JP2003212795A - Diagnostic agent of schizophrenia - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a diagnostic agent of schizophrenia. <P>SOLUTION: This diagnostic agent of the schizophrenia contains an anti-basic fibroblast growth factor antibody as an active ingredient. The schizophrenia is diagnosed by measuring a basic fibroblast growth factor in serum by using the antibody. <P>COPYRIGHT: (C)2003,JPO

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a diagnostic agent for schizophrenia, and more particularly to a diagnostic agent for schizophrenia mainly composed of anti-basic fibroblast growth factor antibody.

[0002]

BACKGROUND OF THE INVENTION Schizophrenia occurs in about 1% of the population and develops from adolescence to early 20s. Symptoms include positive symptoms such as hallucinations and delusions, apathy, demoralization, negative symptoms such as social withdrawal, and cognitive impairment. Due to the peculiarity of the pathological condition, it is desired to establish a comprehensive treatment system such as early diagnosis, treatment, rehabilitation activities, and recurrence prevention. Drug treatment is indispensable for the treatment of schizophrenia, and phenothiazine compounds, butyrophenone compounds, benzamide compounds, iminodibenzyl compounds, thiepine compounds, indole compounds and serotonin / dopamine receptor blockers etc. Is being administered. It is known that genetic factors and environmental factors are involved in the pathological condition of schizophrenia, and a so-called "developmental disorder hypothesis of schizophrenia" has been proposed as a disorder at the developmental stage. Currently, there are no effective biological markers to diagnose schizophrenia. There have been many studies using patient samples such as blood and urine, but none have been established yet.

With respect to many proteins including acidic fibroblast growth factor, the expression levels of the genes of these proteins are changed in patients with schizophrenia, and therefore these genes are used as a diagnostic index for schizophrenia. A method has been proposed (Japanese Patent Laid-Open No. 2001-245661). On the other hand, the basic fibroblast growth factor focused on in the present invention has a growth promoting action not only in fibroblasts but also in a large number of cell lines, and modifies the differentiation function of cells. It is known that the central nervous system also has a neuroprotective action such as an inhibitory action on apoptosis. Basic fibroblast growth factor is overexpressed in many malignant tumors, and it is speculated that it is involved in tumor development. However, the role of basic fibroblast growth factor in psychiatric and neurological diseases such as schizophrenia is completely unknown.

[0004]

[Problems to be Solved by the Invention] Inpatients with schizophrenia account for about 15% of the total number of beds in Japanese hospitals, which is a serious problem from the viewpoint of medical economy. It is desired to develop a diagnostic agent and a diagnostic method for diagnosing schizophrenia at an early stage from the medical field. The object of the present invention is to provide such a diagnostic agent and diagnostic method.

[0005]

The present inventors have found that the level of basic fibroblast growth factor in serum of schizophrenic patients is significantly increased as compared with that of healthy subjects, The present invention has been completed based on these findings. That is,
The present invention is a diagnostic agent for schizophrenia, which comprises an anti-basic fibroblast growth factor antibody as an active ingredient. The anti-basic fibroblast growth factor antibody may be labeled. The present invention also provides a method for diagnosing schizophrenia, which comprises measuring the concentration of basic fibroblast growth factor in the serum of a patient using these diagnostic agents. Further, the present invention is a method for determining a therapeutic drug for schizophrenia using these diagnostic agents. Furthermore, the present invention provides a schizophrenia diagnostic kit comprising an anti-basic fibroblast growth factor antibody or a labeled anti-basic fibroblast growth factor antibody as a main component, and using these schizophrenia diagnostic kits, the bases in the serum of patients. A method for diagnosing schizophrenia, which comprises measuring the concentration of sex fibroblast growth factor. Further, the present invention is a method for assaying a therapeutic drug for schizophrenia using an anti-basic fibroblast growth factor antibody or a labeled anti-basic fibroblast growth factor antibody.

The "anti-basic fibroblast growth factor antibody" in the present invention is an antibody purified by using basic fibroblast growth factor as an antigen, as long as it has the ability to bind to basic fibroblast growth factor. , Polyclonal antibodies, monoclonal antibodies. Further, preferred examples include a polyclonal antibody and a monoclonal antibody that specifically bind to basic fibroblast growth factor. "Labeled anti-basic fibroblast growth factor antibody" refers to basic fibroblast growth factor antibody labeled with peroxidase, β-D-galactosidase, alkaline phosphatase, glucose-6-phosphate dehydratase and other enzymes, and fluorescent labeling such as delphinium. , An antibody designed to bind a radioactive isotope label or an isotope label, biotin, etc. and to quantify the basic fibroblast growth factor. Furthermore, the “labeled anti-basic fibroblast growth factor antibody” also includes anti-basic fibroblast growth factor antibody modified with biotin, 2,4-dinitrophenol or the like. In this case, the labeled anti-basic fibroblast growth factor antibody can be quantified by further using avidin labeled with the labeled anti-2,4-dinitrophenol antibody. “Schizophrenia” is a chronic disease that develops mainly during adolescence and adolescence, and has ego disorders and thought disorders as its roots, and gradually becomes more serious by repeating positive symptoms such as hallucinations and delusions. is there.

