WO2016039426A1

WO2016039426A1 - Method for assisting in selection of therapeutic drug for type-2 diabetes patient, method for predicting effect of therapeutic drug, inspection method, and treatment method

Info

Publication number: WO2016039426A1
Application number: PCT/JP2015/075760
Authority: WO
Inventors: 博文御簾; 周一金子; 有美枝竹下; 俊成篁; 芳郎斎藤; 睦田中
Original assignee: 国立大学法人金沢大学; 学校法人同志社; アルフレッサファーマ株式会社
Priority date: 2014-09-10
Filing date: 2015-09-10
Publication date: 2016-03-17
Also published as: JP6736011B2; JPWO2016039426A1

Abstract

　The present invention addresses the problem of providing a method for assisting in selection of a therapeutic drug for a type-2 diabetes patient, a method for predicting the effect of a therapeutic drug, an inspection method, and a treatment method. The inventors discovered that the effect of administering metformin is high when the selenoprotein P value is higher than a cutoff value in a specimen (particularly blood) originating from a type-2 diabetes patient, and thereby developed the present invention.

Description

Method for assisting selection of therapeutic agent for type 2 diabetes patient, method for predicting effect of therapeutic agent, test method, and therapeutic method

The present invention relates to a method for assisting selection of a therapeutic agent for type 2 diabetes, which comprises selecting metformin hydrochloride administration when a selenoprotein P value is higher than a cutoff value in a specimen derived from a type 2 diabetes patient The present invention relates to a method for predicting an effect of a therapeutic agent, a test method, and a treatment method.
This application claims the priority of Japanese Patent Application No. 2014-184499, which is incorporated herein by reference.

(Type 2 diabetes)
Type 2 diabetes, which continues to increase on a global scale, threatens human QOL and life by promoting arteriosclerotic diseases such as retinal, renal and neurological complications and ischemic heart disease. The development of is urgent. The liver not only plays a major role in glucose and lipid metabolism, but is the largest organ in the body of various biologically active substances including angiogenic factors. In type 2 diabetes, the action of insulin to suppress sugar release from the liver is attenuated, and this phenomenon is called insulin resistance. Insulin resistance leads to hyperglycemia due to increased glucose release from the liver and hyperlipidemia due to increased lipid production, and promotes arteriosclerotic diseases. Furthermore, the liver is the largest producing organ in vivo of various physiologically active substances including angiogenic factors that lead to the risk of arteriosclerosis (see: Patent Document 1).

(Selenoprotein P (SeP))
Selenoprotein P (SeP) is an extracellular protein rich in selenium and is the major selenoprotein that accounts for 53% of plasma selenium. Six types of monoclonal antibodies (BD1, BD3, BF2, AE2, AH5, and AA3) are known for SeP, and an enzyme-linked immunosorbent assay (ELISA) using two types of monoclonal antibodies has been developed as a method for measuring SeP. . SeP undergoes degradation with plasma kallikrein to produce N-terminal and C-terminal fragments, and among the above six monoclonal antibodies, BD1, BD3, BF2, AE2 and AH5 become N-terminal fragments. In contrast, AA3 reacts specifically with the C-terminal fragment. As confirmed by Western blot analysis using AA3, in plasma, there may be a full-length protein of SeP and a fragment protein produced by cleavage by degradation with plasma kallikrein. In order to promote clinical research, there is a need for a method that can easily measure the presence or degree of SeP degradation by plasma kallikrein in vivo (see: Patent Document 2).

