WO2014104305A1

WO2014104305A1 - Diagnosis of inflammatory bowel disease by measuring presepsin

Info

Publication number: WO2014104305A1
Application number: PCT/JP2013/085128
Authority: WO
Inventors: 白川　嘉門
Original assignee: 持田製薬株式会社
Priority date: 2012-12-28
Filing date: 2013-12-27
Publication date: 2014-07-03
Also published as: JPWO2014104305A1; JP6450594B2

Abstract

A method for the detection of an inflammatory bowel disease being unresponsive to biological preparations; a method for selecting a patient of an inflammatory bowel disease being unresponsive to biological preparations; a method for determining the completion timing of the administration of an antibiotic, etc. to a patient of an inflammatory bowel disease, said patient being under the administration of the antibiotic etc.; or a method for treating a patient of an inflammatory bowel disease being unresponsive to biological preparations, said method comprising administering an antibiotic, etc., each method using the concentration of sCD14-ST (also known as presepsin) in a blood specimen as an index.

Description

Diagnosis of inflammatory bowel disease by preceptin measurement

The present invention relates to a method for detecting inflammatory bowel disease unresponsive to a biologic by sCD14-ST (also known as preceptin), a method for selecting a patient with inflammatory bowel disease unresponsive to a biologic, an antibiotic, etc. (An antibiotic and / or a new therapeutic agent for inflammation (for example, anti-CD14 antibody fusion protein, particularly MR1007). The same shall apply hereinafter.) A method for selecting a patient with inflammatory bowel disease to be administered, Alternatively, the present invention relates to a method for determining the end timing of administration of the antibiotics or the like in patients with inflammatory bowel disease to which antibiotics or the like are administered.

Inflammatory bowel disease (IBD: Inflammatory Bowel Disease) is a general term for chronic diseases that cause inflammation of unknown origin mainly in the gastrointestinal tract. A distinction is made between CD (Chron Disease) and ulcerative colitis (UC) where the lesion is confined to the large intestine (Non-patent Document 1).

This disease has the characteristic that inflammation is chronically repeated, and as a result of repeated remission and relapse, it causes trait abnormalities such as colon cancer and colon perforation. In this state, medical treatment at the inflammatory stage is the most important issue because the large intestine excision by surgery is finally necessary.

Common medical treatments include nutrition therapy and drug therapy, which may be used in combination. In pharmacotherapy, 5-ASA (5-aminosalicylic acid) preparations (Pentasa ^(R) , Asacol ^(R) , Salazopine ^(R), etc.), which are anti-inflammatory agents, are the first-line drugs. In the case where a steroid cannot be obtained, a steroid (prednisolone) having a stronger anti-inflammatory action may be used in combination. In severe cases and intractable cases that cannot be clearly improved by these treatments, biological preparations (anti-TNFα antibody Remicade ^(R) , Humira ^(R) ; soluble TNF receptor embrel ^(R) ) are used. .

However, there are patients who do not respond to biologics and have no therapeutic effect, and patients who have been highly effective at the start of treatment, but the effect decreases with long-term use. Infliximab (Remicade ^(R)) or adalimumab (Humira ^(R)) Anti-TNFα antibody therapy for a long time (2-90 months, median 15 months) using the cumulative loss rate of the reactive when done at 1 year There is a report that it was 17%, 38% in 3 years, 59% in 5 years (Non-patent Document 2). In addition, in patients who used Humira for a long time, 70% (16 of 23 cases) eventually stopped using (6 invalid cases, 8 diminished effects, 1 side effect, 1 remission after 6 months) There is also a report that the cumulative cancellation rate was 50% at 6 months, 65% at 12 months, and 72% at 24 months (Non-patent Document 3).

出現 As a cause of the diminished therapeutic effect of biological products, the appearance of neutralizing antibodies is considered, but it is also considered that it is not the only cause. In such patients, treatment with a biological product may be performed by switching from a biological product that has no or no therapeutic effect to another biological product.

Biological products that are used in the treatment of IBD and that have been tried to be used in clinical trials, etc. target cytokines such as TNF-α and α4 integrins. There is. In particular, in patients who have not achieved remission or who have repetitive relapses and relapses, bacterial translocation of intestinal bacteria and endotoxins through the mucosal barrier and transfer into the body due to the breakdown of the barrier function of the intestinal mucosa. It easily causes location (BT: Bacterial Translocation) and causes complications due to bacterial infection.

Antibiotics are not a common choice in the treatment of IBD, but are used for patients with inflammation around the anus and as a combination therapy. Antibiotics are also used in IBD patients suspected of being caused by luminal bacteria. Although antibiotics with a broad antibacterial spectrum are used especially when the causative bacteria are not specified, there is a report that it worsens the intestinal bacterial flora and promotes diarrhea, which is one of the main symptoms of IBD ( Non-patent document 4).

Therefore, there is a need for a method for appropriately selecting a patient to be administered an antibiotic in an IBD patient and further optimizing the administration period of the antibiotic.
In addition to the removal of bacteria by antibiotics, there is a need for new treatments for inflammation associated with infection.

The initial symptom of IBD patients is inflammation, and general inflammatory markers (CRP, IL-6) are increased. At this stage, intestinal abnormalities are present but not accompanied by bacterial infection, so an increase in these markers serves as an indicator of the degree of inflammation but does not indicate bacterial infection.

SCD14-ST (soluble CD14 antigen subtype, also known as preceptin) has been reported to be useful as a diagnostic marker for sepsis, which is a systemic inflammatory syndrome with bacterial infection (WO 2005/108429). In addition, sCD14-ST is produced in the process of phagocytic cells phagocytosing and digesting foreign microorganisms and foreign substances, and in diseases such as arthritis where local autoimmune reactions and phagocytosis associated with infection occur. It has been reported that it is possible to detect an increase in the sCD14-ST concentration (WO 2009/142303). However, whether sCD14-ST is useful as a marker to be used in a method for appropriately selecting patients who should be administered antibiotics in IBD patients and further optimizing the period of administration of antibiotics was investigated. It never happened.

An object of the present invention is to appropriately select an IBD patient who needs a new treatment for antimicrobial removal or inflammation (including treatment with anti-CD14 antibody fusion protein, particularly MR1007) in addition to the usual treatment. Moreover, it is providing the method of optimizing the administration period of antibiotics. That is, it aims at solving the subject of proper use of antibiotics etc. in IBD.

The present inventors, for a normal human, normal inflammatory bowel disease (IBD) patients and biologicals (Remicade ^(R), the common name infliximab) does not react to (unresponsiveness) 3 groups of IBD patients, blood sample SCD14-ST concentrations were measured and compared. Then, there is no difference in the sCD14-ST concentration between the normal group and the normal IBD patient group, but between the normal group and the normal IBD patient group and the IBD patient group that does not respond to the biologics. A difference was observed in the sCD14-ST concentration in the blood sample, and it was also found that the preceptin concentration in the remicade-unresponsive IBD patient was as high as that in the sepsis patient. This is considered to indicate the possibility of severe bacterial invasion and severe inflammation (phagocytosis) in the intestinal mucosa even if systemic infection does not occur in patients who do not respond to remicade. In such patients, it is considered necessary to remove bacteria by antibiotics and new treatments for inflammation (including treatment with anti-CD14 antibody fusion protein, particularly MR1007) in addition to normal treatment. Presepsin is useful as a new indicator for selecting a subject to be used such as antibiotics among IBD patients, when to end treatment, and for selecting other treatments. In an embodiment of the present invention, using the sCD14-ST concentration in a blood sample as an indicator, detection of inflammatory bowel disease unresponsive to a biological product, selection of an inflammatory bowel disease patient unresponsive to a biological product, The present invention provides a method for selecting an inflammatory bowel disease patient to be administered with an antibiotic or the like, or determining the administration end timing of the antibiotic or the like in an inflammatory bowel disease patient receiving an antibiotic or the like.
More specifically, the present invention is as described below, for example.

