JP2009132636A - Pharmaceutical composition for treating gastrointestinal stromal tumor (gist), and kit and method for estimating prognosis of patient suffering from gastrointestinal stromal tumor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a tumor marker for discriminating a prognostic malignant case from a prognostic benign case at a high discrimination ratio in gastrointestinal stromal tumor cases. <P>SOLUTION: The pharmaceutical composition for treating a gastrointestinal stromal tumor of a patient defined based on the expression of DDX39 among patients suffering from the gastrointestinal stromal tumor comprises as an active ingredient 4-(4-methylpiperazin-1-ylmethyl)-N-[4-methyl-3-(4-pyridin-3-yl)pyrimidin-2-ylaminophenyl]-benzamide represented by formula (I) or a pharmacologically acceptable salt thereof. The kit and the method for estimating prognosis of a patient suffering from the gastrointestinal stromal tumor are also provided. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a pharmaceutical composition for treating a gastrointestinal stromal tumor (GIST), and a kit and method for predicting the prognosis of a patient suffering from a gastrointestinal stromal tumor.

  Gastrointestinal stromal tumors are resistant to chemotherapy and radiation therapy, and surgery has been the only complete remission therapy. However, in recent years, a tyrosine kinase inhibitor (imatinib mesylate, trade name: Gleevec) has come to be used for treatment (Patent Document 1: Special Table 2004-512328). Imatinib mesylate has been reported to be effective in many cases of GI stromal tumors but also cause serious side effects. For this reason, imatinib mesylate is currently prescribed mainly for relapsed and advanced cases after surgery.

  If patients with poor prognosis can be identified in advance, treatment with imatinib mesylate can be started in advance. As such prognostic prediction markers, the relationship between clinical pathology and prognosis (Non-patent document 1: Hasegawa T et al. (2002 Jun)), the relationship between mutation in Kit exon 11 and prognosis (Non-patent document 2: Andersson) J et al. (2006)) has been reported. However, these are not sufficient at the level of practical use. Thus, no tumor markers or clinicopathological factors have been established for gastrointestinal stromal tumors that are reliable with sufficient accuracy to determine treatment strategies and can predict prognosis at a level that can be put to practical use in clinical settings. . In addition, there has been no attempt to identify a poor prognosis factor by mRNA or protein expression analysis.

Japanese translations of PCT publication No. 2004-512328 and WO 02/34727 Japanese Patent Application Laid-Open No. 6-87834, Japanese Patent No. 2706682, European Patent Application No. 0564409, US Pat. No. 5,521,184 Japanese translations of PCT publication No. 2001-510192, Japanese Patent No. 3276359, International Publication No. 99/03854 International Publication No. 2007/043295 Pamphlet International Publication No. 2004/022778 Pamphlet Hasegawa T, Matsuno Y, Shimoda T, Hirohashi S., “Gastrointestinal stromal tumor: consistent CD117 immunostaining for diagnosis, and prognostic classification based on tumor size and MIB-1 grade.”, Human Pathology, 2002 Jun; 33 (6): p.669-676 Andersson J, Bumming P, Meis-Kindblom JM, Sihto H, Nupponen N, Joensuu H, Oden A, Gustavsson B, Kindblom LG, Nilsson B., “Gastrointestinal stromal tumors with KIT exon 11 deletions are associated with poor prognosis.”, Gastroenterology, 2006 May; 130 (6): p.1573-1581 T Sugiura et al., “Intracellular characterization of DDX39, a novel growth-associated RNA helicase”, Experimental cell research, 2007; 313: p.782-790 Joon T. Park et al., “Notch3 Gene Amplification in Ovarian Cancer”, Cancer Res, 2006; 66: p.6312-6318 I. Gashaw et al., “Gene signatures of testicular seminoma with emphasis on expression of ets variant gene 4”, CMLS, Cell. Mol. Life Sci, 2005; 62: p.2359-2368 J. Takita et al., “Gene expression profiling and identification of novel prognostic marker genes in neuroblastoma”, Genes, Chromosomes And Cancer 40: 120-132,2004 Skye H. C et al., “Genomic Prediction of Locoregional Recurrence After Mastectomy in Breast Cancer”, J Clin Oncol, 2006; 28: p.4594-4601 Hasegawa T, Yokoyama R, Lee YH, et al., "Prognostic relevance of a histological grading system using MIB-1 for adult soft-tissue sarcoma.", Oncology, 2000; 58: p.66-74 Hasegawa T, Yamamoto S, Nojima T, et al., “Validity and reproducibility of histologic diagnosis and grading for adult soft tissue sarcomas.”, Human Pathology, 2002 Jan; 33 (1): p.111-115

