WO2022075318A1 - Evaluation marker reflecting efficacy of cancer treatment composition - Google Patents

Abstract

The present invention addresses the problem of providing: a method of detecting an evaluation marker that reflects the efficacy of a cancer treatment composition on pancreatic tumor; a cancer treatment composition; a method of using the evaluation marker; the evaluation marker; a test reagent; and an evaluation device that reflects the efficacy of a cancer treatment composition.　This problem is solved by a method of detecting an evaluation marker that reflects the efficacy of a cancer treatment composition on a pancreatic tumor patient with peritoneal dissemination, said method comprising a step for detecting CXCL12 in an area of interest including pancreatic tumor cells in a tissue that is collected from the target patient, wherein the cancer treatment composition comprises a paclitaxel compound.

Description

Evaluation markers that reflect the effectiveness of cancer therapeutic compositions

The present specification describes a method for detecting an evaluation marker that reflects the effectiveness of a composition for treating cancer, a composition for treating cancer, a method for using the evaluation marker, an evaluation marker, a test reagent, and a composition for treating cancer. An evaluation device that reflects the effectiveness of the above is disclosed.

Normally, conversion surgery is not applicable to patients with pancreatic cancer who have peritoneal dissemination. However, as described in Non-Patent Documents 1 to 3, among pancreatic cancer patients with peritoneal dissemination, intraperitoneal paclitaxel administration therapy is remarkably effective, conversion surgery is possible, and the prognosis is significantly improved. exist.

There is no known index for distinguishing between patients who are effective with intraperitoneal paclitaxel administration therapy and those who are not.

The present invention relates to a method for detecting an evaluation marker that reflects the effectiveness of a composition for treating cancer for pancreatic tumor, a composition for treating cancer, a method for using the evaluation marker, an evaluation marker, a test reagent, and a composition for treating cancer. The subject is to provide an evaluation device that reflects the effectiveness of an object.

As a result of intensive studies, the present inventor has found that paclitaxel-based compounds are effective in pancreatic tumor patients with a low abundance of CXCL12 (SDF-1) in pancreatic tumor cells.
The present invention has been completed based on the findings and includes the following aspects.
Item 1.
Including the step of detecting CXCL12 in a region of interest containing pancreatic tumor cells in a tissue collected from a subject.
A method for detecting an evaluation marker that reflects the effectiveness of a composition for treating cancer in patients with pancreatic tumors having peritoneal dissemination.
A method for detecting the evaluation marker, wherein the composition for treating cancer contains a compound represented by the following general formula (I).

(here,
R ¹ has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent.
^R2 has (2-1) a linear or branched alkyl group of C1-4 on the phenyl ring, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, and the like. A phenylcarbonyl group which may have at least one selected from the group consisting of a carboxy group and a cyano group as a substituent, or an alkenylcarbonyl group having (2-2) an alkenyl moiety having 2 to 6 carbon atoms. Indicates an amino group that may have.
R ³ , R ⁴ and R ⁵ represent the same or different hydrogen atom or hydroxyl group.
R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ represent the same or different linear alkyl groups of C1-3.
R ¹² has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent. ).
Item 2.
Further comprising the step of presenting a label indicating that the composition for treating cancer is effective when the score indicating the abundance of CXCL12 in the region of interest is equal to or less than the reference value.
The score is based on the ratio of (1) the staining intensity of CXCL12 protein in the pancreatic tumor cells and (2) the number of pancreatic tumor cells positive for CXCL12 protein staining when the number of pancreatic tumor cells is 100%. 1. The method for detecting the evaluation marker according to Item 1.
Item 3.
Item 2. The method for detecting an evaluation marker according to Item 1 or 2, further comprising a step of presenting a label indicating that the conversation surgery is applicable when the score in the region of interest falls below a reference value.
Item 4.
The compound represented by the general formula (I) is paclitaxel.
Item 8. The method for detecting an evaluation marker according to any one of Items 1 to 3.
Item 5.
A composition for treating cancer for use in patients with pancreatic tumors having peritoneal dissemination, which comprises a compound represented by the following general formula (I).
The cancer therapeutic composition is used for administration to the subject having a CXCL12 score of less than or equal to a reference value in an area of interest containing pancreatic tumor cells in a tissue collected from the subject.
The score is obtained based on the ratio of (1) the staining intensity of CXCL12 in the pancreatic tumor cells and (2) the number of pancreatic tumor cells positive for CXCL12 staining when the number of pancreatic tumor cells is 100%. The composition for treating cancer:

(here,
R ¹ has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent.
^R2 has (2-1) a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, and a nitro group on the phenyl ring. A phenylcarbonyl group which may have at least one selected from the group consisting of a carboxy group and a cyano group as a substituent, or an alkenylcarbonyl group having (2-2) an alkenyl moiety having 2 to 6 carbon atoms. Indicates an amino group that may have.
R ³ , R ⁴ and R ⁵ represent the same or different hydrogen atom or hydroxyl group.
R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ represent the same or different linear alkyl groups of C1-3.
R ¹² has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent. ).
Item 6.
Item 6. The composition for treating cancer according to Item 6, wherein the compound represented by the general formula (I) is paclitaxel.
Item 7.
A method of using CXCL12 in pancreatic tumor cells in a tissue collected from a subject as a marker for evaluating the efficacy of the composition for treating cancer according to Item 5 or 6, wherein the subject is the subject. , Said method, comprising peritoneal dissemination of a pancreatic tumor.
Item 8.
Item 5. An evaluation marker for evaluating the efficacy of the composition for treating cancer according to Item 5 or 6, which comprises CXCL12 and comprises a subject having peritoneal dissemination of a pancreatic tumor.
Item 9.
A detection reagent for detecting CXCL12 in pancreatic tumor cells in a tissue collected from a subject as a marker for evaluating the effectiveness of the cancer therapeutic composition according to Item 5 or 6, wherein the detection reagent is used. Contains an antibody for detecting CXCL12,
Here, the subject has a peritoneal dissemination of a pancreatic tumor.
The detection reagent.
Item 10.
A device for evaluating the effectiveness of a composition for treating cancer, which comprises a processing unit.
The processing unit
A score of CXCL12 in the area of interest containing pancreatic tumor cells in the tissue collected from the subject was obtained.
Compare the obtained score with the standard value and
When the obtained score falls below the reference value, a label indicating that the cancer therapeutic composition according to Item 5 or 6 is effective is output, and / or the obtained score sets the reference value. When it falls below, it outputs a label indicating the applicability of conversion surgery, where the subject has a peritoneal dissemination of a pancreatic tumor.
The evaluation device.

