Method for supporting stratification of brain function level, stratification device, and stratification program

Abstract

To stratify the brain function level of a subject.SOLUTION: A method for supporting stratification of the brain function level of a subject includes: acquiring a measured value of diamine and / or polyamine in urine or in a blood sample collected from the subject; searching a plurality of reference ranges set according to the brain function level for a reference range to which the measured value belongs; and suggesting a brain function level which corresponds to the reference range to which the measured value belongs as the brain function level of the subject.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention discloses a method, a stratification device, and a stratification program for assisting stratification of brain function levels. [Background technology]

[0002] The decline in brain function often occurs mainly with aging, and is accompanied by various symptoms such as learning disabilities, memory disorders, speech disorders, anxiety disorders, insomnia, and loss of motivation. In today's world, where medical advances are leading to an increase in average life expectancy, prevention and measures against the decline in brain function are considered important for extending healthy life expectancy.

[0003] Brain dysfunction is broadly categorized into three types: "healthy" where no abnormalities are found, "dementia" where cognitive decline interferes with daily life, and "mild cognitive impairment (MCI)" where cognitive decline is present but does not interfere with daily life. There is no fundamental cure for dementia, and the main treatments are symptom relief and suppression of progression. Therefore, early detection at the MCI stage and prevention at the "pre-disease" stage are important.

[0004] Currently, neuropsychological tests such as the Hasegawa Dementia Scale-R (HSD-R) and the Mini Mental State Examination (MMSE) and brain imaging tests such as CT and MRI are used to evaluate brain function. Neuropsychological tests require a medical interview, so it is difficult to include them in general examination items such as health checkups. In the case of a medical interview, there is a possibility that the subject himself may give answers that are different from the facts, and there is a problem that objectivity cannot be guaranteed. In addition, it is undeniable that brain imaging tests impose a burden on the subject in terms of time and money, and from the perspective of prevention and early detection, a faster, lower-cost, and scientifically based testing method is desirable.

[0005] Diamines are linear fatty acid hydrocarbon compounds having two amines. Polyamines are linear fatty acid hydrocarbon compounds having three or more amines. The mechanism of action of diamines and / or polyamines is known to be widely involved in the synthesis of nucleic acids and proteins and the regulation of enzyme activity (Non-Patent Document 1). Furthermore, Non-Patent Document 2 reports that the amount of polyamines excreted in urine increases in cancer patients, and a testing method for using this as a marker for early cancer diagnosis is being researched. [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Publication No. 2021 / 200216 [Non-patent literature]

[0007] [Non-Patent Document 1] Shunsuke Sakai et al., Journal of Urology (1986), 32(3): 343-350 [Non-Patent Document 2] Russell DH, et al. Nature New Biol. (1971) 233(39):144-145. Summary of the Invention [Problem to be solved by the invention]

[0008] An objective of the present invention is to stratify the brain function level of a subject quickly and at low cost using a urine or blood sample from the subject, without imposing a time or financial burden on the subject. [Means for solving the problem]

[0009] In view of the above circumstances, the inventors conducted intensive research and discovered that by obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample, searching for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to brain function levels, and suggesting the brain function level corresponding to the reference range to which the measurement values ​​belong as the brain function level of the subject, it is possible to quickly and low-costly stratify the brain function level of a subject, thereby completing the present invention.

[0010] The present invention may include the following aspects. Item 1. A method for assisting in stratifying a subject's brain function level, the method comprising: obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from the subject; searching for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to brain function levels; and suggesting the brain function level corresponding to the reference range to which the measurement values ​​belong as the brain function level of the subject. Item 2. The method according to Item 1, wherein the diamine and / or polyamine includes an acetylated form. Item 3. The method according to Item 1, wherein the measurement value is obtained by an enzymatic method. Item 4. The method according to Item 1, wherein the measurement value is obtained by an enzyme-linked immunosorbent assay method, a sensor method, high performance liquid chromatography method, gas chromatography-mass spectrometry method, liquid chromatography-mass spectrometry method, or capillary electrophoresis-mass spectrometry method. Item 5. A method for supporting the recommendation of diamine and / or polyamine intake for a subject, the method comprising: acquiring a measurement value of diamine and / or polyamine in a urine or blood sample collected from the subject; searching for a reference range to which the measurement value belongs from a plurality of reference ranges set according to brain function levels; and presenting a recommended level of diamine and / or polyamine intake corresponding to the reference range to which the measurement value belongs; A method comprising: Item 6. A stratification device for assisting in stratifying the brain function level of a subject, the stratification device including a processing unit, which obtains measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from the subject, searches for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to the brain function level, and outputs a label of the brain function level corresponding to the reference range to which the measurement values ​​belong as a label indicating the brain function level of the subject. Item 7. The stratification device according to Item 6, further outputting a recommended level of diamine and / or polyamine intake corresponding to a reference range to which the measured value belongs. Item 8. A stratification program for assisting in stratifying the brain function level of a subject, which, when executed by a computer, causes the computer to execute the steps of: obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from a subject; searching for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to brain function levels; and outputting a label of the brain function level corresponding to the reference range to which the measurement values ​​belong as a label indicating the brain function level of the subject. Item 9. Further, the step of outputting a recommended level of diamine and / or polyamine intake corresponding to the reference range to which the measured value belongs, Item 8. The stratification program according to item 8. Item 10. A test kit for carrying out the method according to item 1 or the method according to item 5, the test kit comprising an oxidase that oxidizes diamines and / or polyamines, a peroxidase, and a color-developing reagent. Item 11. The test kit according to Item 10, further comprising acylpolyamine amidohydrolase. Item 12. A test kit for carrying out the method according to item 5, comprising an oxidase that oxidizes diamines and / or polyamines, a peroxidase, and a color-developing reagent. Effect of the Invention