(Prior Patent Literature)
Patent Document 1 discloses “a method for detecting type 2 diabetes, including measuring selenoprotein P”. However, Patent Document 1 does not disclose or suggest a “method for selecting a therapeutic agent for type 2 diabetes using selenoprotein P value as an index”.
Patent Document 2 states that “a method for separately measuring a long-chain type and a short-chain type of a substance to be measured (selenoprotein P) in a sample includes (a) the sample, the long-chain type, and the short-chain type. Mixing microparticles bound to a first specific binding substance capable of binding and microparticles bound to a second specific binding substance that can bind to the long chain type but not to the short chain type, and A step of measuring an agglutination reaction of microparticles; (b) a microparticle having a third specific binding substance bound to the specimen, and a third specific binding substance capable of binding to the long chain type and the short chain type at different recognition sites; A step of mixing the microparticles bound with the specific binding substance and measuring the aggregation reaction of the microparticles; (c) determining the amount of the long chain type using the measurement value of (a); And (d) determining the amount of the short chain using a value obtained by subtracting the measured value of (a) from the measured value of (b). Is disclosed. However, Patent Document 2 does not disclose or suggest a “method for selecting a therapeutic agent for type 2 diabetes using selenoprotein P value as an index”.

(Non-patent literature)
Non-Patent Document 1 states that “selenoprotein P expression is suppressed by treatment with metformin, but this suppression is inhibited by co-administration of an AMPK inhibitor or FoxO3a siRNA. Metformin is mediated through an AMPK inhibitor / FoxO3a pathway. In other words, the expression of selenoprotein P is suppressed. " However, Non-Patent Document 1 does not disclose or suggest a “method for selecting a therapeutic agent for type 2 diabetes using selenoprotein P value as an index”.

International Publication No. 2008/013324 JP 2014-52338 A

In the treatment of type 2 diabetes, it is left to the discretion of the attending physician whether metformin is the first-line treatment or a DPP4 (dipeptidyl peptidase-4) inhibitor is the first-line treatment. However, patients with type 2 diabetes have large individual differences in treatment responsiveness depending on the therapeutic agent to be selected, and there are many cases in which there is no effect.
Metformin has little evidence of hypoglycemia, and there is evidence of life extension. However, metformin has been reported to have anticancer effects, gastrointestinal side effects (diarrhea, decreased appetite), and the like, and there are considerable individual differences in treatment response.
In addition, it has been reported that DPP4 inhibitors have an effect of stimulating insulin secretion and there are individual differences in blood glucose lowering action.
Based on the above, there has been a demand for a method that can objectively determine whether metformin is a first-line treatment or a DPP4 inhibitor is a first-line treatment.
That is, an object of the present invention is to provide a method for assisting selection of a therapeutic drug for type 2 diabetes patients, a method for predicting the effect of the therapeutic drug, a test method, and a therapeutic method.

In order to solve the above-mentioned problems, the present inventors have described that “in specimens (especially blood) derived from type 2 diabetes, the effect of metformin administration is high when the selenoprotein P value is higher than the cutoff value. And the present invention has been completed.

That is, this invention consists of the following.
1.2 A method for assisting selection of a therapeutic agent for type 2 diabetes, wherein in a specimen derived from a type 2 diabetes patient, metformin hydrochloride administration is selected when the selenoprotein P value is higher than the cutoff value.
2. 2. The method for assisting selection of a therapeutic agent according to item 1, wherein the specimen is blood.
3. 3. The method for assisting selection of a therapeutic agent according to

item

1 or 2, wherein the cutoff value is 3.5 μg / mL to 4.7 μg / mL.
4. Predicting the therapeutic effect of type 2 diabetes, characterized in that metformin hydrochloride administration is judged to be effective when the selenoprotein P value is higher than the cut-off value in specimens derived from type 2 diabetes patients how to.
5. 5. The method for predicting a therapeutic effect according to item 4, wherein the specimen is blood.
6). 6. The method for predicting a therapeutic effect according to

item

4 or 5, wherein the cut-off value is 4.5 μg / mL or more.
7. Type 2 for selecting a therapeutic agent for type 2 diabetes, characterized by selecting metformin hydrochloride administration when the selenoprotein P value is higher than the cut-off value in a sample derived from a type 2 diabetes patient A method for testing specimens from diabetic patients.
8). 8. The inspection method according to item 7, wherein the specimen is blood.
9. 9. The inspection method according to item 7 or 8, wherein the cutoff value is 3.5 μg / mL to 4.7 μg / mL.
10. A method for treating type 2 diabetes comprising the following steps:
(1) a step of determining whether a selenoprotein P value is higher than a cut-off value in a sample derived from a type 2 diabetes patient; and (2) metformin if the selenoprotein P value is higher than a cut-off value. Administering hydrochloride to the patient.
11. The treatment method according to item 10, wherein the cut-off value is 3.5 μg / mL to 4.7 μg / mL.