  An object of the present invention is to provide a tumor marker that distinguishes a poor prognosis case and a good prognosis case with a high discrimination rate in gastrointestinal stromal tumor cases.

According to one aspect of the present invention, a pharmaceutical composition for treating a gastrointestinal stromal tumor of a patient defined based on the expression of DDX39 among patients suffering from a gastrointestinal stromal tumor comprising:
Formula (I):
4- (4-methylpiperazin-1-ylmethyl) -N- [4-methyl-3- (4-pyridin-3-yl) pyrimidin-2-ylamino) phenyl] -benzamide or a pharmaceutically acceptable salt thereof A pharmaceutical composition comprising the salt thereof as an active ingredient is provided.

  According to another aspect of the present invention, a kit for predicting the prognosis of a patient suffering from a gastrointestinal stromal tumor, comprising a means for detecting or quantifying the expression of DDX39 in a sample obtained from the patient Is provided.

According to another aspect of the present invention, a method for predicting the prognosis of a patient suffering from a gastrointestinal stromal tumor comprising:
Providing a sample obtained from the patient;
Detecting or quantifying the expression of DDX39 in the sample;
Predicting the prognosis of the patient based on the expression of DDX39.

  As will be described in detail below, according to the present invention, a tumor marker that distinguishes a poor prognosis case and a good prognosis case with a high discrimination rate in gastrointestinal stromal tumor cases is provided.

  Hereinafter, embodiments of the present invention will be described. However, the present invention is not limited to the embodiments described below.

  The inventors surprisingly found that the expression of the protein DDX39 was significantly higher in the primary tumor tissue of postoperative metastatic cases of gastrointestinal stromal tumor than in the primary tumor tissue of non-metastatic cases I found The present inventors have developed a tumor marker for predicting the prognosis of patients with gastrointestinal stromal tumors by proteome analysis. As described in detail below, there were a group of cases with no metastasis for more than 3 years and low pathological malignancy, and a group of cases with early metastasis (within 1 year) and high pathological malignancy. Compared. A comprehensive expression analysis of the protein was performed on the surgical specimen by fluorescent two-dimensional electrophoresis, and DDX39 was found as a specific protein capable of distinguishing both groups from about 5000 protein spots.

  Thus, according to one aspect of the present invention, kits and methods for predicting the prognosis of a patient suffering from a gastrointestinal stromal tumor are provided. That is, according to the present invention, by using DDX39 as a tumor marker, a poor prognosis group can be found in advance among gastrointestinal stromal tumor cases. Furthermore, in such cases, it is expected that treatment results for gastrointestinal stromal tumors will be improved by starting administration of a therapeutic agent for gastrointestinal stromal tumors from an early stage. In addition, the prediction of the prognosis of the gastrointestinal stromal tumor according to the present invention can be performed at any time point after or before the operation of the gastrointestinal stromal tumor. For example, the prediction of the prognosis of gastrointestinal stromal tumor according to the present invention can be used when examining the necessity of chemotherapy such as administration of imatinib mesylate before and / or after surgery. In the present specification, poor prognosis means that gastrointestinal stromal tumors metastasize after surgery or that the disease state of gastrointestinal stromal tumors deteriorates.