According to the present invention, it is possible to identify a pancreatic tumor patient to which a paclitaxel-based compound is significantly effective. In addition, it is possible to identify a patient to whom the conversion surgery can be applied from the patients having peritoneal dissemination of a pancreatic tumor, which was conventionally considered to be not applicable to the conversion surgery.

An example of the overview diagram of the evaluation system 1000 is shown. An example of the block diagram of the evaluation device 10 is shown. An example of the processing of the evaluation program 104b is shown. The score distribution of CXCL12 in the group to which the conversation surgery could not be applied and the group to which the conversation surgery could be applied is shown. The score distribution of CXCR4 in the group to which the conversation surgery could not be applied and the group to which the conversation surgery could be applied is shown.

1. 1. Explanation of Terms (1) Evaluation Marker CXCL motif ligand 12 (CXCL12) is a small molecule protein and is a kind of chemokine. CXCL12 is also called SDF-1 (Stromal Derived Factor-1), and is typically expressed from a gene registered in the National Center for Biotechnology Information with Gene ID: 6387. The evaluation marker consisting of CXCL12 includes a protein expressed from the gene or mRNA. In addition, the evaluation marker includes a protein or mRNA expressed from the gene, as well as a variant thereof.

(2) Pancreatic tumor Pancreatic tumor is not limited as long as it is a tumor derived from cells existing in the pancreas. Pancreatic tumors may include malignant tumors and benign tumors, but are preferably malignant tumors, more preferably cancers. Pancreatic cancer may include pancreatic ductal adenocarcinoma, mucinous cystadenocarcinoma, serous cystadenocarcinoma and the like. It is preferably pancreatic duct adenocarcinoma. The pancreatic tumor may be present in the pancreas, but may also be present in lymph nodes, portal veins, arteries, duodenum, bile duct, liver, peritoneum, lungs and the like. In addition, the pancreatic tumor may be initial or relapsed. It is preferably the first time.
Subjects are not limited as long as they have a pancreatic tumor. Preferably, the person has peritoneal dissemination of a pancreatic tumor.

(3) Composition for cancer treatment The composition for cancer treatment contains a paclitaxel-based compound as an active ingredient. The paclitaxel-based compound is represented by the compound represented by the following general formula (I):

(here,
R ¹ has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent.
^R2 has (2-1) a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, and a nitro group on the phenyl ring. A phenylcarbonyl group which may have at least one selected from the group consisting of a carboxy group and a cyano group as a substituent, or an alkenylcarbonyl group having (2-2) an alkenyl moiety having 2 to 6 carbon atoms. Indicates an amino group that may have.
R ³ , R ⁴ and R ⁵ represent the same or different hydrogen atom or hydroxyl group.
R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ represent the same or different linear alkyl groups of C1-3.
R ¹² has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent. ).

In the present specification, "C" in the description of "C1-4", "C1-3", "C3-4", etc. relating to the compound represents the number of carbon atoms. "C1-4" indicates that the number of carbon atoms is 1 to 4, "C1-3" indicates that the number of carbon atoms is 1 to 3, and "C3-4" indicates that the number of carbon atoms is 3 to 4. Show that.

In the substituents on the phenyl ring at R ¹ , R ² , and R ¹² , the linear or branched alkyl group of C1-4 is the linear alkyl group of C1-4, and the branched chain of C3-4. Indicates an alkyl group. The linear alkyl group of C1-4 is one selected from, for example, a methyl group, an ethyl group, a propyl group, and an n-butyl group. The branched chain alkyl group of C3-4 is one selected from an isopropyl group, a sec-butyl group, an isobutyl group, and a tert-butyl group.

In the substituents on the phenyl ring in R ¹ , R ² , and R ¹² , the linear or branched alkoxy group of C1-4 is the linear alkoxy group of C1-4, and the branched chain of C3-4. Indicates an alkoxy group. The alkyl moiety of the linear alkoxy group of C1-4 is one selected from, for example, a methyl group, an ethyl group, a propyl group, and an n-butyl group. The alkyl moiety of the branched chain alkoxy group of C3-4 is one selected from an isopropyl group, a sec-butyl group, an isobutyl group, and a tert-butyl group.

Regarding the amino group which may have an alkenylcarbonyl group having 2 to 5 carbon atoms in the alkenyl moiety in ^R2 , the alkenyl moiety having 2 to 5 carbon atoms is, for example, a linear or branched alkenyl group. Can be mentioned. Examples of the linear alkenyl group having 2 to 5 carbon atoms include a vinyl group, a 1-propenyl group, a 2-propenyl group, a 1-butenyl group, a 2-butenyl group, a 3-butenyl group and a 1-pentynyl group. , 2-pentynyl group, 3-pentynyl group, 4-pentynyl group can be raised. Examples of the alkenyl group of the branched chain having 3 to 5 carbon atoms include a methyl-propenyl group, a methyl-butenyl group, an ethyl-propenyl group and the like.