[0011] According to the present invention, the brain function level of a subject can be quickly stratified at low cost, and the brain function of a subject can be improved. [Brief description of the drawings]

[0012] [Figure 1] 2 shows the hardware configuration of the stratification device 30. [Diagram 2] 13 is a flowchart showing the flow of processing of the stratification program 3042a. [Diagram 3] 2 shows the hardware configuration of a presentation device 30'. [Figure 4] 13 is a flowchart showing the flow of processing of a presentation program 3042b. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0013] 1. Methods to assist in stratifying brain function levels One embodiment of the present invention relates to a method for assisting in stratifying a subject's brain function level.

[0014] The method of the present invention includes obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from a subject, and searching for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to brain function levels.

[0015] Brain function decline often occurs with age, and can lead to a variety of symptoms, including learning disabilities, memory disorders, speech disorders, anxiety disorders, insomnia, and decreased motivation. A typical disease accompanied by a decline in brain function is dementia, but its precursor is mild cognitive impairment (MCI). MCI is intermediate between healthy individuals and dementia, and refers to a state in which cognitive function declines but does not interfere with daily life. The disease rate is 10-20% in people over 65 years old, and it has been reported that if left untreated, approximately 70% of these people will progress to dementia (Suzuki Yutaka, Nihon University Medical Journal (2012) 71(6):385-389).

[0016] For example, the diagnostic criteria for MCI, according to the evaluation criteria of Petersen et al. (Yamamoto Yasuji, Seishin Shinkeigaku Zasshi (2011) Vol. 113, No. 6), are as follows: 1. Complaint of forgetfulness by the patient or family member (caregiver); 2. Presence of memory impairment that cannot be explained solely by the effects of aging; 3. Independent ability to carry out daily activities; 4. General cognitive function is normal; 5. No evidence of dementia. Dementia treatment is more likely to be effective if started at an earlier stage, and early detection and treatment at the MCI stage can improve cognitive function and halt the progression to dementia. The above further emphasizes the importance of early detection and prevention at the pre-disease stage.

[0017] The US ADNI study established the following criteria for the MMSE neuropsychological test, which is used as a cognitive function test in clinical settings: 27-30 points: normal (does not rule out MCI), 24-26 points: borderline (may be normal, MCI, or have dementia), and 20-23 points: suspected early dementia.

[0018] The subject may include a person who is not suspected of brain function decline, a person who is suspected of brain function decline, and a person who has been determined to have brain function decline. In the evaluation criteria of Pertersen et al., a person who is determined to have MCI may correspond to a person who is suspected of brain function decline. In the evaluation criteria of Pertersen et al., a person who is determined to have dementia may correspond to a person who has been determined to have brain function decline. Furthermore, in the evaluation criteria of Pertersen et al., a person who is determined to be healthy or robust may correspond to a person who is not suspected of brain function decline. It is preferable that the subject is not a person who has been diagnosed with Alzheimer's disease.

[0019] Since the risk of brain function decline is higher in elderly people, the age of the subject is preferably 50 years or older, and more preferably 60 years or older, 70 years or older, or 80 years or older.

[0020] In this specification, a diamine is a straight-chain fatty acid hydrocarbon compound having two amines. A polyamine is a straight-chain fatty acid hydrocarbon compound having three or more amines. There are two types of diamines and two types of polyamines in the human body. In addition, diamines and polyamines may include acetylated forms such as monoacetylated and diacetylated forms. Specific examples of diamines present in the human body are putrescine and cadaverine. Examples of acetylated forms of diamines include acetylputrescine and acetylcadaverine. Specific examples of polyamines present in the human body are spermidine and spermine. Examples of acetylated polyamines include N, N-acetylated and N-acetylated polyamines. 1 -Acetylspermidine, N 8- Acetylspermidine, N 1 ,N 8 -Diacetylspermidine, acetylspermine, N 1 ,N 12 In general, diamines and polyamines are present mainly as acetylated polyamines in urine or blood samples. Therefore, when measuring diamines or polyamines using urine or blood samples, acetylputrescine, acetylcadaverine, N 1 -Acetylspermidine, N 8- Acetylspermidine, N 1 ,N 8 -Diacetylspermidine, acetylspermine, N 1 ,N 12 In addition, when measuring diamines or polyamines using urine or blood samples, it is preferable to target acetylputrescine, N 1 -Acetylspermidine, N 8- Acetylspermidine, N 1 ,N 8 -Diacetylspermidine, acetylspermine, N 1 ,N 12 -Diacetylspermine is preferred as a target.

[0021] The sample from which the diamine or polyamine measurements are taken is a urine or blood sample. The urine sample may be naturally voided urine or catheterized urine. The urine sample may be first early morning urine, random urine, or pooled urine. Preferably, the urine sample is naturally voided first early morning urine.

[0022] The measured values ​​of diamines or polyamines can be obtained according to a standard method. Here, the measurement value is not limited to being measured in the facility visited by the subject. It also includes sending the subject's urine sample to another facility, and obtaining the measured values ​​measured at the facility as data from another computer or the like via a network or a storage medium.

