Renal disease evaluation assistance method, renal disease evaluation system, and renal disease evaluation program

Abstract

To provide renal disease markets for accurately evaluating and determining a wide range of renal disease in subjects.SOLUTION: A method of assisting in evaluation of renal disease is provided, the method involving the use of a combination of a ratio of reabsorption and excretion of D-serine and / or D-asparagine in the kidney of a subject and the blood D-serine level and / or D-asparagine level as an indicator.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to a method for assisting in the evaluation of a renal pathology, a system for evaluating a renal pathology, and a program for evaluating a renal pathology. [Background technology]

[0002] The kidney is an important organ that maintains homeostasis of the biological environment by excreting and absorbing components in the body, and is responsible for the excretion of waste products, regulating blood pressure, and regulating the volume of body fluids and ions, as well as the production of blood and bones. A representative index of renal function is the glomerular filtration rate (GFR). Glomerular filtration rate indicates the amount of fluid filtered from blood by the glomerulus per minute, and measurement of inulin clearance is considered to be the international standard (gold standard). However, measurement of inulin clearance requires continuous infusion of inulin for two hours and multiple urine and blood collections, which places a heavy burden on the subject and the person performing the measurement. Therefore, in daily clinical practice, measurement of inulin clearance is limited to limited situations, such as donors in living donor kidney transplants, and is replaced by measurement of other markers such as creatinine. In addition, inulin clearance is difficult to apply in cases where renal pathology changes in a short period of time, such as acute kidney injury. The values ​​of many markers are significantly different from the actual glomerular filtration rate, such as inulin clearance, which is the gold standard, and this is an obstacle to accurate diagnosis of kidney disease.

[0003] Creatinine is commonly measured in clinical settings as an indicator of renal function. Creatinine is the final metabolite of creatine, which is necessary for muscle contraction. Creatine produced in the liver is taken up by muscle cells, where a portion is metabolized to become creatinine, which is then transported to the kidneys via the blood, filtered by the glomerulus, and excreted in the urine without being reabsorbed by the renal tubules. When glomerular filtration capacity is reduced, excretion is impaired, and the creatinine remains in the blood and increases in value, making it a useful indicator of uremic toxin accumulation, and therefore it is used to evaluate renal function. However, the amount of creatinine in the blood does not show clearly abnormal values ​​unless the GFR is reduced by 50% or more, and it cannot be said to be a sensitive marker.

[0004] Cystatin C is a protein with a molecular weight of 13.36 kDa that is produced at a constant rate from nucleated cells throughout the body, and since it is all filtered in the glomerulus, reabsorbed in the renal tubules, and then broken down in the kidney, it is thought to be removed from the blood according to the amount of filtration, and the amount in the blood is an indicator of GFR. However, when renal function is severely impaired, the increase in the amount of cystatin C in the blood slows down, making it difficult to accurately evaluate renal function in end-stage renal disease.

[0005] As described above, there were no biomarkers that could adequately meet the clinical demand for accurate measurement of individual patients' renal pathology over a wide range from early to end stages using only blood or samples that can be collected non-invasively, without placing a heavy burden on subjects and patients.

[0006] D-amino acids, which were previously thought not to exist in the body of mammals, are now known to exist in various tissues and play a physiological role. It has been shown that the amounts of D-serine, D-alanine, D-proline, D-glutamic acid, and D-aspartic acid in the blood fluctuate in patients with renal failure and correlate with creatinine, and thus can be markers for renal failure (Non-Patent Documents 1, 2, 3, and 4). Furthermore, it has been disclosed that amino acids selected from the group consisting of D-serine, D-threonine, D-alanine, D-asparagine, D-allothreonine, D-glutamine, D-proline, and D-phenylalanine are used as pathological indicators for kidney disease (Patent Document 1). It has also been disclosed that urinary D-serine, D-histidine, D-asparagine, D-arginine, D-allo-threonine, D-glutamic acid, D-alanine, D-proline, D-valine, D-allo-isoleucine, D-phenylalanine, and D-lysine change sharply due to renal impairment, and that parameters related to these amino acids are used as pathological index values ​​for kidney disease (Patent Document 2). In recent years, urinary LFABP, blood NGAL, urinary KIM-1, and the like have been developed as markers for kidney disease, but they are not related to glomerular filtration capacity. [Prior art documents] [Patent documents]

[0007] [Patent Document 1] International Publication No. 2013 / 140785 [Patent Document 2] Patent No. 5740523 [Non-patent literature]

[0008] [Non-Patent Document 1] Fukushima, T. et al., Biol. Pharm. Bull. 18: 1130(1995) [Non-Patent Document 2] Nagata.Y Viva Origino Vol.18(No.2) (1990) Abstracts of the 15th Academic Conference [Non-Patent Document 3] Ishida et al., Kitasato Medicine 23:51-62 (1993) [Non-Patent Document 4] Yong Huang et al. Biol. Pharm. Bull. 21:(2)156-162(1998) Summary of the Invention [Problem to be solved by the invention]

[0009] There is a need for a method to evaluate and determine a subject's renal pathology more broadly and accurately than the kidney disease markers currently known. [Means for solving the problem]

[0010] The present inventors focused on the dynamics of filtration, reabsorption, and excretion of D-serine and D-asparagine in the kidney and analyzed the relationship between their excretion rates and renal pathology. They found that this provided new pathological information useful for the evaluation and assessment of renal pathology, and thus completed the present invention.

[0011] Thus, the present invention relates to the following: [1] A method for assisting in the evaluation of renal pathology using a combination of the reabsorption and excretion rate of D-serine and / or D-asparagine in the kidney of a subject and the amount of D-serine and / or D-asparagine in the blood as an index. [2] The method according to item 1, wherein the rate is the excretion rate of D-serine into the urine of the subject (subject D-serine excretion rate) and / or the excretion rate of D-asparagine into the urine of the subject (subject D-asparagine excretion rate). [3] The method according to item 2, wherein the excretion rate of D-serine and / or the excretion rate of D-asparagine are calculated by correcting the excretion rate of D-serine and / or the excretion rate of D-asparagine using a correction factor derived from blood and / or urine. [4] The method according to item 3, wherein the correction factor is one or more correction factors selected from the group consisting of glomerular filtration rate and urine volume. [5] The method according to item 3, wherein the correction factor is one or more correction factors selected from the group consisting of inulin clearance and creatinine clearance. [6] The method according to item 3, wherein the correction factor is one or more correction factors selected from the group consisting of creatinine amount and L-amino acid amount. [7] The method according to item 3, wherein the correction factor is L-serine and / or L-asparagine. [8] The excretion rate of D-serine is calculated based on the following formula:

number

number

[10] The method according to item 9, wherein the step of evaluating the renal pathology is to evaluate a renal disease or a risk of developing renal disease in the object, or to predict the induction or prognosis of a renal disease, when the first object coordinates are not included in the first standard.

[11] The method according to item 10, wherein the kidney disease is chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis, polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease, or minimal change nephrotic syndrome.

[12] The method according to any one of items 9 to 11, wherein the first criterion is a range of the mean value of the plot of the non-renal disease coordinates ± standard deviation × coefficient Z.

[13] The method according to item 12, wherein the coefficient Z is a value between 1.0 and 3.0.

[14] The method according to item 12 or 13, wherein the coefficient Z is 1.96.

[15] A method for assisting evaluation of kidney pathology based on the relationship between a regression equation calculated by regression analysis of the plot of the non-renal disease coordinates and a target coordinate.

[16] The method according to any one of items 2 to 8, comprising a step of comparing a second object coordinate obtained by plotting the logarithmically transformed D-serine excretion rate (subject D-serine LN excretion rate) and / or the logarithmically transformed D-asparagine excretion rate (subject D-asparagine LN excretion rate) of the subject and the logarithmically transformed amount of D-serine and / or D-asparagine in blood with a second standard calculated from non-renal disease coordinates obtained by plotting the logarithmically transformed urinary D-serine excretion rate (non-renal disease subject D-serine LN excretion rate) and / or D-asparagine excretion rate (non-renal disease subject D-asparagine LN excretion rate) of a plurality of non-renal disease subjects and the logarithmically transformed amount of D-serine and / or D-asparagine in blood, and evaluating a renal pathological condition based on the relationship between the second object coordinate and the second standard.

[17] The method according to item 16, wherein the step of evaluating the renal pathology is to evaluate the renal disease or the risk of developing renal disease in the subject, or to predict the induction or prognosis of renal disease, when the second object coordinate is not included in the second criterion.

[18] The method according to item 17, wherein the renal disease is chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis, polycystic kidney disease, or renal disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease, or minimal change nephrotic syndrome.

[19] The method according to any one of items 16 to 18, wherein the second criterion is the range of the mean value of the plot of the non-renal disease coordinates ± standard deviation × coefficient Z.

[20] The method according to item 19, wherein the coefficient Z is a value between 1.0 and 3.0.

[21] The method according to item 19 or 20, wherein the coefficient Z is 1.96.

[22] The method according to Item 16, wherein the second criterion is that the distance from the average value of the plot of the non-renal disease coordinates is 0.6 or less.

[23] A method for assisting evaluation of renal pathology from the relationship between a regression equation calculated from a regression line of the plot of the non-renal disease coordinates based on the logarithmic transformed values ​​and a target coordinate based on the logarithmic transformed values.

[24] A method for monitoring renal pathology, comprising measuring the excretion rate of D-serine (subject D-serine excretion rate) and / or the excretion rate of D-asparagine (subject D-asparagine excretion rate) into the urine of a subject and the amount of D-serine and / or D-asparagine in the blood over time, and using the fluctuations in the target D-serine excretion rate and / or the target D-asparagine excretion rate and the amount of D-serine and / or D-asparagine in the blood as indicators.

[25] The method according to item 24, for monitoring renal pathology due to chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis, or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease, or minimal change nephrotic syndrome.

