‰” calculated by drawing a graph with the horizontal axis representing the time from administration of a predetermined dose of ¹³ C-labeled carbonic acid compound to the collection of exhalation and Δ ¹³ C (‰) on the vertical axis. ) —Area under the curve (AUC).

“Δ ¹³ C (‰)” and “Δ ¹³ C (‰) — area under time curve” (AUC) are preferable, and “Δ ¹³ C (‰)” is more preferable. Specifically, the behavior of ¹³ CO ₂ can be measured as described below and according to the following.

After orally administering a predetermined dose of ¹³ C-labeled carbonic acid compound to the test mammal, a conventional ¹³ C breath test method (Hagiwara, RADIOISOTOPESM, 41, 45-48 (1992); Sugawara et al., RADIOISOTOPS, 41, 331- according 334 (1992), etc.), breath was collected, during breath sampling (t) in the ^"13 CO ^_2/12 CO ₂ concentration ratio ([delta] ¹³ C)" ( "[delta] ¹³ C _t" is discharged during exhalation the ratio of ¹³ CO ₂ amount by carbon dioxide ¹² CO ₂ amount, a "t" is ¹³ C-labeled elapsed time after the carbonate compound administration ^means breath sampling time in within 60 minutes from the ¹³ C-labeled carbon compound administration ).

Then, as "[delta] ¹³ C ^{_t", 13} C-labeled carbon compounds criteria were previously measured before the oral administration of the ^"13 CO ^_2/12 CO ₂ concentration ratio ([delta] ¹³ C)" and ( "[delta] ¹³ C _0") From the difference, “Δ ¹³ C (‰)” [Δ ¹³ C (‰) = δ ¹³ C _t −δ ¹³ C ₀ ] is calculated.

In this way, the behavior of ¹³ CO ₂ excreted into the exhalation at an arbitrary time point (t) within 60 minutes after administration of the ¹³ C-labeled carbonic acid compound is expressed by the above equation Δ ¹³ C (‰) = δ ¹³ C _t − It can be obtained from δ ¹³ C ₀ .

Further, the temporal behavior of ¹³ CO ₂ discharged into the exhaled breath can be obtained by tracking the temporal transition of the Δ ¹³ C (‰). Specifically, it can be determined by drawing a graph with the elapsed time after administration of ¹³ C-labeled carbonic acid compound as the horizontal axis and Δ ¹³ C (‰) as the vertical axis.

In addition, the measurement and analysis of the label ( ¹³ CO ₂ ) contained in the collected exhalation are generally used analytical methods such as liquid scintillation counter method, mass spectrometry, infrared analysis, emission analysis, magnetic resonance spectrum, etc. Can be used. In view of measurement accuracy, infrared spectroscopy and mass spectrometry are preferred.

Further, “Δ ¹³ C (‰) — area under the time curve” (AUC) represents the elapsed time (exhalation collection time: t) (min) after administration of ¹³ C-labeled carbonic acid compound obtained as described above on the horizontal axis. Further, it can be obtained by calculating the area under the curve from a graph with Δ ¹³ C (‰) on the vertical axis.

In addition, it is preferable that the breath collection time is usually after 10 seconds or more have elapsed since oral administration of the ¹³ C-labeled carbonic acid compound, and more preferably after 1 minute or more has elapsed. In addition, the breath collection time is preferably within 60 minutes, more preferably within 30 minutes, and even more preferably within 15 minutes from the oral administration of the ¹³ C-labeled carbonic acid compound. For example, after ¹³ C-labeled carbonic acid compound is orally administered, exhalation is collected once or twice within 10 seconds to 60 minutes, preferably 1 minute to 30 minutes, more preferably 1 minute to 15 minutes. The test sample is taken as the test sample, and more preferably, the test sample is taken after one breath is collected within the same time.

4). Measurements ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior and the step of comparing the step (2), said step of ¹³ CO ₂ behavior (measured ¹³ CO ₂ behavior) obtained in (1), a mammal in control a step of comparing the (control mammal) ¹³ CO ₂ behavior corresponding to the obtained above for (reference ¹³ CO ₂ behavior).

In this step, “ ¹³ CO ₂ behavior” is as described above, preferably “Δ ¹³ C (‰)” and “Δ ¹³ C (‰) — area under time curve” (AUC), more preferably “ Δ ¹³ C (‰) ”.