[0007]

BEST MODE FOR CARRYING OUT THE INVENTION Diagnosis of schizophrenia according to the present invention can be performed, for example, as follows. Serum is prepared from human blood and the amount of basic fibroblast growth factor in the serum is quantified by various methods. Desirably, the basic fibroblast growth factor is detected by sandwich ELISA using an antibody highly specific to the basic fibroblast growth factor.
Quantify. Diagnosis of schizophrenia is made by utilizing the fact that the concentration of basic fibroblast growth factor in serum of patients with schizophrenia is significantly higher than that of healthy subjects.

Specific methods for measuring basic fibroblast growth factor in serum include, for example, 1. 1. Immobilizing the anti-basic fibroblast growth factor antibody on a solid phase such as polystyrene, nylon, glass, silicon rubber, sepharose; 2. Adding or contacting the serum of the patient to be diagnosed with the solid phase; 3. Washing the solid phase; 4. adding or contacting a labeled anti-basic fibroblast growth factor antibody; A method comprising a step of measuring the amount of basic fibroblast growth factor using the label can be mentioned.

Further, specific methods for measuring basic fibroblast growth factor in serum include, for example: 1. Immobilizing the anti-basic fibroblast growth factor antibody on a solid phase such as polystyrene, nylon, glass, silicon rubber, sepharose; 2. Adding or contacting the serum of the patient to be diagnosed with the solid phase; 3. Washing the solid phase; 4. Adding or contacting anti-basic fibroblast growth factor antibody modified with biotin or 2,4-dinitrophenol; 5. adding or contacting labeled avidin or labeled 2,4-dinitrophenol antibody; A method comprising a step of measuring the amount of basic fibroblast growth factor using the label can be mentioned.

Further, specific methods for measuring basic fibroblast growth factor in serum include, for example, 1. 1. Immobilizing the anti-basic fibroblast growth factor antibody on a solid phase such as polystyrene, nylon, glass, silicon rubber, sepharose; 2. Adding or contacting the serum of the patient to be diagnosed with the solid phase; 3. Washing the solid phase; 4. Adding or contacting biotin-modified anti-basic fibroblast growth factor antibody; 5. Adding or contacting labeled avidin; A method comprising the step of measuring the amount of basic fibroblast growth factor using the label. The shape of the solid phase includes small spheres, wells, test tubes and the like.

As the basic fibroblast growth factor used as the antigen or ELISA standard, a commercially available product may be used, or the basic fibroblast growth factor may be prepared by the following method.
When using a genetic engineering technique, a gene encoding a basic fibroblast growth factor (GeneBank No .: M34641) is incorporated into an appropriate vector, and this is inserted into an appropriate host for transformation, and the culture of this transformation is performed. The desired recombinant basic fibroblast growth factor can be obtained from the silkworm, and it is suitable for producing a uniform and large amount of basic fibroblast growth factor. The above-mentioned host cells are not particularly limited, and various host cells conventionally used in genetic engineering techniques such as Escherichia coli, Bacillus subtilis, yeast, plant or animal cells can be used. The anti-basic fibroblast growth factor antibody is prepared by immunizing rabbits, chickens, turkeys and the like with the basic fibroblast growth factor as an antigen. The labeled anti-basic fibroblast growth factor antibody can be prepared by reacting the anti-basic fibroblast growth factor antibody with a commercially available kit of a biotinylation reagent or a peroxidase with a crosslinking agent.

The present invention is also useful for determining a therapeutic drug for schizophrenia. That is, a compound having an action of decreasing the basic fibroblast growth factor may be useful as a therapeutic drug for schizophrenia. In addition, a model animal (mouse, rat, etc.) having a high amount of basic fibroblast growth factor is also useful as a model of schizophrenia. Therefore, by using this assay method, it is possible to screen a new therapeutic drug for schizophrenia.

The therapeutic agents found by such methods include:
Drugs that can be administered parenterally or orally can be included. The exact dose and dosing schedule of the therapeutic agent must be determined according to the required dose for each treatment symmetry, the treatment method, the degree of disease or need, and, of course, the judgment of the doctor. When administered parenterally, the dose and frequency of administration vary depending on symptoms, age, weight, dosage form, etc., but when administered subcutaneously or intravenously as an injection, for example, an adult patient weighs 1 k
g, in the range of about 0.1 mg to about 2500 mg per day,
The dose is preferably selected from the range of about 1 mg to about 500 mg, for example, when administered as a spray to the trachea, the adult patient weighs 1 kg, about 0.1 mg to about 2 per day.
Range of 500 mg, preferably about 1 mg to about 500 mg
The dose is selected from the range. The administration schedule includes daily administration, intermittent administration, or a combination thereof. The dose and the number of administrations in the case of oral administration vary depending on the symptoms, age, body weight, administration form, etc., but for example, 1 kg of adult patient's body weight, about 0.5 mg to about 2500 m per day.
The dosage is selected from the range of g, preferably from about 1 mg to about 1000 mg.