[0023] For example, methods for measuring diamines or polyamines include enzyme methods, enzyme-linked immunosorbent assay (ELISA), sensor methods, chromatography methods such as high performance liquid chromatography (HPLC), and mass spectrometry methods such as gas chromatography-mass spectrometry (GC-MS), liquid chromatography-mass spectrometry (LC-MS), and capillary electrophoresis-mass spectrometry (CE-MS). These methods are known (for example, literature: Kensa to Gijutsu, vol. 14, no. 3, March 1986).

[0024] In the enzymatic method, for example, an oxidase is applied to each diamine and / or polyamine to generate hydrogen peroxide, which is then reacted with a redox color-developing reagent (hereinafter, simply referred to as a "color-developing reagent") by the activity of peroxidase to obtain a color corresponding to the amount of diamine and / or polyamine in the urine sample. Examples of oxidases that oxidize diamines and / or polyamines include putrescine oxidase, diamine oxidase, amine oxidase, spermidine dehydrogenase, polyamine oxidase, and the like (reference: THE CHEMICAL TIMES, No. 3, 1985). As the putrescine oxidase, it is preferable to use the putrescine oxidase described in JP-B-2-60314 and JP-B-2-60315. As the diamine oxidase, an enzyme derived from pig kidney or pea seedlings can be used. As the polyamine oxidase, an enzyme derived from molds such as Penicillium chrysogenum and Aspergillus terreus, or from rat liver can be used. Here, the names of the enzymes are common names and do not indicate the substrate specificity of each enzyme. For example, putrescine oxidase can oxidize not only diamines putrescine and cadaverine, but also polyamines spermidine and spermine.

[0025] When the diamine or polyamine is an acetylated product, it is preferable to act an acyldiamine amidohydrolase or an acylpolyamine amidohydrolase on the acetylated product before acting on the diamine oxidase or polyamine oxidase to deacetylate the product, and then act on the product with an oxidase that oxidizes the diamine and / or polyamine. Deacetylation may be performed by acid treatment instead of using an acylamine hydrolase such as acyldiamine amidohydrolase or acylpolyamine amidohydrolase. The deacetylation method by acid treatment is known.

[0026] The peroxidase is not limited as long as it can catalyze the dye oxidation reaction of the chromogen by hydrogen peroxide. For example, any type of peroxidase may be used as long as it catalyzes the reaction between hydrogen peroxide and a redox color-developing reagent, and examples of such peroxidase include peroxidases derived from plants, bacteria, and basidiomycetes. Among these, horseradish, rice, and soybean-derived peroxidases are preferred, and horseradish-derived peroxidases are more preferred, for reasons of purity, availability, price, and the like. As commercially available products, PEO-131 (manufactured by Toyobo), PEO-301 (manufactured by Toyobo), PEO-302 (manufactured by Toyobo), and the like are preferably used. There are no particular limitations on the amount used, the form of addition, and the like.

[0027] The redox coloring reagent used in the biological component measuring method of the present invention may be any type of dye that reacts with hydrogen peroxide to produce a color, such as a combination of a hydrogen donor and a coupler. There are no particular limitations on the amount used or the form of addition. All of these are commercially available.

[0028] A representative example using a hydrogen donor and a coupler is the Trinder method, in which a hydrogen donor and a coupler are oxidatively condensed with hydrogen peroxide in the presence of peroxidase to form a dye.

[0029] The hydrogen donor is not particularly limited as long as it has the property of being able to donate hydrogen and can provide the effects of the present invention. In the biological component measurement method of the present invention, phenol, phenol derivatives, aniline derivatives, naphthol, naphthol derivatives, naphthylamine, naphthylamine derivatives, etc. are used as hydrogen donors for use in the Trinder method, etc. Such hydrogen donors are also called oxidation coloring reagents.

[0030] For example, N-ethyl-N-sulfopropyl-3-methoxyaniline, N-ethyl-N-sulfopropylaniline, N-ethyl-N-sulfopropyl-3,5-dimethoxyaniline, N-sulfopropyl-3,5-dimethoxyaniline, N-ethyl-N-sulfopropyl-3,5-dimethylaniline, N-ethyl-N-sulfopropyl-3-methylaniline, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline, N-ethyl-N-(2-hydroxy-3-sulfopropyl)aniline, N-ethyl-N-(2-hydroxy-3-sulfopropyl)aniline, N-ethyl-N-(2-hydroxy-3-sulfopropyl) 3,5-dimethoxyaniline, N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethylaniline, N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3-methoxyaniline, N-sulfopropylaniline, N-(2-hydroxy-3-sulfopropyl)-2,5-dimethylaniline, N-ethyl-N-(3-methylphenyl)-N'-succinylethylenediamine, and N-ethyl-N-(3-methylphenyl)-N'-acetylethylenediamine are examples thereof.

[0031] These hydrogen donors can be used in combination with couplers.

[0032] Couplers used in the present invention for measuring biological components include aminoantipyrine compounds such as 4-aminoantipyrine (4AA) and aminoantipyrine derivatives, vanillin diamine sulfonic acid compounds such as vanillin diamine sulfonic acid, and methylbenzthiazolinone hydrazone compounds such as methylbenzthiazolinone hydrazone (MBTH) and sulfonated methylbenzthiazolinone hydrazone (SMBTH). It is assumed that aminoantipyrine compounds may contain 4-hydroxyantipyrine as an impurity.