“Measured ¹³ CO ₂ behavior” means the behavior of ¹³ CO ₂ obtained in the step (1).

The “reference ¹³ CO ₂ behavior” means the behavior of ¹³ CO ₂ obtained in the same manner except that the “test mammal” is replaced with the “control mammal” in the step (1). That is, in this step, the subject mammal ¹³ CO ₂ measured in behavior as (measured ¹³ CO ₂ behavior) of ¹³ CO ₂ behavior of the same type employed as a reference ¹³ CO ₂ behavior, if subject mammal is a human subject If the mammal is also a human and the measured ¹³ CO ₂ behavior is Δ ¹³ C (‰), the reference ¹³ CO ₂ behavior is also Δ ¹³ C (‰).

Also, for example, if the measured ¹³ CO ₂ behavior is derived from a breath taken after administration of a ¹³ C-labeled carbonic acid compound when the test mammal is fasted, the reference ¹³ CO ₂ behavior is also ¹³ when the control mammal is fasted. It is preferably derived from exhaled breath collected by administration of a C-labeled carbonic acid compound.

Alternatively, the reference ¹³ CO ₂ behavior may be a ¹³ CO ₂ behavior obtained using a control mammal simultaneously with the measurement of the measured ¹³ CO ₂ behavior or after the measurement of the measured ¹³ CO ₂ behavior, or It may be the ¹³ CO ₂ behavior previously obtained for the control mammal.

The measured ¹³ CO ₂ behavior thus obtained may be compared with the reference ¹³ CO ₂ behavior.

5. The step (3) of determining the presence or absence of gastric mucosa atrophy is a step of determining the presence or absence of gastric mucosa atrophy of the test mammal based on the difference between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior.

The measurement ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior are as described above.

In this step, the presence or absence of gastric mucosal atrophy can be determined based on the difference between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior.

For example, the control mammal may or may not have gastric mucosal atrophy. If the control mammal does not have gastric mucosal atrophy, based on the difference between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior, the measured ¹³ CO ₂ behavior is the same as or higher than the reference ¹³ CO ₂ behavior The test mammal is determined to have no gastric mucosal atrophy, and if the measured ¹³ CO ₂ behavior is lower than the reference ¹³ CO ₂ behavior, it can be determined that the test mammal has gastric mucosal atrophy. Also, if the control mammal has gastric mucosa atrophy, based on the difference between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior, said measuring ¹³ CO ₂ behavior is higher than the reference ¹³ CO ₂ behavior The test mammal is determined to have no gastric mucosal atrophy, and it can be determined that the test mammal has gastric mucosal atrophy when the measured ¹³ CO ₂ behavior is the same or lower than the reference ¹³ CO ₂ behavior.

From the viewpoint of more accurately determining the presence or absence of gastric mucosal atrophy, the control mammal does not have gastric mucosal atrophy. That is, in step (3), preferably, when the measured ¹³ CO ₂ behavior is the same as or higher than the reference ¹³ CO ₂ behavior obtained from a control mammal without gastric mucosal atrophy, the test mammal is treated with gastric mucosal atrophy. When the measured ¹³ CO ₂ behavior is lower than the reference ¹³ CO ₂ behavior, it is determined that the test mammal has gastric mucosal atrophy.

Further, as described above, the ¹³ C-labeled carbonic acid compound reacts with gastric acid in the stomach of the test mammal after oral administration, and is excreted as ¹³ C-labeled carbon dioxide in the exhaled breath. For this reason, the control mammal is preferably exemplified by those that do not have a disease associated with secretion of gastric acid and those that are not affected by preparations such as gastric acid secretion inhibitor / promoter.

They accumulate data on ¹³ CO ₂ behavior obtained by control mammals without gastric mucosal atrophy and / or control mammals with gastric mucosal atrophy, Based on the cut-off value provided based on the ¹³ CO ₂ behavior data (a value that serves as a boundary for the presence or absence of gastric mucosa atrophy), the cut-off value and the measured ¹³ CO ₂ behavior When the measured ¹³ CO ₂ behavior is higher than the cut-off value, it is determined that the test mammal has no gastric mucosal atrophy, and the measured ¹³ CO ₂ behavior is the same or lower than the cut-off value. In the present invention, it is included to determine that the test mammal has gastric mucosal atrophy.