A pharmaceutical composition may be prepared which comprises admixing the therapeutic agent for schizophrenia obtained by the method of the present invention with a pharmaceutically acceptable non-toxic carrier. When such a composition is prepared for parenteral administration (subcutaneous injection, intramuscular injection, or intravenous injection), a solution form or suspension form is particularly preferable, and a vaginal or rectal administration is particularly preferable. Semisolid dosage forms such as creams or suppositories are preferred, especially for nasal administration, especially powders, nasal drops or aerosol dosage forms.

The composition may be administered in a single dosage form, and may be for example Remington's Pharmaceutical Sciences (Mac Publishing Company, Easton, PA, 1970).
Years) and can be prepared by any method well known in the pharmaceutical art. Injectable formulations include, as pharmaceutical carriers, plasma-derived proteins such as albumin,
Amino acids such as glycine and sugars such as mannitol can be added. When used in the form of an injection, a buffering agent, a solubilizing agent, an isotonic agent and the like can be further added. When used as a water-soluble formulation or a freeze-dried formulation, Tween 80 (registered trademark) or Tween 20 is used to prevent aggregation.
It is preferable to add a surfactant such as (registered trademark). Parenteral dosage forms other than injections may also contain distilled water or physiological saline, polyalkylene glycols such as polyethylene glycol, oils of vegetable origin, hydrogenated naphthalene and the like. Formulations for vaginal or rectal administration such as suppositories contain, for example, polyacetylene glycol, petrolatum, cacao oil and the like as common excipients. The vaginal preparation may contain an absorption enhancer such as bile salt, ethylenediamine salt and citrate. Formulations for inhalation may be solid, may contain eg lactose as an excipient and nasal drops may be a water or oil solution.

[0016]

EXAMPLES The present invention will be illustrated below with reference to Examples.
It is not intended to limit the light.Example 1 The test in this example was conducted for schizophrenia patients and healthy individuals.
This was done for each 20 persons in the group. Blood from these patients
After collecting, serum is obtained by a standard method. High sensitivity basic fiber
Fibroblast growth factor measurement kit (R & S Systems Inc. (Minneapo
lis, MN, USA), detection sensitivity: 0.25 pg / mL)
did. That is, the anti-basic fibroblast growth factor antibody is coated.
96-well plate buffer (100 μL)
After that, 150 μL of serum was added. Stander for quantitation
Human basic fibroblast growth factor (1 to 64 pg /
(mL) is used. Let react for 3 hours at room temperature
After washing, wash with buffer 6 times, and then use alkaline phosphater
ZE-labeled anti-basic fibroblast growth factor antibody is added at room temperature at 2:00
React for a while. Contains a stabilizer after washing 6 times with buffer
Add NADPH (50 μL) and react at room temperature for 45 minutes
Let Then add enhancer (50 μL) at room temperature
After reacting for 45 minutes, add stop solution (50 μL)
It Within 30 minutes, the pre-reader Emax (Molecular De
vices, USA) at a wavelength of 490 nm. Calibration
Calculate the concentration of basic fibroblast growth factor in the sample from the line
It As a result, in the schizophrenia group and the healthy group,
The level of basic fibroblast growth factor in serum is 3.6 on average.
1 pg / mL, 3.14 (SD), in healthy subjects, salt
The level of basic fibroblast growth factor is 1.61 pg / m on average
L, 2.31 (SD). The value between these two groups
Non-parametric Wilcoxon rank test
As a result of analysis, it was significant in patients with schizophrenia (z = -2.0
1, p = 0.04).

[0017]Example 2 About untreated patients (9) and patients under treatment (11)
Then, the inspection was performed in the same manner as in Example 1. Example 1
As a result of similar statistical analysis, z = -1.13, p = 0.26,
Basic fibroblast proliferation in untreated and treated patients
No significant change in the factors was observed. In addition, during treatment
Of 11 patients (Chlorpromazine conversion)
As a result of analysis, correlation with chlorpromazine conversion (r
= 0.11, p = 0.74) is not significant, and the correlation with disease duration
The relationship (r = 0.23, p = 0.32) was not significant. Immediately
However, untreated patients and those undergoing treatment have a basic fibroblast growth factor.
Not correlated with offspring levels, disease duration and salt
Not correlated with levels of basic fibroblast growth factor
understood.

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Claims (4)

[Claims] 1. A diagnostic agent for schizophrenia, which comprises an anti-basic fibroblast growth factor antibody as an active ingredient. 2. The diagnostic agent according to claim 1, wherein the anti-basic fibroblast growth factor antibody is labeled. 3. A method for diagnosing schizophrenia, which comprises measuring the concentration of basic fibroblast growth factor in the serum of a patient using the diagnostic agent according to claim 1. 4. A method for determining a therapeutic drug for schizophrenia using the diagnostic drug according to claim 1.