[0033] In addition to the above, examples of the enzymatic method include those described in JP-A-1-174400 and JP-A-1-281099, in which an oxidase is allowed to act on a diamine and / or a polyamine, and the generated aminoalkylaldehyde is further allowed to act on an aminoalkylaldehyde dehydrogenase in the presence of NAD+ or NADP+ to generate NADH or NADPH, which is then measured by absorbance.

[0034] Also, N 1 , N 12 -Diacetylspermine, N 1 , N 8 -Diacetylspermidine can also be detected by the ELISA method. Kits for this purpose are commercially available. In the ELISA method, a color is produced according to the amount of diamine or polyamine in the urine or blood sample by reacting an enzyme labeled with a detection antibody with a color-developing substrate.

[0035] The color intensity obtained by the enzyme method or the ELISA method may itself be used as a measurement value of the diamine or polyamine. Alternatively, the color intensity may be applied to a calibration curve prepared for each diamine or polyamine, converted into the amount of the diamine or polyamine, and this value may be used as the measurement value of the diamine or polyamine. The calibration curve is prepared from the color intensity of a standard sample in which the type and content of each diamine or polyamine are known, and the amount of each diamine or polyamine contained in the standard sample. The amount of the diamine or polyamine may be expressed as mass or weight per unit volume, or may be expressed as the number of moles of the diamine or polyamine per unit volume. In the enzyme method or ELISA method, a luminescent substrate may be used instead of a chromogenic substrate.

[0036] As a sensor method for detecting the amount of diamine or polyamine using a sensor, for example, an extended gate-type organic thin-film transistor (OTFT)-based polyamine sensor described in the literature J. Mater. Chem. C, 2021, 9, 11690-11697 (DOI: 10.1039 / D1TC01542G) can be used. In the sensor method, the value of diamine or polyamine is calculated based on the voltage value.

[0037] When the amount of each diamine or polyamine is quantified by chromatography or mass spectrometry, a spectrum of the diamine or polyamine is obtained from a urine or blood sample and a standard sample. The standard sample is a sample in which the content of each diamine or polyamine is known. Next, the peak area of ​​each diamine or polyamine in the urine or blood sample and the peak area of ​​each diamine or polyamine contained in the standard sample are obtained from the spectrum. The peak area of ​​the diamine or polyamine in the standard sample reflects the amount of each diamine or polyamine contained in the standard sample, so the amount of each diamine or polyamine in the urine or blood sample can be calculated from the peak area of ​​each diamine and / or polyamine in the urine or blood sample and the peak area of ​​the standard sample containing the same diamine or polyamine as each diamine or polyamine. In this case, the measured value of the diamine or polyamine may be a peak area, and the peak area may be applied to a calibration curve prepared for each polyamine to convert it into the amount of the diamine or polyamine, and this value may be the measured value of the polyamine. The calibration curve is prepared from the peak areas of standard samples in which the type and content of each diamine or polyamine is known, and the amount of each diamine or polyamine contained in the standard samples.

[0038] Next, a reference range to which the measured value of diamine or polyamine in the urine or blood sample belongs is searched. Searching for a reference range to which the measured value belongs is intended to compare the measured value of the subject with each of a plurality of reference ranges previously set according to the brain function level. For example, when the brain function level is classified based on the score of the MMSE test (J Psychiatr Res. 1975 Nov;12(3):189-98. doi: 10.1016 / 0022-3956(75)90026-6.), the reference range is set for three categories: healthy, borderline, and suspected early dementia. The reference range is set based on the measured value of diamine or polyamine in the urine or blood sample collected from subjects belonging to each of the subjects classified as healthy, the subjects classified as borderline, and the subjects classified as suspected early dementia. The reference range can be set based on the group of measured values ​​obtained in each category, for example, from a hazard ratio, a confidence interval, an odds ratio, etc. Alternatively, the reference range may be determined from the median and interquartile range calculated from the group of measurement values ​​in each category, or the reference range may be set from the mean, standard deviation (SD), variance (CV), etc. calculated from the group of measurement values ​​in each category.

[0039] If the search results in a determination of a reference range to which the measured value falls, it is suggested that the brain function level corresponding to the reference range to which the measured value falls is the brain function level of the subject.

[0040] The diamine or polyamine measurement values ​​are preferably corrected based on the creatinine concentration in the urine sample of the subject at the time the diamine or polyamine measurement values ​​were collected, and the 24-hour urinary creatinine excretion amount. The diamine or polyamine measurement values ​​may also be corrected based on the 24-hour urinary creatinine excretion amount of 1 g per day. When correction is performed, the reference range is also preferably determined based on the corrected diamine and / or polyamine measurement values. The creatinine concentration in the urine sample can be measured by the Jaffe method, the enzymatic method, etc.

[0041] 2. Methods for supporting the presentation of diamine and / or polyamine intake Certain embodiments of the present invention relate to methods for assisting in the presentation of diamine and / or polyamine intake in a subject.

[0042] The method includes obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from a subject, searching for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to brain function levels, and presenting a recommended intake level of diamines and / or polyamines corresponding to the reference range to which the measurement values ​​belong.

[0043] The explanation of obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from a subject and searching for a reference range to which the measurement values ​​belong from among a plurality of reference ranges set according to brain function levels is incorporated herein by reference to the explanation of 1 above.

