HK1117754A - Oral preparation useful in measuring capacity to metabolize pyridine - Google Patents

Description

Oral preparation for measuring pyrimidine metabolic capability

Technical Field

The present invention relates to an oral preparation capable of efficiently and highly accurately evaluating the occurrence or degree of pyrimidine metabolic capability disorder, the rate of pyrimidine metabolism, and the like. The invention also relates to a method for producing such an oral preparation.

Background

5-Fluorouracil (hereinafter sometimes referred to as "5-FU") and its various derivatives (e.g., tegafur, carmofur, doxifluridine, etc.), and fluorouracil-like drugs have been widely used in the current treatment of cancer. It is known that 5-FU, when taken by a human body, is first degraded by the action of dihydropyrimidine dehydrogenase (hereinafter sometimes referred to as "DPD"), the first enzyme in the pyrimidine metabolic pathway. Therefore, it is believed that the concomitant administration of a drug inhibiting DPD enzyme activity can maintain the efficacy of fluorouracil drugs such as 5-FU and the like. On the other hand, it is known that when a fluorouracil drug such as 5-FU is administered to a subject with DPD enzyme deficiency or reduced enzyme activity, the drug is not metabolized in a normal manner, resulting in abnormally high concentrations of the fluorouracil drug in the blood, thereby causing serious side effects (e.g., bone marrow suppression, digestive tract symptoms, etc.).

Therefore, in order to effectively exert the pharmacological effects of fluorouracil drugs or to prevent side effects thereof, the ability to diagnose pyrimidine metabolism prior to the administration of fluorouracil drugs, i.e., the presence, extent, and the like of pyrimidine metabolic disorders in subjects, is considered to be very important.

A method for diagnosing pyrimidine metabolic activity in a subject has been reported in which an isotope-labeled pyrimidine compound is administered to the subject, and the ability of pyrimidine metabolism, i.e., the presence, degree, and the like of a pyrimidine metabolic disorder in the subject, is determined by measuring the excretion behavior of the isotope-labeled metabolic product excreted in the subject (for example, patent document 1). In the above method, a particle or fine particle containing an isotope-labeled pyrimidine compound and a carrier have been disclosed as a diagnostic agent for pyrimidine metabolizing ability for the above method.

However, despite the large number of isotopically labelled pyrimidine compounds such as¹³The powder of C-uracil is a fine particle of only a few microns, but has low solubility and characteristically high viscosity. Therefore, particles or fine particles prepared from isotope-labeled pyrimidine compounds and the like by standard methods cannot be dissolved rapidly, and in part for this reason, the compounds have disadvantages such as slow and uneven absorption rate in living bodies, and variation in absorption rate due to individual difference. Therefore, in order to achieve more accurate diagnosis of pyrimidine metabolic capability, it is necessary to overcome the above-mentioned drawbacks, thereby reducing variation in excretion time and variation in the number of isotopically labeled metabolites, and reducing inconsistency in diagnosis accuracy due to individual differences.

Patent document 1: international publication No. WO02/072153, single file.

Disclosure of Invention

Problems to be solved by the invention

The object of the present invention is to provide an oral preparation which can be used for diagnosing the presence, degree, etc. of pyrimidine metabolic disorder with high accuracy and with little variation depending on individual differences.

Means for solving the problems

The inventors of the present invention have made intensive studies to solve the above problems, and found that: an oral preparation prepared by mixing (a) an isotopically labeled compound and/or a metabolite thereof with (b) a sugar and/or a sugar alcohol and pulverizing the obtained powdery material enables diagnosis of pyrimidine metabolizing ability with high accuracy and with little variation depending on individual differences. On the basis of this finding, the present invention has been completed by further improvements.

The present invention provides the following oral preparations, and a production method thereof, and the like.

An oral preparation produced by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof and (b) a sugar and/or a sugar alcohol to obtain a powdery material, wherein at least 1 of a carbon atom, an oxygen atom and a nitrogen atom of the pyrimidine compound and/or the metabolite thereof is labeled with an isotope.

The oral preparation according to claim 1, wherein 50% of the powdery material has a particle size of 40 μm or less.

The oral preparation according to claim 3, wherein the oral preparation contains component (a) in a proportion of 5 to 20% by weight.

The oral formulation according to item 4 of claim 1, wherein the component (a) is an isotopically labeled uracil.

The oral preparation according to item 5 or 1, wherein the component (b) is mannitol.

The oral formulation according to item 6, wherein the component (a) is an isotopically labeled uracil and the component (b) is mannitol.

The oral formulation according to claim 7, wherein the oral formulation is a granular formulation.

The oral preparation according to claim 8 or 7, which is produced by granulating a powdery material by extrusion.

The oral formulation according to claim 9 or 7, wherein the mean particle diameter of the granule formulation is 1400 μm or less.

The oral formulation of claim 1, wherein the oral formulation is for diagnosing pyrimidine metabolizing capacity.

The oral formulation of claim 11, wherein the oral formulation is used to determine gastric emptying capacity.

The oral formulation of claim 1, wherein the oral formulation is used for diagnosing dyspepsia.

Item 13 a process for producing an oral preparation, comprising the steps of:

(1) a powdery material produced by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof and (b) a sugar and/or a sugar alcohol, wherein at least 1 of carbon atoms, oxygen atoms, and nitrogen atoms of the pyrimidine compound and/or the metabolite thereof is labeled with an isotope; and

(2) preparing the powdery material obtained in the step (1) into a preparation.

Item 14 the method according to item 13, wherein the particle size of the powdery material produced in the step (1) is 40 μm or less.

The method according to item 15, wherein the oral preparation contains component (a) in a proportion of 5 to 20% by weight.

Item 16 the method according to item 13, wherein the component (a) is an isotopically labeled uracil.

The method according to item 17, wherein the component (b) is mannitol.