The present invention provides, for example, the detection method, selection method, determination method or treatment method described below.
(1-1) A method for detecting inflammatory bowel disease unresponsive to a biologic, using the sCD14-ST concentration in a blood sample derived from a subject as an index.
(1-2) A method for detecting inflammatory bowel disease unresponsive to a biologic, comprising a step of measuring the sCD14-ST concentration in a blood sample derived from a subject.
(1-3) A method for detecting inflammatory bowel disease unresponsive to a biologic, comprising the following steps:
1) a step of measuring the sCD14-ST concentration in a blood sample derived from a subject;
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value.
(1-4) The detection method according to (1-3), further comprising the following steps:
4) A step of determining that an unresponsive inflammatory bowel disease is detected in the biological preparation when the measured value is higher than the reference value.
(1-5) The detection method according to any one of (1-1) to (1-4) above, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.
(1-6) The detection method according to any one of (1-1) to (1-5) above, wherein the biological preparation is an immunosuppressant.
(1-7) The biological product is selected from the group consisting of anti-TNFα antibody, soluble TNF receptor, anti-IL-6 receptor antibody, anti-IL-2 receptor antibody, anti-α integrin antibody and antisense NFκB. The detection method according to any one of (1-1) to (1-6), which is at least one of
(1-8) The detection method according to any one of (1-1) to (1-7), wherein the reference value is a normal value.
(1-9) The detection method according to any one of (1-1) to (1-8) above, wherein the subject is a patient with inflammatory bowel disease.
(1-10) The detection method according to any one of (1-3) to (1-9), wherein the reference value is 500 pg / mL.

(2-1) A method for selecting an inflammatory bowel disease patient to be administered with an antibiotic and / or MR1007, using sCD14-ST concentration in a blood sample derived from the patient as an index.
(2-2) A method for selecting an inflammatory bowel disease patient to be administered with an antibiotic and / or MR1007, comprising a step of measuring the sCD14-ST concentration in a blood sample derived from a patient.
(2-3) A method for selecting an inflammatory bowel disease patient to be administered with an antibiotic and / or MR1007, comprising the following steps:
1) a step of measuring the sCD14-ST concentration in a blood sample derived from a patient;
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value.
(2-4) A method for selecting a patient to be administered an antibiotic and / or MR1007 among patients with inflammatory bowel disease, the method comprising the following steps:
1) a step of measuring the sCD14-ST concentration in a blood sample derived from a patient;
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value.
(2-5) The selection method according to (2-3) or (2-4), further comprising the following steps:
4) A step of selecting the patient as a subject to be administered with an antibiotic and / or MR1007 when the measured value is higher than the reference value.
(2-6) The selection method according to any one of (2-1) to (2-5) above, wherein the inflammatory bowel disease patient is Crohn's disease patient or ulcerative colitis patient.
(2-7) Patients with inflammatory bowel disease to whom the antibiotic and / or MR1007 is administered are patients who have not responded to biological products, patients who have stopped responding to biological products, biological products The selection method according to any one of the above (2-1) to (2-6), which is a patient with a reduced therapeutic effect due to or a patient who has been administered a biologic and is associated with a bacterial infection.
(2-8) The selection method according to the above (2-7), wherein the biological preparation is an immunosuppressant.
(2-9) The biological product is selected from the group consisting of anti-TNFα antibody, soluble TNF receptor, anti-IL-6 receptor antibody, anti-IL-2 receptor antibody, anti-α integrin antibody and antisense NFκB. The selection method according to (2-7) or (2-8), which is at least one of
(2-10) The selection method according to any one of (2-3) to (2-9), wherein the reference value is 500 pg / mL.

(3-1) Timing of completion of administration of the antibiotic and / or MR1007 in an inflammatory bowel disease patient to which an antibiotic and / or MR1007 is administered, using the sCD14-ST concentration in a blood sample derived from the patient as an index Decision method.
(3-2) Completion of administration of the antibiotic and / or MR1007 in an inflammatory bowel disease patient to which an antibiotic and / or MR1007 has been administered, comprising the step of measuring the sCD14-ST concentration in a blood sample derived from a patient How to determine timing.
(3-3) A method for determining the end timing of administration of the antibiotic and / or MR1007 in a patient with inflammatory bowel disease to which an antibiotic and / or MR1007 is administered, comprising the following steps:
1) a step of measuring the sCD14-ST concentration in a blood sample derived from a patient;
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is less than or equal to the reference value.
(3-4) The determination method according to (3-3), further including the following steps:
4) A step of determining the end of administration of the antibiotic and / or MR1007 to the patient when the measured value is not more than the reference value.
(3-5) The determination method according to any one of (3-1) to (3-4) above, wherein the inflammatory bowel disease patient is Crohn's disease patient or ulcerative colitis patient.
(3-6) The patient is not responsive to a biologic, is no longer responsive to a biologic, is not treated with a biologic, or is receiving a biologic The determination method according to any one of (3-1) to (3-5) above, which is a patient with a bacterial infection.
(3-7) The determination method according to the above (3-6), wherein the biological preparation is an immunosuppressant.
(3-8) The biological product is selected from the group consisting of anti-TNFα antibody, soluble TNF receptor, anti-IL-6 receptor antibody, anti-IL-2 receptor antibody, anti-α integrin antibody and antisense NFκB. The determination method according to (3-6) or (3-7), which is at least one of
(3-9) The determination method according to any one of (3-1) to (3-8), wherein the reference value is a normal value.
(3-10) The determination method according to any one of (3-3) to (3-8), wherein the reference value is 500 pg / mL.

(4-1) A method for treating inflammatory bowel disease unresponsive to a biologic, comprising administering an antibiotic and / or MR1007 using the sCD14-ST concentration in a blood sample derived from a patient as an index.
(4-2) A method of treating inflammatory bowel disease, comprising the following steps:
1) measuring sCD14-ST in a blood sample derived from a patient;
2) a step of selecting a patient to be administered with an antibiotic and / or MR1007 using the measured value of sCD14-ST in the sample as an index, and 3) a step of administering an antibiotic and / or MR1007 to the selected patient.
(4-3) The above (4-2), wherein the step of selecting a patient to be administered with an antibiotic and / or MR1007 using the measured value of sCD14-ST in the sample as an index includes the following steps: Method described in:
1) a step of comparing a measured value of sCD14-ST in a sample with a reference value; and 2) a step of selecting a patient as an administration target of an antibiotic and / or MR1007 if the measured value of the sample is higher than the reference value.
(4-4) The method according to any one of (4-1) to (4-3) above, further comprising the following steps:
1) measuring sCD14-ST in a blood sample derived from a patient to whom an antibiotic and / or MR1007 is administered over time,
2) a step of comparing the measured value of sCD14-ST in the sample with a predetermined reference value, and 3) if the measured value of the sample falls below the predetermined reference value, the end of administration of the antibiotic and / or MR1007 is determined. Process.
(4-5) The method according to any one of (4-1) to (4-4) above, wherein the reference value is a normal value.
(4-6) The method according to any one of (4-1) to (4-5) above, wherein the inflammatory bowel disease is ulcerative colitis or Crohn's disease.
(4-7) The method according to any one of (4-1) to (4-6) above, wherein the patient is a patient with inflammatory bowel disease.