In the substituents on the phenyl ring in R ¹ , R ² , and R ¹² , the halogen atom may include, for example, a chlorine atom, a fluorine atom, a bromine atom, an iodine atom and the like.

The substituents on the phenyl ring at R ¹ , R ² , and R ¹² may be at any of the ortho, meta, and para positions with respect to the carbon atom to which the alkylene group is attached.

R ¹ is preferably a phenyl group without a substituent.

^R2 is preferably a phenylcarbonyl group, an alkenylcarbonyl group having 4 carbon atoms in the alkenyl group, or an unsubstituted amino group.

R ³ is preferably a hydrogen atom or a hydroxyl group.

R ⁴ and R ⁵ are preferably hydroxyl groups.

R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ are preferably methyl groups.
R ¹² is preferably a phenyl group without a substituent.

The paclitaxel-based compound is preferably any of the compounds represented by the following formulas (II) to (V).

The paclitaxel-based compound is more preferably any of the compounds represented by the following formulas (II') to (V').

The common name of the compound represented by the above formula (II) or (II') is paclitaxel.

The common name of the compound represented by the above formula (III) or (III') is cephalomanin.

The common name of the compound represented by the above formula (IV) or (IV') is 3'-N-debenzoyltaxol.

The generic name of the compound represented by the above formula (V) or (V') is 3'-N-debenzoyl-2'-deoxytaxel.

The most preferable paclitaxel-based compound is paclitaxel.

In addition to paclitaxel-based compounds, various compositions for treating cancer are generally used as ordinary drugs depending on the dosage form of the pharmaceutical composition, such as excipients, binders, disintegrants, lubricants, etc. It may contain a colorant, a flavoring agent, a flavoring agent, a surfactant and the like.

The composition for cancer treatment can be administered by intravenous drip infusion, intraperitoneal administration, or the like according to a conventionally reported method for administering a paclitaxel-based compound. Intraperitoneal administration or a combination of intraperitoneal administration and intravenous drip administration is preferable. The composition for treating cancer can be determined according to the dose of the paclitaxel-based compound previously reported. For example, when only a paclitaxel-based compound is used as an active ingredient for cancer treatment, in the case of intravenous drip infusion, the dose of the paclitaxel-based compound is 80 mg / m ² (body surface area) once a day for adults. From 210 mg / m ² (body surface area) over 1 to 24 hours, and if necessary, continuously administer for 3 to 6 weeks. After administration, it is preferable to withdraw from the drug for at least 2 to 3 weeks as needed. From the start of medication to the end of medication, or if there is a drug holiday, one course may be from the start of medication to the end of the drug holiday, and the medication course may be repeated as necessary while observing the degree of tumor shrinkage. ..

When intraperitoneally administered, the dose of paclitaxel-based compounds should be about 10 mg / m ² (body surface area) to 40 mg / m ² (body surface area) per day for adults. Can be administered. Intraperitoneal administration can be performed, for example, once or twice a week.

In the case of a combination of intraperitoneal administration and intravenous drip infusion, the dose of paclitaxel-based compound to adults is 30 mg / m ² (body surface area) to 30 mg / m ² per day by intravenous drip infusion. It can be administered to about (body surface area) and intraperitoneally at about 10 mg / m ² (body surface area) to 40 mg / m ² (body surface area) per day. The composition for treating cancer can be administered, for example, once or twice a week.

In addition, the composition for treating cancer may be used in combination with other drugs. As another drug, S-1 which is one of the oral fluoropyrimidine derivatives can be mentioned. When the cancer therapeutic composition is used in combination with S-1, 50 mg / m ² (body surface area) to 100 per day is continued from the start of administration of the cancer therapeutic composition to adults. It can be orally administered at mg / m ² (body surface area) in about 1 to 2 weeks.

2. 2. Method for Detecting Evaluation Marker One aspect of the present disclosure relates to an evaluation marker that reflects the effectiveness of a composition for treating cancer in a patient with pancreatic tumor having peritoneal dissemination and a method for detecting the same. The detection method includes a step of detecting CXCL12 in a region of interest containing pancreatic tumor cells in a tissue collected from a subject. That is, CXCL12 expressed or present in the region of interest containing pancreatic tumor cells in the tissue collected from the examiner is an evaluation that reflects the effectiveness of the cancer therapeutic composition in pancreatic tumor patients with peritoneal dissemination. It can be used as a marker.

Detection of CXCL12 in a pancreatic tumor is performed on an area of interest, which is a target area for detecting CXCL12. The area of interest is not limited as long as it includes pancreatic tumor cells such as pancreatic tumor cells or pancreatic tumor tissue collected from a subject. Pancreatic tumor cells or pancreatic tumor tissue can be collected, for example, by surgical excision or biopsy from the primary or metastatic lesion, endoscopic excision or biopsy, separation from ascites, or the like. Whether it is a tumor tissue or a normal tissue can be determined by macroscopic findings, microscopic observation, or the like. Further, it may be determined whether or not the tissue is a tumor tissue by using the cell proliferation ability and the expression of a tumor marker (for example, CA19-9) as an index. When determining whether or not the tissue is a tumor tissue using the cell proliferation ability as an index, for example, when the labeling index of the BrdU or Ki-67 protein of the tissue to be inspected is higher than that of the normal tissue, the tissue to be inspected is said to be the tissue to be inspected. It can be determined that the tissue is likely to be tumor tissue. When the tumor marker is used as an index, it can be determined that the tissue in which the expression of the tumor marker is positive is highly likely to be the tumor tissue.