The method according to item 18, wherein the component (a) is an isotopically labeled uracil and the component (b) is mannitol.

The method of claim 13, wherein the oral formulation is granulated.

Item 20 the method according to item 19, wherein the step (2) prepares the powdery material obtained in the step (1) into a formulation by extrusion granulation.

The method of item 21 to 19, wherein the oral formulation is a granular formulation having an average particle size of 1400 μm or less.

The method of claim 13, wherein the oral formulation is used to diagnose pyrimidine metabolizing capacity.

The method of claim 13, wherein the oral formulation is used to determine gastric emptying capacity.

The method of claim 24, wherein the oral formulation is used to diagnose dyspepsia.

The use of the powdery material obtained by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof and (b) a sugar and/or a sugar alcohol in the production of a preparation for diagnosing the metabolic capability of pyrimidine, wherein at least 1 of the carbon atom, oxygen atom and nitrogen atom of the pyrimidine compound and/or the metabolite thereof is labeled with an isotope.

The use of the powdery material obtained by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof and (b) a sugar and/or a sugar alcohol in the production of a preparation for measuring the emptying capacity of the stomach, wherein at least 1 of the carbon atom, the oxygen atom and the nitrogen atom of the pyrimidine compound and/or the metabolite thereof is labeled with an isotope.

The use of a powdery material obtained by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof and (b) a sugar and/or a sugar alcohol in the production of a preparation for diagnosing dyspepsia, wherein at least 1 of a carbon atom, an oxygen atom and a nitrogen atom of the pyrimidine compound and/or the metabolite thereof is isotopically labeled.

Advantageous effects of the invention

The oral preparation of the present invention is a preparation made of a powdery material obtained by mixing and pulverizing (a) an isotopically labeled compound and/or a metabolite thereof and (b) a sugar and/or a sugar alcohol. By this formulation design, the oral preparation of the present invention makes it possible to diagnose pyrimidine metabolic ability and gastric emptying ability with high accuracy and with little variation depending on individual differences. As a result, the behavior of the isotopically labeled metabolic products can be determined by 1 or a few measurements after administration for 20 to 30 minutes, so that the time for determination and the number of measurements are reduced, thereby also reducing the burden on the patient.

Brief description of the drawings

FIG. 1 shows the degradation (metabolism) behavior of pyrimidine compounds (uracil, 5-fluorouracil (5-FU), and thymine) by a series of pyrimidine metabolizing enzymes (dihydropyrimidine dehydrogenase (DPD), dihydropyrimidinase (DHPase), and β -ureidopropionase (β -UPase)).

FIG. 2 shows the administration of the granular formulation of example 2 to three healthy subjects (subjects A, B, and C) and observing their exhaled breath¹³CO₂The time variation and the results are compared.

FIG. 3 shows the administration of the granule preparation of comparative example 2 to three healthy subjects (subjects A, B, and C) and observing the exhaled breath of the subjects¹³CO₂The time variation and the results are compared.

FIG. 4 is a graph showing the administration of the preparation of example 1 to 20 suspected patients with gastroparesis in their expired air in test example 5¹³CO₂The change over time.

FIG. 5 shows that the patients were divided into 3 groups (normal gastric emptying capacity, decreased gastric emptying capacity, insufficient gastric emptying capacity) and the preparation of example 1 was administered to them 20 minutes later, 2-¹³C concentration of uracil.

Best mode for carrying out the invention

Hereinafter, the present invention will be described in detail.

The oral preparation of the present invention contains an isotopically labeled pyrimidine compound and/or a metabolite thereof (hereinafter sometimes referred to as "component (a)").

The pyrimidine compound used in the present invention may be any one of a number of compounds having a pyrimidine skeleton, and is preferably a substrate for a pyrimidine-metabolizing enzyme, particularly a substrate for dihydropyrimidine dehydrogenase, which is the first enzyme in the pyrimidine-metabolizing pathway in vivo. Specific examples of such pyrimidine compounds include uracil, thymine, and derivatives thereof. Such uracil and thymine derivatives are not limited as long as they are substrates of DPD and as long as their final metabolites formed by pyrimidine metabolic pathways are excreted into excretions such as exhaled breath, urine, or sweat. Specific examples of such derivatives include halides of uracil, such as 5-fluorouracil, 5-bromouracil, and the like; derivatives of thymine, such as 5-fluorothymine, 5-bromothymine, and the like; and so on. Examples of preferred pyrimidine compounds include uracil, thymine, and 5-fluorouracil.

In addition to the compounds described above as direct substrates for DPD, useful pyrimidine compounds also include compounds that are indirect substrates for enzymes, i.e., precursors (including prodrugs), which are metabolized or degraded in vivo to DPD substrates (e.g., uracil, thymine, and 5-fluorouracil, among others). Examples of such precursors include precursors of uracil, such as cytosine, uridine, and their phosphates (e.g., uridylic acid); precursors of thymine, such as 5-methylcytosine, thymidine, and their phosphates (e.g., thymidylate); and precursors of 5-fluorouracil (prodrugs), such as tegafur, carmofur, doxifluridine, and the like.

The metabolite of the pyrimidine compound corresponds to a metabolic intermediate of the pyrimidine compound, and is a substrate for a pyrimidine-metabolizing enzyme, particularly, a second enzyme in the pyrimidine-metabolizing pathway in the living body, dihydropyrimidinase (hereinafter sometimes referred to as "DHPase"), or a third enzyme, β -ureidopropionase (hereinafter sometimes referred to as "β -UPase"). Specific examples of metabolites of pyrimidine compounds include dihydrouracil, dihydrothymine, and derivatives thereof (e.g., halides of dihydrouracil, such as 5-fluorodihydrouracil, and the like), which are substrates of DHPase; beta-ureidopropionic acid, beta-ureidoisobutyric acid, and derivatives thereof (e.g., halides of beta-ureidopropionic acid, such as fluoro-beta-ureidopropionic acid, and halides of beta-ureidoisobutyric acid), which are substrates for beta-UPase.