Moreover, according to 1 aspect of this invention, the kit described below is provided.
(5) A kit for detecting inflammatory bowel disease unresponsive to a biologic, comprising a device for measuring the concentration of sCD14-ST in a blood sample.
(6) Use of an anti-preceptin antibody and / or recombinant soluble CD14 fragment in the manufacture of a kit comprising a device for measuring sCD14-ST concentration for detecting inflammatory bowel disease unresponsive to biologics.

Furthermore, according to one aspect of the present invention, there is provided the use of the method for measuring the sCD14-ST concentration in a blood sample described below.
(7) A method of using a method for measuring sCD14-ST concentration for detecting inflammatory bowel disease unresponsive to a biologic.
(8) A method of using a method for measuring sCD14-ST concentration for selecting a patient with inflammatory bowel disease unresponsive to a biologic.
(9) A method of using the method for measuring the sCD14-ST concentration for determining the end timing of administration of the antibiotic in a patient with inflammatory bowel disease to which an antibiotic and / or MR1007 is administered.

Moreover, according to one aspect of the present invention, there is provided a method for treating inflammatory bowel disease by administering an antibiotic.
(10-1) A method for treating inflammatory bowel disease, comprising administering an antibiotic, comprising the following steps.
1) measuring sCD14-ST in a blood sample derived from a patient;
2) a step of selecting a patient to be administered an antibiotic using the measured value of sCD14-ST in the sample as an index; and 3) a step of administering an antibiotic to the selected patient.
(10-2) The method according to (10-1) above, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.
(10-3) The inflammatory bowel disease is an inflammatory bowel disease unresponsive to a biological product, and the biological product comprises an anti-TNFα antibody, a soluble TNF receptor, an anti-IL-6 receptor antibody, The treatment method according to (10-1) or (10-2) above, which is at least one selected from the group consisting of an anti-IL-2 receptor antibody, an anti-α integrin antibody and an antisense NFκB.

Furthermore, according to one aspect of the present invention, there is provided a method for treating inflammatory bowel disease by administering MR1007.
(11-1) A method for treating inflammatory bowel disease, comprising administering MR1007, comprising the following steps.
1) measuring sCD14-ST in a blood sample derived from a patient;
2) a step of selecting a patient to be administered an antibiotic using the measured value of sCD14-ST in the sample as an index; and 3) a step of administering MR1007 to the selected patient.
(11-2) The method according to (11-1) above, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.
(11-3) The inflammatory bowel disease is an inflammatory bowel disease unresponsive to a biological product, and the biological product comprises an anti-TNFα antibody, a soluble TNF receptor, an anti-IL-6 receptor antibody, The treatment method according to (11-1) or (11-2) above, which is at least one selected from the group consisting of an anti-IL-2 receptor antibody, an anti-α integrin antibody and an antisense NFκB.

According to the present invention, it is possible to appropriately select an IBD patient to whom an antibiotic or the like is to be administered, and to optimize the administration period of the antibiotic or the like.

FIG. 1 is a diagram showing the measurement results of preceptin concentration in blood samples of 20 normal subjects, 5 inflammatory bowel disease (IBD) patients, and 5 IBD patients who did not respond to remicade treatment.

Hereinafter, the present invention will be described in detail.
[SCD14-ST]
sCD14-ST (soluble CD14 antigen subtype, also known as presepsin) is one of the molecular species of soluble CD14, characterized in that it migrates to a molecular weight of 13 ± 2 kDa in SDS-PAGE under non-reducing conditions, It holds the N end of CD14. In addition, sCD14-ST has an amino acid sequence that is greatly deleted on the C-terminal side compared to full-length CD14, and both are different in three-dimensional structure, and thus show different immunogenicity. Therefore, it is possible to distinguish between the two by the antibody to be bound, and sCD14-ST has the property of specifically binding to an antibody prepared using a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2 as an antigen. . Furthermore, sCD14-ST specifically binds to an antibody that binds to a peptide consisting of the 17th to 26th amino acids of SEQ ID NO: 3, does not bind to 3C10 antibody, and does not bind to MEM-18 antibody. In addition, it additionally has any one or more of the characteristics that it has no LPS binding ability and can be obtained from human blood. As an amino acid sequence, sCD14-ST has a feature that it has the amino acid sequence of SEQ ID NO: 1 at the N-terminal sequence. More specifically, it is specified by the feature that the N-terminus is position 1 of the amino acid sequence shown in SEQ ID NO: 3 and the C-terminus is any one of positions 59 to 90 of the amino acid sequence shown in SEQ ID NO: 3. Can do. sCD14-ST is disclosed in detail in WO 2005/108429. In the present specification, sCD14-ST means human sCD14-ST unless otherwise specified.

Measurement of sCD14-ST in a blood sample can be performed by a known method or a known device. For example, an immunoassay system that specifically detects sCD14-ST disclosed in International Publication No. 2004/044005 or International Publication No. 2005/108429 can be used. An antibody prepared by using a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2 as an antigen is an antibody that specifically detects sCD14-ST. Therefore, sCD14-ST may be measured using this antibody. Specifically, an antibody prepared by using a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2 as an antigen, an antibody that binds to a peptide consisting of positions 17 to 26 of the amino acid sequence described in SEQ ID NO: 3, or the antibody A sandwich immunoassay system using a combination with an antibody that competes with F1106-13-3 antibody or F1031-8-3 antibody is preferable. The antibody produced using a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2 as an antigen is also an antibody that specifically binds to a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2.

As the measured value of the sCD14-ST concentration in the blood sample, any of a quantitative value, a semi-quantitative value, or a qualitative value may be used. When the semi-quantitative value is used, the sCD14-ST concentration can be displayed in stages such as 0, 1, 2, 3 or-, +, ++, +++. Since this stage correlates with the quantitative sCD14-ST concentration, whether or not the sCD14-ST concentration is equal to or higher than a predetermined reference value depends on the correlation between the semi-quantitative step display and the quantitative sCD14-ST concentration. Judge based on Alternatively, in the case of semi-quantitative, less than the reference value may be set to a stage such as 0 or −. When using a qualitative value, it may be set to be negative when the value is less than the reference value and positive when the value is equal to or more than the reference value.

The blood sample is not particularly limited, and any of whole blood, plasma, and serum may be used. The blood sample may be a sample to which an anticoagulant such as EDTA, heparin, or citric acid is added after blood collection.

In the present invention, the sCD14-ST concentration in a blood sample derived from a subject or a patient may be measured over time. This is because detection, selection, or determination can be performed at a more appropriate timing by performing measurement over time.
In particular, when administering antibiotics, it is possible to prevent deterioration of the intestinal bacterial flora of the patient by appropriately selecting or determining the patient to be administered, the start of administration, the end of administration and / or the continuation of administration It becomes. As for the timing of measurement, when it is suspected that the biologic product is unresponsive, or when it is necessary to determine the start, end or continuation of antibiotic administration, from the first sCD14-ST concentration measurement day, every day, What is necessary is just to set suitably every 2nd day or the 3rd day, the 5th day, and / or the 7th day.