  Note that DDX39 (UniProt Accession No .: O00148) is a kind of DEAD box RNA Helicases and has been reported to be hardly expressed in normal cells. RNA Helicases is considered to play an important role in RNA metabolism such as RNA splicing and transport, and DEAD box protein family is thought to be involved in development, spermatogenesis, cell proliferation and differentiation. . In the present specification, DDX39 consists of the amino acid sequence of SEQ ID NO: 1, and in the amino acid sequence of SEQ ID NO: 1, one or several amino acid residues are deleted, substituted, added or inserted. Includes those consisting of arrays.

  So far, DDX39 has been reported to be highly expressed in several tumor tissues (Non-patent Document 3: T Sugiura et al. (2007), Non-patent Document 4: Joon T. Park et al. (2006), Non-patent document 5: I. Gashaw et al. (2005), Non-patent document 6: J Takita et al. (2004)), multiple genetic markers for diagnosing the prognosis of breast and ovarian cancer tumors It was recognized as one of the sets (Non-patent Document 7: Skye H. C et al (2006), Patent Document 4: International Publication No. 2004/022778 pamphlet). However, according to the present invention, the prognosis of the tumor tissue can be predicted by using DDX39 as a single marker. Regarding the prediction of tumor prognosis, the use of DDX39 as a single marker has not been reported for any tumor. In particular, nothing related to the expression of DDX39 in gastrointestinal stromal tumors has been reported. That is, the present inventor has found that the prognosis of a patient suffering from a gastrointestinal stromal tumor can be predicted by using DDX39 as a single marker. As described above, it becomes possible to predict the prognosis with a single marker, which is technically simpler and quicker than when predicting the prognosis with multiple markers. In terms of cost, it becomes practical.

  The sample for detecting or quantifying the expression of DDX39 is preferably derived from a diseased tissue or blood of a patient suffering from a gastrointestinal stromal tumor. The sample to be used for the test may be the sample itself collected from the patient, or may be a sample obtained by processing the sample according to the test. For example, when a sample is subjected to an immunostaining method, a paraffin section prepared from a specimen obtained from a patient can be used as a sample to be subjected to the test. For example, when a sample is subjected to Western blotting or RT-PCR, a protein extract or mRNA extract prepared from a specimen obtained from a patient can be used as a sample to be tested.

  The expression of DDX39 in the sample can be detected or quantified by any method as exemplified below. The detection or quantification of DDX39 expression may be simply detection of the presence or absence of DDX39 expression, or may determine relative or absolute expression level of DDX39. DDX39 expression may also be detected or quantified at the protein level or may be detected or quantified at the mRNA level.

  Detection or quantification at the protein level includes, for example, immunostaining methods (including fluorescent antibody methods, enzyme antibody methods, heavy metal-labeled antibody methods, and radioisotope-labeled antibody methods), electrophoresis separation and fluorescence, enzymes, and radioisotopes. It can be carried out by a combination with detection or quantification by an element or the like (including Western blotting and fluorescence two-dimensional electrophoresis), enzyme immunoassay adsorption (ELISA), dot blotting and the like. In addition, detection or quantification at the mRNA level can be performed by, for example, RT-PCR (preferably real-time RT-PCR), Northern blotting, Branched DNA assay, and the like.

  The results of detection or quantification of DDX39 expression can be classified into two stages (positive and negative) or more than two stages. The classification of DDX39 expression is preferably performed by a pathologist, clinician, laboratory technician or laboratory with sufficient experience depending on the detection or quantification method. For example, the classification of DDX39 expression can be performed by a pathologist when using immunostaining, and can be performed by a laboratory technician when using RT-PCR.

  The classification of DDX39 expression is preferably performed by comparing the expression level of DDX39 in the sample from the patient with the expression level of DDX39 in the control. Depending on the number of stages that classify the results of DDX39 expression, it is preferable to use multiple controls. For example, when the results of DDX39 expression are classified into two types (positive and negative), it is preferable to use two types of controls (DDX39 positive control and DDX39 negative control) corresponding to each step. Moreover, when classifying the result of DDX39 expression into three types of stages, it is preferable to use three types of controls corresponding to the respective stages. In addition, as one of the controls, it is preferable to use a control derived from a healthy person or a gastrointestinal stromal tumor patient with a good prognosis. The control may be prepared in advance or tested at the same time as the sample from the patient.