According to the present invention, the presence or absence of gastric mucosa atrophy can be determined in this way. In addition, since the present invention can determine the presence or absence of gastric mucosal atrophy based on the measured ¹³ CO ₂ behavior, it is also useful for the diagnosis of atrophic gastritis. That is, the present invention includes the steps (1) to (3). It can also be said that it provides a diagnostic method for the presence or absence of atrophic gastritis. Also, from this, the measured ¹³ CO ₂ behavior is also useful as a precancerous marker.

Further, according to the present invention, since the presence or absence of gastric mucosa atrophy can be determined in this way, the therapeutic effect can be judged by treating the gastric mucosa atrophy and obtaining the measured ¹³ CO ₂ behavior in the same manner. For example, in a patient with gastric mucosal atrophy, if the measured ¹³ CO ₂ behavior after treatment is higher than the measured ¹³ CO ₂ behavior before treatment, it can be said that there was a therapeutic effect. From this, it can be said that the present invention also provides a method for evaluating the therapeutic effect on gastric mucosal atrophy, comprising the steps (1) to (3). As a therapeutic means for mammals having gastric mucosal atrophy, eradication is exemplified when Helicobacter pylori is positive, and excision of a cancerous part is exemplified after follow-up because H.pylori is negative in a precancerous state.

  According to the present invention, it is useful in that the presence or absence of gastric mucosa atrophy can be determined non-invasively by collecting a few breaths (preferably one breath collection) without restraining the test mammal for a long time. . Moreover, according to the present invention, it is useful in that the presence or absence of gastric mucosa atrophy can be easily determined as described above. Therefore, according to the present invention, the presence or absence of gastric mucosa atrophy can be determined non-invasively and simply.

Further, according to the present invention, since the behavior of ¹³ CO ₂ can be grasped, the degree of gastric mucosal atrophy can also be determined. In this case, compared to the reference ¹³ CO ₂ behavior, it can be said that the lower the value of the measured ¹³ CO ₂ behavior, the higher the degree of gastric mucosal atrophy.

  Hereinafter, the present invention will be described in more detail with reference to examples, but the present invention is not limited thereto.

Test example 1
・ Production
A preparation containing ¹³ C-labeled carbonate compound was produced as follows. ¹³ C-labeled calcium carbonate ( ¹³ C-CaCO ₃ ) (MW: 101, manufactured by Cambridge Isotopology) 200 mg as an active ingredient, D-mannitol, low-substituted hydroxypropylcellulose, hydroxypropylcellulose and light anhydrous silicic acid Granules (Example 1) were produced according to a conventional method. The content of ¹³ C-labeled calcium carbonate in the preparation is 50% by weight.

・ Experiment 1
A total of 40 male and female subjects were orally administered 200 mg of the preparation on an empty stomach. This test was fasted after 22:00 on the day before the test (only water may be taken), and the following test was started at 8-9 am the next day. In addition, 200 mg of the preparation was mixed with water to make a 200 ml solution, and then orally administered to each subject.

For all subjects, exhalation was collected before administration of the preparation (before ¹³ C-CaCO ₃ administration) and 15 minutes after administration (after ¹³ C-CaCO ₃ administration), and Δ ¹³ C (‰) in the exhalation was obtained. It was determined using a mass spectrometer for breath analysis (ABCA: manufactured by SerCon). Δ ¹³ C (‰) ^is, 13 C-CaCO ₃ pre-dose and ¹³ CO ^_2/12 CO ₂ concentration ratio in the breath after administration ([delta] ¹³ C value (the pre-dose [delta] ¹³ C value _0, the post-dose [delta] ¹³ shows the C value ₁₅₎₎ were measured, respectively, calculates the difference [delta] ¹³ C value ₀ before administration and [delta] ¹³ C value ₁₅ after administration (delta13C value ₁₅ -Deruta13C value ₀₎ from the delta ¹³ C (‰) And asked.

・ Experiment 2
Further, the ratio of pepsinogen I / pepsinogen II (PGI / PGII ratio) was determined for the 40 subjects according to the pepsinogen method. In this experiment, a PGI / PGII ratio of less than 2.5 was judged to have gastric mucosa atrophy, and a PGI / PGII ratio of 2.5 or more was judged to have no gastric mucosa atrophy. The standard was in accordance with the standard described in M. Kabuto1, J. Epidemiol, 1993, Vol. 3, No. 1, pp. 35-39. As a result, it was determined that 14 cases had gastric mucosal atrophy and the remaining 26 cases had no gastric mucosal atrophy.