In the present invention, component (a) is preferably a pyrimidine compound, more preferably uracil, thymine, or 5-fluorouracil, still more preferably 5-fluorouracil.

At least one of a carbon atom, an oxygen atom and a nitrogen atom in a molecule of the pyrimidine compound and/or its metabolite used in the present invention is isotopically labeled. Specific examples of isotopes include but are not limited to,¹³C、¹⁴C、¹⁸o and¹⁵n. isotopes may or may not be radioactive, but from a safety point of view, are non-radioactive¹³C、¹⁸O or¹⁵N is preferred.

The pyrimidine compound and/or its metabolite used in the present invention may be labeled with 1 isotope in the molecule, or may be labeled with 2 or more isotopes of the same or different elements. Although not limited, preferably, the carbon atom or oxygen atom in the pyrimidine compound or its metabolite is labeled so that at least CO is produced via the pyrimidine metabolic pathway₂Is isotopically labelled. Examples of such pyrimidine compounds include those having an isotopically labeled carbon atom at the 2-position of the pyrimidine backbone. Specific examples include 2-¹³C-labelled uracil and 2-¹³C-labeled fluorouracil.

The method of labeling pyrimidine compounds and/or metabolites thereof with Isotopes as described above is not limited, and various conventional methods (Sasaki, "5.1 Application of Stable Isotopes in Clinical diagnostics"; Kagaku no Ryoiki (Journal of Japanese chemistry)107, "Application of Stable Isotopes in Medicine, Pharmacy, and Biology", Nankodo, pp.149-163 (1975); Kajiwara, "RadiISOTOPES", 41, 45-48(1992), etc.) can be utilized. Some of these isotopically labeled pyrimidine compounds and their metabolites are commercialized, and these commercial products are convenient to use.

The proportion of the component (a) in the oral preparation of the present invention is, for example, usually 5 to 20% by weight, preferably 6 to 18% by weight, more preferably 8 to 15% by weight.

The oral preparation in the present invention contains, in addition to the component (a), a sugar or sugar alcohol (hereinafter sometimes referred to as "component (b)").

The sugar used in the present invention may be any pharmaceutically acceptable one. Examples of such sugars include: monosaccharides such as glucose, galactose, fructose, xylose, arabinose, and mannose; disaccharides such as maltose, isomaltose, cellobiose, lactose, sucrose, and trehalose; and so on. Among them, preferred are glucose and sucrose.

The sugar alcohol used in the present invention may be any pharmaceutically acceptable one. Specific examples of the sugar alcohol include erythritol, mannitol, xylitol, sorbitol, maltitol, reduced isomaltulose (paratinose), lactitol; and so on. Among them, mannitol, xylitol and erythritol are preferable, and mannitol is most preferable.

In the present invention, the component (b) is preferably a sugar alcohol, more preferably mannitol, xylitol or erythritol, and still more preferably mannitol.

The proportion of component (b) in the oral preparation of the present invention is, for example, usually 80 to 95% by weight, preferably 82 to 94% by weight, more preferably 85 to 92% by weight, based on the total weight of the preparation.

The weight ratio of component (b) to component (a) in the oral preparation of the present invention is, for example, 400-1900 parts by weight, preferably 450-1550 parts by weight, more preferably 550-1150 parts by weight, for 100 parts by weight of component (a). The combined use of the component (a) and the component (b) in such a ratio further improves the accuracy of the diagnosis of pyrimidine metabolic disorders.

The oral preparation of the present invention is produced by formulating a powdery material containing the components (a) and (b) into a preparation. The powdery material for use in the preparation of the oral preparation in the present invention is obtained by mixing the components (a) and (b) in the above-mentioned ratio and pulverizing the resulting mixture.

The oral preparation of the present invention may contain the same constituent components as those of the powdery material after pulverization, or may contain other components in addition to the powdery material. Therefore, the ratio of the components (a) and (b) in the powdery material is appropriately selected depending on the ratio of the components (a) and (b) in the final form of the oral preparation, the preparation step of the oral preparation, and the like.

The powdery material can be obtained by mixing and grinding the pharmaceutically acceptable additives and the components (a) and (b) together as long as the effects of the present invention are not impaired. Such additives may be those which can be added when formulating the pulverulent material. Specific examples of such additives will be given below.

The particle size of the powdery material is not limited as long as the particle size is the result of mixing and grinding the components (a) and (b), but in order to improve the accuracy of the pyrimidine metabolizable capacity diagnosis, it is desirable that 50% of the particles have a particle size of 40 μm or less, preferably 30 μm or less, more preferably 5 to 20 μm.

Examples of the preferred powdery material are those in which 50% of the particle diameter of the powdery material is 40 μm or less and 90% of the particle diameter is 200 μm or less; more preferably, the powdery material has a particle size distribution such that 50% of the particles have a particle size of 30 μm or less and 90% of the particles have a particle size of 100 μm or less; still more preferably, the powdery material has a particle size distribution such that 50% of the particles have a particle size of 5 to 20 μm and 90% have a particle size of 10 to 70 μm. The oral preparation made using such a powdery material having a particle size distribution enables the component (a) to be absorbed by the living body at a rapid and uniform rate, thereby making it possible to diagnose the pyrimidine metabolism with higher accuracy.

The meaning of 50% particle size and 90% particle size of the pulverulent material mentioned here is as follows: starting from the particle with the smallest particle size, the particle volumes of the powdery material are accumulated until the accumulated volume accounts for 50% or 90% of the total volume of the powdery material particles, and the particle size of the last accumulated particle is 50% or 90%. The particle diameters of 50% and 90% can be measured by a dry laser method (measurement conditions: focal length 100mm, average measurement 10 times, average interval 5 msec, and air pressure 0.4 MPa).