The present invention relates to a method for assisting selection of a therapeutic agent for type 2 diabetes, which comprises selecting metformin administration when a selenoprotein P value is higher than a cut-off value in a specimen derived from a type 2 diabetes patient. A drug effect prediction method, a test method, and a treatment method can be provided.
As a result, the therapeutic effect of metformin administration can be predicted before administration, and a high therapeutic effect can be obtained by administering metformin from an early stage to patients with type 2 diabetes who are expected to have an effect of metformin administration. it can.

Schematic diagram showing cleavage of selenoprotein P (SeP) by plasma kallikrein. A graph (calibration curve) showing the relationship between the SeP concentration and the amount of change in absorbance in the FL-SeP measurement system. The vertical axis represents the amount of change in absorbance, and the horizontal axis represents the SeP concentration (μg / mL). The graph which shows the change of a blood glucose level in a metformin administration group, and the change of a HbA1c value (using the Pearson product-moment correlation coefficient). The graph which shows the change of the blood glucose level in a DPP4 inhibitor administration group, and the change of a HbA1c value (using the Pearson product-moment correlation coefficient). The graph which shows the change of the selenoprotein P value in a metformin administration group and a DPP4 inhibitor administration group (using the Pearson product-moment correlation coefficient). Changes in blood glucose levels in the metformin administration group and the DPP4 inhibitor administration group (using Pearson product moment correlation coefficient). Change in selenoprotein P level in patients with high pre-dose SeP level (4.55 μg / mL or more) in the metformin group. Distribution map of selenoprotein P level in serum of type 2 diabetic patients before treatment. Changes in blood glucose levels in the metformin administration group and the DPP4 inhibitor administration group in cases with a selected cut-off value or higher.

According to the present invention, “in a specimen derived from a type 2 diabetes patient, when the selenoprotein P value is higher than the cut-off value, the administration of metformin hydrochloride is selected. Method "," Method for predicting therapeutic effect, characterized in that metformin hydrochloride administration is judged to be effective when the selenoprotein P value is higher than the cut-off value in a specimen derived from a type 2 diabetic patient " “In a specimen derived from a type 2 diabetic patient, if the selenoprotein P value is higher than the cut-off value, the administration of metformin hydrochloride is selected. Method for testing specimens from type 2 diabetic patients "and" In the case of specimens from type 2 diabetic patients, if the selenoprotein P value is higher than the cutoff value, metformin A therapeutic method comprising administering hydrochloride to the patient. This will be described in detail below.

(Assistive method for selection of treatment for type 2 diabetes)
The method for assisting selection of a therapeutic agent for type 2 diabetes according to the present invention includes the following steps.
(1) A step of determining whether a selenoprotein P value is higher than a cutoff value in a sample derived from a type 2 diabetes patient.
(2) A step of determining that metformin hydrochloride administration is selected when the selenoprotein P value is higher than the cutoff value.

(Method for predicting the therapeutic effect of metformin hydrochloride administration on type 2 diabetes)
The method for predicting the therapeutic effect of metformin hydrochloride administration of type 2 diabetes according to the present invention includes the following steps.
(1) A step of determining whether a selenoprotein P value is higher than a cutoff value in a sample derived from a type 2 diabetes patient.
(2) A step of determining that metformin hydrochloride administration is effective when the selenoprotein P value is higher than the cut-off value.
“Effective” means that administration of metformin hydrochloride has an effect of improving blood glucose level, improving HbA1c level, lowering SeP level in blood, and / or treating diabetes.

(Test method for specimens from patients with type 2 diabetes to select therapeutic agents for type 2 diabetes)
The method for examining a specimen derived from a type 2 diabetes patient for selecting a therapeutic agent for type 2 diabetes according to the present invention includes the following steps.
(1) A step of determining whether a selenoprotein P value is higher than a cutoff value in a sample derived from a type 2 diabetes patient.
(2) A step of determining that metformin hydrochloride administration is selected when the selenoprotein P value is higher than the cutoff value.