[Biologics]
In the present invention, the biological preparation is not particularly limited as long as it can be used for the treatment of inflammatory bowel disease. Examples of the biological preparation include Remicade ^(R) (generic name: Infliximab: Mitsubishi Tanabe Pharma, St. Core), Humira ^(R) (generic name: adalimumab: Abbott Laboratories), Simpony ^(R) (generic name: Golimumab: Janssen Pharma) ), Simdia ^(R) (generic name: Centrizumab Pegor: UCB Pharma) and other TNFα antibodies; Embrel ^(R) (generic name: Etanercept: Weiss) and other soluble TNF receptors; Actemra ^(R) (generic name: Tocilizumab: Chugai pharmaceutical) anti-IL-6 receptor antibody such as; Simulect ^(R) (generic name basiliximab: anti IL-2 receptor antibody of Novartis Pharma), and the like; TYSABRI ^(R) (generic name natalizumab: Elan Pharma International) anti such α4 integrin antibody; Orencia ^(R (Generic name abatacept: Bristol-Myers Squibb) CD28-CD80 / 86 interaction inhibitors such as; Rituxan ^(R): the anti-CD20 antibody (common name rituximab Chugai) and the like; and, NFKB decoy oligonucleotide (AnGes MG) such Examples include antisense NFκB.

As the biological preparation, an immunosuppressive agent is preferable, and it is selected from the group consisting of anti-TNFα antibody, soluble TNF receptor, anti-IL-6 receptor antibody, anti-IL-2 receptor antibody, anti-α integrin antibody and antisense NFκB. More preferred is at least one selected. The anti-TNFα antibody is preferably an anti-human TNFα monoclonal antibody or a PEGylated derivative thereof. Specific examples of the PEGylated derivative include a derivative obtained by binding polyethylene glycol (PEG) to a Fab ′ fragment excluding the Fc region of the anti-human TNFα monoclonal antibody. Soluble TNF receptors include TNFR derivatives, and specifically, a fusion protein of the extracellular domain of human TNFR-II and the Fc region of human IgG1 is preferred. Among them, a biological product for which IBD is an applicable disease or a clinical trial for IBD is preferred, and from the viewpoint of a large number of users, anti-TNFα antibodies include Remicade ^(R) (generic name: infliximab; Human TNFα monoclonal antibody formulation), Humira ^(R) (generic name adalimumab; human anti-human TNFα monoclonal antibody formulation) or Simdia ^(R) (generic name centrizumab pegol; PEGylated anti-human TNFα monoclonal antibody formulation) are preferred, soluble TNF The receptor is preferably embrel ^(R) (generic name etanercept; fully human soluble TNFα / LTα receptor preparation), and the anti-α4 integrin antibody is preferably tysabri ^(R) (generic name: natalizumab).

[Inflammatory bowel disease]
In the present invention, the inflammatory bowel disease is preferably Crohn's disease or ulcerative colitis.

(Diagnosis and severity of Crohn's disease)
Crohn's disease may be diagnosed according to conventionally known diagnostic criteria. However, longitudinal ulcers, paving stones, or non-drying characteristics are confirmed by X-ray examinations (barium small bowel and barium enema examinations) and endoscopic examinations. Preferably, characteristic findings such as epithelioid granuloma are observed.
The method for evaluating the severity of Crohn's disease is not particularly limited, but it is preferable to use an IOIBD assessment score or CDAI (Crohn's disease activity index). In the IOIBD assessment score, (1) abdominal pain, (2) diarrhea or mucous stool more than 6 times a day, (3) anal lesion, (4) fistula, (5) other complications, (6) abdominal mass, (7) Weight loss, (8) Fever of 38 ° C. or higher, (9) Abdominal tenderness, and (10) 10 g / dl hemoglobin of each of 10 items are summed, and the total score is used as a score. The higher the score, the more severe it is. It is preferable to evaluate remission as a state where the score is 1 or 0 and the red blood cell level and CRP are normalized in the blood test. CDAI consists of (1) number of watery or loose stools, (2) degree of abdominal pain, (3) subjective general condition, (4) presence of extraintestinal complications, (5) use of antidiarrheal drugs, (6 It is an index obtained by scoring and adding the weight of abdominal mass, (7) hematocrit, and (8) body weight. Preferably less than 150 is remission, 150-220 is mild, 220-300 is moderate, 300-450 is severe, and 450 or more is severe.

(Diagnosis and severity of ulcerative colitis)
Diagnosis of ulcerative colitis may be performed according to conventionally known diagnostic criteria, but other enteritis is based on clinical symptoms, endoscopic findings, enema X-ray findings, pathological findings (cell examination). It is preferably performed by excluding (infectious enteritis, drug enteritis, radiation enteritis, ischemic enteritis, Crohn's disease, etc.).
The method for evaluating the severity of ulcerative colitis is not particularly limited, but it is preferable to make a determination based on, for example, the degree of bloody stool and diarrhea, the presence or absence of fever, tachycardia and anemia, and blood tests. Mild bloody stool and diarrhea, mild symptoms without systemic symptoms, moderate disease between mild and severe, (1) diarrhea more than 6 times a day, (2) strong bloody stool, (3) 37. Fever of 5 ° C or higher, (4) tachycardia with 90 or more pulses per minute, (5) anemia with hemoglobin value of 10 g / dl or less, (6) increased blood sedimentation of 30 mm / hour or more (inflammation by blood test) Severe, those who satisfy 4 or more of 6 items of the grade of), more than 15 bloody diarrhea, fever of 38 ° C or more, 10000 / mm ³ or more leukocytosis, strong abdominal pain all When satisfying, it is preferable to determine as fulminant type.

[Standard value]
In the present invention, the reference values are a group of patients with inflammatory bowel disease unresponsive to biological products, and normal and normal IBD patients (not “people with inflammatory bowel disease unresponsive to biological products”). This is the sCD14-ST concentration in the blood sample that is set in advance to distinguish it from the population of IBD patients).
The reference value may be set to a value with good screening efficiency so that a balance between false positives and false negatives can be achieved, and the frequency of occurrence of inflammatory bowel disease patients who have not responded to biologics has rapidly increased. The starting value may be set, a pathologically or physiologically theoretical value may be set, or a statistically determined value may be set.
More specifically, the reference value is obtained, for example, by calculating an average value and a standard deviation (SD) of measured values of sCD14-ST concentration in a blood sample of a medically healthy person, and a range of the average value +0.5 SD to +5 SD Specifically, the average value + SD, the average value + 2SD, the average value + 3SD, etc. may be set, or the average value of 5 to 95, 10 to 90, 15 to 85, or 25 to 75 percentile value may be set. May be. The reference value is preferably set to the average value + 2SD or the average value + 3SD. Specific numerical values include 500 pg / mL or 600 pg / mL, preferably 600 pg / mL, but are not limited thereto.

[Antibiotic agents]
Antibiotics are not particularly limited, for example, penicillin, penem, carbapenem, cephem, monobactam, fosfomycin, glycopeptide, amino acid glycoside, macrolide, ketolide, lincomycin, Antibiotics such as tetracyclines, new quinolones, sulfonamides, oxazolidinones, streptogramins, and azoles can be used, and one or more of these antibiotics can be appropriately selected and used.

Further, instead of or together with an antibiotic, an anti-CD14 antibody fusion protein (see International Publication No. 2006/1229849), preferably MR1007 (Nakamura et al., Critical Care 11 (Supp. 4): P4, 2012 ) And / or anti-IL-6 antibodies (Mudter & Neurath, Inflammatory & Bowel & Disease < 13 (8) > 1016-1023, 2007).

The anti-CD14 antibody fusion protein is a fusion protein containing an anti-CD14 antibody and a proteolytic enzyme inhibitor, and the MR1007 comprises an anti-CD14 antibody and a modified light chain (serine protease inhibitory active fragment) of an inter-α inhibitor. It is a fusion protein. The anti-CD14 fusion protein may have an effect of preventing the severity of bacterial infection by targeting membrane-bound CD14 and blood coagulation protease.