  According to the present invention, a kit for predicting the prognosis of a patient suffering from a gastrointestinal stromal tumor is provided. The kit includes means for detecting or quantifying DDX39 expression in a sample obtained from a patient. That is, in the case of performing detection or quantification at the protein level, the means for detecting or quantifying expression preferably includes a means for performing immunostaining or the like. For example, anti-DDX39 antibody (monoclonal and polyclonal antibodies, Including a labeled antibody, a chimeric antibody, a humanized antibody, and binding-binding fragments thereof). In addition, in the case of performing detection or quantification at the mRNA level, the means for detecting or quantifying expression preferably includes means for performing the above-described RT-PCR and the like, for example, including a probe for DDX39 mRNA. It is preferable. Furthermore, as will be apparent to those skilled in the art, means for detecting or quantifying expression include reagents such as buffers, chromogenic substrates, secondary antibodies, blocking agents, instruments required for testing, controls, and instruction manuals and evaluation instructions. (Including a recording medium such as a CD-ROM containing electronic data having the same content).

  As described in detail in the Examples, the inventors have found that the expression of DDX39 correlates with poor patient prognosis. That is, it can be predicted that the higher the expression of DDX39, the higher the possibility of a poor prognosis.

  It should be noted that the judgment criteria based on the expression of DDX39 can be individually set according to the stage of treatment and treatment means (administration of imatinib mesylate, etc.). For example, the judgment standard for examining the necessity of administration of imatinib mesylate before surgery and the judgment standard for examining the necessity of administration of imatinib mesylate after surgery can be different. In addition, imatinib mesylate has problems of large side effects and high medical costs, but the above criteria are based on the administration of imatinib mesylate before and / or after surgical removal of gastrointestinal stromal tumors. It is preferable to use it when examining the necessity of this.

  As described above, according to the present invention, the prognosis of a gastrointestinal stromal tumor can be predicted with a higher discrimination rate, and accordingly, a more appropriate treatment plan can be made based on this, and more appropriate treatment is performed. It becomes possible. For example, when predicting the prognosis of a patient to investigate the necessity of administration of imatinib mesylate before and / or after surgical removal of gastrointestinal stromal tumors, the prognosis may be poor as described above When it is determined that the drug is high, it can be determined that it is necessary to receive chemotherapy with imatinib mesylate. In addition, when it is determined that the possibility of poor prognosis is low, it can be determined that there is no need for chemotherapy with imatinib mesylate.

  As will be described in detail in the Examples, according to the present invention, the prognosis of a patient can be predicted with very high sensitivity and specificity. In addition, imatinib mesylate has problems of large side effects and high medical costs.According to the present invention, administration of the therapeutic agent to patients who do not require treatment with imatinib mesylate is avoided. Thus, the above-mentioned problems can be avoided, and further, treatment with the therapeutic agent can be started more rapidly for patients who require treatment with imatinib mesylate. Thus, according to the present invention, a prognosis prediction method and kit having a very high diagnostic value are provided. On the other hand, the prediction according to the present invention can be performed inexpensively by a normal technical level pathologist or the like with a commonly used simple technique such as an immunostaining method without requiring a special apparatus or the like. it can.

  According to another aspect of the present invention, there is provided a pharmaceutical composition for treating a gastrointestinal stromal tumor of a patient defined based on the expression of DDX39 among patients suffering from a gastrointestinal stromal tumor. The pharmaceutical composition according to the present invention comprises 4- (4-methylpiperazin-1-ylmethyl) -N- [4-methyl-3- (4-pyridin-3-yl) pyrimidine- represented by the above formula (I). 2-ylamino) phenyl] -benzamide or a pharmaceutically acceptable salt thereof as an active ingredient.