Pancreatic tumor cells or pancreatic tumor tissue collected from the subject are pretreated according to the detection method of CXCL12.

CXCL12 in pancreatic tumors can be detected as a protein or as mRNA.

Examples of the method for detecting CXCL12 as a protein include known methods such as immunostaining and Western blotting. Further, as a method for detecting CXCL12 as mRNA, known methods such as in situ hybridization, RT-PCR (including quantitative RT-PCR), microarray, RNA-Seq and the like can be mentioned.

When performing immunostaining or in situ hybridization using pancreatic tumor tissue, as a pretreatment, the pancreatic tumor tissue is fixed with a known fixing solution such as formalin and paraformaldehyde, and then the paraffin-embedded block is placed. To make. Alternatively, the pancreatic tumor tissue is fixed or not fixed, and then embedded in a resin for making a frozen block such as OCT compound (registered trademark) to prepare a frozen block. Next, the prepared paraffin-embedded block or frozen block is sliced to prepare a tissue section, which is subjected to immunostaining or in situ hybridization. Here, the pancreatic tumor tissue embedded in the block may be only the tumor portion, but may include, for example, a normal region or the like.

When immunostaining or in situ hybridization is performed using pancreatic tumor cells, the cells are smeared or collected on a slide glass as a pretreatment, and fixed with formalin, paraformaldehyde, ethanol, or the like.

Here, when CXCL12 is immunostained, a positive signal appears on the cell surface, that is, on the cell membrane.

When CXCL12 is detected as a protein by Western blotting or the like, pancreatic tumor cells or pancreatic tumor tissue are lysed with a predetermined lysis buffer as a pretreatment. The sample dissolved in the dissolution buffer is used as the inspection sample.

When CXCL12 is detected as mRNA by RT-PCR, microarray, RNA-Seq, etc., total RNA or mRNA is extracted from pancreatic tumor cells or pancreatic tumor tissue as a pretreatment. Further, if necessary, the extracted total RNA or mRNA may be used as a template for reverse transcription to synthesize complementary DNA (cDNA). Total RNA or mRNA, or cDNA is used as a test sample.

The primary antibody for detecting CXCL12 by immunostaining or Western blotting is not limited as long as CXCL12 can be detected. For example, Anti-SDF-1 rabbit polyclonal antibody (ab9797, abcam) and the like can be mentioned. The primary antibody bound to CXCL12 can be detected by the reaction between the enzyme-labeled secondary antibody bound to the primary antibody and the enzyme and the substrate. In the case of immunostaining, tissue sections may be treated with a proteolytic enzyme such as trypsin after deparaffinization and water immersion treatment, and then immunostaining may be performed.

A method for producing a probe used for in situ hybridization is known. Alternatively, a commercially available probe may be used.

As the primer used for RT-PCR (in the case of quantitative RT-PCR, a probe may be included), a commercially available primer can be used. In addition, commercially available microarrays can also be used.

For RNA-Seq, the number of reads of CXCL12 mRNA can be obtained using a next-generation sequencer (for example, manufactured by Illumina).

When CXCL12 is detected by immunostaining or in situ hybridization, the presence or absence of CXCL12 is detected by observing a tissue specimen subjected to immunostaining or in situ hybridization by a human using a microscope or a slide scanner. be able to. When a signal of immunostaining or in situ hybridization is confirmed in the tumor cells in the tissue specimen, it can be determined (determined) that CXCL12 has been detected. When at least one tumor cell having CXCL12 in the cell is detected in the tumor portion in the tissue specimen, it may be determined that "CXCL12 is detected" or "expression of CXCL12 is positive". Alternatively, for example, when the number of tumor cells existing in a predetermined section of a microscope or a slide scanner is 100%, the number of tumor cells having CXCL12 in the cells is 10% or more, preferably 5% or more, more preferably. May be determined that "CXCL12 was detected" or "CXCL12 expression is positive" when 1% or more is present.

When CXCL12 is detected by Western blotting, RT-PCT, RNA-Seq, "CXCL12 is detected" or "Expression of CXCL12" when CXCL12 is detected in a sample extracted from tumor cells or tumor tissue. Is positive. " Alternatively, the amount of protein of CXCL12 in the test sample derived from tumor cells or tissue is compared with the amount of protein of CXCL12 derived from normal cells or normal tissue or the amount of mRNA of CXCL12 from the test sample derived from tumor cells or tissue. , Or "CXCL12 was detected" or "CXCL12 expression is positive" when the amount of CXCL12 mRNA is higher than the amount of CXCL12 protein derived from normal cells or tissues, or the amount of CXCL12 mRNA. May be decided. Further, when the amount of protein of CXCL12 or the amount of mRNA of CXCL12 in the test sample derived from tumor cells or tissue is similar to the amount of protein of CXCL12 derived from normal cells or normal tissue or the amount of mRNA of CXCL12, " It may be determined that "CXCL12 is not detected" or "the expression of CXCL12 is negative". Here, "showing a high value" can be exemplified as a case where a value showing a value 1.2 times or more, preferably 1.5 times or more, more preferably 2 times or more, still more preferably 5 times or more is shown. "Same degree" can be exemplified by about 0.8 to 1.1 times. Before comparing the amount of protein of CXCL12 or the amount of mRNA of CXCL12, the amount of protein or RNA in each test sample is adjusted to the amount of protein derived from housekeeping genes such as GAPDH, β2-microglobulin, and β-actin. Alternatively, it may be normalized by the amount of mRNA. The amount of protein may be expressed by mass or concentration, but may also be expressed by the emission intensity of the substrate or the like. The amount of mRNA may be the number of copies or reads of mRNA, but may be expressed by fluorescence intensity or the like.