IL-6 is an inflammatory cytokine similar to TNFα, and has various actions such as activation of T cells and B cells and induction of C-reactive protein (CRP). If inflammation is present, the concentration of IL-6 will increase along with CRP. In active IBD patients, serum and intestinal mucosa, especially intestinal mucosa, have increased IL-6 concentrations, which have been suggested for some time to be associated with disease states. In addition, when IL-6 binds to a receptor, a signal is transmitted into the cell through the JAK-STAT pathway, but it has been proved that dysregulation of the JAK-STAT pathway is involved in inflammation and carcinogenesis. Therefore, inhibition of excessive signal transduction with an anti-IL-6 antibody may prevent not only the improvement of IBD but also the occurrence of colorectal cancer.

Similarly, when an anti-CD14 fusion protein or MR1007 or anti-IL-6 antibody is administered instead of or together with an antibiotic, selection of these administration subjects using the sCD14-ST concentration as an index, Administration start, administration end and / or continuation of administration can be determined.

Antibiotics and MR1007 are preferably formulations with the following information:
1) A preparation that is administered to a patient with inflammatory bowel disease that is unresponsive to a biological preparation, using the sCD14-ST concentration in a blood sample derived from a subject as an index.
2) A preparation for detecting an inflammatory bowel disease patient who is unresponsive to a biological preparation, using the sCD14-ST concentration in a blood sample derived from the subject as an index, and administering it to the patient.
3) A preparation for measuring a sCD14-ST concentration in a blood sample derived from a patient with inflammatory bowel disease, selecting a patient with inflammatory bowel disease to be administered, and administering it to the patient.
The above information can be provided in the package insert, interview form, brochure, instructions, etc. of the formulation, and can be provided simultaneously with the formulation or separately.

[Detection method, selection method, determination method and treatment method]
1. Method for Detecting Inflammatory Bowel Disease Unresponsive to Biological Product According to one embodiment of the present invention, an inflammatory bowel unresponsive to a biologic agent, which uses sCD14-ST concentration in a blood sample derived from a subject as an index A method for detecting a disease is provided.

According to this detection method, it is possible to perform efficient screening by detecting inflammatory bowel disease unresponsive to biological products using sCD14-ST concentration in a blood sample derived from a subject as an index. Inflammatory bowel disease unresponsive to biologics can be detected earlier. As a result, appropriate treatment can be performed earlier and an improvement in the overall outcome can be expected.

The subject is not particularly limited, and inflammatory bowel disease may or may not be diagnosed. A subject who has been diagnosed with inflammatory bowel disease may be referred to as a patient. In addition, when inflammatory bowel disease is diagnosed, it is preferable that the patient is a patient who needs treatment with a biological product or is actually receiving treatment with a biological product. Calprotectin measurement may be used to identify patients with inflammatory bowel disease.

The inflammatory bowel disease unresponsive to a biological product is not particularly limited as long as it is an inflammatory bowel disease unresponsive to at least one biological product, and may not respond from the start of treatment. However, the reaction may have occurred at the start of the treatment, but the effect may be reduced due to the continuation of treatment and the reaction may stop. In addition, an inflammatory bowel disease that is unresponsive to a biological product is treated with a biological product, even if it is not actually treated with the biological product. Inflammatory bowel disease, which may be unresponsive to active formulations. It may also be an inflammatory bowel disease with bacterial infection.

The method for detecting inflammatory bowel disease unresponsive to the biological preparation of the present invention preferably comprises a step of measuring the sCD14-ST concentration in a blood sample derived from a subject. 1) In a blood sample derived from a subject measuring the sCD14-ST concentration; 2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value. More preferably, further comprising 4) a step of determining that an unresponsive inflammatory bowel disease is detected in the biological product when the measured value is higher than the reference value. . A step of preparing a blood sample derived from the subject may be provided before the step of measuring the sCD14-ST concentration in the blood sample derived from the subject. The detection method of the present invention can be performed in vitro.

By comparing the measured value of sCD14-ST in the specimen with a predetermined reference value, it can be determined whether or not the subject is an IBD patient who has not responded to the biological product. As the reference value used in the method for detecting IBD that does not respond to the biologic of the present invention, a normal value set based on the measured value of sCD14-ST in a blood sample of a healthy person can be used. As the normal value, an average value of sCD14-ST measurement results in a blood sample of a healthy person or a value obtained by standardizing the measurement results by setting a range, for example, can be used. If the measurement value of a sample from a healthy person is almost equivalent to the background value of the measurement system, standardize the background value by setting an average value or a range, for example, of the background value in the measurement system The value may be used. The background value in the measurement system is a measurement value when a buffer or an assay solution is added to the measurement system instead of a specimen. The values obtained by standardizing the measured values of the specimens derived from the subjects are the average value of healthy individuals +0.5 SD to +5 SD (SD is the standard deviation), and the measured values of healthy persons 5 to 95, 10 to 90, 15 to 85, or 25 to A 75th percentile value or the like can be used. Preferably, it is the average value of healthy individuals + SD, + 2SD or + 3SD.

The method for detecting IBD that is unresponsive to the biological product of the present invention can be paraphrased as a method for selecting an IBD patient who is unresponsive to the biological product to which antibiotics and / or MR1007 are administered. A subject in whom an IBD that is unresponsive to a biologic is detected is a patient likely to be an IBD that is unresponsive to the biologic and is a suitable subject for antibiotic and / or MR1007 administration. Among IBD patients, antibiotics and / or MR1007 should be administered if the IBD is unresponsive to the biologic. By using the measured value of sCD14-ST in a blood sample as an indicator, it is possible to detect whether or not the IBD is unresponsive to a biological product, and the biological agent to which an antibiotic and / or MR1007 is administered It became possible to select IBD patients who did not respond to the formulation. That is, according to one aspect of the present invention, an IBD patient who does not respond to a biological product to which an antibiotic and / or MR1007 is administered is characterized by measuring sCD14-ST in a blood sample derived from a patient. A method is provided. In the method for selecting an IBD patient who does not respond to a biological product to which the antibiotic of the present invention and / or MR1007 is administered, the embodiment in the method for detecting IBD not responding to a biological product can be applied as it is.

2. Method for selecting inflammatory bowel disease patients to be administered antibiotics etc. According to one embodiment of the present invention, antibiotics and the like (antibiotics and / or antibiotics) using sCD14-ST concentration in a blood sample derived from a patient as an index. Provided is a method for selecting an inflammatory bowel disease patient to be administered with a therapeutic agent against inflammation (for example, anti-CD14 antibody fusion protein, particularly MR1007).

According to this selection method, by selecting sCD14-ST concentration in a blood sample derived from a patient as an index, by selecting an inflammatory bowel disease patient to be administered an antibiotic or the like from inflammatory bowel disease patients, Subjects to which antibiotics and the like should be administered can be appropriately selected. As a result, patients who need treatment with antibiotics are treated. Survival rate, incidence of complications, incidence of infectious diseases and side effects, occurrence of surgical procedures (colectomy, etc.) Improvements in rates, patient functional health, patient QOL, treatment satisfaction, etc. can be expected.

The patient is not particularly limited as long as it is a patient with inflammatory bowel disease, and may or may not be treated with a biologic. Preferably, the patient is a patient in need of treatment with a biologic or is actually undergoing treatment with a biologic.