  4- (4-Methylpiperazin-1-ylmethyl) -N- [4-methyl-3- (4-pyridin-3-yl) pyrimidin-2-ylamino) phenyl] -benzamide or a pharmaceutically acceptable salt thereof And the use thereof as an anti-tumor agent is described in Patent Document 2 (Embodiment 21 of European Patent Application No. 0564409, as well as other corresponding applications and corresponding patents such as Japanese Patent No. 2706682). U.S. Pat. No. 5,521,184).

  Also, 4- (4-methylpiperazin-1-ylmethyl) -N- [4-methyl-3- (4-pyridin-3-yl) pyrimidin-2-ylamino) phenyl] -benzamide or pharmaceutically acceptable The salt is preferably a pharmaceutically acceptable acid addition salt, such as a salt with an inorganic acid such as hydrochloric acid, sulfuric acid or phosphoric acid, or a suitable organic carboxylic acid or sulfonic acid such as trifluoro Such as acetic acid, acetic acid, propionic acid, glycolic acid, succinic acid, maleic acid, fumaric acid, hydroxymaleic acid, mono- or di-carboxylic acid such as malic acid, tartaric acid, citric acid or oxalic acid, or arginine or lysine Amino acids, benzoic acid, 2-phenoxy-benzoic acid, 2-acetoxy-benzoic acid, salicylic acid, 4-aminosalicylic acid Aromatic carboxylic acids such as, aromatic-aliphatic carboxylic acids such as mandelic acid or cinnamic acid, heterocyclic aromatic carboxylic acids such as nicotinic acid or isonicotinic acid, methane-, ethane- or 2-hydroxy Salts with aliphatic sulfonic acids such as ethane-sulfonic acid or aromatic sulfonic acids such as benzene-, p-toluene- or naphthalene-2-sulfonic acid.

  Also, monomethanesulfonic acid addition salt of 4- (4-methylpiperazin-1-ylmethyl) -N- [4-methyl-3- (4-pyridin-3-yl) pyrimidin-2-ylamino) phenyl] -benzamide And preferred crystal forms thereof are described in Patent Document 3 (International Patent Application No. 99/03654 pamphlet and other corresponding applications and corresponding patents such as Japanese Patent No. 3276359).

  The effective dosage of the pharmaceutical composition according to the present invention is, for example, about 100 to 1000 mg per day as imatinib for a patient having a body weight of about 70 kg, depending on the age, weight, medical condition, mode of administration and clinical picture of the patient. Preferably it is 200-600 mg, More preferably, it is 400 mg.

  The pharmaceutical composition according to the present invention may further comprise a pharmaceutically acceptable carrier. As the pharmaceutically acceptable carrier, any standard carrier, buffer and excipient (for example, phosphate buffered saline, 5% aqueous dextrose solution) can be used. The preferred carrier depends on the mode of administration. Typical modes of administration include enteral (eg, oral) or parenteral (eg, subcutaneous, intramuscular, intravenous or intraperitoneal injection, or topical, transdermal, or transmucosal administration). Is preferred. For example, for oral administration, it can be administered in solid dosage forms such as capsules, tablets and powders, or liquid dosage forms such as elixirs, syrups and suspensions.

  As described above, the pharmaceutical composition according to the present invention is intended for patients defined based on the expression of DDX39. Specifically, as described above, the pharmaceutical composition according to the present invention is preferably intended for patients predicted to have a poor prognosis based on the expression of DDX39. As described above, according to the present invention, there is provided a pharmaceutical composition for treating a gastrointestinal stromal tumor of a patient defined based on the expression of DDX39 among patients suffering from a gastrointestinal stromal tumor.

  In addition, imatinib mesylate, which is a therapeutic agent for gastrointestinal stromal tumor, is sold as Gleevec (registered trademark) by Novartis Pharma Co., Ltd. The attached document clearly states that patients diagnosed with KIT (CD117) -positive gastrointestinal stromal tumors by immunohistological examination are targeted for the therapeutic agent. Thus, the target patient group of the pharmaceutical composition according to the present invention and the target patient group of Gleevec (registered trademark) can be clearly distinguished by those skilled in the art, and the pharmaceutical composition according to the present invention is The target patient, i.e. in medical use, is distinct from Gleevec®.