[26] Treatment of a subject with kidney disease. A method for monitoring the therapeutic effect of a renal pathology, comprising measuring the urinary excretion rate of D-serine (target D-serine excretion rate) and / or the excretion rate of D-asparagine (target D-asparagine excretion rate) before and after an intervention and the amount of D-serine and / or D-asparagine in blood over time, and using the fluctuations in the target D-serine excretion rate and / or the target D-asparagine excretion rate and the amount of D-serine and / or D-asparagine in blood as indicators.

[27] The method according to item 26, wherein the renal disease is chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or a renal disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease, or minimal change nephrotic syndrome.

[28] A method for assisting in the evaluation of renal pathology using the amount of D-serine and / or D-asparagine in the blood of a subject from whom urine cannot be collected as an index.

[29] The method according to item 28, which assists in the evaluation of renal pathology due to chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease or minimal change nephrotic syndrome.

[30] A method for assisting in the determination of systemic lupus erythematosus when the amount of D-serine in the blood of a subject is 9 nmol / mL or more.

[31] A system for evaluating a renal pathology comprising a memory unit, an input unit, an analysis and measurement unit, a data processing unit, and an output unit, wherein the memory unit stores a threshold value and a calculation formula for a urinary D-serine excretion rate and / or a calculation formula for a D-asparagine excretion rate input from the input unit, the analysis and measurement unit quantifies the amount of D-serine and / or D-asparagine in a blood sample and / or a urine sample, and the data processing unit calculates a urinary D-serine excretion rate and / or a D-asparagine excretion rate generated from an element including the quantified amount of D-serine and / or D-asparagine in the blood sample and / or the urine sample and the calculation formula for the D-serine excretion rate and / or the calculation formula for the D-asparagine excretion rate stored in the memory unit, The data processing unit evaluates a renal pathology based on a comparison between a threshold value stored in the storage unit and a combination of the urinary D-serine excretion rate and / or the D-asparagine excretion rate and the blood D-serine amount and / or the D-asparagine amount, and the output unit outputs the evaluation result of the renal pathology of the subject.

[32] The D-serine excretion rate is calculated based on the following formula:

number

number

[33] A program for causing an information processing device including an input unit, an output unit, a data processing unit, and a storage unit to evaluate a renal pathology, the program comprising: storing in the storage unit a threshold value for evaluating a renal pathology, a calculation formula for the urinary D-serine excretion rate and / or a calculation formula for the D-asparagine excretion rate, and variables required for the calculation, which are input from the input unit; storing in the storage unit variables required for calculating the amount of D-serine and / or the amount of D-asparagine in a blood sample and / or a urine sample and the amount of D-serine and / or the amount of D-asparagine excretion rate in the urine, which are input from the input unit; The program includes instructions for causing the information processing device to execute the following: to cause the data processing device to call up the formula for calculating the urinary D-serine excretion rate and / or the formula for calculating the excretion rate of D-asparagine stored in advance in the storage device, and the amount of D-serine and / or the amount of D-asparagine in the blood sample and / or the urine sample stored in the storage device, and to substitute the variables into the formula for calculating the urinary D-serine excretion rate and / or the formula for calculating the excretion rate of D-asparagine; to cause the data processing device to evaluate a renal pathology based on a comparison between the threshold value stored in the storage device and a combination of the D-serine excretion rate and / or the D-asparagine excretion rate in urine and the amount of D-serine and / or the amount of D-asparagine in blood; and to output the evaluation result of the renal pathology of the subject to an output device.

[34] The program includes instructions for causing the information processing device to execute the following:

number

number

[0012] The analysis of the dynamics (reabsorption, excretion rate) of D-serine and / or D-asparagine in the kidney of the present invention provides a method for determining the renal pathology of a subject more broadly and accurately than currently known renal disease markers. [Brief description of the drawings]

[0013] [Figure 1] FIG. 1 shows the D-serine excretion rate and D-asparagine excretion rate in non-renal disease subjects and their logarithmic values. [Diagram 2] FIG. 2 shows the D-serine excretion rate and D-asparagine excretion rate in subjects with renal disease and their logarithmic values. [Diagram 3] FIG. 3 is a histogram of the logarithm of the D-serine excretion rate calculated from the amounts of D-serine and creatinine measured in the blood and urine of a subject. [Figure 4] FIG. 4 is a plot of the logarithm of the amount of D-serine in blood and the D-serine excretion rate measured in non-renal subjects and patients with renal disease. [Diagram 5] FIG. 5 is a histogram of the logarithm of the D-asparagine excretion rate calculated from the amounts of D-asparagine and creatinine measured in the blood and urine of subjects. [Figure 6] FIG. 6 is a plot of the logarithm of the amount of D-asparagine in blood and the rate of D-asparagine excretion measured in non-renal subjects and patients with renal disease. [Figure 7] FIG. 7 is a plot of the logarithm of the amount of D-serine in blood and the rate of D-serine excretion measured in non-renal subjects and patients with renal disease. [Figure 8]FIG. 8 is a plot of the logarithm of the amount of D-asparagine in blood and the rate of D-asparagine excretion measured in non-renal subjects and patients with renal disease. [Figure 9] Figure 9 is a chart showing the treatment and medication details and progress of a patient with systemic lupus erythematosus. [Figure 10] FIG. 10 is a plot of the amount of D-serine in blood and the D-serine excretion rate measured over time before and after therapeutic intervention in patients with systemic lupus erythematosus. [Figure 11] FIG. 11 shows a configuration diagram of the renal pathology evaluation system of the present invention. [Figure 12] FIG. 12 is a flow chart showing an example of the operation for evaluating a renal pathology by the program of the present invention. [Figure 13] FIG. 13 is a plot of the amount of D-serine in blood and the rate of D-serine excretion measured in patients diagnosed with kidney disease. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0014] The present invention relates to a method for determining renal pathology by analyzing the dynamics (reabsorption, excretion) of D-serine and / or D-asparagine in the kidney. The present inventors have found that the dynamics (reabsorption, excretion) of D-serine and D-asparagine in the kidney reflect renal pathology, respectively, and can be used to determine renal pathology in a subject. Therefore, the present invention may be a method for determining renal pathology by analyzing the dynamics (reabsorption, excretion) of D-serine in the kidney, a method for determining renal pathology by analyzing the dynamics (reabsorption, excretion) of D-asparagine in the kidney, or a method for determining renal pathology by analyzing the dynamics (reabsorption, excretion) of D-serine and D-asparagine in the kidney. Renal pathology can be diagnosed using the results of analysis of the dynamics (reabsorption, excretion) of D-serine or D-asparagine in the kidney, respectively. However, using the results of analysis of the dynamics (reabsorption, excretion) of both D-serine and D-asparagine in the kidney increases the accuracy of the evaluation and also makes it possible to determine false negatives and false positives.

[0015] In this specification, terms such as "first", "second", etc. are used to distinguish one element from other elements. For example, a first element may be expressed as a second element, and similarly, a second element may be expressed as a first element, without departing from the scope of the present invention.

[0016] In this specification, "urinary excretion rate of D-serine in a subject" may be written as "D-serine excretion rate in a subject", and "urinary excretion rate of D-serine in a non-renal disease subject" may be written as "D-serine excretion rate in a non-renal disease subject", and each of these terms has the same meaning when used interchangeably. In this specification, "urinary excretion rate of D-asparagine in a subject" may be written as "D-asparagine excretion rate in a subject", and "urinary excretion rate of D-asparagine in a non-renal disease subject" may be written as "D-asparagine excretion rate in a non-renal disease subject", and each of these terms has the same meaning when used interchangeably.

[0017] In this specification, "logarithmically transformed D-serine excretion rate of subject" may be written as "D-serine LN excretion rate of subject", and "logarithmically transformed value of D-serine excretion rate into urine in non-renal disease subject" may be written as "D-serine LN excretion rate of non-renal disease subject", and each of these terms has the same meaning when used interchangeably.In addition, in this specification, "logarithmically transformed D-asparagine excretion rate of subject" may be written as "D-asparagine LN excretion rate of subject", and "logarithmically transformed value of D-asparagine excretion rate into urine in non-renal disease subject" may be written as "D-asparagine LN excretion rate of non-renal disease subject", and each of these terms has the same meaning when used interchangeably.

[0018] As used herein, the term "subject" refers to all mammals, preferably humans, regardless of whether they have kidney disease. As used herein, the term "non-renal disease subject" refers to a subject who does not have kidney disease or has never been diagnosed with kidney disease, and is preferably a subject who does not suffer from kidney disease or other diseases that induce kidney damage.

[0019] In one embodiment, the present invention provides a method for assisting in the evaluation of renal pathology, using the combination of the reabsorption and excretion rate of D-serine and / or D-asparagine in the kidney of a subject and the amount of D-serine and / or D-asparagine in blood as an index. The reabsorption and excretion rate of D-serine and D-asparagine can be calculated by quantifying the amount of D-serine and D-asparagine in blood and the amount of D-serine and D-asparagine in urine, respectively. Therefore, in one embodiment, the "reabsorption and excretion rate of D-serine and / or D-asparagine in the kidney of a subject" in the present invention may be the "excretion rate of D-serine into the urine of a subject" ("subject D-serine excretion rate") and / or the "excretion rate of D-asparagine into the urine of a subject" ("subject D-asparagine excretion rate").