(Method for treating type 2 diabetes)
The method for treating type 2 diabetes according to the present invention includes the following steps.
(1) A step of determining whether a selenoprotein P value is higher than a cutoff value in a sample derived from a type 2 diabetes patient.
(2) A step of administering metformin hydrochloride to the patient when the selenoprotein P value is higher than the cutoff value.
Furthermore, the following processes are included as needed.
(3) A step of changing to DPP4 inhibitor administration or a combination of DPP4 inhibitor when metformin hydrochloride administration does not improve patient symptoms.

(Sample)
The specimen of the present invention is not particularly limited as long as it is derived from a type 2 diabetic patient and selenoprotein P is present. Examples thereof include whole blood, blood (peripheral mononuclear cells), serum, plasma, urine, cerebrospinal fluid, ascites, lymph, liver-derived fluid, and milk. In particular, peripheral mononuclear cells, serum and the like are most preferable in terms of easy handling such as collection and low invasiveness. The method for obtaining a sample from a type 2 diabetes patient is not particularly limited, and a method known per se can be used.

(Selenoprotein P)
Selenoprotein P (SeP) is a protein containing 10 residues of selenocysteine. Selenoprotein P acts as an enzyme having glutathione peroxidase-like activity that reduces hydrogen peroxide and lipid peroxide to detoxify and controls intracellular redox.
In addition, the present inventors have disclosed that selenoprotein P is a detection marker for type 2 diabetes (see: Patent Document 1).
Selenoprotein P is contained in human serum and can be isolated and purified from human serum according to the method described in the literature “Saito Y. et al., J Biol chem 274: 2866-2871, 1999”. it can.
Selenoprotein P (SeP) is cleaved by plasma kallikrein (see: FIG. 1). The upper part of FIG. 1 represents full-length SeP, and the middle and lower parts represent SeP fragments generated by plasma kallikrein cleavage. In FIG. 1, “N” and “C” represent the N-terminal side and the C-terminal side, respectively, and “Sec” represents a selenocysteine residue. Kallikrein consists of arginine at position 235 (“R235” in FIG. 1) and glutamine at position 236 (“Q236” in FIG. 1), and arginine at position 242 (“R242” in FIG. 1) and asparagine at position 243. Cleaves with the acid (“D243” in FIG. 1), resulting in three fragments including an N-terminal fragment of SeP (amino acid residues 1 to 235) and a C-terminal fragment (amino acid residues 243 to 361) Obtain (Figure 1). The “long chain type” is an uncut full length SeP (amino acid residues 1 to 361: sometimes referred to as “FL-SeP” in this specification), and the “short chain type” is generated by cleavage. It is a SeP fragment (sometimes referred to herein as “S-SeP”), and the SeP fragment can be, for example, an N-terminal fragment (amino acid residues 1 to 235).

(Metformin hydrochloride)
Metformin hydrochloride (Chemical name: 1,1-Dimethylbiguanide monohydrochloride) is “Metgluco Tablets”, “Glycolane Tablets”, “Metformin Hydrochloride Tablets”, and other brand names from several pharmaceutical companies as a treatment for type 2 diabetes. It is sold (hereinafter referred to as “Metogluco Tablet”).
In general, for adults, for adults, start with metformin hydrochloride at 500 mg per day, orally in 2 to 3 divided doses, orally before or after a meal. The maintenance dose is determined while observing the effect, but is usually 750 mg to 1,500 mg per day. The dose may be adjusted according to the patient's condition, but the maximum daily dose is 2,250 mg.