Evaluation Evaluation FIG. 1 shows Δ ¹³ C (‰) obtained in the above-described Experiment 1. In the figure, the vertical axis indicates Δ ¹³ C (‰), and the horizontal axis indicates “no atrophy” and “with atrophy”. The results of subjects who were judged as having no gastric mucosal atrophy in Experiment 2 were plotted in the “no atrophy” column, and the results of subjects judged as having gastric mucosal atrophy in Experiment 2 were plotted in the “with atrophy” column. As a result, high in the "no atrophy" column Δ ^{13 C (‰)} value subject, "atrophy yes" in the column Δ ^{13 C (‰)} low value subjects localized.

From this, it was found that the experiment 1 and the experiment 2 have a high correlation. In addition, it can be seen that Δ ¹³ C (‰) values obtained from subjects with gastric mucosal atrophy tend to be lower than Δ ¹³ C (‰) values obtained from subjects without gastric mucosal atrophy. I understood.

Thus, it was confirmed that the presence or absence of gastric mucosal atrophy can be determined non-invasively based on the behavior of ¹³ CO _{2 in} exhaled air discharged after oral administration of a ¹³ C-labeled carbonic acid compound. In addition, as described above, since the presence or absence of mucosal atrophy can be determined simply by analyzing the exhaled air that is excreted after oral administration of a ¹³ C-labeled carbonic acid compound, the method is simple and rapid. It was confirmed that the presence or absence of can be determined.

Test example 2
Forty subjects in Test Example 1 were classified as having no atrophy (PGI / PGII ratio of 2.5 or more and with atrophy (PGI / PGII ratio of less than 2.5)), and 5, 10, 15, 20, 25 after administration of the preparation. , 30, 40, 50, 60, 80, 100 and 120 minutes, exhaled breath was collected in the same manner as described above, and the value of ¹³ C (‰) was obtained.

The results are shown in FIG. In the figure, the vertical axis indicates ¹³ C (‰), and the horizontal axis indicates time after administration of the preparation. There was a difference in ¹³ C (‰) value between the absence of atrophy and the presence of atrophy, and in particular, there was a large difference in values up to about 60 minutes after administration. From this, it was confirmed that this method can determine the presence or absence of gastric mucosal atrophy even at one point within 60 minutes after administration of the preparation.

Test example 3
Each plot of Δ ¹³ C (‰) and PGI / PGII ratio measured in Test Example 1 is shown in FIG. In the figure, the vertical axis represents Δ ¹³ C (‰) 15 minutes after administration of the preparation, and the horizontal axis represents the PGI / PGII ratio. As is clear from FIG. 3, since these showed a positive correlation, it was confirmed that the presence or absence of gastric mucosa atrophy can be evaluated by expiration.

Test example 4
In Test Example 1, 40 subjects were divided into 4 groups having a PGI / PGII ratio of 3 or more, a PGI / PGII ratio of 2.5 to less than 3, a PGI / PGII ratio of 2 to less than 2.5, and a PGI / PGII ratio of less than 2. Each Δ ¹³ C (‰) was plotted. The results are shown in FIG. In the figure, the vertical axis represents Δ ¹³ C (‰) 15 minutes after administration of the preparation, and the four groups were provided on the horizontal axis. As apparent from FIG. 4, since the value of Δ ¹³ C (‰) was correlated with PGI / PGII ratio, the value of the atrophy progresses Δ ¹³ C (‰) is showing a declining trend, delta ¹³ C ( It was suggested that the severity of atrophy can be determined from the value of ‰).