[0044] In this embodiment, further, a recommended level of diamine and / or polyamine intake, which is preset in correspondence with the plurality of reference ranges, is presented. For example, when the brain function level is classified based on the score of the MMSE test, the recommended level is set for three categories: healthy, borderline, and suspected early dementia. When the measured value falls within the healthy reference range, for example, a label indicating that "intake of diamine and / or polyamine is recommended" is presented as the recommended level. When the measured value falls within the borderline reference range, for example, a label indicating that "intake of diamine and / or polyamine is strongly recommended" is presented as the recommended level. When the measured value falls within the reference range of suspected early dementia, for example, a label indicating that "intake of diamine and / or polyamine is very strongly recommended" is presented as the recommended level. These labels may be text, symbols, or numerical values. In addition, the daily intake of diamine and / or polyamine may be presented more specifically according to the reference range.

[0045] Another embodiment of the present invention relates to a method for suggesting that the intake of diamines and / or polyamines is effective for subjects who are not suspected of brain function decline, subjects who are suspected of brain function decline, or subjects who have been determined to have brain function decline. The method includes obtaining a measurement value of diamines and / or polyamines in a urine sample collected from a subject who is not suspected of brain function decline, subjects who are suspected of brain function decline, or subjects who have been determined to have brain function decline, and suggesting that the intake of diamines and / or polyamines is effective when the measurement value is below a reference value. For example, the control subject is preferably 50 years old or older, 60 years old or older, 70 years old or older, or 80 years old or older. The subject who is suspected of brain function decline, or the subject who has been determined to have brain function decline, is a subject who is suspected of brain function decline, or a subject who has been determined to have brain function decline, based on the above-mentioned evaluation criteria.

[0046] As the reference value of the measured value of diamine and / or polyamine, a value capable of distinguishing between the high value group and the low value group of the measured value can be selected. The reference value can be calculated from the group of measured values ​​of diamine and / or polyamine in the urine sample of the control. For example, it can be set from the hazard ratio, confidence interval, odds ratio, etc. calculated from the group of measured values ​​of diamine and / or polyamine in the urine sample of the control. Alternatively, the reference range may be determined from the median and interquartile range calculated from the group of measured values ​​of diamine and / or polyamine in the urine sample of the control. Alternatively, it can be determined from the average value, standard deviation (SD), variance (CV), etc. calculated from the group of measured values ​​of each category. The control is a person who falls into an age group that may cause brain function decline. For example, the control is preferably 50 years old or older, 60 years old or older, 70 years old or older, or 80 years old or older.

[0047] 3. Stratification device to assist in stratifying brain function levels An embodiment of the present invention relates to a stratification device 30 (hereinafter, also referred to simply as "stratification device 30") for assisting in stratifying the brain function level of a subject. The hardware configuration of the stratification device 30 will be described with reference to FIG.

[0048] The stratification device 30 may be connected to an input device 311, an output device 312, and a media drive 313. The stratification device 30 may be communicably connected to an analysis device 90 via a wired or wireless network to constitute a stratification system 3000 for stratifying the progress of brain function levels. The analysis device 90 may be an absorbance meter, a microplate reader, a mass spectrometer, or the like.

[0049] In the stratification device 30, a processing unit 301, a memory 302, a ROM (read only memory) 303, a storage device 304, a communication interface (I / F) 305, an input interface (I / F) 306, an output interface (I / F) 307, and a media interface (I / F) 308 are connected to each other via a bus 309 so as to be able to communicate data with each other. The memory 302 and the storage device 304 may be collectively referred to simply as a storage unit. The storage unit stores the measurement values ​​and reference values ​​in a volatile or non-volatile manner.

[0050] The processing unit 301 is a CPU of the stratification device 30, and is also called a calculation device. The processing unit 301 may cooperate with a GPU. The processing unit 301 cooperates with an operating system (OS) 3041 stored in the storage device 304 or the ROM 303 to execute a disease information stratification program 3042a (hereinafter, sometimes simply referred to as the "stratification program 3042a") described later, and processes the acquired data, whereby the computer functions as the stratification device 30.

[0051] The ROM 303 stores a stratification program 3042a executed by the processing unit 301 and data used therein. The processing unit 301 may be the MPU 101. The ROM 303 stores a boot program executed by the processing unit 301 when the stratification device 30 is started up, and programs and settings related to the operation of the hardware of the stratification device 30.

[0052] The memory 302 is used to read out the stratification programs 3042a recorded in the ROM 303 and the storage device 304. The memory 302 is also used as a working area when the processing unit 301 executes the stratification programs 3042a.

[0053] The storage device 304 is configured with a hard disk, etc. The storage device 304 stores various stratification programs 3042a, such as an operating system and application programs, to be executed by the processing unit 301, and various setting data used for executing the stratification programs 3042a. Specifically, the storage device 304 stores a reference range database (DB) DB2 in a non-volatile manner.

[0054] The communication I / F 305, under the control of the processing unit 301, receives data from the analysis device 90 or other external devices, and transmits or displays information stored or generated by the stratification device 30 to the analysis device 90 or externally as necessary. The communication I / F 305 may communicate with the analysis device 90 or other external devices via a network.

[0055] The input I / F 306 accepts character input, clicks, voice input, and the like from the input device 311. The accepted input contents are stored in the memory 302 or the storage device 304.

[0056] The input device 311 is composed of a touch panel, a keyboard, a mouse, a pen tablet, a microphone, etc., and is used to input characters or voice to the stratification device 30. The input device 311 may be connected to the stratification device 30 from outside, or may be integrated with the stratification device 30.