The pulverization treatment for preparing the powdery material is not limited, but the pulverization treatment is preferably carried out by a dry mill. Specific examples of drying mills include: hammer mills, pin mills, jet mills, and the like.

If necessary, the oral preparation of the present invention can be produced by adding additives such as excipients, binders, pH adjusters, disintegrants, absorption enhancers, lubricants, coloring agents, corrigents, flavoring agents, etc. to the powdery materials and formulating the resulting mixture by granulation or other shaping processes selected according to the formulation form of the preparation. When the oral preparation of the present invention is a powdery preparation, such powdery material may be used as a final form of the oral preparation.

Specific examples of additives that may be used in the formulation include: lactose, starch, refined white sugar, dextrin, mannitol, xylitol, sorbitol, erythritol, calcium dihydrogen phosphate, sodium chloride, glucose, calcium carbonate, kaolin, crystalline cellulose, silicate, etc.; binders such as water, ethanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethyl cellulose, sodium carboxymethyl cellulose, shellac, methyl cellulose, hydroxypropyl cellulose, polyvinylpyrrolidone, polyvinyl alcohol, gelatin, dextrin, pullulan, etc.; pH regulators such as citric acid, citric anhydride, sodium citrate dihydrate, anhydrous disodium hydrogen phosphate, anhydrous sodium dihydrogen phosphate, disodium hydrogen phosphate, and anhydrous disodium hydrogen phosphate; disintegrating agents such as carboxymethylcellulose calcium, low-substituted hydroxypropylcellulose, carboxymethylcellulose, croscarmellose sodium, sodium carboxymethyl starch, and crospovidone; absorption promoters such as polysorbate 80, quaternary ammonium hydroxide and sodium lauryl sulfate; pure talcum powder, stearate, polyethylene glycol, colloidal silicic acid, sucrose fatty acid, hydrogenated oil and other lubricants; colorants such as iron oxide yellow, iron oxide yellow sesquioxide, iron sesquioxide, β -carotene, titanium dioxide, food color (e.g., food color blue No. 1), copper chlorophyll, and riboflavin; corrigents such as ascorbic acid, aspartame, sweet hydrangea leaf (sweet hydrangea leaf), and sodium chloride; and so on.

The dosage form of the oral preparation in the present invention is not limited as long as it is a solid preparation, and fine granules, powders, tablets (including naked tablets and sugar-coated tablets), capsules, pills, and other dosage forms can be selected as needed. Among them, in order to enhance the effect of the present invention, granular formulations such as fine particles and granules, particularly granular formulations produced by extrusion granulation, are preferable.

When the oral preparation in the present invention is a granular preparation, the average particle diameter of the preparation is, for example, usually 1400 μm or less, preferably 50 to 1200 μm, and more preferably 100 to 1000 μm. When the granular preparation has such a particle size, the granular preparation enables more accurate diagnosis of pyrimidine metabolizing ability. The particle size of the preparation can be measured by a vibration sieve method (specifically, the measuring instrument used is a Robot Shifter RPS-95 (manufactured by Seishin Enterprise Co., Ltd.) with a vibration level of 5, a vibration time of 5 minutes, and a pulse interval of 1 second).

After administration of the oral formulation of the present invention, the ability of pyrimidine metabolism, i.e., whether or not a subject has a pyrimidine metabolic disorder and its degree, the rate of pyrimidine metabolism, etc., can be evaluated by measuring the excretion behavior of isotope-labeled metabolites excreted by the body. Thus, the oral formulation of the present invention is a formulation that can be used to determine the metabolic capacity of pyrimidines. Further, as described hereinafter, since the gastric emptying ability can also be evaluated based on the result of the evaluation of pyrimidine metabolizing ability, specifically, the result of the measurement of pyrimidine metabolizing rate, the oral preparation of the present invention can also be a preparation for measuring the gastric emptying ability. Specific embodiments of the agent for measuring pyrimidine metabolizing ability and the agent for measuring gastric emptying ability will be described below in detail.

Preparation for measuring pyrimidine metabolizing ability

Since the oral preparation of the present invention can be used for determining pyrimidine metabolic capacity, relating to the presence and extent of pyrimidine metabolic disorders, the preparation can be used for detecting, measuring and diagnosing pyrimidine metabolic disorders. Specific conditions, methods and the like for measuring pyrimidine metabolizable ability using the oral preparation of the present invention are as follows.

As shown in FIG. 1, when the oral preparation of the present invention is administered to a test subject having a normal pyrimidine metabolizing ability, a series of pyrimidine metabolizing enzymes (DPD, DOHase, and. beta. -UPase) function normally in vivo (hereinafter sometimes referred to as a healthy test subject) as the ingredient (a) contained in the preparation is metabolized and degraded into. beta. -alanine, fluoro-. beta. -alanine,. beta. -aminoisobutyric acid, NH-₃、CO₂And so on.

Final metabolite CO produced by metabolism₂Is excreted by the exhaled breath, whereas beta-alanine, fluoro-beta-alanine or beta-aminoisobutyric acid is mainly excreted with the urine. Among the excreted final metabolites, at least CO differs depending on the isotopically labeled sites of the pyrimidine compound and/or its metabolite as the component (a)₂Or one selected from the group consisting of beta-alanine, fluoro-beta-alanine and beta-aminoisobutyric acid is isotopically labeled. Such isotopic labeling is used to measure the excretion behavior (the amount of excretion or the rate of excretion over time) of these final metabolites when CO is present₂When labeled, using exhaled breath as the test sample, or when beta-alanine, fluoro-beta-alanine, beta-aminoisobutyric acid or ammonia is labeledUrine was used as a test sample.

The ability of the test subject to metabolize pyrimidine can be determined by the thus measured excretion behavior (the amount of excretion or the expression of excretion rate over time) of the isotopically labeled metabolic product.