The method for selecting an inflammatory bowel disease patient to be administered with the antibiotic of the present invention preferably comprises a step of measuring the sCD14-ST concentration in a patient-derived blood sample. 1) A patient-derived blood sample Measuring the sCD14-ST concentration therein; 2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value It is more preferable to include a step, and 4) it is further preferable to include a step of selecting the patient as a subject to be administered an antibiotic or the like when the measured value is higher than the reference value. A step of preparing a patient-derived blood sample may be provided before the step of measuring the sCD14-ST concentration in the patient-derived blood sample.

Since the measured value of sCD14-ST in the sample is an indicator of whether or not the IBD patient is unresponsive to the biological product, the measured value of the sCD14-ST concentration is compared with a predetermined reference value. In addition, it is possible to select a patient with inflammatory bowel disease to which an antibiotic or the like is administered. More specifically, when the measured value is higher than the reference value, the patient can be selected as an inflammatory bowel disease patient to be administered with an antibiotic or the like.

1 above. The normal value set based on the measured value of sCD14-ST in the blood sample of a healthy person described in the embodiment can be used as the reference value. For example, the average value of healthy individuals + SD, + 2SD, or + 3SD. If the measured value of sCD14-ST falls within the normal range, it can be considered that the IBD has been cured or ameliorated to a level equivalent to that of a healthy person. Preferably, the measured value of sCD14-ST in the blood sample of a healthy person, an IBD patient who is not responsive to the biological product, and an IBD patient who is not responsive to the biological product, A cut-off value set so that the sensitivity and / or specificity of detection can be optimized can be used as a reference value. For example, 500 pg / mL or 600 pg / mL is mentioned, Preferably it is 600 pg / mL, However It is not limited to these.

The method for selecting an inflammatory bowel disease patient to be administered with the antibiotic or the like of the present invention is the above-mentioned “4) When the measured value is higher than a reference value, the patient is administered with the antibiotic or the like. Instead of “the step of selecting as a target to be performed”, “when the measured value is higher than the reference value, the step of selecting the patient as a patient to start administration of an antibiotic etc.” Can be paraphrased as a method for selecting an IBD patient to start administration. In addition, the same description is “the step of determining the start of administration of antibiotics to the patient when the measured value is higher than the reference value”, so that the timing of the start of administration of antibiotics, etc. in IBD patients It can be paraphrased as a determination method.

3. Method for determining timing of completion of administration of antibiotics in patients with inflammatory bowel disease to which antibiotics or the like are administered According to one aspect of the present invention, antibiotics using sCD14-ST concentration in a blood sample derived from a patient as an index Etc. (refers to antibiotics and / or therapeutic agents for inflammation (for example, anti-CD14 antibody fusion protein, particularly MR1007)) in patients with inflammatory bowel disease A determination method is provided.

According to this determination method, it is possible to determine an appropriate timing for terminating antibiotic administration in patients with inflammatory bowel disease to which antibiotics or the like are administered, using the sCD14-ST concentration in the blood sample derived from the patient as an index. As a result, treatment with antibiotics is terminated at an appropriate time, and unnecessary antibiotics are not administered, resulting in the survival rate, incidence of complications, incidence of infections and side effects in the patient. Improvements in the incidence of surgical procedures (such as colectomy), functional health of patients, patient QOL, treatment satisfaction, etc. can be expected.

The patient is not particularly limited as long as it is an inflammatory bowel disease patient to whom an antibiotic or the like is administered, and may or may not be treated with a biologic. The patient is preferably a patient who has been administered antibiotics and needs to be treated with a biologic or is actually receiving treatment with a biologic.

Preferably, the method for determining the end timing of administration of an antibiotic or the like in an inflammatory bowel disease patient to which an antibiotic or the like of the present invention is administered comprises a step of measuring the sCD14-ST concentration in a blood sample derived from the patient. 1) a step of measuring the sCD14-ST concentration in a blood sample derived from a patient; 2) a step of comparing the measured value of the sCD14-ST concentration with a reference value; and 3) the measured value is less than or equal to the reference value It is more preferable to include a step of determining whether or not, and 4) a step of determining the end of administration of the antibiotic or the like to the patient when the measured value is lower than the reference value. Is more preferable. A step of preparing a patient-derived blood sample may be provided before the step of measuring the sCD14-ST concentration in the patient-derived blood sample.

The measured value of sCD14-ST in the sample is an indicator of whether or not the IBD patient is unresponsive to the biological product. Therefore, by comparing the measured value of the specimen with a predetermined reference value, it is possible to determine the end timing of administration of the antibiotic or the like in the inflammatory bowel disease patient who is being treated with the antibiotic or the like. More specifically, when the measured value falls below the reference value, it can be determined that an IBD that has not responded to a biologic is in remission, severity is no longer severe, or no longer active, such as antibiotics The end of administration can be determined. The reference value used in the method for determining the end timing of administration of the antibiotic of the present invention may be any value that can confirm the success of the antibiotic. For example, a value such as 1/2, 1/5, or 1/10 of the measured value of sCD14-ST measured from before administration of antibiotics to 24 hours after the start of administration can be set as the reference value. In addition, the above 1. The normal value set based on the measured value of sCD14-ST in the blood sample of a healthy person described in the embodiment can be used as the reference value. For example, the average value of healthy individuals + SD, + 2SD, or + 3SD. If the measured value of sCD14-ST falls within the normal range, it can be considered that the IBD has been cured or ameliorated to a level equivalent to that of a healthy person. Preferably, the measured value of sCD14-ST in the blood sample of a healthy person, an IBD patient who is not responsive to the biological product, and an IBD patient who is not responsive to the biological product, A cut-off value set so that the sensitivity and / or specificity of detection can be optimized can be used as a reference value. For example, 500 pg / mL or 600 pg / mL is mentioned, Preferably it is 600 pg / mL, However It is not limited to these.

The method for selecting an inflammatory bowel disease patient to be administered with the antibiotic or the like of the present invention is the above-mentioned “4) The antibiotic for the patient when the measured value is lower than the reference value. Instead of “the step of determining the end of administration such as”, “when the measured value is lower than the reference value, the step of selecting the patient as a patient to end the administration of antibiotics” In other words, it can be paraphrased as a method for selecting a patient with inflammatory bowel disease to finish administration of an antibiotic or the like. Moreover, the administration of antibiotics and the like is continued by setting the same description as “a step of selecting the patient as a patient to continue administration of antibiotics when the measured value is higher than the reference value”. In other words, it can be paraphrased as a method of selecting patients with inflammatory bowel disease.

4). Method of treating IBD by administering an antibiotic or the like According to one embodiment of the present invention, a method of treating IBD is provided. This treatment method uses a measured value of sCD14-ST in a blood sample derived from a subject as an index, in addition to the usual treatment, a new treatment method (anti-CD14 antibody fusion protein, particularly MR1007) for removal of bacteria by antibiotics and inflammation. A method of treating IBD by administering an antibiotic or the like to the selected patient. Specifically, as described in 2. above. An IBD patient to be administered an antibiotic or the like is selected by the method described in the embodiment, and the antibiotic or the like is administered to the selected patient. In addition, the 3. By determining the end timing of administration of antibiotics or the like by the method described in the embodiment, more efficient treatment can be performed.