[0057] The output I / F 307 outputs information generated by the processing unit 301 to the output device 312. The output I / F 307 outputs information generated by the processing unit 301 and stored in the storage device 304 to the output device 312.

[0058] The output device 312 is composed of, for example, a display, a printer, etc., and displays the measurement results transmitted from the analysis device 90, various operation windows in the stratification device 30, analysis results, etc.

[0059] The media I / F 308 reads, for example, application software etc. stored in the media drive 313. The read application software etc. is stored in the memory 302 or the storage device 304. The media I / F 308 also writes information generated by the processing unit 301 to the media drive 313. The media I / F 308 writes information generated by the processing unit 301 and stored in the storage device 304 to the media drive 313.

[0060] The media drive 313 is configured with a flexible disk, a CD-ROM, a DVD-ROM, etc. The media drive 313 is connected to the media I / F 308 by a flexible disk drive, a CD-ROM drive, a DVD-ROM drive, etc. The media drive 313 may store application programs and the like for the computer to execute operations.

[0061] The processing unit 301 may acquire application software and various settings necessary for controlling the stratification device 30 via a network instead of reading them from the ROM 303 or the storage device 304. It is also possible that the application program is stored in a storage device of a server computer on the network, and the stratification device 30 accesses this server computer to download the stratification program 3042a and store it in the ROM 303 or the storage device 304.

[0062] An operating system that provides a graphical user interface environment, such as Windows (registered trademark) manufactured and sold by Microsoft Corporation, is installed in the ROM 303 or the storage device 304. The application program according to the second embodiment runs on the operating system. That is, the stratification device 30 may be a personal computer or the like.

[0063] The stratification system 3000 does not need to be installed in one place, and the stratification device 30 and the analysis device 90 may be located in different places and connected by a network. In addition, the stratification device 30 may be a device that does not require an operator by omitting the input device 311 and the output device 312.

[0064] 4. Processing stratification program to assist in stratifying brain function levels The process executed by the stratification program 3042a for supporting stratification of the brain function level of a subject will be described with reference to FIG.

[0065] When an operator inputs a command to start the process from the input device 311, the processing unit 301 of the stratification device 30 starts the stratification process for supporting stratification of brain function levels.

[0066] In step S31, processing unit 301 obtains the measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from a subject from analytical device 90. The details of obtaining the measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from a subject are the same as those described in 1 above, and therefore the description of 1 above is incorporated herein by reference.

[0067] Next, in step S32, the processing unit 301 searches for a reference range to which the measurement value belongs from a plurality of reference ranges set according to brain function levels. The plurality of reference ranges are stored in a reference range database DB2 in association with a label indicating the brain function level.

[0068] Next, in step S33, the processing unit 301 outputs a label associated with the brain function level corresponding to the reference range to which the measurement value belongs as a label indicating the brain function level. The label indicating the brain function level may be text in Japanese, English, or the like, such as "healthy", "borderline", "suspected early dementia", etc. Furthermore, the label associated with the brain function level and the label to be output do not necessarily have to be the same, as long as they have the same clinical meaning. Furthermore, if the measurement value belongs to the reference range of "healthy", step S33 may not be performed and the process may end.

[0069] Furthermore, in step S34, the processing unit 301 outputs the recommended intake levels of diamines and / or polyamines corresponding to the reference ranges to which the measured values ​​found in step S32 belong. The recommended intake levels of diamines and / or polyamines are linked to the respective reference ranges and stored in the reference range database DB2. The explanation of the recommended intake levels of diamines and / or polyamines is incorporated herein by reference in the explanation of 2 above.

[0070] Furthermore, when executing step S34, the processing unit 301 does not need to execute step S33.

[0071] Another form of this embodiment relates to a presentation device (hereinafter also referred to simply as the presentation device) 30' that suggests that intake of diamines and / or polyamines is effective for a subject who is not suspected of brain function decline, a subject who is suspected of brain function decline, or a subject who has been determined to have brain function decline.

[0072] The presentation device 30' may be connected to an input device 311, an output device 312, and a media drive 313. The stratification device 30 may be communicably connected to the presentation device 30' via a wired or wireless network to configure a presentation system 3000' that suggests that ingestion of diamines and / or polyamines is effective for a subject who is not suspected of brain function decline, a subject who is suspected of brain function decline, or a subject who has been determined to have brain function decline. The analysis device 90 may be an absorbance meter, a microplate reader, a mass spectrometer, or the like.

[0073] FIG. 3 shows the hardware configuration of the presentation device 30'. The configuration of the presentation device 30' is the same as that of the stratification device 30, except for the following points. The storage device 304 of the presentation device 30' stores a presentation program (hereinafter simply referred to as the "presentation program") 3042b, instead of the stratification program 3042a, that suggests that the intake of diamines and / or polyamines is effective for subjects who are not suspected of brain function decline, subjects who are suspected of brain function decline, or subjects who have been determined to have brain function decline. In addition, the reference range database DB2 stores reference values ​​calculated from a group of measured values ​​of diamines and / or polyamines in urine samples of control subjects.

[0074] The process executed by the presentation program 3042b will be described with reference to FIG. When an operator inputs a command to start the process from the input device 311, the processing unit 301 of the presentation device 30' starts a stratification process for supporting stratification of brain function levels.