When the oral preparation of the present invention is used for measuring the pyrimidine metabolizing ability, the dosage of the oral preparation of the present invention is not limited, but a preferable amount is the component (a) corresponding to 1 to 2000mg, and a preferable amount is 10 to 300 mg.

When the oral formulation of the present invention is used to determine the ability of pyrimidine metabolism, it is preferred to use CO that causes isotopic labeling₂A pyrimidine compound and/or a metabolite thereof as a component (a) which is excreted into exhaled breath as a result of metabolism. Using this formulation, the pyrimidine metabolizing ability of the test subject can be measured by isotopically labeled CO₂Is determined by administering the present formulation to a subject and measuring the isotopically labeled CO in the exhaled breath of the subject₂And (4) finishing.

When the active ingredient contained in the preparation is a pyrimidine compound, the formed isotopically-labeled compound is not isotopically-labeled CO₂And beta-alanine, fluoro-beta-alanine, beta-aminoisobutyric acid, etc., the excretion such as urine, sweat, etc. is used instead of the expired air as the test sample.

When using exhaled breath as a test sample, isotope-labeled CO in exhaled breath is measured₂Depending on whether the isotope used is radioactive or not. Conventional analytical methods are available including liquid scintillation counting, mass spectrometry, infrared spectroscopy, emission spectroscopy, magnetic resonance spectroscopy, and the like. From the viewpoint of measurement accuracy, infrared spectroscopy and mass spectrometry are preferable. When excreta such as urine or sweat is used as a test sample, an isotopically labeled pyrimidine compound (or isotopically labeled pyrimidine metabolite), an isotopically labeled metabolic intermediate, and an isotopically labeled metabolite contained in the test sampleThe products can be separated and analyzed simultaneously by using liquid chromatography, gas chromatography and other separation techniques. So that the excretion behavior of the isotopically labeled metabolic products can be selectively measured.

The pyrimidine metabolizing ability of a test subject can be evaluated, for example, by comparing the excretion behavior (the amount of excretion or the expression of excretion rate over time) of an isotope-labeled metabolite in the test subject measured by the above-described method with the excretion behavior of an isotope-labeled metabolite of a healthy test subject having a normal pyrimidine metabolizing ability measured in the same manner. In particular isotopically labelled CO in exhaled breath₂Is measured as an isotopically labeled metabolite, the isotopically labeled CO is measured at a predetermined time point after administration of the oral formulation₂Amount of gas, CO₂Delta (‰) value of gas (isotopically labeled in exhaled breath samples collected before and after administration of the present oral formulation)¹³CO₂/¹²CO₂Difference in concentration ratio), or isotopically-labeled CO in exhaled breath₂The initial rate of excretion of the gas can be used as an indicator of the excretion behavior of the isotopically labeled metabolic products. For example, with the CO of a healthy subject₂The value of the gas or the initial rate, CO₂Subjects with lower Δ (‰) values of gas or lower initial rates are diagnosed with a decreased pyrimidine metabolizing capacity.

Further, instead of or in addition to the excretion behavior of the isotopically labeled metabolite, the area under the curve (AUC), the excretion rate (particularly the initial excretion rate), the maximum excretion concentration (Cmax), or other parameters in the subject, preferably pharmacokinetic parameters, may be used for comparison with corresponding data for healthy subjects.

The absence or presence of a pyrimidine metabolizing enzyme (at least one of DPD, DHPase and β -UPase) can be determined based on the presence or absence of excretion of isotopically labeled metabolites without comparison to the excretion behavior of healthy subjects. The decrease or increase in pyrimidine metabolic capacity (pyrimidine metabolic disorder) and its extent (extent of pyrimidine metabolic disorder) can be determined by comparing the excretory behavior of the subject or parameters obtained therefrom with those of a healthy subject.

Preparation for measuring gastric emptying capacity

When the oral formulation of the present invention is used to measure gastric emptying capacity, it is preferable to be able to label the isotope with CO₂Is excreted into exhaled breath as a result of metabolism of the pyrimidine compound and/or its metabolite component (a).

After oral ingestion by the subject, the oral formulation of the present invention enters the stomach and eventually passes through the contraction of the stomach and peristalsis to be excreted via the pylorus. After excretion from the pylorus, component (a) is rapidly absorbed by the duodenum and downstream parts of the gastrointestinal tract (duodenum, jejunum, ileum, etc.), metabolized, and isotopically labeled CO₂The gas is discharged in the form of exhaled air. The component (a) used in the oral preparation of the present invention is not or hardly absorbed into the stomach, but after being discharged from the stomach, the component is rapidly absorbed, metabolized, and isotopically labeled CO₂The gas is discharged in the form of exhaled air. Thus, isotopically labelled CO in exhaled breath₂The excretion behavior of gases (which expression is, for example, isotopically-labelled CO in exhaled air)₂Gas and¹²CO₂ratio of (isotopically labeled CO)₂/¹²CO₂) Depending on the gastric emptying rate (gastric emptying time) of component (a) contained in the oral formulation of the invention.

The dosage of the oral formulation of the present invention may be the same as that when the oral formulation of the present invention is used for determining pyrimidine metabolizing ability.

Isotopically labelled CO in exhaled breath₂The measurement method can be the same as that in the case where the oral preparation of the invention is used for determining the pyrimidine metabolizing ability.

The gastric emptying capacity of a subject can be determined by an index of gastric emptying capacity: the oral preparation is administered with isotopically labeled CO after a predetermined time₂Amount of gas, CO₂Delta (‰) value of gas (isotopically labeled CO in exhaled breath samples collected before and after administration of the present oral formulation)₂/¹²CO₂Difference in concentration ratio), or isotopically-labeled CO₂Initial rate of gas discharge. For example, with the CO of a healthy subject₂The value of the gas or the initial rate, CO₂Subjects with lower Δ (‰) values of gas or lower initial rates are diagnosed with decreased gastric emptying.