Test Example 5
In Test Example 1, in place of the Δ ^{13 C (‰),} the exhaled breath - it went tested as addition to obtain the "Δ ^{13 C (‰)} time area under the curve" (AUC). More ^{specifically,} 13 C-CaCO ₃ pre-dose and ¹³ CO ^_2/12 CO ₂ concentration ratio in the breath after administration ([delta] ¹³ C value (the pre-dose [delta] ¹³ C value _0, the post-dose [delta] ¹³ C the shown value ₁₅₎ were measured, in 0-15 min administration - were calculated "Δ ¹³ C (‰) times the area under the curve" _{(AUC 15-0).}

The results are shown in FIG. In the figure, the vertical axis represents “Δ ¹³ C (‰) — area under the time curve” (AUC _15-0 ), and “no atrophy” and “with atrophy” are provided on the horizontal axis as in FIG. The results of subjects who were determined to have no gastric mucosal atrophy in Experiment 2 were plotted in the “no atrophy” column and the results of subjects determined to have gastric mucosal atrophy were plotted in the “with atrophy” column. As a result, a subject with a high “Δ ¹³ C (‰) — area under the time curve” (AUC _15-0 ) value appears in the “no atrophy” column, and “Δ ¹³ C (‰) — A subject with a low area under the time curve (AUC _15-0 ) value was localized.

Therefore, - even when the reference to "Δ ^{13 C (‰)} times the area under the curve" (AUC), high correlation and pepsinogen method was observed. In addition, the “area under the Δ ¹³ C (‰)-time curve” (AUC) value obtained from the subject with gastric mucosal atrophy is “Δ ¹³ C (‰) — time obtained from the subject without gastric mucosal atrophy. It was found that a tendency to be lower than the “area under the curve” (AUC) value was observed.

Therefore, even when “Δ ¹³ C (‰) — area under time curve” (AUC) is used, the amount of ¹³ CO _{2 in} exhaled breath discharged after oral administration of ¹³ C-labeled carbonic acid compound It was confirmed that the presence or absence of gastric mucosa atrophy can be determined non-invasively by the behavior.

Thus, since the presence or absence of mucosal atrophy can be determined simply by analyzing the exhaled air that is excreted after oral administration of the ¹³ C-labeled carbonic acid compound, the method is simple and rapid, and whether or not there is gastric mucosal atrophy. It was confirmed that can be determined. Furthermore, it was confirmed that the degree of mucosal atrophy can also be determined based on the level of the calculated value.

Claims (7)

A method for determining the presence or absence of gastric mucosal atrophy in a mammal,
(1) a predetermined dose of ¹³ breath test ^{mammal C-labeled} carbon compound is discharged at any time within 60 minutes after being orally administered as a test sample, the ¹³ CO ₂ emitted into the air the call Measuring the behavior,
(2) the step of ¹³ CO ₂ behavior (measured ¹³ CO ₂ behavior) obtained in (1), against a mammal (control mammal) ¹³ CO ₂ behavior corresponding to the obtained above for ( method having steps, and (3) determining the presence or absence of gastric mucosal atrophy mammalian subject based on the difference of the between the measured ¹³ CO ₂ behavior and the reference ¹³ CO ₂ behavior is compared with the reference ¹³ CO ₂ behavior) . The determination method according to claim 1, wherein the test sample is a breath collected once at an arbitrary time point within 60 minutes after oral administration of the ¹³ C-labeled carbonate compound. The determination according to claim 1 or 2, wherein the behavior of ¹³ CO ₂ is Δ ¹³ C (‰) obtained from exhaled breath collected at any time within 60 minutes after oral administration of the ¹³ C-labeled carbonic acid compound. Method.   The determination method according to claim 1, wherein the predetermined dose is 10 mg to 5 g. The ¹³ C-labeled carbonate compound is selected from the group consisting of an alkali metal carbonate, an alkaline earth metal carbonate, ammonium carbonate, an alkali metal bicarbonate, an alkaline earth metal bicarbonate, and an ammonium bicarbonate. The determination method according to claim 1, wherein the determination method is at least one carbonate compound. Step (3) determines that the test mammal has no gastric mucosal atrophy when the measured ¹³ CO ₂ behavior is the same as or higher than a reference ¹³ CO ₂ behavior obtained from a control mammal without gastric mucosal atrophy. and, if the measured ¹³ CO ₂ behavior is lower than the reference ¹³ CO ₂ behavior is a step of determining the gastric mucosa atrophy is in a mammalian subject, the determination method according to claim 1. Step (3) determines that the test mammal has no gastric mucosal atrophy when the measured ¹³ CO ₂ behavior is higher than the cut-off value, and the measured ¹³ CO ₂ behavior is the same or lower than the cut-off value In this case, the determination method according to any one of claims 1 to 6, which is a step of determining that the test mammal has gastric mucosal atrophy.