[0075] In step S331, processing unit 301 obtains the measurement values ​​of diamines and / or polyamines in a urine sample collected from a subject from analytical device 90. The details of obtaining the measurement values ​​of diamines and / or polyamines in a urine sample collected from a subject are the same as those described in 1 above, and therefore the description of 1 above is incorporated herein by reference.

[0076] Next, in step S332, the processing unit 301 compares the measured value acquired in step S331 with the reference values ​​of diamines and / or polyamines. The reference values ​​are stored in the reference range database DB2.

[0077] In step S332, when the measured value is equal to or lower than the reference value (when "YES"), the processing unit 301 proceeds to step S333 and outputs a label recommending the subject to take diamines and / or polyamines. In addition, in step S332, when the measured value exceeds the reference value (when "NO"), the processing unit 301 ends the processing. In step S332, when the measured value exceeds the reference value (when "NO"), the processing unit 301 may output a label indicating that the subject does not need to take diamines and / or polyamines.

[0078] 5. Storage medium storing the program An embodiment of the present invention relates to a program product, such as a media drive, that stores the stratification program 3042a. That is, the stratification program 3042a can be stored in a media drive, such as a hard disk, a semiconductor memory device such as a flash memory, or an optical disk. The media drive may also be a computer, such as a server device. The format of recording the program in the media drive is not limited as long as each device can read the program. It is preferable that the recording in the media drive is non-volatile.

[0079] 6. Test kits to help stratify brain function levels An embodiment of the present invention relates to a test kit for assisting in stratifying the brain function level of a subject or for assisting in indicating the intake of diamines and / or polyamines. The test kit is not limited as long as it can obtain the measurement value of diamines and / or polyamines in urine or blood samples.

[0080] For example, when detecting diamines and / or polyamines by an enzymatic method, the test kit may contain polyamine oxidase, peroxidase, a buffer satisfying the above conditions for each enzyme to be suitably active, and a substrate for the peroxidase. If necessary, the test kit may contain acylpolyamine amidohydrolase, acylpolyamine amidohydrolase, and a buffer satisfying the above conditions for the enzyme to be suitably active. Regarding each enzyme and substrate, the explanation in 1. above is incorporated herein by reference. EXAMPLES

[0081] The present invention will be described in more detail below with reference to examples, but the present invention should not be construed as being limited to these examples.

[0082] 1. Subject of study In this study, subjects who met all of the following conditions were selected as subjects, and the effect of improving cognitive function when polyamine-containing capsules (hereinafter referred to as "the test food") were continuously taken for 12 weeks was evaluated. - The purpose and contents of this study have been fully explained, and you have the ability to consent Those who fully understand the information, voluntarily apply, and give written consent to participate Healthy Japanese subjects aged 40 years or older at the time of obtaining informed consent Those who are aware of forgetfulness MMSE score of 24 or higher at screening

[0083] 2. Test Method (1) Study design a. Randomization: Randomized comparison b. Blinding: Double-blind c. Control: Placebo control d. Allocation: Parallel group comparison In this study, 60 subjects were assigned to each of the following study groups, 30 subjects each. Group I: Subjects took 9 tablets of test food A per day for 12 weeks, 3 tablets each after breakfast, lunch, and dinner. Group II: Subjects took nine tablets of test food B daily for 12 weeks, three tablets each after breakfast, lunch, and dinner.

[0084] (2) Overview of the foods used in this study Test food Test foods A and B were either polyamine-containing capsules (2700 mg, 9 tablets, containing 1.1 mg of polyamine as an active ingredient) or placebo (2700 mg, 9 tablets). Each test food was mixed with excipients and placed in a dark-colored capsule so that the contents of the capsule could not be seen from the outside. b. Ingredients / raw material display Polyamine-containing capsule: Wheat germ extract 409 mg, starch hydrolysate 372 mg, sodium citrate 149 mg, dextrin 1734 mg, calcium stearate 36 mg Placebo: sodium citrate 149 mg, dextrin 2515 mg, calcium stearate 36 mg

[0085] (3) Evaluation items The evaluation items were as follows: a.Primary endpoint Cognitive function test (Cognitrax) b. Secondary endpoint ·Mini-Mental Status Examination (MMSE) - Visual Analogue Scale (VAS) questionnaire for cognitive function Urinary polyamines Blood polyamines c. Safety evaluation items Subjective findings (daily diary) Objective findings (questionnaire)

[0086] (4) Allocation method This study was conducted using a randomized comparative method to ensure that differences in medical background did not occur between the study groups due to allocation. The allocation manager assigned 30 participants to groups I and II, 30 in each group, based on the MMSE scores and median ages at screening.

[0087] (5) Test flow and schedule, and sample and information acquisition method a. Sample (information) and acquisition method On each test day, information was collected using the following methods. Consent of study subjects: Name of consenter, date of consent Background of test subjects: age (date of birth), gender, etc. Questionnaire at the time of the test (exercise and meals the day before, physical condition and meals on the day, etc.) Physical examination: height (only at pre-intake test), weight, BMI Urine test: Urinary polyamine content Blood test: Blood polyamine levels Daily diary (including daily test food intake and physical condition during the study period) b. Schedule The above tests were performed according to the following schedule.