(DPP4 inhibitor)
DPP4 inhibitor is an antidiabetic drug, which means a drug that inhibits the DPP4 (dipeptidyl peptidase-4) enzyme, and the generic names sitagliptin, vildagliptin, alogliptin, linagliptin, teneligliptin and anagliptin are known .
For example, the dosage and administration timing can be exemplified as follows, but are not particularly limited.
○ Sitagliptin Normally, for adults, 50 mg of sitagliptin is orally administered once a day. If the effect is insufficient, 100 mg can be increased once a day while observing the progress sufficiently.
○ Vildagliptin Usually, for adults, 50 mg of vildagliptin is orally administered twice a day in the morning and evening. Depending on the patient's condition, 50 mg can be administered once a day in the morning.
○ Alogliptin Usually, for adults, 25 mg of alogliptin is orally administered once a day, preferably after breakfast.
○ Linagliptin Usually, for adults, 5 mg of linagliptin is orally administered once a day.
○ Tenerigliptin Usually, for adults, 20 mg of tenerigliptin is orally administered once a day. If the effect is insufficient, 40 mg can be increased once a day while observing the progress sufficiently.
○ Anagliptin Normally, for adults, 100 mg of anagliptin is orally administered twice a day in the morning and evening. If the effect is insufficient, the single dose can be increased to 200 mg while observing the progress sufficiently.

(Cutoff value for selection of treatment for type 2 diabetes patients)
The range of the cut-off value for selecting a therapeutic agent for patients with type 2 diabetes according to the present invention is 3.5 μg / mL to 4.7 μg / mL, preferably 3.7 μg in the specimen (particularly serum) from Example 4 below. / mL to 4.5 μg / mL, more preferably 3.9 μg / mL to 4.3 μg / mL, and most preferably about 4.1 μg / mL.

(Cutoff value for determining that metformin hydrochloride administration is effective)
The range of the cut-off value for judging that the administration of metformin hydrochloride of the present invention is effective is 4.5 μg / mL or more, for example, 4.5 μg / mL in the sample (particularly in serum) from Example 3 below. It is ˜6.0 μg / mL, more preferably 4.5 μg / mL to 5.5 μg / mL, and most preferably 4.5 μg / mL to 5.0 μg / mL. One indicator that metformin hydrochloride administration was effective is a significant decrease in blood selenoprotein P level.

(Measurement method of selenoprotein P value)
The method for assisting selection of a therapeutic agent for type 2 diabetes patients, the method for predicting the effect of a therapeutic agent, the method for measuring the selenoprotein P value in the test method and the therapeutic method of the present invention are not particularly limited. Alternatively, selenoprotein P mRNA may be calculated by measuring mRNA of selenoprotein P.
As a method of directly measuring selenoprotein P, it can be performed by an immunoassay using an anti-selenoprotein P antibody that specifically recognizes and binds to selenoprotein P. Anti-selenoprotein P antibody can be prepared by a known method. Examples of the immunological measurement method include a method using a carrier on which an anti-selenoprotein P antibody is immobilized, western blotting, and the like. Examples of the method using a solid-phase support include, but are not limited to, ELISA using a solid-phase microtiter plate, and an aggregation method using solid-phase particles. Adopted, blood selenoprotein P can be measured.
The selenoprotein P mRNA can be measured by Northern blotting, RT-PCR, a method using a DNA chip (DNA microarray), or the like. These methods can also be performed by a known method.

The method for measuring selenoprotein P is preferably an enzyme-linked immunosorbent method using two types of monoclonal antibodies {Reference: Saito, Y. et al., J. Health Sci. (2001) 47, 346-352}, Examples thereof include a measurement method using an antigen-antibody reaction using a gold colloid shown in the following Examples (see Patent Document 2).

The present invention will be specifically described with reference to the following examples, but the present invention is not limited thereto. The following cases have been implemented with the approval of the Kanazawa University Medical Ethics Committee.

(Construction of selenoprotein P level measurement system in specimen)
A measurement system for measuring the selenoprotein P value of the specimen was constructed. Details are as follows.

(Preparation Example 1: Preparation of colloidal gold solution)
To 1 L of distilled water at 95 ° C., 2 mL of a 10 w / v% aqueous chloroauric acid solution was added with stirring, and after 1 minute, 10 mL of a 2 w / v% aqueous sodium citrate solution was added, stirred for another 20 minutes, and then cooled to 30 ° C. After cooling, the pH was adjusted to 7.1 with a 0.1 w / v% aqueous potassium carbonate solution.