[0020] In the present invention, the excretion rate is an index showing the extent to which a target component is excreted in urine after passing through the renal tubule regulatory functions of reabsorption and secretion out of the amount of the target component filtered by the glomerulus, and is expressed in any unit such as a ratio or percentage. By correcting the amount of water, a value that excludes the influence of water reabsorption and concentration can be calculated, and the amount is sometimes expressed as fractional excretion (FE). Since the concentration rate of urine may not be constant, a "correction factor" that corrects the concentration rate of urine may be used to correct the rate of reabsorption and excretion of D-serine and / or D-asparagine in the kidney of the subject. For example, in one embodiment of the present invention, the subject D-serine excretion rate and / or the subject D-asparagine excretion rate may be corrected with a correction factor derived from blood and / or urine. The excretion rate is most simply expressed as a ratio obtained by dividing the amount of the target component in urine by the glomerular filtration rate of the target component, and the glomerular filtration rate obtained from inulin clearance or the like, the actually measured urine volume, and the amount of the target component in blood and / or urine may be used for the calculation. The amount of L-amino acid in urine (preferably the amount of L-serine and / or L-asparagine) can be used as a urine volume correction factor to calculate the D-amino acid excretion rate. As a correction factor, creatinine clearance calculated from the amount of creatinine in urine or blood can be used, and for example, the excretion rate of D-serine is expressed by the following formula: This may be multiplied by 100 to express it as a percentage (%).

number

[0021] For example, the excretion rate of D-asparagine can be expressed by the following formula: This may be multiplied by 100 to express it as a percentage (%).

number

[0022] In kidney disease, the partial excretion rate of sodium is used to distinguish whether it is due to dehydration or renal disorder. In addition, the partial excretion rate of potassium and the partial excretion rate of urea nitrogen are also used clinically as indicators of pathological condition. In general, the excretion rate is understood according to the principle of homeostasis, that if the intake or biosynthesis of the target component is high, the amount of excretion in urine increases, and if the intake is low and the amount of biodegradation is high, the amount of excretion decreases. Therefore, disorders and pathological changes in the kidney, which are the main homeostasis of the components in the body, can affect the change in the excretion rate. While creatinine, a conventional kidney disease marker, is excreted in its entirety and cystatin C is reabsorbed in its entirety, D-serine and D-asparagine are thought to be strictly regulated in terms of excretion and reabsorption in the renal tubules, like electrolytes, and it was thought that they could be more sensitive and accurate pathological markers.

[0023] In the present invention, D-serine and D-asparagine used in the analysis are optical isomers of L-serine and D-asparagine, which are protein-constituting amino acids. The amounts of D-serine and D-asparagine are strictly controlled in each tissue and body fluid by metabolic enzymes such as serine racemase and D-amino acid oxidase, and transporters, while the amounts of D-serine and D-asparagine in blood and urine fluctuate when renal dysfunction occurs.

[0024] In the present invention, "the amount of D-serine and / or D-asparagine in blood and urine" may refer to the amount of D-serine and / or D-asparagine in a specific blood volume and / or urine volume, and may be expressed as a concentration. The amount of D-serine and / or D-asparagine in blood and / or urine is measured as the amount in a sample obtained by centrifuging, sedimenting, or pretreating for analysis in collected blood and / or urine. Therefore, the amount of D-serine and / or D-asparagine in blood and / or urine can be measured as the amount in a blood sample derived from collected whole blood, serum, plasma, etc., or the amount in a urine sample derived from whole urine or urine excluding solid components, proteins, etc. As an example, in the case of analysis using HPLC, the amount of D-serine contained in a predetermined amount of blood and / or urine is represented by a chromatogram, and the height, area, shape, and size of the peak can be quantified by comparison with a standard or analysis by calibration. By comparing with samples with known concentrations of D-serine and / or D-asparagine, the concentrations of D-serine and / or D-asparagine in blood and urine can be measured, and the concentrations of D-serine and / or D-asparagine in blood and urine can be used as the amounts of D-serine and / or D-asparagine in blood and urine. In the enzymatic method, the amino acid concentrations can be calculated by quantitative analysis using a calibration curve of a standard.

[0025] The amounts of D- and L-amino acids, for example, D-serine and / or D-asparagine and L-serine and / or L-asparagine, can be measured by any method, for example, chiral column chromatography, enzyme method, or immunological method using monoclonal antibodies that distinguish optical isomers of amino acids. The amount of D-serine and L-serine in a sample in the present invention can be measured by any method known to those skilled in the art. For example, chromatography and enzyme methods (Y. Nagata et al., Clinical Science, 73 (1987), 105. Analytical Biochemistry, 150 (1985), 238., A. D'Aniello et al., Comparative Biochemistry and Physiology Part B, 66 (1980),319. Journal of Neurochemistry, 29 (1977), 1053., A. Berneman et al., Journal of Microbial & Biochemical Technology, 2 (2010), 139., WG Gutheil et al., Analytical Biochemistry, 287 (2000), 196., G. Molla et al., Methods in Molecular Biology, 794 (2012), 273., T. Ito et al., Analytical Biochemistry, 371 (2007), 167., etc.), antibody methods (T. Ohgusu et al., Analytical Biochemistry, 357 (2006), 15., etc.), gas chromatography (GC) (H. Hasegawa et al., Journal of Mass Spectrometry, 46 (2011), 502., MC Waldhier et al., Analytical and Bioanalytical Chemistry, 394 (2009), 695., A. Hashimoto, T. Nishikawa et al., FEBS Letters, 296 (1992), 33., H. Bruckner and A. Schieber, Biomedical Chromatography, 15 (2001), 166., M. Junge et al., Chirality, 19 (2007), 228., M. C. Waldhier et al., Journal of Chromatography A, 1218 (2011), 4537, etc.), capillary electrophoresis (CE) (H. Miao et al., Analytical Chemistry, 77 (2005), 7190., D. L. Kirschner et al., Analytical Chemistry, 79 (2007), 736., F. Kitagawa, K. Otsuka, Journal of Chromatography B, 879 (2011), 3078., G. Thorsen and J. Bergquist, Journal of Chromatography B, 745 (2000), 389, etc.), high performance liquid chromatography (HPLC) (N. Nimura and T. Kinoshita, Journal of Chromatography, 352 (1986), 169., A. Hashimoto et al., Journal of Chromatography, 582 (1992), 41., H. Bruckner et al., Journal of Chromatography A, 666 (1994), 259., N. Nimura et al., Analytical Biochemistry, 315(2003), 262., C. Muller et al., Journal of Chromatography A, 1324 (2014), 109.,S. Einarsson et al., Analytical Chemistry, 59 (1987), 1191., E. Okuma and H. Abe, Journal of Chromatography B, 660 (1994), 243., Y.Gogami et al., Journal of Chromatography B, 879 (2011), 3259., Y. Nagata et al., Journal of Chromatography, 575 (1992), 147., S. A. Fuchs et al., Clinical Chemistry, 54 (2008), 1443., D. Gordes et al., Amino Acids, 40 (2011), 553., D. Jin et al., Analytical Biochemistry, 269 (1999), 124., J. Z. Min et al., Journal of Chromatography B, 879 (2011), 3220., T. Sakamoto et al., Analytical and Bioanalytical Chemistry, 408 (2016), 517., W. F. Visser et al., Journal of Chromatography A, 1218 (2011), 7130., Y. Xing et al., Analytical and Bioanalytical Chemistry, 408 (2016), 141., K. Imai et al., Biomedical Chromatography, 9 (1995), 106., T. Fukushima et al., Biomedical Chromatography, 9 (1995), 10., R. J. Reischl et al., Journal of Chromatography A, 1218 (2011), 8379., R. J. Reischl and W. Lindner, Journal of Chromatography A, 1269 (2012), 262., S. Karakawa et al., Journal of Pharmaceutical and Biomedical Analysis, 115 (2015), 123., etc.).

[0026] The optical isomer separation and analysis system of the present invention may combine a plurality of separation and analysis. More specifically, the method for analyzing optical isomers includes the steps of passing a sample containing components having optical isomers through a first column packing material as a stationary phase together with a first liquid as a mobile phase to separate the components of the sample, individually retaining each of the components of the sample in a multi-loop unit, supplying each of the components of the sample individually retained in the multi-loop unit through a flow path to a second column packing material having an optically active center as a stationary phase together with a second liquid as a mobile phase to separate the optical isomers contained in each of the components of the sample, and detecting the optical isomers contained in each of the components of the sample (Patent No. 4291628). In HPLC analysis, D- and L-amino acids may be derivatized in advance with a fluorescent reagent such as o-phthalaldehyde (OPA) or 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F), or diastereomerized using N-tert-butyloxycarbonyl-L-cysteine ​​(Boc-L-Cys) or the like (Kenji Hamase and Kiyoshi Zaitsu, Analytical Chemistry, Vol. 53, 677-690 (2004)). Alternatively, D-amino acids can be measured by immunological techniques using monoclonal antibodies that distinguish optical isomers of amino acids, for example, monoclonal antibodies that specifically bind to D-serine, L-serine, D-asparagine, or L-asparagine. In addition, when the total amount of D- and L-isomers is used as an index, it is not necessary to separate and analyze D- and L-isomers, and amino acids can be analyzed without distinguishing between D- and L-isomers. In this case, separation and quantification can be performed by enzymatic methods, antibody methods, GC, CE, and HPLC.

[0027] The amount of D-serine and D-asparagine in blood is more strongly correlated with glomerular filtration rate than creatinine, a conventional marker. This is because the amount of creatinine in blood is strongly influenced by muscle mass, and therefore shows high values ​​in athletes, patients with acromegaly, and those who consume a large amount of meat, and shows low values ​​in patients with neuromuscular diseases (muscular dystrophy, etc.), emaciation, long-term bed rest, frailty, sarcopenia, locomotive syndrome, anputation, and those who restrict protein intake, and therefore cannot accurately reflect renal function. In healthy individuals without pathological findings, the amount of D-serine in blood is kept within a very small range of about 1-2% of the total serine amount, in contrast to the amount in urine, which reaches 30-60%. Interestingly, unlike L-serine, which is reabsorbed by the renal tubules, about 50-80% of D-serine is excreted. In addition, in healthy individuals without pathological findings, the amount of D-asparagine in the blood is maintained in a very small range of about 0.1-0.6% of the total asparagine amount, whereas the amount in the urine ranges from 20-50%. Interestingly, unlike L-asparagine, which is reabsorbed in the renal tubules at about 99%, D-asparagine is excreted at about 50-80%.

[0028] The excretion rates of D-serine and D-asparagine proposed in the present invention have been shown to be uncorrelated with the glomerular filtration rate, unlike the amounts of D-serine and D-asparagine in the blood, by chiral amino acid metabolomics and multivariate analysis of related parameters (OPLS). Since it has been suggested that the reabsorption of the optical isomers of asparagine in the renal tubules is strictly controlled, 15 healthy volunteers were recruited as a study population to analyze the excretion rates of D-serine and D-asparagine in non-renal disease subjects in order to investigate the physiological significance of D-serine and D-asparagine. The study protocol was approved by the ethics committee of the National Institutes of Biomedical Innovation, Health and Nutrition, and written informed consent was obtained from all subjects. The non-renal disease subjects had a mean age of 44, 80% males, mean height of 1.70 m, mean weight of 68.9 kg, and mean BSA of 1.80 m.2 , average BMI22.6kg / m 2 , the mean serum creatinine was 0.75 mg / dL.