As another embodiment, when the reference value of the CXCL12 protein amount or the RNA amount is determined in advance and the CXCL12 protein amount or the RNA amount in the test sample derived from the tumor cell or the tumor tissue is higher than the reference value, " It may be determined that "CXCL12 has been detected" or "the expression of CXCL12 is positive". Further, when the amount of CXCL12 protein or RNA in the test sample derived from the tumor cell or tumor tissue is lower than the reference value, it is determined that "CXCL12 is not detected" or "the expression of CXCL12 is negative". May be good. The reference value is not limited as long as it is a value that can determine whether the amount of protein of CXCL12 or the amount of mRNA of CXCL12 is detected or the expression is positive, and can be determined by a known method. The value that can determine whether the amount of protein of CXCL12 or the amount of mRNA of CXCL12 is detected or the expression is positive is the ROC (receiver operating characteristic curve) curve, discriminant analysis method, mode method, Kittler method, 3σ method. , P-tile method or the like. Further, as reference values, sensitivity, specificity, negative predictive value, positive predictive value, first quartile, and the like can be exemplified.

The evaluation marker detection method may further include a step of determining that the cancer therapeutic composition is effective when CXCL12 is not detected. Alternatively, it may include a step of determining that the cancer therapeutic composition is not effective when CXCL12 is detected.

As another embodiment, the method for detecting the evaluation marker may include a step of acquiring a score indicating the abundance of CXCL12 in the region of interest.

The score is based on the ratio of (1) the staining intensity of CXCL12 protein in the pancreatic tumor cells and (2) the number of pancreatic tumor cells positive for CXCL12 protein staining when the number of pancreatic tumor cells is 100%. Is obtained.

In (1), the CXCL12 protein in pancreatic tumor cells is stained by, for example, immunostaining, and the staining intensity is determined.
No expression: level 0,
Weak: Level 1,
Medium: Level 2 or Strong: Level 3
It is classified into 4 stages.
In (2), the ratio of the number of pancreatic tumor cells positive for CXCL12 protein staining when the number of pancreatic tumor cells is 100% is determined.
0%: Level 0,
1 to 5%: Level 1,
From 5% to 25%: Level 2,
Higher than 25 and up to 50%: Level 3,
Higher than 50 up to 75%: Level 4 or Higher than 75 up to 100%: Level 5
It is classified into 5 stages.
Then, the value obtained by multiplying the values of each level obtained in (1) and (2) is used as the score.

The evaluation marker detection method compares the score of the region of interest obtained by the above method with the reference value of the predetermined score, and when the score of the region of interest falls below the reference value, the composition for cancer treatment is described. It can be determined that the thing is valid. Further, when the score of the region of interest is equal to or higher than the reference value, it can be determined that the composition for treating cancer is not effective. Alternatively, the evaluation marker detection method compares the score of the region of interest obtained by the above method with the reference value of the predetermined score, and when the score of the region of interest falls below the reference value, the cancer treatment is described. Suggests that the composition for use is effective. Further, when the score of the region of interest is equal to or higher than the reference value, it is suggested that the composition for treating cancer is not effective.

The reference value is not limited as long as it is possible to distinguish between a group in which the composition for treating cancer is effective and a group in which the composition for cancer treatment is not effective. For example, a score of 6 can be used as a reference value.

As a method for detecting the evaluation marker, the cancer therapeutic composition is effective when CXCL12 is not detected in the area of interest, or when the score indicating the abundance of CXCL12 in the area of interest falls below the reference value. It may include a step of presenting a label indicating that. Further, as a method for detecting the evaluation marker, the cancer therapeutic composition is effective when CXCL12 is detected in the area of interest, or when the score indicating the abundance of CXCL12 in the area of interest is equal to or higher than the reference value. It may include a step of presenting a label indicating that it is not.

Further, the evaluation marker detection method may be applicable to conversion surgery when CXCL12 is not detected in the area of interest, or when the score indicating the abundance of CXCL12 in the area of interest falls below the reference value. It may include a step of presenting a label indicating. In addition, the evaluation marker detection method is not applicable to conversion surgery when CXCL12 is detected in the area of interest, or when the score indicating the abundance of CXCL12 in the area of interest is equal to or higher than the reference value. It may include a step of presenting a label indicating.

The presentation of whether or not the composition for cancer treatment is effective and the presentation of whether or not the combination surgery is applicable may be either one or both.

3. 3. Test Reagent Another aspect of the present disclosure relates to a test reagent for detecting CXCL12 present in pancreatic tumor cells collected from a subject as a marker for evaluating the effectiveness of the above-mentioned cancer therapeutic composition.

The test reagent may include a reagent for detecting CXCL12 protein and / or a reagent for detecting CXCL12 mRNA.

The reagent for detecting CXCL12 protein contains at least one or more antibodies (eg, primary antibody) capable of binding to a part of CXCL12 protein. As the "antibody", any of a polyclonal antibody, a monoclonal antibody, and a fragment thereof (for example, Fab, F (ab'), F (ab) 2, etc.) can be used. The immunoglobulin class and subclass are not particularly limited. Further, the antibody may be one screened from an antibody library, a chimeric antibody, scFv, or the like.
Further, the antibody does not necessarily have to be purified, and may be an antiserum containing the antibody, ascites, an immunoglobulin fraction fractionated from these, or the like.

The antibody contained in the test reagent may be in a dry state or may be dissolved in a buffer such as phosphate buffered saline. Further, the test reagent is a stabilizer such as β-mercaptoethanol and DTT; a protective agent such as albumin; a surfactant such as polyoxyethylene (20) sorbitan monolaurate and polyoxyethylene (10) octylphenyl ether. It may contain at least one preservative such as sodium azide.