[0088] [Table 1] During the study, subjects were advised to pay attention to the following points: Do not make any major changes to your lifestyle, such as your diet or exercise habits. Refrain from taking any new medicines or health foods (foods with specified health uses, foods with functional claims, and foods with nutrient functions) that may affect your cognitive function. During the study period, participants will refrain from taking polyamine-containing supplements other than the test foods, natto (one or more packs per day), and soy milk (one or more packs (approximately 200 mL) per day). The day before the test, avoid strenuous exercise and finish eating by 9 p.m. Also, avoid overeating, drinking a lot of alcohol, smoking, and staying up late. - On the day of the test, avoid excessive exercise until the test is completed. - If you have an acute illness such as fever, diarrhea, or vomiting on the day of the test, the test will be cancelled. Do not disclose any information you learn about this test to a third party. In particular, do not provide any information via social media (Facebook, Twitter, Instagram, LINE, etc.).

[0089] (6) Exclusion criteria Subjects who met the following criteria were excluded from the study. 1) Those who have a chronic illness and are receiving drug treatment, or those with a history of serious illness 2) Those suffering from cancer, familial adenomatous polyposis, or Helicobacter pylori infection 3) People who have allergies to the test food (especially people who have allergies to wheat, as the test food contains wheat germ extract) 4) People who regularly take large amounts of polyamine-containing supplements 5) People who consume one or more packs of natto every day 6) Those who consume more than one pack (approximately 200 mL) of soy milk every day 7) Those who regularly take medicines or functional foods (foods for specified health uses, foods with functional claims, and foods with nutrient functions) that may affect cognitive function 8)Those who have participated in other studies within one month prior to the start of this study, or who plan to participate in other studies after consenting to this study 9) Those who are judged ineligible by the study director and study representative 10) Those who are breastfeeding, pregnant, or planning or hoping to become pregnant during the study period

[0090] (7) Statistical analysis method As a general rule, the Shapiro-Wilk test was used to examine the normality of the obtained evaluation results in the case of continuous data, and paired t-test was used if normality was indicated, and Wilcoxon signed-rank test was used if it was not. For unpaired data, variance was tested using F-test if normality was indicated, followed by Student's t-test (with homogeneity of variance) or Aspin-Welch's t-test (without homogeneity of variance), and if normality was not indicated, Mann-Whitney U-test was used. For ordinal data with many stages, Wilcoxon signed-rank test was used for paired data, and Mann-Whitney U-test was used for unpaired data. For ordinal data and nominal data with few stages, McNemar test was used for paired data, and Fisher's exact test or χ2 test was used for unpaired data depending on the number of data. 2 Tests were performed. If multiple comparisons were required, the Bonferroni method, Tukey method, Dunnett method, etc. were used depending on the format of the data. Analysis of variance, correlation coefficient tests, and subgroup analyses were also performed as necessary. Regarding safety evaluation items, in principle, only intra-group comparisons of values ​​before and after intake and between-group comparisons of values ​​before and after intake were performed, and between-group comparisons of differences before and after intake were not performed. In a two-sided test, a significance level of less than 5% was judged to be "statistically significant difference present," and a level between 5% and less than 10% was judged to be "tendency present." Statistical analysis was mainly performed using R, but IBM SPSS Statistics (ver. 25) was also used when necessary.

[0091] 3. Test Results 3-1.Efficacy evaluation (primary endpoint) Of the 60 study participants, 11 dropped out for personal reasons and 1 person was found to have deviated from compliance with the study protocol, and the remaining 48 participants were included in the efficacy analysis. Table 2 shows the background of the subjects in the efficacy analysis.

[0092] [Table 2]

[0093] The analysis results of the cognitive function tests are shown in Table 3. The polyamine group showed significant improvements in overall memory, cognitive function speed, simple attention, and motor speed compared to the placebo group. Additionally, the polyamine group showed a significant tendency toward improvement in verbal memory and visual memory compared to the placebo group.

[0094] [Table 3]

[0095] 3-2.Efficacy evaluation (secondary endpoint) As with the primary endpoint, of the 60 study participants, 11 dropped out for personal reasons and 1 participant was observed to have deviated from compliance with the study protocol, and the remaining 48 participants were included in the efficacy analysis.

[0096] The analysis results of the VAS questionnaire are shown in Table 4. Regarding questions about cognitive function speed, the polyamine intake group showed a significant improvement compared to the placebo intake group.

[0097] [Table 4]

[0098] 3-3. Safety evaluation items The results of the safety evaluation analysis are shown in Table 5. Significant decreases in weight and BMI were observed in both the polyamine intake group and the placebo intake group, but no significant differences were observed when comparing the groups.

[0099] [Table 5]

Claims

1. A stratification device for assisting in stratifying the brain function level of a subject, the stratification device including a processing unit, which obtains measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from the subject, searches for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to the brain function level, and outputs a label of the brain function level corresponding to the reference range to which the measurement values ​​belong as a label indicating the brain function level of the subject.

2. The stratification device according to claim 1 , further comprising: outputting a recommended level of diamine and / or polyamine intake corresponding to a reference range to which the measured value belongs.

3. A stratification program for assisting in stratifying the brain function level of a subject, which, when executed by a computer, causes the computer to execute the steps of: obtaining measurement values ​​of diamines and / or polyamines in a urine or blood sample collected from the subject; searching for a reference range to which the measurement values ​​belong from a plurality of reference ranges set according to brain function levels; and outputting a label of the brain function level corresponding to the reference range to which the measurement values ​​belong as a label indicating the brain function level of the subject.

4. and outputting a recommended level of diamine and / or polyamine intake corresponding to the reference range to which the measured value belongs. The stratification program according to claim 3.

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