[0029] Using the following formulas based on the quantitative analysis values ​​of D-serine and D-asparagine in the subjects' blood and urine, the average excretion rate of D-serine was calculated to be 62.76%, with its average logarithm value being 4.12, and the average excretion rate of D-asparagine was calculated to be 64.12%, with its average logarithm value being 4.16 (Figure 1).

number

number

[0030] Regarding D-serine, when the logarithmic data was plotted in 6 quantiles, a normal distribution-like shape was observed (Figure 3). A Shapiro-Wilk Normality Test was performed on this data, and a value of P=0.395 was obtained, so the null hypothesis was not rejected and the possibility of a normal distribution was supported. Therefore, the reference value for these non-renal disease subjects was calculated to be 42.46-89.66% from the mean ± 1.96 standard deviation, and the logarithmic value was calculated to be 3.75-4.50. Subjects falling outside this range can be used to help predict the pathology, risk, or prognosis of kidney disease and renal impairment.

[0031] In addition, when the logarithmic data for D-asparagine was plotted in 6 quantiles, a normal distribution-like shape was observed (Figure 5). A Shapiro-Wilk Normality Test was performed on this data, with a value of P=0.243, so the null hypothesis was not rejected and the possibility of a normal distribution was supported. Therefore, the reference value for these non-renal disease subjects was calculated to be 51.65-78.74% from the mean ± 1.96 standard deviation, and the logarithmic value was calculated to be 3.95-4.37. Subjects falling outside this range can be used to help predict kidney disease, the pathology of kidney disease, its risk, or prognosis.

[0032] Since the amount of D-serine and D-asparagine in blood strongly correlates with the glomerular filtration rate, their analysis has been shown to be applicable to the severity classification (G1-5) of chronic kidney disease (CKD) defined in the guidelines of the Japanese Society of Nephrology. However, the D-serine excretion rate, which is analyzed by adding the amount of D-serine in urine, and the D-asparagine excretion rate, which is analyzed by adding the amount of D-asparagine in urine, can assist in the evaluation of renal pathology through a completely different mechanism that is not correlated with the glomerular filtration rate, and are therefore highly clinically useful in differential diagnosis, pathology, and prognosis diagnosis, which were difficult with conventional markers.

[0033] In one embodiment, the present invention may provide a method comprising the step of comparing first object coordinates obtained by plotting the object D-serine excretion rate and / or object D-asparagine excretion rate in the object and the amount of D-serine and / or D-asparagine in the blood with a first standard calculated from non-renal disease coordinates obtained by plotting the excretion rates of D-serine into urine (non-renal disease subject D-serine excretion rate) and / or the excretion rates of D-asparagine (non-renal disease subject D-asparagine excretion rate) in a plurality of non-renal disease subjects and the amount of D-serine and / or D-asparagine in the blood, and evaluating a renal pathological condition from the relationship between the first object coordinates and the first standard.

[0034] Therefore, for example, in the first embodiment of the above invention, the method may include a step of comparing first object coordinates obtained by plotting the object D-serine excretion rate and the amount of D-serine in blood in the object with a first standard calculated from non-renal disease coordinates obtained by plotting the excretion rates of D-serine into urine in a plurality of non-renal disease subjects (non-renal disease subject D-serine excretion rates) and the amount of D-serine in blood, and evaluating the renal pathology from the relationship between the first object coordinates and the first standard.

[0035] Furthermore, for example, in a second embodiment of the above invention, the method may include a step of comparing first object coordinates obtained by plotting the object D-asparagine excretion rate and the amount of D-asparagine in the blood in the object with a first standard calculated from non-renal disease coordinates obtained by plotting the excretion rates of D-asparagine into urine in a plurality of non-renal disease subjects (non-renal disease object D-asparagine excretion rates) and the amount of D-asparagine in the blood, and evaluating the renal pathological condition from the relationship between the first object coordinates and the first standard.

[0036] In addition, in order to evaluate renal pathology, the method of the first embodiment and the method of the second embodiment described above may be used in combination. In this case, not only is the accuracy of the evaluation of renal pathology improved, but false positives and false negatives can also be determined.

[0037] In the present specification, the "first standard" refers to a standard calculated from coordinates (referred to as "non-renal disease coordinates") plotting the excretion rate of D-serine into urine in a plurality of non-renal disease subjects (D-serine excretion rate in non-renal disease subjects) and / or the excretion rate of D-asparagine into blood in a plurality of non-renal disease subjects, and is used to evaluate the renal pathology of a subject. In one embodiment, the first standard that can be used in the present invention may be calculated from non-renal disease coordinates plotting the excretion rate of D-serine into urine in a plurality of non-renal disease subjects (D-serine excretion rate in non-renal disease subjects) and the amount of D-serine in blood. In addition, in one embodiment, the first standard that can be used in the present invention may be calculated from non-renal disease coordinates plotting the excretion rate of D-asparagine into urine in a plurality of non-renal disease subjects (D-asparagine excretion rate in non-renal disease subjects) and the amount of D-asparagine in blood. The number of "non-renal disease subjects" used to calculate the first criterion is preferably a number sufficient to calculate a statistically significant criterion, and for example, a number of 3, 5, 10, 15, 20, 30, 50, 100 or more can be used in the present invention.

[0038] In the present specification, the "first object coordinate" refers to a coordinate obtained by plotting the target D-serine excretion rate and / or the target D-asparagine excretion rate and the blood D-serine amount and / or the blood D-asparagine amount in a subject to be evaluated for renal pathology. For example, in one embodiment, the first object coordinate that can be used in the present invention may be a coordinate obtained by plotting the target D-serine excretion rate and the blood D-serine amount in a subject to be evaluated for renal pathology. Also, for example, in one embodiment, the first object coordinate that can be used in the present invention may be a coordinate obtained by plotting the target D-asparagine excretion rate and the blood D-asparagine amount in a subject to be evaluated for renal pathology. In the present invention, the renal pathology in a subject can be evaluated by comparing the first object coordinate with a first standard.

[0039] In one embodiment, the first criterion in the present invention may be in the range of the mean value of the plot of non-renal disease coordinates ± standard deviation × coefficient Z. In this specification, "coefficient Z" is a coefficient used to calculate a confidence interval used in statistics, and is preferably, for example, a value between 1.0 and 3.0, more preferably 1.96. In one embodiment, the first criterion is preferably in the range of 0.4 to 0.9.

[0040] In one embodiment, the step of assessing renal pathology included in the present invention may be assessing a subject's renal disease or risk of developing renal disease, or predicting the induction or prognosis of renal disease, when the first object coordinate is not included in the first criterion.

[0041] In one embodiment, the kidney disease that can be evaluated in the present invention may be, for example, chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease or minimal change nephrotic syndrome.

[0042] In another embodiment, the present invention may provide a method for assisting in the evaluation of renal pathology from the relationship between the regression equation calculated by regression analysis of the plot of non-renal disease coordinates and the target coordinates. The position or distance on the coordinates between the plot of the analysis target and the regression equation enables the evaluation of the fluctuation of D-serine and / or D-asparagine kinetics in non-renal disease patients. For example, if the excretion rate axis fluctuates to the positive side, excretion is enhanced, and if it fluctuates to the negative side, reabsorption is enhanced in the renal pathology, and the greater the distance, the greater the degree of the renal pathology.

[0043] In another embodiment, the present invention may provide a method comprising the steps of: comparing second object coordinates obtained by plotting a logarithmically transformed subject D-serine excretion rate (subject D-serine LN excretion rate) and / or a logarithmically transformed subject D-asparagine excretion rate (subject D-asparagine LN excretion rate) and a logarithmically transformed blood D-serine amount and / or D-asparagine amount with a second standard calculated from non-renal disease coordinates obtained by plotting a logarithmically transformed urinary D-serine excretion rate (non-renal disease subject D-serine LN excretion rate) and / or D-asparagine excretion rate (non-renal disease subject D-asparagine LN excretion rate) and a logarithmically transformed blood D-serine amount and / or D-asparagine amount in a plurality of non-renal disease subjects, and evaluating a renal pathology based on a relationship between the second object coordinates and the second standard.

[0044] Therefore, for example, in a first embodiment of the above-mentioned invention, a method may be provided which includes a step of comparing second object coordinates on which the logarithmically transformed subject D-serine excretion rate (subject D-serine LN excretion rate) and the logarithmically transformed amount of D-serine in the blood are plotted with a second standard calculated from non-renal disease coordinates on which the logarithmically transformed urinary D-serine excretion rates (non-renal disease subject D-serine LN excretion rates) and the logarithmically transformed amount of D-serine in the blood are plotted in a plurality of non-renal disease subjects, and evaluating the renal pathology from the relationship between the second object coordinates and the second standard.

[0045] Furthermore, for example, in a second embodiment of the above invention, the present invention may provide a method comprising the steps of comparing second object coordinates obtained by plotting the logarithmically transformed subject D-asparagine excretion rate (subject D-asparagine LN excretion rate) and the logarithmically transformed amount of D-asparagine in blood with a second standard calculated from non-renal disease coordinates obtained by plotting the logarithmically transformed urinary D-asparagine excretion rate (non-renal disease subject D-asparagine LN excretion rate) and the logarithmically transformed amount of D-asparagine in blood in a plurality of non-renal disease subjects, and evaluating the renal pathology from the relationship between the second object coordinates and the second standard.

[0046] In addition, in order to evaluate renal pathology, the method of the first embodiment and the method of the second embodiment described above may be used in combination. In this case, not only is the accuracy of the evaluation of renal pathology improved, but false positives and false negatives can also be determined.