The antibody that binds to CXCL12 may be labeled with an enzyme or a fluorescent dye. The antibody that binds to CXCL12 may be immobilized on a microplate, magnetic beads, or the like.
The CXCL12 protein detection reagent may be provided as a test kit that includes a package insert that describes the test reagent and how to use the reagent or the URL of a web page that describes how to use the reagent. When the antibody that binds to CXCL12 is an unlabeled primary antibody, the test kit may contain a secondary antibody labeled with an enzyme or a fluorescent dye. In addition, the test kit may include a substrate that reacts with the enzyme.

The CXCL12 mRNA detection reagent contains a nucleic acid that hybridizes with all or part of CXCL12 mRNA or CXCL12 cDNA. The nucleic acid is preferably a detection nucleic acid (DNA or RNA) having a function as a primer and / or a probe. The length of the detection nucleic acid is not particularly limited.

If the detection nucleic acid is a primer used in the PCR reaction, the sequence that hybridizes with CXCL12 mRNA or CXCL12 cDNA is preferably 50 mer or less, more preferably 30 mer or less, and further preferably 15 It is about ~ 25 mer. The primer may contain a sequence that does not hybridize with CXCL12 mRNA or CXCL12 cDNA. Further, the primer may be labeled with a fluorescent dye or the like.

In addition to primers, RT-PCR can also use a quantification probe that is degraded during the PCR reaction and is used for real-time quantification of PCR products. The quantification probe is also not limited as long as it hybridizes with CXCL12 mRNA or CXCL12 cDNA. The quantification probe is preferably a nucleic acid of about 5 to 20 mer, which contains a sequence that hybridizes with CXCL12 mRNA or CXCL12 cDNA. Further, it is preferable that one end of the quantification probe is labeled with a fluorescent dye, and the other end of the quantification probe is labeled with a quencher of the fluorescent dye.

If the detection nucleic acid is used as a capture probe in a microarray or the like, the sequence that hybridizes with CXCL12 mRNA or CXCL12 cDNA is preferably about 100 mer, more preferably about 60 mer, and further preferably. Is about 20 to 30 mer. The capture probe may contain a sequence that does not hybridize to CXCL12 mRNA or CXCL12 cDNA. Further, the capture probe is preferably immobilized on the chip.

When the detection nucleic acid is a probe for in situ hybridization, the detection nucleic acid may be an oligonucleotide having a sequence hybridizing with CXCL12 mRNA of about 15 to 100 mer, and a polynucleotide exceeding 100 mer. May be. The polynucleotide may be DNA or RNA. Labeling substances such as digoxigenin and fluorescent dyes can be bound to the probe for in situ hybridization. The probe may contain a sequence that does not hybridize to CXCL12 mRNA.

The CXCL12 mRNA detection reagent may be provided as a test kit containing a test reagent and an attachment containing the URL of a web page that describes how to use the reagent or how to use the reagent. When CXCL12 mRNA or CXCL12 cDNA is detected by RT-PCR, the test kit contains a nucleic acid amplification reagent (including polymerase, buffer, dNTP, etc. even if it is heat-resistant DNA), reverse transcriptase, etc. May be good. The nucleic acid amplification reagent may contain a dye such as SYBER GREEN (registered trademark), if necessary. When CXCL12 mRNA or CXCL12 cDNA is detected by a microarray, the test kit may include a hybridization buffer, a washing buffer, and the like. When CXCL12 mRNA is detected by in situ hybridization, the test kit may include a proteolytic enzyme such as proteinase K, a hybridization buffer, a washing buffer, and the like.

4. Evaluation device for the effectiveness of the composition for cancer treatment 4-1. Configuration of Evaluation Device for Efficacy of Cancer Treatment Composition One embodiment of the present disclosure is an evaluation system 1000 for the effectiveness of a cancer treatment composition in a patient with pancreatic tumor having peritoneal dissemination (hereinafter referred to as “evaluation system 1000”). (Abbreviation)) and the evaluation device 10 for evaluating the effectiveness of the composition for treating cancer (hereinafter, abbreviated as “evaluation device 10”).

FIG. 1 is an overview diagram of the evaluation system 1000, and the evaluation system 1000 may include an analysis device 5a or an analysis device 5b in addition to the evaluation device 10 as one aspect.
FIG. 2 shows a block diagram of the evaluation device 10. The evaluation device 10 may be connected to the input unit 111, the output unit 112, and the storage medium 113.

In the evaluation device 10, the processing unit 101, the main storage unit 102, the ROM (read only memory) 103, the auxiliary storage unit 104, the communication interface (I / F) 105, and the input interface (I / F) 106 The output interface (I / F) 107 and the media interface (I / F) 108 are connected to each other by a bus 109 so as to be capable of data communication. The main storage unit 102 and the auxiliary storage unit 104 may be collectively referred to as a storage unit. The storage unit volatilely or non-volatilely stores the measurement result, the score in the area of interest, the reference value, and the like.

The processing unit 101 is the CPU of the evaluation device 10. The processing unit 101 may cooperate with the GPU. The evaluation device 10 functions by the processing unit 101 executing the operation system 104a and the evaluation program 104b stored in the auxiliary storage unit 104 and processing the acquired data.

The ROM 103 is composed of a mask ROM, a PROM, an EPROM, an EEPROM, and the like, and records a computer program executed by the processing unit 101 and data used for the computer program. The processing unit 101 may be the MPU 101. The ROM 103 stores the boot program executed by the processing unit 101 when the evaluation device 10 is started, and the programs and settings related to the operation of the hardware of the evaluation device 10.