The oral preparation of the present invention may be taken alone or may be taken at the same time as or immediately before or after the test meal. Preferably, the composition for measuring gastric emptying capacity of the present invention is taken immediately after ingestion of the test meal. The test meal is not limited as long as the effect of measuring gastric emptying ability of the formulation of the present invention is not impaired, and may be a solid food, a fluid food, or a liquid food.

The main causes of dyspepsia (non-ulcerative upper gastrointestinal syndrome) are gastrointestinal motility disorders, and a partial reduction in gastric emptying capacity. Therefore, the oral formulation of the present invention can be an effective formulation for diagnostic tests of dyspepsia, especially dyspepsia mainly caused by insufficient gastric emptying ability (e.g., hypomotic dyspepsia).

Furthermore, the use of the oral formulation of the present invention for determining gastric emptying capacity makes it possible to determine the efficacy or therapeutic effect of gastrointestinal drugs, especially drugs associated with gastrointestinal motility functions, on individual subjects. In particular, the determination can be made by measuring the gastric emptying capacity of the oral formulation of the invention used before and after the administration of gastrointestinal drugs, in particular drugs related to gastrointestinal motor function, and comparing the two sets of measurements. The efficacy of the drug itself can thus be evaluated. In addition, the drug can be evaluated for individual testeesThe present oral formulation may also be used to screen for drugs appropriate for individual subjects. Examples of the drugs related to gastrointestinal motility function include drugs which control the peristalsis of the stomach by enhancement or inhibition, such as gastrointestinal motility function-promoting agents, gastrointestinal motility function-enhancing agents, gastrointestinal motility function-activating agents (specifically, acetylcholine agonists, dopamine receptor antagonists, dopamine D)₂Receptor antagonists, 5-hydroxytryptamine receptor agonists, opioid agonists, and traditional Chinese medicines (Liu Jun ZITang, Ban Xia Kie Xin Tang, and An Zhong San), and gastrointestinal motor function inhibitors (anticholinergic agents, muscarinic receptor antagonists, etc.), and the like. The same assay can also be performed on patients with dyspepsia, especially those with dyspepsia mainly caused by hypomotility of the stomach (patients with dyskinetic dyspepsia) as the test subject. In this case, the therapeutic effect of the drug in a single dyspepsia patient can be measured, thereby making it possible to screen for suitable drugs related to gastrointestinal motor function (such as the above-mentioned gastrointestinal motor function-promoting agent, gastrointestinal motor function-enhancing agent, or gastrointestinal motor function-activating agent).

Examples

The following examples and test examples illustrate the invention and show production examples of formulations and evaluation of the properties of the formulations. However, the scope of the present invention is not limited to these examples and test examples.

< formulation production example >

Example 1

20 g of the powder¹³Uracil labeled C and 380 g of D-mannitol (manufactured by Mannit, Kyowa Hakko Kogyo Co., Ltd.) were mixed, put into a sample mill (KIIWG-1F, manufactured by Fuji Paudal Co., Ltd.), and mixed and pulverized (pulverization conditions: the rotation speed of a pulverizing rotor was 12800rpm, and the rotation speed of a rotor at a sample feed port was about 10rpm, using a sieve having a hole diameter of 1 mm) to prepare a powdery material. The resulting powdery material was weighed out to 200 g and put into a speed kneader (NSK-150, Oka)da Seiko Co., Ltd.), 20 g of pure water was added and kneaded. The obtained wet powder was subjected to extrusion granulation by an extrusion granulator (Dome Gran DG-L, manufactured by Fuji Paudal Co.) having a hole with a diameter of 1mm on a Dome-shaped die, and dried by a blower (SPHH-200, manufactured by Espec Co.) set at 60 ℃. Of these dried granules, those which passed through a sieve having a pore size of 1400 μm and which failed to pass through a sieve having a pore size of 355 μm were regarded as granules containing 5% by weight of the active ingredient¹³A granular formulation of C-labeled uracil.

Thus obtained containing 5% by weight of¹³The particle size of the granular preparation of C-labeled uracil was measured by a vibration sieve method (specifically, a measuring instrument used was Robot Shifter RPS-95 (manufactured by Seishin Enterprise Co., Ltd.) with a vibration level of 5, a vibration time of 5 minutes, and a pulse interval of 1 second). The results are shown in Table 1.

[ TABLE 1 ]

Particle size	Ratio (wt.%)
		Greater than or equal to 1400 μm	2.09
Greater than or equal to 1000 μm and less than 1400 μm	7.29
		850 μm or more and 1000 μm or less	22.07
More than or equal to 710 μm and less than 850 μm	59.04
		More than or equal to 500 μm and less than 710 μm	8.99
355 μm or more and 500 μm or less	0.09
		250 μm or more and 355 μm or less	0.00
Greater than or equal to 150 μm and less than 250 μm	0.09
		Less than 150 μm	0.34
Total of	100.0

Comparative example 1

Mixing 10 g¹³The C-labeled uracil and 190 g of D-mannitol (manufactured by Mannit, Kyowa Hakko Kogyo Co., Ltd.) were put into a speed kneader (NSK-150, manufactured by Okada Seiko Co., Ltd.) and mixed, and then kneaded after adding 20 g of pure water without pulverization. Followed by granulation, drying and the particle size was adjusted by sieving under the same conditions as in example 1 to obtain a powder containing 5% by weight¹³A granular formulation of C-labeled uracil. Thus obtained containing 5% by weight of¹³Particles of C-labeled uracilThe particle size of the formulation was measured by the same method as used in example 1. The results are shown in Table 2.