[0047] In this specification, the term "logarithmically transformed value" refers to a value obtained by converting a target value into a logarithm. For example, the term may refer to a value obtained by converting a target value into a natural logarithm, or may refer to a value obtained by converting a target value using any base, such as common logarithm.

[0048] As used herein, the "second criterion" refers to the logarithmically transformed target D-serine excretion rate (target D-serine LN excretion rate) and / or It is calculated from the coordinate (referred to as "non-renal disease coordinate") of the logarithmic transformation of the target D-serine excretion rate (target D-asparagine LN excretion rate) and the logarithmic transformation of the blood D-serine amount and / or D-asparagine amount, and refers to a criterion used to evaluate the renal pathology of a subject. In one embodiment, the second criterion that can be used in the present invention may be calculated from the non-renal disease coordinate of the logarithmic transformation of the target D-serine excretion rate (target D-serine LN excretion rate) and the logarithmic transformation of the blood D-serine amount. In one embodiment, the second criterion that can be used in the present invention may be calculated from the non-renal disease coordinate of the logarithmic transformation of the target D-asparagine excretion rate (target D-asparagine LN excretion rate) and the logarithmic transformation of the blood D-asparagine amount. The number of "non-renal disease subjects" used to calculate the second criterion is preferably a number sufficient to calculate a statistically significant criterion, and for example, a number of 3, 5, 10, 15, 20, 30, 50, 100 or more can be used in the present invention.

[0049] In one embodiment, the second criterion that can be used in the present invention may be in the range of the mean value of the plot of non-renal disease coordinates ± standard deviation × coefficient Z. In this case, the coefficient Z is preferably a value between 1.0 and 3.0, more preferably 1.96. In one embodiment, the second criterion is preferably in the range of 3.5 to 5.0.

[0050] In one embodiment, a second criterion that may be used in the present invention may be a distance of 0.6 or less from the mean value of the plot of non-renal disease coordinates.

[0051] In one embodiment, the step of assessing the renal pathology of the present invention may be to assess the subject's renal disease or risk of developing renal disease, or to predict the induction or prognosis of renal disease, when the second object coordinate is not included in the second criterion.

[0052] In another embodiment, the present invention may be a method for assisting in the evaluation of renal pathology from the relationship between the regression equation calculated by the regression line of the plot of non-renal disease coordinates based on the logarithmic transformed values ​​and the target coordinates based on the logarithmic transformed values. The position or distance on the coordinates between the plot of the analysis target and the regression equation enables the evaluation of the fluctuation of D-serine and / or D-asparagine kinetics in non-renal disease patients. For example, if the excretion rate axis fluctuates to the positive side, excretion is enhanced, and if it fluctuates to the negative side, reabsorption is enhanced in the renal pathology, and the greater the distance, the greater the degree of the renal pathology.

[0053] When the pathological condition is determined by the method of the present invention, a treatment plan can be determined based on the determination. A treatment method can be appropriately selected according to each pathological condition. For example, the above-mentioned first object coordinate or second object coordinate may be controlled while being monitored over time so that it falls within the reference range for non-renal subjects (for example, the range of the above-mentioned first standard or second standard). Therapeutic intervention includes lifestyle improvement, dietary guidance, blood pressure management, anemia management, electrolyte management, uremic toxin management, blood glucose management, immune management, and lipid management, which are independently or in combination. As lifestyle improvement, smoking cessation and weight loss to a BMI value of less than 25 are recommended. As dietary guidance, salt reduction and protein restriction are performed. Among these, blood pressure management, anemia management, electrolyte management, uremic toxin management, blood glucose management, immune management, and lipid management can be treated by medication. As blood pressure management, blood pressure is controlled to be 130 / 80 mmHg or less, and hypertension drugs can be administered in some cases.Examples of antihypertensive drugs include diuretics (thiazide diuretics, e.g., trichlormethiazide, benzylhydrochlorothiazide, hydrochlorothiazide, thiazide-like diuretics, e.g., methiclane, indabamide, tribamide, mefruside, loop diuretics, e.g., furosemide, potassium-sparing diuretics / aldosterone antagonists, e.g., triamterene, spironolactone, eplerenone, etc.), calcium antagonists (dihydropyridine-based, e.g., nifedipine, amlodipine, efonidipine, cilnidipine, nicardipine, nisoldipine, nitrendipine, nilvadipine, barnidipine, felodipine, benidipine, manidipine, azelnidipine, aranidipine, benzothiazepine-based, diltiazem, etc.), angiotensin-converting enzyme inhibitors (captopril, enalapril, Acela Pril, delapril, cilazapril, lisinopril, benazepril, imidapril, temocapril, quinapril, trandolapril, belindopril erbumine, etc.), angiotensin receptor antagonists (angiotensin II receptor antagonists, e.g., losartan, candesartan, valsartan, telmisartan, olmesartan, irbesartan, azilsartan, etc.), sympatholytic agents (β-blockers, e.g., atenolol, bisoprolol, betaxolol, metoprolol, aceptolol, celiprolol, propranolol, nadolol, carteolol, pindolol, nipradilol, amosulalol, arotinolol, carvedilol, labetalol, bevantolol, urapidil, terazosin, brazosin, doxazosin, bunazosin, etc.), etc. may be used. Erythropoietin preparations, iron preparations, HIF-1 inhibitors, etc. are used as anemia treatment drugs. Calcium receptor agonists (cinacalcet, etelcalcetide, etc.) and phosphate binders are used as electrolyte regulators. Activated charcoal is used as a uremic toxin binder. Blood glucose levels are controlled to be less than Hba1c 6.9%, and hypoglycemic drugs are administered in some cases.As hypoglycemic drugs, SGLT2 inhibitors (ipragliflozin, dapagliflozin, luseogliflozin, tofogliflozin, canagliflozin, empagliflozin, etc.), DPP4 inhibitors (sitagliptin phosphate, vildagliptin, saxagliptin, alogliptin, linagliptin, teneligliptin, trelagliptin, anagliptin, omarigliptin, etc.), sulfonylureas (tolbutamide, acetohexamide, etc.), Treatments include thiazolidinedione (pioglitazone, etc.), biguanides (metformin, buformin, etc.), α-glucosidase inhibitors (acarbose, voglibose, miglitol, etc.), glinides (nateglinide, mitiglinide, repaglinide, etc.), insulin preparations, and NRF2 activators (bardoxolone methyl, etc.). Immunosuppressants (steroids, tacrolimus, anti-CD20 antibodies, cyclohexamide, mycophenolate mofetil (MMF), etc.) are used for immune management. In lipid management, LDL-C is controlled to be less than 120 mg / dL, and in some cases, dyslipidemia treatment drugs such as statin drugs (rosuvastatin, pitavastatin, atorvastatin, cerivastatin, fluvastatin, simvastatin, pravastatin, lovastatin, mevastatin, etc.), fibrate drugs (clofibrate, bezafibrate, fenofibrate, clinofibrate, etc.), nicotinic acid derivatives (tocorrel nicotinate, nicomol, niceritrol, etc.), cholesterol transporter inhibitors (ezetimibe, etc.), PCSK9 inhibitors (evolocumab, etc.), EPA preparations, etc. are used. Each drug may be in the form of a single drug or a combination drug. Depending on the degree of decline in renal function, renal replacement therapy such as peritoneal dialysis, hemodialysis, continuous hemofiltration dialysis, hemoapheresis (plasma exchange, plasma adsorption, etc.) or kidney transplantation is performed.

[0054] Therefore, in one embodiment, the present invention may be a method for monitoring a renal pathology, which comprises measuring the excretion rate of D-serine (subject D-serine excretion rate) and / or the excretion rate of D-asparagine (subject D-asparagine excretion rate) into the urine of a subject and the amount of D-serine and / or D-asparagine in the blood over time, and using the fluctuations in the subject D-serine excretion rate and / or the subject D-asparagine excretion rate and the amount of D-serine and / or D-asparagine in the blood as indicators. For example, in one embodiment, the present invention may be a method for monitoring renal pathology by measuring the excretion rate of D-serine into a subject's urine (subject D-serine excretion rate) and the amount of D-serine in the blood over time, and using the fluctuations in the target D-serine excretion rate and the amount of D-serine in the blood as indicators.For example, in one embodiment, the present invention may be a method for monitoring renal pathology by measuring the excretion rate of D-asparagine into a subject's urine (subject D-asparagine excretion rate) and the amount of D-asparagine in the blood over time, and using the fluctuations in the target D-asparagine excretion rate and the amount of D-asparagine in the blood as indicators, or a method for monitoring renal pathology by combining both.

[0055] In another embodiment, the present invention may be a method for monitoring the therapeutic effect of a renal pathology by measuring the urinary excretion rate of D-serine (subject D-serine excretion rate) and / or the excretion rate of D-asparagine (subject D-asparagine excretion rate) and the blood amount of D-serine and / or D-asparagine over time in a subject with kidney disease before and after therapeutic intervention, and using the fluctuations in the target D-serine excretion rate and / or the target D-asparagine excretion rate and the blood amount of D-serine and / or D-asparagine as indicators. For example, in one embodiment, the present invention may be a method for monitoring the therapeutic effect of a renal pathology by measuring the urinary excretion rate of D-serine (subject D-serine excretion rate) and the amount of D-serine in the blood of a subject with renal disease before and after therapeutic intervention over time, and using the fluctuations in the target D-serine excretion rate and the amount of D-serine in the blood as indicators. For example, in one embodiment, the present invention may be a method for monitoring the therapeutic effect of a renal pathology by measuring the urinary excretion rate of D-asparagine (subject D-asparagine excretion rate) and the amount of D-asparagine in the blood of a subject with renal disease before and after therapeutic intervention over time, and using the fluctuations in the target D-asparagine excretion rate and the amount of D-asparagine in the blood as indicators, or may be a method for monitoring the therapeutic effect of a renal pathology by combining both.

[0056] The methods of the present invention can be used to assess kidney disease in a subject, for example, chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease, or minimal change nephrotic syndrome.