The main storage unit 102 is composed of a RAM (Random access memory) such as a SRAM or a DRAM. The main storage unit 102 is used to read out the computer program recorded in the ROM 103 and the auxiliary storage unit 104. Further, the main storage unit 102 is used as a work area when the processing unit 101 executes these computer programs.

The auxiliary storage unit 104 is composed of a hard disk, a semiconductor memory element such as a flash memory, an optical disk, or the like. The auxiliary storage unit 104 stores an operation system 104a, an evaluation program 104b described later, and a reference value database (DB) 104c for storing reference values. The evaluation program 104b cooperates with the operation system 104a to perform evaluation processing.

The communication I / F 105 includes a serial interface such as USB, IEEE1394, RS-232C, a parallel interface such as SCSI, IDE, IEEE1284, an analog interface including a D / A converter, an A / D converter, and a network interface controller ( It is composed of Network interface controller (NIC) and the like. Under the control of the processing unit 101, the communication I / F 105 receives data from the analyzers 5a and 5b or other external devices, and stores or generates information as needed by the evaluation device 10 in the analyzer 5a, 5b or send or display to the outside. The communication I / F 105 may communicate with the analyzers 5a, 5b or other external devices via the network.

The input I / F 106 is composed of, for example, a serial interface such as USB, IEEE1394, RS-232C, a parallel interface such as SCSI, IDE, and IEEE1284, and an analog interface including a D / A converter and an A / D converter. To. The input I / F 106 accepts character input, click, voice input, and the like from the input unit 111. The received input contents are stored in the main storage unit 102 or the auxiliary storage unit 104.

The input unit 111 is composed of a touch panel, a keyboard, a mouse, a pen tablet, a microphone, etc., and inputs characters or voices to the evaluation device 10. The input unit 111 may be connected from the outside of the evaluation device 10 or may be integrated with the evaluation device 10.

The output I / F 107 is composed of an interface similar to that of the input I / F 106, for example. The output I / F 107 outputs the information generated by the processing unit 101 to the output unit 112. The output I / F 107 outputs the information generated by the processing unit 101 and stored in the auxiliary storage unit 104 to the output unit 112.

The output unit 112 is composed of, for example, a display, a printer, or the like, and displays measurement results transmitted from the analyzers 5a and 5b, various operation windows in the evaluation device 10, analysis results, and the like.

The media I / F 108 reads, for example, application software stored in the storage medium 113. The read application software and the like are stored in the main storage unit 102 or the auxiliary storage unit 104. Further, the media I / F 108 writes the information generated by the processing unit 101 into the storage medium 113. The media I / F 108 writes the information generated by the processing unit 101 and stored in the auxiliary storage unit 104 to the storage medium 113.

The storage medium 113 is composed of a flexible disk, a CD-ROM, a DVD-ROM, or the like. The storage medium 113 is connected to the media I / F 108 by a flexible disk drive, a CD-ROM drive, a DVD-ROM drive, or the like. The storage medium 113 may store an application program or the like for the computer to execute an operation.

The processing unit 101 may acquire the evaluation program 104b and various settings necessary for controlling the evaluation device 10 via the network instead of reading from the ROM 103 or the auxiliary storage unit 104. The application program is stored in the auxiliary storage unit of the server computer on the network, and the evaluation device 10 may access the server computer to download the computer program and store it in the ROM 103 or the auxiliary storage unit 104. It is possible.

Further, an operation system that provides a graphical user interface environment such as Windows (registered trademark) manufactured and sold by Microsoft Corporation in the United States is installed in the ROM 103 or the auxiliary storage unit 104. The application program according to the second embodiment shall operate on the operating system. That is, the evaluation device 10 may be a personal computer or the like.

The evaluation system 1000 does not have to be installed in one place, and the evaluation device 10 and the analysis devices 5a and 5b may be arranged in different places and connected by a network. Further, the evaluation device 10 may be a device that does not require an operator who omits the input unit 111 and the output unit 112.

The analyzer 5a is a device for measuring the amount or concentration of protein, and includes a sample storage area 51, a reaction unit 52, and a detection unit 53. The cytolytic solution set in the sample storage area 51 is dispensed and incubated into a microplate on which the antigen-capturing antibody placed in the reaction unit 52 is immobilized. After removing the unreacted antigen as needed, the detected antibody is dispensed into microplates and incubated. After removing the unreacted antigen as needed, the substrate for detecting the detection antibody is dispensed to the microplate, the microplate is moved to the detection unit 53, and the signal generated by the reaction of the substrate is measured. Will be done. It was

Another aspect of the analyzer 5a is an apparatus for measuring the expression level of mRNA by microarray analysis, in which the reverse transcriptase set in the sample storage area 51 is divided onto the microarray chip set in the reaction unit 52. After pouring, hybridization, and washing, the sample is moved to the detection unit 53 to detect the signal.

Further, another aspect of the analyzer 5a is an apparatus for measuring the expression level of mRNA by RT-PCR, in which the reverse transcription reagent set in the sample storage 51 is placed in a microtube set in the reaction unit 52. Dispense, then dispense the quantitative PCR reagent into a microtube. While performing the PCR reaction in the reaction unit 52, the detection unit 53 detects the signal in the tube.

The analyzer 5b is an apparatus for measuring the expression level of mRNA by the RNA-Seq method, and includes a sequence analysis unit 54. The sample subjected to the reaction for RNA-Seq is set in the sequence analysis unit 54, and the base sequence is analyzed in the sequence analysis unit 54.