[ TABLE 2 ]

Particle size	Ratio (wt.%)
		Greater than or equal to 1400 μm	1.24
Greater than or equal to 1000 μm and less than 1400 μm	5.80
		850 μm or more and 1000 μm or less	30.39
More than or equal to 710 μm and less than 850 μm	54.87
		More than or equal to 500 μm and less than 710 μm	6.01
355 μm or more and 500 μm or less	0.20
		250 μm or more and 355 μm or less	0.10
Greater than or equal to 150 μm and less than 250 μm	0.20
		Less than 150 μm	1.19
Total of	100.0

Example 2

20 g of the powder¹³Uracil labeled C and 180 g of D-mannitol (manufactured by Mannit, Kyowa Hakko Kogyo Co., Ltd.) were mixed, put into a sample mill (KIIWG-1F, manufactured by Fuji Paudal Co., Ltd.), and mixed and pulverized (the rotation speed of the grinding rotor was 12800rpm, and the rotation speed of the rotor at the sample feed port was about 10rpm, using a sieve having a pore diameter of 1 mm) to prepare a powdery material. The resulting powdery material was weighed out 144 g and put into a speed kneader (NSK-150, manufactured by Okada Seiko Co., Ltd.), 14.4 g of pure water was added and kneaded. The obtained wet powder was extruded through an extrusion granulator (Dome Gran DG-L, manufactured by FujiPaudal) having a hole with a diameter of 1mm on a Dome-shaped die, and dried by a blower (SPHH-200, manufactured by Espec) set at 60 ℃. Of these dried granules, granules which passed through a sieve having a pore size of 1400 μm and which could not pass through a sieve having a pore size of 355 μm were defined as containing 10% by weight of the total amount of the granules¹³Granular formulations of C-labeled uracil.

Comparative example 2

20 g of the powder¹³The C-labeled uracil and 180 g of D-mannitol (manufactured by Mannit, Kyowa Hakko Kogyo Co., Ltd.) were thoroughly mixed, and placed in a speed kneader (NSK-150, manufactured by Okada Seiko Co., Ltd.). Adding 20 g of pure water and kneading. Followed by granulation, drying and the particle size was adjusted by sieving under the same conditions as in example 2 to obtain a powder containing 10% by weight¹³A granular formulation of C-labeled uracil.

Comparative example 3(tablet)

100 g¹³C-labeled uracil, 60 g of lactose (manufactured by H.M.S.), 25 g of corn starch (manufactured by Shokuhin Kako Co.), 10 g of crystalline cellulose (manufactured by Asahi Kasei Co.), and 4 g of hydroxypropyl cellulose (HPC-L purified powder, manufactured by Nippon Soda Co.) were put into a speed kneader (NSK-150, manufactured by Okada Seiko Co.) and mixed. 40 g of purified water was added and kneaded. The resulting kneaded powder was then extruded through an accelerated mill (ND-02, manufactured by Okada Seiko corporation) having a sieve with a hole of 3mm in diameter, and dried by a blower (SPHH-200, manufactured by Espec corporation) set at 70 ℃. The dried granules were subjected to particle size adjustment through a 16-mesh sieve, and 1 g of magnesium stearate was added to 199 g of the granules obtained after the particle size adjustment for the preparation of tablets. The granules were compressed into tablets weighing 200mg per tablet by a single-punch tablet press (model: 2B, manufactured by Kikusui Seisakusho Co., Ltd.) with punches and a die having a diameter of 8mm and rounded corners.

Example 3

20 g of the powder¹³C-labeled uracil and 180 g of D-mannitol (manufactured by Mannit, Kyowa Nara machinery Co., Ltd.) were thoroughly mixed, and put into a sample mill (SAM, manufactured by Okada Seiko Co., Ltd.), and mixed and pulverized (shape of grinding blade: needle shape; rotation speed of rotor: 4000rpm, sieving: sieve having a pore diameter of 3 mm) to obtain a powdery preparation.

Comparative example 4

20 g¹³The C-labeled uracil was sieved through a No.30 sieve to prepare a powdery formulation.

Comparative example 5

200 g of¹³The C-labeled uracil was put into a sample mill (SAM, manufactured by Nara Machinery Co., Ltd.) and pulverized under the same conditions as in example 3 to prepare a powdery formulation.

< evaluation of formulation Properties >

Test example 1Measurement of particle size distribution

The particle size distribution of the powdery formulations in example 3 and comparative examples 4 and 5 was measured by a dry particle size distribution measuring instrument (LDSA-1SOOA, manufactured by Tohnichi Computer) under the following conditions: focal length 100mm, average number of times 10, average interval 5 ms, and air pressure 0.4 MPa. From the measured particle size distribution, particle diameters of 10% (10% D), 50% (50% D) and 90% (90% D) were calculated. The results are shown in Table 3.

[ TABLE 3 ]

	10％D(μm)	50％D(μm)	90％D(μm)
				Example 3	5.74	14.95	56.58
Ratio ofComparative example 4	6.46	75.58	235.00
				Comparative example 5	6.01	52.60	260.57

As shown in table 3, only by sieving¹³Powdery preparation of comparative example 4 obtained from C-labeled uracil, and pulverization alone¹³The powdery preparation of comparative example 5 obtained with uracil labeled C did not have a reduced particle size, indicating insufficient pulverization effect, while the powdery preparation of example 3 obtained by mixing and pulverization had a reduced particle size, indicating sufficient pulverization effect.

Test example 2Evaluation of solubility of the preparation

100 ml of tap water was added to each 200 ml beaker at room temperature. Then, 2000mg of the granular formulations of example 1 and comparative example 1 were added to each beaker while performing magnetic bar stirring (RCN-7D, product EYELA) at 200rpm, and then the time required for dissolution of the formulations was visually observed. Further, the undissolved formulation residue was visually assessed 3 minutes after addition of the granular formulation.

The results are shown in Table 4. As a result, it is apparent that the dissolution time of the formulation of comparative example 1 is long and there is a large amount of undissolved residual formulation, as opposed to the dissolution time of the formulation of example 1 being short and there is only a small amount of undissolved residual formulation.