  In addition, the pharmaceutical composition according to the present invention is also preferably intended for patients diagnosed as KIT (CD117) -positive gastrointestinal stromal tumors by immunohistological examination, as in Gleevec (registered trademark). As widely performed by those skilled in the art, KIT (CD117) positive confirmation is performed by, for example, immunization using an anti-human c-Kit (CD117) rabbit polyclonal antibody (code number: A4502, manufactured by Dako Japan Co., Ltd.). It can be performed by a staining method. The immunostaining method can be performed according to the method described in the manufacturer's instruction manual. Confirmation of KIT (CD117) positivity can be performed by immunostaining using the antibody, as described in the attached document of the anti-human c-Kit (CD117) rabbit polyclonal antibody.

  Examples of the present invention will be described below. However, the present invention is not limited to the examples described below.

[Materials and methods]
[1. sample〕
(1-1. Surgical specimen)
Surgical specimens obtained from patients with gastrointestinal stromal tumors with written consent to use surgical specimens for research were used. The study was reviewed and approved by the National Cancer Center's Genetic Ethics Committee for the use of surgical specimens for gastrointestinal stromal tumor cases.

(1-2. Protein recovery)
Tumor tissue was collected from the surgical specimen of the case and frozen with liquid nitrogen. The frozen tumor tissue was crushed with a multi-bead shocker (Yasui Kikai, Osaka) to form a powder. Protein extraction buffer (6M urea, 2M thiourea, 3% CHAPS, 1% TritonX-100) was added to the powdered tissue to extract the protein.

[2. (Fluorescence two-dimensional electrophoresis)
(2-1. Sample adjustment)
The extracted protein was centrifuged at 15000 rpm, and the supernatant was collected. Proteins contained in the supernatant were quantified with a protein quantification kit (DC protein assay kit, Bio-Rad, USA). 5 μg of the extracted protein was labeled with a fluorescent dye (saturation dye Cy5, GE Healthcare Biosciences). The labeling was performed as follows. (1) Tris buffer at pH 8.0 was added to a final concentration of 30 mM, then (2) 1 nmol of TECP (Tris (2-carboxyethyl) phosphine hydrochloride, Sigma) was added, and (3) 37 degrees For 60 minutes. Next, (4) 4 nmol of Cy5 fluorescent dye was added and treated at 37 degrees for 60 minutes. An equal amount of protein sample was collected from the protein sample used in this experiment and mixed to obtain an internal standard sample. 5 μg of the internal standard sample was labeled with a fluorescent dye (saturation dye Cy3, GE Healthcare Biosciences) as described above. Individual samples labeled with Cy5 and internal standard samples labeled with Cy3 were mixed and brought to a final volume of 420 μl with urea solubilization solution. At that time, DTT was added so that the final concentration was 65 mM, and ampholine (GE Healthcare Biosciences) was added so as to be 2%. A sample in which an individual sample labeled with Cy5 and an internal standard sample labeled with Cy3 were mixed was run on a single two-dimensional electrophoresis gel.

(2-2. Electrophoresis)
First, immobiline gel (24 cm, pI4-7, GE Healthcare Biosciences) and Multiphor II (GE Healthcare Biosciences) were used for the first dimension electrophoresis. Immobiline gels were swollen overnight at room temperature with the protein samples to be run. Electrophoresis was performed at 40,000 Vh. The second-dimension electrophoresis used a 9-15% polyacrylamide gradient gel and GiantGelRunner (Biocraft). Electrophoresis was carried out at 20 degrees for 17 hours at 18 W per electrophoresis apparatus.

(2-3. Protein detection)
After completion of the electrophoresis, for the purpose of detecting the protein, the gel in a state of being sandwiched between the glass plates was placed on a laser scanner (Typhoon Trio, GE Healthcare Biosciences) and scanned.

(2-4. Expression analysis)
The read image was analyzed by image analysis software Progenesis (Non-linear Dynamic).