The 1. of the present invention. ~ 3. Antibiotics and the like used in the embodiment described in 1) are not particularly limited, but antibiotics that can be used at least for IBD are suitable. Examples include penicillins, penems, carbapenems, cephems, monobactams, fosfomycins, glycopeptides, amino acid glycosides, macrolides, ketolides, lincomycins, tetracyclines, new quinolones, sulfonamides And antibiotics such as oxazolidinone, streptogramin, and azole. One or more of these antibiotics can be appropriately selected and used. Preference is given to the new quinolone series used in IBD, in particular ciprofloxacin (Ciproxan ^(R) ), or the azole series, especially metronidazole (Frazil ^(R) ).

Further, instead of or together with an antibiotic, an anti-CD14 antibody fusion protein (see International Publication No. 2006/1229849), preferably MR1007 (Nakamura et al., Critical Care 11 (Supp. 4): P4, 2012 ) And / or anti-IL-6 antibodies (Mudter & Neurath, Inflammatory Bowel Disease 13 (8): 1016-1023, 2007).
The anti-CD14 antibody fusion protein is a fusion protein containing an anti-CD14 antibody and a proteolytic enzyme inhibitor, and the MR1007 comprises an anti-CD14 antibody and a modified light chain (serine protease inhibitory active fragment) of an inter-α inhibitor. It is a fusion protein. The anti-CD14 fusion protein may have an effect of preventing the severity of bacterial infection by targeting membrane-bound CD14 and blood coagulation protease.

[Detection kit]
5. Kit comprising a device for measuring the concentration of sCD14-ST in a blood sample According to one embodiment of the present invention, detection of inflammatory bowel disease unresponsive to a biological product comprising a device for measuring the concentration of sCD14-ST in a blood sample A kit is provided.
The detection kit of the present invention comprises, as desired, an antibody (anti-preceptin antibody) prepared using a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2 as an antigen, from the 17th to 26th amino acid sequences described in SEQ ID NO: 3. And at least one selected from the group consisting of an antibody that binds to the peptide, a multiwell plate, and a spectrophotometer. This kit preferably measures the sCD14-ST concentration in a blood sample by a sandwich immunoassay.

6). Use of anti-preceptin antibody and / or recombinant soluble CD14 fragment in the manufacture of a kit comprising a device for measuring sCD14-ST concentration for detecting inflammatory bowel disease unresponsive to a biologic Includes a device for measuring sCD14-ST concentration in a blood sample in the manufacture of a kit including a device for measuring sCD14-ST concentration for detecting inflammatory bowel disease unresponsive to a biologic. It relates to the use of anti-presepsin antibodies and / or recombinant soluble CD14 fragments in a “kit”.

[Use of sCD14-ST concentration measurement method in blood sample]
7). Use of Detecting Inflammatory Bowel Disease Unresponsive to Biological Product This embodiment is a method for detecting sCD14-ST concentration in a blood sample in “1. Method for detecting inflammatory bowel disease unresponsive to biological product”. This is a usage method (use) of the measurement method.

8). Use for selecting inflammatory bowel disease patients who are unresponsive to biological products In this aspect, the antibiotics described in “1. Method for detecting inflammatory bowel disease unresponsive to biological products” are administered. This is a method (use) of using a method for measuring the concentration of sCD14-ST in a blood sample in a method for selecting an IBD patient who does not respond to a biologic.

9. Use of determining timing of completion of administration of the antibiotic etc. in patients with inflammatory bowel disease to which antibiotics and the like are administered This aspect is the above-mentioned "3. Antibiotics in patients with inflammatory bowel disease to which antibiotics etc. are administered. The method for using (measuring) the method of measuring the concentration of sCD14-ST in a blood sample in “Determination Method of Administration End Timing of Agents”.

The methods described above are useful for the treatment of Crohn's disease (CD) or ulcerative colitis (UL), which are inflammatory bowel diseases.
For example, ulcerative colitis requires attention to responsiveness to medical treatment, complications due to drugs, etc. for severe cases and intermediate cases with systemic disorders. Thus, if inflammatory bowel disease unresponsive to a biologic can be detected by the method of the present invention, unnecessary drug therapy can be suppressed, which is useful. In the elderly, a fatal course may occur due to side effects (such as opportunistic infections such as MRSA infection) caused by therapeutic agents having an immunosuppressive effect. Therefore, it is highly useful to perform treatment effect determination etc. at an early stage by the method of the present invention.

In addition, there is still no treatment method to completely cure Crohn's disease, and the severity is important in determining the treatment method. Therefore, it is useful to detect inflammatory bowel disease that does not respond to biological products by the method of the present invention. Some drug therapies involve immunosuppression, and it is necessary to perform treatment while paying attention to complications such as infection. However, administration of an excessive amount of biological preparation can be suppressed by the method of the present invention.

The present invention will be described in further detail with reference to the following examples, but the present invention should not be construed as being limited to these examples.

1. 1. Preparation of blood preceptin measurement system 1-1. Preparation of peroxidase-labeled antibody To produce a sandwich ELISA system, F1106-13-3 antibody (see WO 2005/108429) was labeled with peroxidase.
(1) First, F1106-13-3 antibody was cleaved with lysyl endopeptidase ^(R) (Wako Pure Chemical Industries) to prepare F (ab ′) ₂ . F1106-13-3 antibody was dialyzed against 50 mM Tirs-HCl (pH 8.5), and antibody and lysyl endopeptidase were mixed at a molar ratio of antibody: lysyl endopeptidase ^(R) = 10: 1 and 1 at 37 ° C. Reacted for hours. After 1 hour, TLCK (N _α -tosyl-L-lysine chloromethyl ketone hydrochloride, CAS No. 4272-74-6) was added to a final concentration of 30 mM to stop the reaction. To remove fc, the reaction liquid was added Protein A column ^{(PROSEP (R)} -A, Millipore) to, to recover the unadsorbed fraction. The collected non-adsorbed fraction was concentrated and purified by gel filtration (Superdex 75, GE Healthcare) to separate the contained F (ab ′) ₂ and Fab. The obtained F (ab ′) ₂ was concentrated and dialyzed against 10 mM carbonate buffer (pH 9.5).
(2) Next, according to the method of Nakane et al. (J. Histochem. Cytochem., 22, 1084, 1974), 1 mg of peroxidase (Toyobo) was dissolved in distilled water, and 100 mM periodic acid dissolved in distilled water was dissolved. The mixture was added and reacted at 25 ° C. for 15 minutes. After completion of the reaction, 1.5% ethylene glycol was added, reacted at 25 ° C. for 20 minutes, and dialyzed against 1 mM acetate buffer (pH 4.4). The next day, 1 mg of F1106-13-3F (ab ′) ₂ antibody was mixed with 0.8 mg of peroxidase activated by adding 0.2 M carbonate buffer (pH 9.5) and reacted at 25 ° C. for 2 hours. I let you. 4 mg / mL sodium borohydride was added and allowed to react for an additional 2 hours at 4 ° C. The reaction solution was dialyzed against PBS (pH 7.4) to obtain a peroxidase-labeled antibody. The amount of the solution was measured, and the antibody concentration was calculated from the amount of antibody used.