[ TABLE 4 ]

	Dissolution time	Undissolved residue
			Example 1	1 minute and 10 seconds	And small amount
Comparative example 1	3 minutes or longer	A large number of

Test example 3Evaluation of solubility of the preparation

The 6 tablets obtained in comparative example 3 were subjected to disintegration test according to the disintegration test in the general test procedure of the pharmacopoeia of japanese 14 th edition. The results showed that the disintegration time was 15 minutes or more for all tablets.

Test example 4Evaluation of accuracy of pyrimidine metabolic capability disorder diagnosis

After 3 healthy subjects (subjects A, B and C) were administered the formulations of example 2 and comparative example 2, the expired air from the subjects over time was collected and measured in the expired air with a GC-MS analyzer (ABCA-G, manufactured by Europa Scientific)¹³C-labelled CO₂The concentration of the gas.

FIG. 2 shows the exhalation after taking the formulation of example 2¹³C-labelled CO₂A change in concentration of the gas; FIG. 3 shows the exhalation after taking the formulation of comparative example 2¹³C-labelled CO₂The change in concentration of the gas. In fig. 2 and 3, the ordinate represents Δ¹³C value (‰), showing delta of exhaled breath collected before taking the preparation for measuring pyrimidine metabolizing ability¹³C value ([ permillage ]), (¹³CO₂And¹²CO₂concentration ratio) of the exhaled breath collected during each period after the administration of the preparation and delta¹³The C values (‰) are different. The abscissa represents the time (minutes) for collecting exhaled breath after taking the formulation. One of three subjects took the preparation of comparative example 2¹³C-labelled CO₂The concentration of the gas varied very little, indicating the difference between the testees (see fig. 3). In contrast, when the formulation of example 2 was administered to the same three subjects, the subjects were each other¹³C-labelled CO₂The concentration changes of the gases are similar, showing that the differences between the testees are small. These results show that by taking the preparation of example 2, the exhaled breath collected 20-30 minutes after the administration of the preparation¹³C-labelled CO₂The concentration of the gas is used as an index for diagnosing the pyrimidine metabolic capacity disorder, the pyrimidine metabolic capacity disorder can be rapidly and accurately diagnosed, and the difference is small among different individuals (see figure 2).

Test example 5Diagnosis of gastric emptying capacity

To patients suspected of having postoperative gastroparesis (20 patients), the preparation of example 1 was orally administered at a dose equivalent to 100mg within 20 days after the gastrotomy¹³C-uracil. The patients' exhaled breath was collected 10, 20, 30, 40, 50 and 60 minutes after taking the drug, respectively, and the obtained exhaled breath samples as well as the exhaled breath samples (pre) taken in the same manner before taking the drug¹³CO₂Concentrations were measured by GC/MS. Subsequent calculation of exhaled breath¹³CO₂Change in concentration (. DELTA.)¹³C value(% o)). The results are shown in FIG. 4.

As shown in FIG. 4, the exhaled breath test using the preparation of the present invention enables the patients (20 cases) to be classified into those with normal gastric emptying ability (normal type: solid line), those with diminished gastric emptying ability (delayed gastric emptying type: broken line), and those with insufficient gastric emptying ability (deficient type: dotted line). When these patients were measured for 20 minutes after taking the preparation 2-¹³C uracil concentration, 2-¹³The decrease in the concentration of C-uracil is consistent with gastric emptying capacity. This indicates that the expired air test using the oral formulation of the present invention can effectively reflect the gastric emptying ability.

Claims (18)

1. An oral preparation prepared using a powdery material obtained by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof, and (b) a sugar and/or a sugar alcohol, wherein at least 1 of a carbon atom, an oxygen atom, and a nitrogen atom of the pyrimidine compound and/or the metabolite thereof is labeled with an isotope.

2. The oral formulation of claim 1, wherein 50% of the particle size of the powdery material is 40 μm or less.

3. The oral preparation according to claim 1, wherein the oral preparation comprises component (a) in a proportion of 5 to 20% by weight.

4. The oral formulation according to claim 1, wherein the component (a) is an isotopically labeled uracil.

5. The oral formulation according to claim 1, wherein the component (b) is mannitol.

6. The oral formulation according to claim 1, wherein the component (a) is an isotopically labeled uracil and the component (b) is mannitol.

7. The oral formulation of claim 1, wherein the oral formulation is a granular formulation.

8. The oral formulation according to claim 7, wherein the oral formulation is produced by extrusion granulation of a powdery material.

9. The oral formulation of claim 7, wherein the granular formulation has an average particle size of 1400 μm or less.

10. A method for producing an oral formulation comprising the steps of:

(1) preparing a powdery material by mixing and pulverizing (a) a pyrimidine compound and/or a metabolite thereof, and (b) a sugar and/or a sugar alcohol, wherein at least 1 of a carbon atom, an oxygen atom and a nitrogen atom of the pyrimidine compound and/or the metabolite thereof is labeled with an isotope; and

(2) preparing the powdery material obtained in the step (1) into a preparation.

11. The method according to claim 10, wherein 50% of the powdery material produced in the step (1) has a particle size of 40 μm or less.

12. The method according to claim 10, wherein the oral formulation comprises component (a) in a proportion of 5-20% by weight.

13. The method of claim 10, wherein component (a) is an isotopically labeled uracil.

14. The method according to claim 10, wherein the component (b) is mannitol.

15. The method of claim 10, wherein component (a) is an isotopically labeled uracil and component (b) is mannitol.

16. The method of claim 10, wherein the oral formulation is in the form of granules.

17. The method according to claim 16, wherein the step (2) prepares the powdery material obtained in the step (1) into a formulation by extrusion granulation.

18. The method of claim 16, wherein the oral formulation is a granular formulation having an average particle size of 1400 μ ι η or less.