[3. Protein identification
(3-1. In-gel digestion method)
Spots were collected from the gel into a 96-well plate using a fully automatic spot collection device ProHunter (AsOne). The gel was thoroughly washed with methanol and treated with proteolytic enzyme (trypsin) at 37 degrees overnight. This treatment results in peptide formation of the protein. The obtained peptide was recovered by washing the gel with 60% acetonitrile.

(3-2. Mass spectrometry)
LTQ (Thermo) was used to measure the mass of the peptide. MasCot was used to search the database for protein identification.

[4. (Western blotting)
10 μg of protein extracted from the surgical specimen by the method described above was separated by SDS-PAGE and transferred to a nitrocellulose membrane. The antibody used was an anti-DDX39 antibody (Gene way). The dilution concentration of the antibody was 1000 times, and an ECL kit (GE Healthcare Biosciences) was used for detection.

[6. Pathological judgment)
(6-1. Presence or absence of metastasis)
If there are no obvious lesions appearing anywhere in the body, such as lungs, lymph nodes, abdominal cavity, etc., based on examination findings and routine imaging (CT, MRI, simple radiograph) during outpatient follow-up It was judged.

(6-2. Pathological grade (risk classification))
In accordance with the method described in Non-Patent Document 1 (Hasegawa H et al. (2002)), pathological malignancy was determined as described in Table 1 and the above description related thereto. A pathologist determined the score for each of tumor differentiation, tumor necrosis and MIB-1.

[Example 1]
Proteins were extracted from surgical specimens of gastrointestinal stromal tumors and analyzed for proteins involved in metastasis and prognosis using fluorescence two-dimensional electrophoresis and mass spectrometry. 1) 9 cases with metastasis within 1 year after surgery and high degree of pathological malignancy (risk classification) 2) No metastasis and moderate pathological malignancy for 3 years after surgery Eight cases of surgical specimens were used. From about 5000 protein spots, 38 protein spots having different concentrations between the two groups were selected (P value of Wilcoxon test was 0.01 or less and the average value was 2 times or more).

  As a result of protein identification using a mass spectrometer, one protein spot (DDX39 / spot) derived from the protein DDX39 was included in 38 spots. Depending on the concentration of DDX39 / spot, the above 9 cases with poor prognosis and 8 cases with good prognosis have sensitivity (true positive rate, probability that a patient with poor prognosis will be positive), specificity (true negative rate) The probability that a patient with a good prognosis would have a negative test) could be identified at 100% (see FIG. 1). Even when Western blotting using a specific antibody against DDX39 is performed on the same surgical specimen, 9 cases with poor prognosis can be distinguished from 8 cases with good prognosis with both accuracy and specificity of 100%. (See FIG. 2).

It is a graph which shows the density | concentration of DDX39 * spot obtained by the fluorescence two-dimensional electrophoresis method in 9 cases with a bad prognosis and 8 cases with a good prognosis. (A) It is a photograph showing the results of Western blotting in 9 cases with poor prognosis and 8 cases with good prognosis.

Claims (5)

A pharmaceutical composition for treating a gastrointestinal stromal tumor of a patient defined based on expression of DDX39 among patients suffering from a gastrointestinal stromal tumor, comprising:
Formula (I):
4- (4-methylpiperazin-1-ylmethyl) -N- [4-methyl-3- (
A pharmaceutical composition comprising 4-pyridin-3-yl) pyrimidin-2-ylamino) phenyl] -benzamide or a pharmaceutically acceptable salt thereof as an active ingredient.   The pharmaceutical composition according to claim 1, wherein the patient is a DDX39-positive patient among patients suffering from gastrointestinal stromal tumors.   A kit for predicting the prognosis of a patient suffering from a gastrointestinal stromal tumor, comprising a means for detecting or quantifying the expression of DDX39 in a sample obtained from the patient. A method for predicting the prognosis of a patient suffering from a GI stromal tumor,
Providing a sample obtained from the patient;
Detecting or quantifying the expression of DDX39 in the sample;
Predicting the prognosis of the patient based on the expression of DDX39.   The method according to claim 8, wherein the predicting step determines that the prognosis is poor when DDX39 is positive.