(Preparation Example 2: Preparation of various anti-selenoprotein P antibody-binding gold colloid reagents)
Anti-human selenoprotein P rat monoclonal antibodies AH5 and AA3 were obtained according to the procedure described in the literature {Saito, Y. et al., J. Health Sci. (2001) 47, 346-352}. AH5 is a monoclonal antibody that recognizes the N-terminal side of selenoprotein P generated by kallikrein cleavage of selenoprotein P, and AA3 is a monoclonal antibody that recognizes the C-terminal side of selenoprotein P.

For anti-selenoprotein P monoclonal antibodies AH5 and AA3, antibody-bound gold colloid reagents were prepared as follows. The antibody was diluted with 10 mM 2- [4- (2-hydroxyethyl) -1-piperazinyl] ethanesulfonic acid (hereinafter “HEPES”) buffer (pH 7.1) containing 0.05 w / v% sodium azide. To a concentration of 50 μg / mL. 100 mL of the obtained antibody solution was added to about 1 L of the gold colloid solution prepared in Preparation Example 1 and stirred for 2 hours under refrigeration. Further, 110 mL of 10 mM HEPES buffer (pH 7.1) containing 5.46 w / v% mannitol, 0.5 w / v% bovine serum albumin (BSA), and 0.05% sodium azide was added, and the temperature was changed to 37 ° C. And stirred for 90 minutes. Next, the mixture was centrifuged at 8000 rpm for 40 minutes, and the supernatant was removed. Next, about 5 mM HEPES buffer (pH 7.5) (A solution) containing 3 w / v% mannitol, 0.1 w / v% BSA, and 0.05 w / v% sodium azide was added to the resulting precipitate. 1 L was added to disperse the antibody-bound colloidal gold particles, followed by centrifugation at 8000 rpm for 40 minutes to remove the supernatant. Further, A solution containing 0.9% sodium dextran sulfate was added to the precipitate to disperse the antibody-bound gold colloidal particles to a total volume of 240 mL, which was recovered as an antibody-bound gold colloid reagent.

The colloidal gold reagent AH5 and colloidal gold reagent AA3 were used as colloidal gold reagent 1 and colloidal gold reagent 2, respectively.

(Preparation Example 3: Preparation of selenoprotein P measurement reagent)
5 w / v% sodium chloride, 1.0 w / v% ethylenediaminetetraacetic acid dihydrogen disodium dihydrate, 0.1 w / v% alkylphenyl disulfonic acid sodium salt, and 0.5 w / v% polyoxyethylene lauryl ether 0.25M 2-amino-2-hydroxymethyl-1,3-propanediol hydrochloric acid buffer solution (pH 7.8) containing polyethylene glycol in an amount of 2.4 w / v% for the following “FL-SeP measurement” To obtain a reagent.

(Design of FL-SeP measurement system)
For measurement of full-length SeP ("FL-SeP"), measurement using gold colloid particles bound with antibodies recognizing the N-terminal side of selenoprotein P and gold colloid particles bound with antibodies recognizing the C-terminal side A system was designed (also referred to herein as “FL-SeP measurement system”). In the FL-SeP measurement system, only FL-SeP can participate in the agglutination reaction.

(Preparation of calibration curve for FL-SeP measurement system)
Purified SeP (FL-SeP) was purified from human plasma as described in the literature {Saito, Y. et al., J. Biol. Chem. (1999) 274, 2866-2871}.
The purified SeP was added to a standard matrix (3 w / v) at concentrations of 0.0 μg / mL, 0.75 μg / mL, 1.5 μg / mL, 3.0 μg / mL, 6.0 μg / mL, and 9.0 μg / mL, respectively. FL-SeP standard solution prepared in 50 mM 2- [bis (2-hydroxyethyl) amino] -2- (hydroxymethyl) propane-1,3-diol buffer (pH 6.5)) containing% BSA did. 170 μL of the FL-SeP measurement reagent of Preparation Example 3 is dispensed into 3 μL of the obtained FL-SeP standard solution, heated at 37 ° C. for about 5 minutes, and then gold colloid reagent 1 and gold colloid reagent 2 are added. 85 μL of the mixture was dispensed and reacted at 37 ° C. Next, the obtained reaction solution was subjected to a Hitachi 7070 automatic analyzer, and the change in absorbance from the photometry point 18 to 31 was measured at a main wavelength of 505 nm and a sub wavelength of 660 nm, and the change in absorbance at 505 nm was changed to the change in absorbance at 660 nm. The value obtained by adding the absolute value was determined and used as the amount of change in absorbance.
A calibration curve showing the relationship between the SeP concentration and the amount of change in absorbance in the FL-SeP measurement system was prepared (see FIG. 2).