[0057] In another embodiment, the present invention provides a method for assisting in the evaluation of renal pathology using the amount of D-serine and / or D-asparagine in the blood of a subject from whom urine cannot be collected as an index. As used herein, the term "subject from whom urine cannot be collected" refers to, for example, a subject with chronic renal failure or acute renal failure whose renal function is extremely reduced and who is eligible for renal replacement therapy (dialysis, plasma exchange, kidney transplantation, etc.).

[0058] In another embodiment, the present invention provides a method for assisting in determining whether a subject has systemic lupus erythematosus when the D-serine level in the subject's blood is 9 nmol / mL or more.

[0059] Another aspect of the present invention may relate to a system and a program for executing the above-mentioned method for assisting in the evaluation of renal pathology. Fig. 11 is a configuration diagram of a renal pathology evaluation system of the present invention. A sample analysis system 10 shown in Fig. 11 is configured to be able to execute the method for assisting in the evaluation of renal pathology of the present invention. Such a sample analysis system 10 includes a memory unit 11, an input unit 12, an analysis and measurement unit 13, a data processing unit 14, and an output unit 15, and is capable of analyzing a blood sample and / or a urine sample, and outputting a calculated excretion rate and pathology information.

[0060] More specifically, in the sample analysis system 10 of the present invention, the memory unit 11 stores a combination of the excretion rate and the amount of D-serine and / or D-asparagine in blood samples and urine samples input from the input unit 12, which are calculated from the amounts of D-serine and / or D-asparagine, and a reference value / pathological condition information correspondence table or graph. The analysis measurement unit 13 separates and quantifies D-serine and / or D-asparagine in the blood samples and / or urine samples. The data processing unit 14 calculates the excretion rate and the amount of D-serine and / or D-asparagine in blood samples and urine samples. The amount of serine and / or D-asparagine can be substituted into an equation obtained from the reference values ​​and pathological condition information, or read out from a correspondence table or graph, to determine the pathological condition, and the output unit 15 can output the pathological condition information.

[0061] In a more preferred embodiment, the renal pathology evaluation system of the present invention may further include a step in which the memory unit 11 stores the reference value input from the input unit 12, and a step in which the data processing unit 14 compares the combination of the excretion rate calculated from the separated and quantified D-serine and / or D-asparagine amounts and the D-serine and / or D-asparagine amounts in blood with the reference value. In this case, when the combination of the D-serine excretion rate and / or D-asparagine excretion rate and the D-serine and / or D-asparagine amounts in blood is outside the reference range, the output unit 15 outputs a suspicion of kidney disease.

[0062] The storage unit 11 has a memory device such as a RAM, a ROM, a flash memory, a fixed disk device such as a hard disk drive, or a portable storage device such as a flexible disk or an optical disk. The storage unit stores data measured by the analysis and measurement unit, data and instructions input from the input unit, calculation results performed by the data processing unit, computer programs used for various processes of the information processing device, databases, etc. The computer program may be installed from a computer-readable recording medium such as a CD-ROM or a DVD-ROM, or via the Internet. The computer program is installed in the storage unit using a known setup program, etc. The storage unit stores data on a formula, a correspondence table, or a graph calculated from the association between the combination of the D-serine excretion rate and the amount of D-serine in blood input in advance from the input unit 12 and the pathology. It can also store a renal pathology classification according to the excretion rate.

[0063] The input unit 12 is an interface or the like, and also includes an operation unit such as a keyboard, a mouse, etc. This allows the input unit to input data measured by the analysis and measurement unit 13, instructions for arithmetic processing to be performed by the data processing unit 14, etc. Furthermore, when the analysis and measurement unit 13 is external, for example, the input unit 12 may include an interface unit that can input the measured data, etc. via a network or a storage medium, separate from the operation unit.

[0064] The analysis and measurement unit 13 performs a step of measuring D-serine and / or D-asparagine in a blood sample and / or a urine sample. Therefore, the analysis and measurement unit 13 has a configuration that enables separation and measurement of D- and L-isomers of amino acids. Amino acids may be analyzed one by one, or some or all types of amino acids may be analyzed together. The analysis and measurement unit 13 is not intended to be limited to the following, but may be, for example, a chiral chromatography system equipped with a sample introduction unit, an optical resolution column, and a detection unit, preferably a high performance liquid chromatography system. From the viewpoint of detecting only the amount of a specific amino acid, quantification may be performed by an enzyme method or an immunological method. The analysis and measurement unit 13 may be configured separately from the renal pathology evaluation system, and the measured data, etc. may be input via the input unit 12 using a network or a storage medium.

[0065] The data processing unit 14 can evaluate and determine renal pathology by calculating the excretion rate from the measured D-serine amount and / or D-asparagine amount, substituting the calculated rate into a formula calculated from the relationship between the excretion rate and the combination of the D-serine amount and / or D-asparagine amount in blood, or by reading out the corresponding table or graph. If the formula, the corresponding table or graph calculated from the relationship between the D-serine excretion rate and / or D-asparagine excretion rate and the combination of the D-serine amount and / or D-asparagine amount in blood, requires other correction values, such as age, weight, sex, height, etc., such information is input in advance from the input unit and stored in the storage unit. When the data processing unit calculates the excretion rate and pathology information, it can call up such information and substitute it into the formula or read it out from the corresponding table or graph to calculate the excretion rate and pathology information. The data processing unit 14 can also determine kidney disease and kidney pathology classification from the determined excretion rate, the amount of D-serine and / or D-asparagine in blood, and pathology information. The data processing unit 14 executes various arithmetic processing on the data measured by the analysis and measurement unit 13 and stored in the memory unit 11 according to a program stored in the memory unit. The arithmetic processing is performed by a CPU included in the data processing unit. This CPU includes functional modules that control the analysis and measurement unit 13, the input unit 12, the memory unit 11, and the output unit 15, and can perform various controls. Each of these units may be composed of an independent integrated circuit, microprocessor, firmware, etc.

[0066] The output unit 15 is configured to output a combination of the excretion rate and the amount of D-serine and / or D-asparagine in blood, which are the results of the calculation performed by the data processing unit, and pathological condition information. The output unit 15 may be an output means such as a display device such as a liquid crystal display that directly displays the results of the calculation, or a printer, or may be an interface unit for outputting to an external storage device or via a network. The D-serine excretion rate and / or D-asparagine excretion rate, the amount of D-serine and / or D-asparagine in blood, and / or renal pathological condition classification may be output together with or independently of the glomerular filtration capacity.

[0067] 12 is a flow chart showing an example of the operation for determining the excretion rate and pathological condition information by the program of the present invention. Specifically, the program of the present invention is a program for causing an information processing device including an input unit, an output unit, a data processing unit, and a storage unit to evaluate a renal pathological condition. The program of the present invention performs the following operations: storing in the storage unit a threshold value for evaluating a renal pathological condition, a calculation formula for the urinary D-serine excretion rate and / or a calculation formula for the D-asparagine excretion rate, and variables required for calculation, which are input from the input unit; storing in the storage unit variables required for calculating the amount of D-serine and / or the amount of D-asparagine in a blood sample and / or a urine sample and the D-serine excretion rate and / or the D-asparagine excretion rate into urine, which are input from the input unit; The program includes instructions to cause the information processing device to execute the following: to call the calculation formula of the D-serine excretion rate into urine and / or the calculation formula of the D-asparagine excretion rate stored in advance in the storage unit, and the amount of D-serine and / or the amount of D-asparagine in the blood sample and / or the urine sample stored in the storage unit, and to substitute the variables into the calculation formula of the D-serine excretion rate into urine and / or the calculation formula of the D-asparagine excretion rate; to cause the data processing device to evaluate a renal pathology based on a comparison between the threshold value stored in the storage unit and a combination of the D-serine excretion rate into urine and / or the D-asparagine excretion rate and the amount of D-serine and / or the amount of D-asparagine in blood; and to output the evaluation result of the renal pathology of the subject to the output unit. The program of the present invention may be stored in a storage medium or provided via a telecommunication line such as the Internet or a LAN.

[0068] When the information processing device is equipped with an analytical measurement unit, instead of inputting values ​​of the D-serine and / or D-asparagine amounts from the input unit, the analytical measurement unit may include instructions for causing the information processing device to measure the values ​​from a blood sample and / or a urine sample and store them in the memory unit.

[0069] All documents mentioned herein are incorporated by reference in their entirety.

[0070] The following examples of the present invention are for illustrative purposes only and do not limit the technical scope of the present invention. The technical scope of the present invention is limited only by the claims. The present invention can be modified, for example, by adding, deleting, and replacing the constituent elements of the present invention, without departing from the spirit of the present invention. EXAMPLES

[0071] Study population: From a cohort of patients with kidney disease admitted to the Department of Nephrology, Osaka University Hospital between 2016 and 2017 for diagnostic and / or therapeutic purposes, patients with primary aldosteronism (PA), myeloma nephropathy (IGAN), diabetic nephropathy (DM), and immunoglobulin A nephropathy (IGAN) were used in this retrospective study. Subjects with IgA nephropathy had blood pressure above the reference range, so they were administered angiotensin II receptor blockers (ARBs) as antihypertensive drugs. Separately, 15 healthy volunteers were recruited as non-renal disease subjects at the National Institute of Biomedical Innovation, Health and Nutrition, as mentioned above. The study protocol was approved by the ethical committees at each institution, and written informed consent was obtained from all subjects.

[0072] Measurement of D-serine and D-asparagine in blood and urine Sample preparation Sample preparation from human plasma and urine was performed as follows: 20 volumes of methanol were added to the plasma and mixed thoroughly. After centrifugation, 10 μL of the supernatant obtained from the methanol homogenate was transferred to a brown tube and dried under reduced pressure. 20 μL of 200 mM sodium borate buffer (pH 8.0) and 5 μL of fluorescent labeling reagent (40 mM 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) in anhydrous MeCN) were added to the residue, and then heated at 60° C. for 2 min. The reaction was stopped by adding 75 μL of 0.1% TFA aqueous solution (v / v), and 2 μL of the reaction mixture was subjected to two-dimensional HPLC.