1-2. Preparation of sandwich ELISA system (1) S68-peptide polyclonal antibody (see International Publication No. 2005/108429) was diluted to 5 μg / mL with D-PBS (Dulbecco phosphate buffered saline, pH 7.4), and immunoplate (MaxSorp ^{(R), NUNC)} was 50μL added to each well of (here, S68- peptide polyclonal antibody, a peptide consisting of 16 amino acid residues described in SEQ ID NO: 2 was prepared as an antigen, sCD14-ST specific antibodies .) After overnight reaction at 4 ° C., washed 5 times with ion-exchanged water, and ^{0.1% StabilGuard (R) (SurModics} ), 200μL added D-PBS containing 0.1% ^{Tween (R)} 20 to each well And blocked.
(2) Next, using D-PBS (pH 7.4) containing 1% CD14-absorbed serum and 0.1% BSA as a diluent, concentrations 0, 9.2, 18.4, 36.9, 73.8 147.5, 295 pg / mL preceptin protein standard dilution series was prepared. As a sCD14-ST protein standard, a recombinant soluble CD14 fragment described in WO 2005/108429, specifically, rsCD14-ST was used. A standard dilution series was added at 50 μL per well and allowed to react at 25 ° C. for 1 hour. After completion of the reaction, it was washed five times with physiological saline containing ^{0.05% Tween (R) 20,} 2% rat serum, 1% mouse serum, PBS containing ^{0.1% Tween (R) 20 (} pH7.4 F1106-13-3 F (ab ′) ₂ antibody labeled with peroxidase diluted to 0.65 μg / mL with 50 μL was added to each well. After reacting at 25 ° C. for 2 hours, the well was washed 5 times in the same manner, and 50 μL of tetramethylbenzidine solution (TMB, BioFix) was added to each well. After 20 minutes of reaction at room temperature, the reaction was stopped with 50 μL of 0.5 M sulfuric acid solution. . Absorbance at 450/630 nm was measured with a plate spectrophotometer (Multiscan JX, Thermoelectron). It was confirmed that the standard dilution series could be measured without problems.

2. 2. Measurement of blood preceptin concentration in normal persons and patients with inflammatory bowel disease 2-1. Measurement Method 20 normal persons (ProMed DX), 5 patients with normal inflammatory bowel disease (IBD) (hereinafter also simply referred to as “IBD patients”) (ProMed DX, No. 100004915 to 100004919) and Remicade treatment Blood samples of 5 IBD patients (Bioreclamation, No. BRH318495-BRH318499) who had not been received (normal people were plasma, IBD patients and IBD patients who did not respond to remicade treatment were serum, all without pretreatment) . Table 1 shows the specimen number, diagnosis (diagnostic disease name), symptom, condition (disease activity), and IBD treatment history for 5 IBD patients and 5 IBD patients who did not respond to Remicade.
Using the sandwich ELISA system prepared in “1. Preparation of blood presepsin measurement system”, each purchased specimen was diluted 20 times and the preceptin concentration was measured.

2-2. Measurement results The measurement results are shown in FIG. Table 1 shows the preceptin concentrations of 5 IBD patients and 5 IBD patients who did not respond to remicade.
The preceptin concentration of 465 ± 105 pg / mL (mean value ± SD) of IBD patients (5 cases) was comparable to the preceptin concentration of 346 ± 109 pg / mL (mean value ± SD) of normal patients (20 cases).
In contrast, the preceptin concentration of Remicade unresponsive patients (5 cases) was as high as 13441 ± 23623 pg / mL (mean value ± SD, range 513 to 55562 pg / mL).
Based on the above, in severe IBD patients, such as patients who do not respond to remicade, there is a possibility that the intestinal barrier function is broken, and the increase of preceptin is caused by bacterial translocation through the intestinal barrier in these patients. Indicates what is happening.

Antibiotic treatment in IBD patients is not used as a general treatment method. However, by measuring the preceptin concentration in the specimen, for example, by adopting a cutoff value of 600 pg / mL as reported by Endo et al. (Infect Chemother. 2012 Jun 13) as a reference value, in the diagnosis of infection in IBD patients, and It has been shown that it can be used to select, use and / or discontinue treatment methods such as antibiotics.

3. 3. Measurement of blood preceptin concentration in clone patients and ulcerative colitis patients 3-1. Measurement Method Blood samples (serum without pretreatment) of 46 patients with Crohn's disease (CD) (Bioreclamation) and 34 patients with ulcerative colitis (UC) (Bioreclamation) were purchased.
Using the sandwich ELISA system prepared in “1. Preparation of blood presepsin measurement system”, each purchased specimen was diluted 20 times and the preceptin concentration was measured.

3-2. Measurement results Among CD patients (46 cases) and UC patients (34 cases), patients with preceptin concentrations of 600 pg / mL or more were 3/46 (6.5%) and 2/34 (5.9%), respectively. )Met. In 3 CD patients and 2 UC patients with a preceptin concentration of 600 pg / mL or more, the stage was relapsed, the severity was severe, and the condition (disease state) was at least one of the active.
From the above, in CD patients and UC patients, the concentration of presepsin is generally low, but by measuring the blood presepsin concentration, in IBD that is not classified as an infectious disease, severe infection caused by infection It could be used to detect IBD patients and to select IBD patients whose bacterial infection is related to aging, relapse of disease or activity.

According to the above-described embodiments or examples, an IBD patient to be administered an antibiotic or the like (refers to an antibiotic and / or a therapeutic agent for inflammation (for example, anti-CD14 antibody fusion protein, particularly MR1007)). Can be appropriately selected, and the administration period of antibiotics and the like can be optimized. This is because antibiotics and the like should be administered to IBD patients with bacterial infection. In addition, the administration period of antibiotics can be shortened by determining the timing of the end of administration of antibiotics etc. using the sCD14-ST concentration in the blood sample as an index. Proper use of antibiotics in IBD using the measured value of sCD14-ST in blood as an index, without compromising safety and efficacy, or without worsening intestinal bacterial flora, remission of IBD, severe IBD This is useful in that it can reduce the degree of activity, suppress the activity of IBD, shorten the treatment period, reduce the amount of antibiotics used, and reduce medical costs.

Claims

Detecting inflammatory bowel disease unresponsive to biological preparations, using sCD14-ST concentration in blood samples derived from patients with inflammatory bowel disease as an index.
A method for detecting inflammatory bowel disease unresponsive to a biologic comprising the following steps:
1) measuring the sCD14-ST concentration in a blood sample derived from a patient with inflammatory bowel disease,
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value.
The detection method according to claim 2, further comprising the following steps:
4) A step of determining that an unresponsive inflammatory bowel disease is detected in the biological preparation when the measured value is higher than the reference value.
The detection method according to claim 1, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.
The biologic is at least one selected from the group consisting of anti-TNFα antibody, soluble TNF receptor, anti-IL-6 receptor antibody, anti-IL-2 receptor antibody, anti-α integrin antibody and antisense NFκB. The detection method according to claim 1, wherein
A method for selecting an inflammatory bowel disease patient to be administered with an antibiotic, comprising the following steps:
1) measuring the sCD14-ST concentration in a blood sample derived from a patient with inflammatory bowel disease;
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value.
A method for selecting an inflammatory bowel disease patient to be administered MR1007, comprising the following steps:
1) measuring the sCD14-ST concentration in a blood sample derived from a patient with inflammatory bowel disease;
2) comparing the measured value of the sCD14-ST concentration with a reference value; and 3) determining whether the measured value is higher than the reference value.
A method of treating inflammatory bowel disease by administering an antibiotic comprising the following steps:
1) a step of measuring sCD14-ST in a blood sample derived from an inflammatory bowel patient,
2) a step of selecting a patient to be administered an antibiotic using the measured value of sCD14-ST in the sample as an index; and 3) a step of administering an antibiotic to the selected patient.
A method for treating inflammatory bowel disease by administering MR1007 comprising the following steps:
1) a step of measuring sCD14-ST in a blood sample derived from an inflammatory bowel patient,
2) a step of selecting a patient to which MR1007 is administered using the measured value of sCD14-ST in the specimen as an index; and 3) a step of administering MR1007 to the selected patient.