(Therapeutic drug administration for type 2 diabetes patients)
The background of patients with type 2 diabetes and the method of administering each therapeutic agent in this example are as follows.

(Background of type 2 diabetes patients)
The background of type 2 diabetes patients is shown in the table below.

(Method of administration of therapeutic drugs for patients with type 2 diabetes)
Type 2 diabetics 79 patients were assigned to DPP4 inhibitor Alogliptin (Neshina ^TM) 25 mg / after breakfast oral group with metformin hydrochloride (Metoguruko ^®) 1,000 mg / breakfast and after dinner oral group randomly. At the discretion of the attending physician, it was possible to increase the amount of Metogluco up to 2,250 mg / day. Changes in blood glucose levels and HbA1c (hemoglobin, A1 sea) levels before and after oral administration of each therapeutic drug for 3 months are measured by a method known per se, and changes in serum selenoprotein P levels are measured by the methods described below. did.

Serum 3 μL obtained from 79 patients with type 2 diabetes by a method known per se is subjected to the FL-SeP measurement system described in Example 1, and the seleno in the serum is obtained with reference to a calibration curve (see FIG. 2). Protein P value was measured.

(Analysis of selenoprotein P value in specimens of patients with type 2 diabetes)
The change in blood glucose level, the change in HbA1c value, and the change in serum selenoprotein P value were analyzed before and after oral administration of each therapeutic drug obtained in Example 2 for 3 months. Details are as follows.

(Changes in blood glucose levels and changes in HbA1c levels in the metformin administration group)
FIG. 3 shows changes in blood glucose level and changes in HbA1c value in the metformin administration group. As is apparent from the graph shown in FIG. 3, in the metformin group, the SeP level lowering degree and the blood glucose level lowering degree and the SeP level lowering degree and the HbA1c level lowering degree for three months tended to correlate. Furthermore, in the metformin group, the blood glucose level tended to improve well in cases where the SeP level decreased well.

(Change in blood glucose level and change in HbA1c level in the DPP4 inhibitor administration group)
FIG. 4 shows changes in blood glucose level and changes in HbA1c value in the DPP4 inhibitor administration group. As is clear from the graph shown in FIG. 4, in the DPP4 inhibitor administration group, the SeP value lowering degree and the blood sugar level lowering degree and the SeP value lowering degree and the HbA1c lowering degree for three months were not correlated.

(Changes in selenoprotein P levels in the metformin administration group and the DPP4 inhibitor administration group)
FIG. 5 shows changes in SeP values in the metformin administration group and the DPP4 inhibitor administration group. As is apparent from the graph shown in FIG. 5, in the metformin administration group, the SeP value before administration correlated with the degree of decrease in SeP value for 3 months after administration. However, in the DPP4 inhibitor administration group, the SeP value before administration did not correlate with the degree of decrease in SeP value for 3 months after administration. That is, in cases where the SeP value before the start of treatment was high, the SeP value was further reduced by administration of metformin.

(Changes in blood glucose levels in the metformin administration group and the DPP4 inhibitor administration group)
FIG. 6 shows changes in blood glucose levels in the metformin administration group and the DPP4 inhibitor administration group. As is clear from the graph shown in FIG. 6, in the metformin administration group, the SeP value before administration correlated with the blood glucose level decrease for 3 months after administration. However, in the DPP4 inhibitor administration group, the SeP level before administration did not correlate with the degree of blood glucose decrease for 3 months after administration. That is, the blood SeP value before administration of the therapeutic agent was able to predict the blood glucose level lowering level only after metformin administration.