[0073] Quantification of Amino Acid Optical Isomers by Two-Dimensional HPLC Amino acid optical isomers were quantified using the following two-dimensional HPLC system. NBD-derivatives of amino acids were separated and eluted with a mobile phase (5-35% MeCN, 0-20% THF, and 0.05% TFA) using a reversed-phase column (KSAA RP, 1.0 mm i.d. × 400 mm; Shiseido Co., Ltd.). The column temperature was set at 45°C, and the flow rate of the mobile phase was set at 25 μL / min. The separated amino acid fractions were collected using a multi-loop valve and optically resolved continuously using a chiral column (KSAACSP-001S, 1.5 mm i.d. × 250 mm; Shiseido Co., Ltd.). As the mobile phase, a mixture of MeOH-MeCN containing citric acid (0-10 mM) or formic acid (0-4%) was used depending on the retention of the amino acid. NBD-amino acids were detected by fluorescence at 530 nm using 470 nm excitation light. The retention times of NBD-amino acids were identified using standards of amino acid optical isomers and quantified using a calibration curve.

[0074] Calculation of D-serine excretion rate and D-asparagine excretion rate The amounts of D-serine, D-asparagine and creatinine in blood and urine were substituted into the following formula to calculate the excretion rate.

number

number

[0075] Evaluation and assessment of pathological condition The logarithmic transformation values ​​of blood D-serine levels and logarithmic transformation values ​​of D-serine excretion rates in subjects with renal disease and subjects without renal disease were plotted on a two-axis coordinate. The non-renal disease group formed a cluster, with the logarithmic mean value of blood D-serine levels being 0.40 and the logarithmic mean value of D-serine excretion rates being 4.12. In this case, the reference range of distance from the mean can be set to the mean value ± 1.96 standard deviations to 0.558. In the renal disease patient group, IGAN was within the reference range, but PA, MGRS, and DM were outside the reference range. DM was found to be useful in differentiation because blood D-serine levels were well separated from the reference range. In addition, in this two-axis plot, the non-renal disease group had a correlation coefficient R 2 = 0.601, which shows high linearity, indicating that regression analysis can be used for pathological analysis (Figure 7). Although it will be necessary to increase the variation of pathological conditions and the number of subjects in the future to improve the accuracy of the analysis, it was confirmed that the combination of the amount of D-serine in blood and the rate of reabsorption and excretion of D-serine in the kidney as an indicator is useful for elucidating pathological mechanisms, drug discovery and treatment research, and as an aid to pathological and differential diagnosis in clinical practice. Equivalent results were also obtained with logarithmic transformed values.

[0076] The logarithmic transformation values ​​of blood D-asparagine levels and logarithmic transformation values ​​of D-asparagine excretion rates in renal disease subjects and non-renal disease subjects were also plotted on a two-axis coordinate (Figure 6). The non-renal disease group formed a cluster, with the logarithmic mean value of blood D-asparagine levels being -1.95 and the logarithmic mean value of D-asparagine excretion rate being 4.16. In this case, the reference range of the distance from the mean can be set to 0.515 from the mean ± 1.96 standard deviations. In the renal disease patient group, IGAN was within the reference range, but PA, MGRS, and DM were outside the reference range. DM was well separated from the reference range of blood D-asparagine levels, indicating its usefulness in differentiation (Figure 6). Furthermore, in this two-axis plot, the non-renal disease group had a correlation coefficient R 2= 0.0002, which shows high linearity, indicating that regression analysis can be used for pathological analysis (Figure 8). Although it will be necessary to increase the variation of pathological conditions and the number of subjects in the future to improve the accuracy of the analysis, it was confirmed that the combination of the amount of D-asparagine in blood and the rate of reabsorption and excretion of D-asparagine in the kidney as an indicator is useful for elucidating pathological mechanisms, drug discovery and treatment research, and as an aid to pathological and differential diagnosis in clinical practice. Equivalent results were also obtained with logarithmic transformed values.

[0077] Monitoring of therapeutic effects The D-serine excretion rate of IGAN administered ARB for hypertension changed from 64.56% to 25.73%, a value below the reference value (Figure 2). In addition, the D-asparagine excretion rate of IGAN administered ARB also changed from 45.71% to 35.39%, a value below the reference value (Figure 2). It was suggested that pathological changes such as blood pressure reduction affect the excretion rate during therapeutic intervention such as drug administration, and the usefulness of D-serine excretion rate and D-asparagine excretion rate in assisting policy decisions such as continuation or discontinuation of treatment in research aimed at elucidating pharmacological mechanisms and drug discovery, as well as in monitoring the effects of therapeutic intervention, was demonstrated. EXAMPLES

[0078] Subject information: A 36-year-old woman was admitted to Osaka University Hospital with systemic lupus erythematosus. After obtaining written informed consent under the ethical approval of the university, blood and urine samples were collected over time. 90 days before admission, serum creatinine level had rapidly deteriorated from 0.57mg / dL to 11.68mg / dL, and urinary protein concentration had rapidly deteriorated from 0.5g / gCre to 4.0g / gCre. Blood pressure was 122 / 65mmHg, heart rate was 64bpm, percutaneous arterial oxygen saturation was 100% (room air), and body temperature was 36.5℃. Mouse ulcers, hair loss, and retinal hemorrhage were noted, but no abnormal lung sounds, heart sounds, or lower limb edema were observed. Laboratory tests showed a blood hemoglobin of 4.6 g / dL, normal levels of complement C3: 88 mg / dL, C4: 21 mg / dL, positive anti-dsDNA antibodies 13.0 IU / mL1, and P-ANCA 182.0 U / mL. Rapidly progressive glomerulonephritis was suspected, so a session of plasma exchange (PE) followed and a renal biopsy was performed. 79% of the glomeruli showed cellular crescents and 13% showed cellular fibrous crescents, and glomerular capillaries were thickened with bubbles and spikes, but no glomerulosclerosis was observed. Interstitial areas showed moderate diffuse infiltration of inflammatory cells, but only slight fibrillation. Tubular atrophy was focal and mild. Immunofluorescence staining was granular and overall glomerular capillary wall positive for IgG, IgA, IgM, C3, C4, and C1q. A diagnosis of crescentic glomerulonephritis potentially associated with ANCA and lupus nephritis class V was made. Pulse prednisolone therapy (1 g for 3 days) was administered, followed by oral prednisolone (40 mg / day) and intermittent pulse intravenous cyclophosphamide therapy (500 mg / m 2 The patient was treated with 500 mg / day of mycophenolate mofetil (MMF) and 8-series plasma exchange. In response to these treatments, the serum creatinine level decreased to 0.72 mg / dL, but the urinary protein level persisted. A follow-up renal biopsy showed regression of the glomerular cellular crescents, but 30% of the glomeruli remained grossly sclerotic, and capillary thickening persisted.

[0079] D-Serine Excretion Rate The collected blood and urine samples were prepared and quantified in the same manner as in Example 1, and the D-serine excretion rate was calculated.

[0080] Evaluation and assessment of pathology, monitoring of therapeutic effects The blood D-serine concentration immediately after admission of an SLE patient was 17.06 nmol / mL, which was an order of magnitude higher than the value of the non-renal group, and this value alone was enough to determine whether the patient's pathology was different. Immediately after the start of treatment, it was 0 (below the standard), 8 days later it was 0 (below the standard), 12 days later it was 0 (below the standard), 16 days later it was 0 (below the standard), 22 days later it was 0 (below the standard), 29 days later it was 58.9% (within the standard), 34 days later it was 87.6% (above the standard), and 48 hours later it was 41.7% (within the standard). As the creatinine level returned to the normal range due to treatment, the D-serine excretion rate transiently increased and fell within the standard calculated in Example 1.

[0081] In renal damage caused by systemic lupus erythematosus, the excretion rate of D-serine was observed to pass through the reference range and rise above the reference range during the progression and regression of acute renal damage (Figure 10). This suggests that the kidney is controlling the excretion rate to defend itself against risk or damage caused by some cause. To further improve the accuracy of the evaluation of the pathology, progress, or improvement or deterioration, information on the blood D-serine level was referenced. This confirmed the usefulness of monitoring the two-axis plot of D-serine excretion rate and blood D-serine level in assisting in pathology, differential diagnosis, evaluation, and treatment decision-making, as to whether D-serine excretion is in a state of combating risk or damage to the disease or in a state of sedation (Figure 4). This information can also be used in research aimed at elucidating pathological and pharmacological mechanisms and developing drugs and treatments. EXAMPLES

[0082] A cohort of patients with renal disease admitted to the Department of Nephrology, Osaka University Hospital for diagnostic and / or therapeutic purposes between 2016 and 2017, including patients with interstitial nephritis (TIN), benign prostatic hyperplasia (BPH), Fabry disease, and minimal change nephrotic syndrome (MCNS), was used in this retrospective study. The study protocol was approved by the ethical committee at Osaka University, and written informed consent was obtained from all subjects.

[0083] D-Serine Excretion Rate The collected blood and urine samples were prepared and quantified in the same manner as in Example 1, and the D-serine excretion rate was calculated.

[0084] The amount of D-serine in the blood and the D-serine excretion rate of each subject were plotted on a two-axis coordinate system together with the renal disease subjects of Example 1 (FIG. 13). The plots had higher resolution than the information on the amount of D-serine in the blood and the D-serine excretion rate of each pathology alone, demonstrating their usefulness in assisting in the differentiation of causes and the evaluation and determination of disease progression.

Claims

1. A method for assisting in the evaluation of renal pathology using as an index a combination of the rate of reabsorption and excretion of D-serine and / or D-asparagine in the kidney of a subject and the amount of D-serine and / or D-asparagine in the blood.

2. The method according to claim 1, wherein the ratio is the excretion rate of D-serine (subject D-serine excretion rate) and / or the excretion rate of D-asparagine (subject D-asparagine excretion rate) into the urine of the subject.

3. The method according to claim 2, wherein the excretion rate of D-serine and / or the excretion rate of D-asparagine are calculated by correcting using a correction factor derived from blood and / or urine.

4. The method of claim 3, wherein the correction factor is one or more correction factors selected from the group consisting of glomerular filtration rate and urine volume.