(Changes in selenoprotein P level in patients with high pre-dose SeP levels in the metformin group)
FIG. 7 shows changes in the SeP value in cases where the pre-administration SeP value in the metformin administration group was high (4.55 μg / mL or more). As is clear from the graph shown in FIG. 7, the blood SeP value was significantly reduced by administration of metformin for 3 months.

From the results of the graphs shown in FIGS. 3 to 7, when the SeP level in the blood of type 2 diabetes patients is high, the blood glucose level and HbA1c level are improved by the administration of metformin, and type 2 diabetes in the blood is further improved. It was confirmed that the SeP value, which is a detection marker, was significantly reduced.

(Set cutoff value to select metformin administration for type 2 diabetes patients)
A cut-off value was set for selecting metformin administration for patients with type 2 diabetes. Details are as follows.

(Selection from selenoprotein P value distribution in serum of type 2 diabetic patients before treatment)
The SeP value distribution in the serum of type 2 diabetic patients before treatment is shown in FIG. From the distribution chart shown in FIG. 8, 4.1 μg / mL, which is higher than the median, was selected as the cutoff value.

(Verify selected cutoff value)
FIG. 9 shows changes in blood glucose levels in the metformin administration group and the DPP4 inhibitor administration group in cases with a selected cutoff value of 4.1 μg / mL or more. As is apparent from the results of the graph shown in FIG. 9, the blood glucose level was significantly improved in the metformin administration group. That is, the SeP value in the serum of type 2 diabetic patients before treatment is 4.5 μg / mL or more, for example, 4.5 μg / mL to 6.0 μg / mL, more preferably 4.5 μg / mL to 5.5 μg / mL, most preferably In the case of 4.5 μg / mL to 5.0 μg / mL, it was confirmed that metformin administration improved blood glucose levels (diabetes treatment effect).

(General)
According to the above examples, when the selenoprotein P level in the serum of type 2 diabetic patients before treatment is higher than the cutoff value, metformin administration improves blood glucose level, improves HbA1c level, and decreases blood SeP level. Furthermore, it can be predicted that there is the effectiveness of diabetes treatment.

In specimens from patients with type 2 diabetes, if the selenoprotein P value is higher than the cutoff value, metformin administration is selected. Providing methods, testing methods and treatment methods.

Claims

A method for assisting selection of a therapeutic agent for type 2 diabetes, wherein in a sample derived from a type 2 diabetes patient, if the selenoprotein P value is higher than the cut-off value, metformin hydrochloride administration is selected.
The method for assisting selection of a therapeutic agent according to claim 1, wherein the specimen is blood.
The method for assisting selection of a therapeutic agent according to claim 1 or 2, wherein the cutoff value is from 3.5 µg / mL to 4.7 µg / mL.
A method for predicting the therapeutic effect of type 2 diabetes characterized by determining that metformin hydrochloride administration is effective when a selenoprotein P value is higher than a cutoff value in a sample derived from a type 2 diabetes patient .
The method for predicting a therapeutic effect according to claim 4, wherein the specimen is blood.
The method according to claim 4 or 5, wherein the cutoff value is 4.5 µg / mL or more.
In a sample derived from a type 2 diabetes patient, if the selenoprotein P value is higher than the cut-off value, the administration of metformin hydrochloride is selected. A method for testing specimens of origin.
The test method according to claim 7, wherein the specimen is blood.
The inspection method according to claim 7 or 8, wherein the cutoff value is 3.5 μg / mL to 4.7 μg / mL.
A method for treating type 2 diabetes comprising the following steps:
(1) a step of determining whether a selenoprotein P value is higher than a cut-off value in a sample derived from a type 2 diabetes patient; and (2) metformin if the selenoprotein P value is higher than a cut-off value. Administering hydrochloride to the patient.
The treatment method according to claim 10, wherein the cut-off value is 3.5 μg / mL to 4.7 μg / mL.