5. The method of claim 3 , wherein the correction factor is one or more correction factors selected from the group consisting of inulin clearance and creatinine clearance.

6. The method according to claim 3, wherein the correction factor is one or more correction factors selected from the group consisting of creatinine amount and L-amino acid amount.

7. The method of claim 3, wherein the correction factor is L-serine and / or L-asparagine.

8. The excretion rate of the D-serine is calculated by the following formula: [0010] [In the formula, U Ｄ－Ｓｅｒ represents the amount of D-serine in urine, P Ｄ－Ｓｅｒ represents the amount of D-serine in the blood, and U ｃｒｅ represents the amount of creatinine in the urine, and P ｃｒｅ represents the amount of creatinine in the blood.], and / or the excretion rate of D-asparagine is calculated from the following formula: [0025] [In the formula, U Ｄ－Ａｓｎ represents the amount of D-asparagine in urine, P Ｄ－Ａｓｎ represents the amount of D-asparagine in the blood, and U ｃｒｅ represents the amount of creatinine in the urine, and P ｃｒｅ The method according to claim 2 or 3, wherein the creatinine concentration is calculated from the above formula:

9. The method according to any one of claims 2 to 8, comprising the step of comparing first object coordinates obtained by plotting the object D-serine excretion rate and / or object D-asparagine excretion rate in the object and the amount of D-serine and / or D-asparagine in the blood with a first standard calculated from non-renal disease coordinates obtained by plotting the urinary excretion rates of D-serine (non-renal disease object D-serine excretion rate) and / or the excretion rate of D-asparagine (non-renal disease object D-asparagine excretion rate) in a plurality of non-renal disease subjects and the amount of D-serine and / or the amount of D-asparagine in the blood, and evaluating a renal pathological condition based on the relationship between the first object coordinates and the first standard.

10. 10. The method of claim 9, wherein the step of assessing the renal pathology is to assess the subject's renal disease or risk of developing renal disease, or to predict the induction or prognosis of renal disease, when the first object coordinate is not included in the first criterion.

11. The method according to claim 10, wherein the kidney disease is chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease or minimal change nephrotic syndrome.

12. The method according to any one of claims 9 to 11, wherein the first criterion is the range of the mean value ± standard deviation × coefficient Z of the plot of the non-renal disease coordinates.

13. The method of claim 12, wherein the coefficient Z is a value between 1.0 and 3.

0.

14. 14. The method of claim 12 or 13, wherein the coefficient Z is 1.

96.

15. A method for assisting in the evaluation of renal pathology based on the relationship between a regression equation calculated by regression analysis of the plot of the non-renal disease coordinates and the target coordinates.

16. The method according to any one of claims 2 to 8, comprising: comparing a second object coordinate obtained by plotting the logarithmically transformed D-serine excretion rate (subject D-serine LN excretion rate) and / or the logarithmically transformed D-asparagine excretion rate (subject D-asparagine LN excretion rate) of the subject and the logarithmically transformed D-serine amount and / or D-asparagine amount in blood with a second standard calculated from a non-renal disease coordinate obtained by plotting the logarithmically transformed urinary D-serine excretion rate (non-renal disease subject D-serine LN excretion rate) and / or D-asparagine excretion rate (non-renal disease subject D-asparagine LN excretion rate) of a plurality of non-renal disease subjects and the logarithmically transformed D-serine amount and / or D-asparagine amount in blood, and evaluating a renal pathology based on the relationship between the second object coordinate and the second standard.

17. 17. The method of claim 16, wherein the step of assessing the renal pathology is to assess the subject's renal disease or risk of developing renal disease, or to predict the induction or prognosis of renal disease, if the second object coordinate is not included in the second criterion.

18. The method according to claim 17, wherein the kidney disease is chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease or minimal change nephrotic syndrome.

19. The method according to any one of claims 16 to 18, wherein the second criterion is the range of the mean value of the plot of the non-renal disease coordinates ± standard deviation × coefficient Z.

20. 20. The method of claim 19, wherein the coefficient Z is a value between 1.0 and 3.

0.

21. 21. The method of claim 19 or 20, wherein the coefficient Z is 1.

96.

22. 17. The method of claim 16, wherein the second criterion is a distance from the mean value of the plot of the non-renal disease coordinates of 0.6 or less.

23. A method for assisting in the evaluation of renal pathology based on the relationship between a regression equation calculated from the regression line of a plot of non-renal disease coordinates based on the logarithmically transformed values ​​and target coordinates based on the logarithmically transformed values.

24. A method for monitoring a renal pathology, comprising measuring the excretion rate of D-serine (subject D-serine excretion rate) and / or the excretion rate of D-asparagine (subject D-asparagine excretion rate) into the urine of a subject, and the amount of D-serine and / or the amount of D-asparagine in the blood over time, and using the fluctuations in the subject D-serine excretion rate and / or the subject D-asparagine excretion rate and the amount of D-serine and / or D-asparagine in the blood as indicators.

25. 25. The method of claim 24, wherein renal pathology due to chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or renal disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease or minimal change nephrotic syndrome is monitored.

26. A method for monitoring the therapeutic effect of a renal pathology, comprising measuring the urinary excretion rate of D-serine (subject D-serine excretion rate) and / or the excretion rate of D-asparagine (subject D-asparagine excretion rate) of a subject with kidney disease before and after therapeutic intervention and the amount of D-serine and / or D-asparagine in the blood over time, and using the fluctuations in the subject D-serine excretion rate and / or the subject D-asparagine excretion rate and the amount of D-serine and / or D-asparagine in the blood as indicators.

27. 27. The method of claim 26, wherein the kidney disease is chronic kidney disease, myeloma kidney, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or kidney disease caused by systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease, or minimal change nephrotic syndrome.

28. A method for assisting in the evaluation of renal pathology using as an index the amount of D-serine and / or D-asparagine in the blood of a subject from whom urine cannot be collected.

29. 29. The method of claim 28, which aids in the assessment of renal pathology due to chronic kidney disease, myeloma nephropathy, diabetic nephropathy, IgA nephropathy, interstitial nephritis or polycystic kidney disease, or renal disease due to systemic lupus erythematosus, primary aldosteronism, benign prostatic hyperplasia, Fabry disease or minimal change nephrotic syndrome.

30. A method for assisting in determining whether a subject has systemic lupus erythematosus when the D-serine level in the subject's blood is 9 nmol / mL or more.

31. A system for evaluating a renal pathology comprising a memory unit, an input unit, an analysis and measurement unit, a data processing unit, and an output unit, wherein the memory unit stores a threshold value and a calculation formula for a urinary D-serine excretion rate and / or a calculation formula for a D-asparagine excretion rate input from the input unit, the analysis and measurement unit quantifies the amount of D-serine and / or D-asparagine in a blood sample and / or a urine sample, and the data processing unit calculates a urinary D-serine excretion rate and / or a D-asparagine excretion rate generated from an element including the quantified amount of D-serine and / or D-asparagine in the blood sample and / or the urine sample and the calculation formula for the D-serine excretion rate and / or the D-asparagine excretion rate stored in the memory unit, The evaluation system is characterized in that a data processing unit evaluates a renal pathology based on a comparison between a threshold value stored in a memory unit and a combination of the urinary D-serine excretion rate and / or the D-asparagine excretion rate and the blood D-serine amount and / or the D-asparagine amount, and an output unit outputs an evaluation result of the renal pathology of the subject.

32. The D-serine excretion rate is calculated by the following formula: [0030] [In the formula, U Ｄ－Ｓｅｒ represents the amount of D-serine in urine, P Ｄ－Ｓｅｒ represents the amount of D-serine in the blood, and U ｃｒｅ represents the amount of creatinine in the urine, and P ｃｒｅ represents the amount of creatinine in the blood.], and / or the formula for calculating the D-asparagine excretion rate is the following formula: [0045] [In the formula, U Ｄ－Ａｓｎ represents the amount of D-asparagine in urine, P Ｄ－Ａｓｎ represents the amount of D-asparagine in the blood, and U ｃｒｅ represents the amount of creatinine in the urine, and P ｃｒｅ represents the amount of creatinine in the blood.

33. A program for causing an information processing device including an input unit, an output unit, a data processing unit, and a storage unit to evaluate a renal pathology, the program comprising: storing in the storage unit a threshold value for evaluating a renal pathology, a calculation formula for a urinary D-serine excretion rate and / or a calculation formula for a D-asparagine excretion rate, and variables required for the calculation, which are input from the input unit; storing in the storage unit variables required for calculating the amount of D-serine and / or the amount of D-asparagine in a blood sample and / or a urine sample and the D-serine excretion rate and / or the D-asparagine excretion rate in urine, which are input from the input unit; the data processing unit to call up a formula for calculating the urinary D-serine excretion rate and / or a formula for calculating the D-asparagine excretion rate, which are stored in advance in the storage unit, and the amounts of D-serine and / or D-asparagine in the blood sample and / or urine sample, and the variables, which are stored in the storage unit, and substitute the variables into the formula for calculating the urinary D-serine excretion rate and / or the formula for calculating the D-asparagine excretion rate; the data processing unit to evaluate a renal pathology based on a comparison of the threshold value stored in the storage unit with a combination of the D-serine excretion rate and / or D-asparagine excretion rate into the urine and the amount of D-serine and / or D-asparagine in the blood; and the program comprising instructions for causing the information processing device to execute the following:

34. The calculation formula for the D-serine excretion rate is the following formula: [0050] [In the formula, U Ｄ－Ｓｅｒ represents the amount of D-serine in urine, P Ｄ－Ｓｅｒ represents the amount of D-serine in the blood, and U ｃｒｅ represents the amount of creatinine in the urine, and P ｃｒｅ represents the amount of creatinine in the blood.], and / or the formula for calculating the D-asparagine excretion rate is the following formula: [006] [In the formula, U Ｄ－Ａｓｎ represents the amount of D-asparagine in urine, P Ｄ－Ａｓｎ represents the amount of D-asparagine in the blood, and U ｃｒｅ represents the amount of creatinine in the urine, and P ｃｒｅ represents the amount of creatinine in the blood.

Images