The analyzer 5b is a fully automated Western blotting device for measuring the amount of protein by Western blotting, and includes a chemiluminescence signal detection unit 54. A sample of pancreatic tumor cells or pancreatic tumor tissue lysed with a lysis buffer is set in a predetermined position on an automatic Western blotting device, and SDS-PAGE, blotting to a membrane, antibody reaction, chemiluminescence is performed, and a chemiluminescence signal detector is performed. Analysis is performed at 54 to quantify the signal intensity.

The analyzers 5a and 5b are connected to the evaluation device 10 by wire or wirelessly. The analyzer 5a A / D-converts the measured value of protein or the measured value of mRNA and transmits it as digital data to the evaluation device 10. Similarly, the analyzer 5b A / D-converts the measured value of mRNA and sends it to the evaluation device 10 as digital data. As a result, the evaluation device 10 can acquire the measured value of the protein or the measured value of mRNA as digital data that can be calculated.

4-2. Processing of the evaluation program FIG. 3 shows an example of a flowchart of the processing executed by the evaluation program 104b.

The processing unit 101 of the evaluation device 10 starts processing for evaluating the effectiveness of the cancer therapeutic composition by inputting a processing start request from the input unit 111 by the operator.

In step S11, the processing unit 101 acquires the score of CXCL12 in the region of interest including the pancreatic tumor cells in the tissue input by the operator from the input unit 111. Alternatively, the amount of protein of CXCL12 in pancreatic tumor cells collected from the subject from the analyzer 5a or the analyzer 5b, or the amount of mRNA of CXCL12 or a value reflecting these is obtained as a measured value of CXCL12. Alternatively, the operator inputs from the input unit 111 whether the expression of CXCL12 by immunostaining or in situ hybridization is positive or negative (or whether CXCL12 is detected), and the processing unit 101 inputs this input. It may be acquired as a measured value of CXCL12.

Next, in step S12, the processing unit 101 compares the reference value of the score of CXCL12 or the reference value of CXCL12 stored in the storage unit with the value acquired in step S11.

If the obtained score or value is lower than the reference value in step S13, the processing unit 101 proceeds to step S14 (YES), determines that the cancer therapeutic composition is effective, and determines the determination result. The indicated label is output to the output unit 112 (step S16). Alternatively, in step S16, it is determined that the conversation surgery is applicable, and a label indicating that the conversion surgery is applicable is output. Further, in step S13, when the acquired score or value is equal to or higher than the reference value, the process proceeds to step S15 (NO), it is determined that the cancer therapeutic composition is not effective, and a label indicating the determination result is shown. Is output to the output unit 112 (step S16). Alternatively, in step S16, it is determined that the conversation surgery is not applicable, and a label indicating that the conversion surgery is not applicable is output.

The label indicating the effectiveness of the cancer therapeutic composition contains information indicating that "the cancer therapeutic composition is effective" or "the cancer therapeutic composition may be effective". .. The label indicating that the cancer therapeutic composition is not effective contains information indicating that "the cancer therapeutic composition is ineffective" or "the cancer therapeutic composition may be ineffective". Is done.

The label indicating the applicability of the conversion surgery includes information indicating that "conversion surgery is valid" or "conversion surgery may be valid". The label indicating that the conversion surgery is not applicable includes information indicating that "conversion surgery is invalid" or "conversion surgery may be invalid". The information may be a mark such as x, 〇, exclamation mark or the like.
For details on the reference value, comparison method, judgment method, etc., refer to 2. above. The explanation of is used here.

5. A storage medium in which a program is stored Further, an embodiment of the present embodiment relates to a program product such as a storage medium in which the evaluation program 104b is stored. That is, the computer program can be stored in a hard disk, a semiconductor memory element such as a flash memory, or a storage medium such as an optical disk. The recording format of the program on the storage medium is not limited as long as the evaluation device 10 can read the program. Recording on the storage medium is preferably non-volatile.

The present disclosure will be described in more detail with reference to examples below, but the present disclosure is not construed as being limited to the examples.
This study was conducted with the approval of the Institutional Review Board of Kansai Medical University based on the Declaration of Helsinki.

1. 1. Patient selection The subjects were pancreatic ductal adenocarcinoma patients with peritoneal dissemination who visited Kansai Medical University Hospital from January 2006 to December 2018.

2. 2. Histological examination A surgically resected tissue sample was fixed with formalin to prepare a tissue specimen. For tissue specimens, all fixed tissue samples were embedded in paraffin to prepare sliced sections.

3. 3. Tissue Microarray For each patient's tissue, select the most morphologically characteristic part of the hematoxylin-eosin-stained tissue specimen and select two tissue masses (2 mm in diameter) from the paraffin-embedded block. Was collected with a punch. The punched tissue mass was further arranged and embedded in a paraffin block, and sliced to prepare a tissue section.

4. Immunostaining Immunostaining was performed using an automatic staining device (Discovery, Roche Diagnostics, Basel, Switzerland) according to the protocol attached to the device. Anti-SDF-1 rabbit polyclonal antibody (ab9797, abcam) was diluted 1,000-fold and used as the primary antibody for staining CXCL12. As the secondary antibody, a peroxidase-labeled anti-rabbit immunoglobulin antibody was used, and diaminobenzidine (DAB) was used for color development. Expression of CXCL12 was positive when there was a signal on the surface (on the cell membrane) of pancreatic tumor cells. Above 2. The score of each section was obtained according to the scoring method described in 1.

5. Anti-cancer drug therapy Paclitaxel is administered to 50 mg / m ² by intravenous drip infusion and 20 mg / m ² intraperitoneally on day 1 and day 8 with the start date of anti-cancer drug treatment as day 1. bottom. At the same time, S-1 was orally administered at 80 mg / m ² / d from day 1 for 14 consecutive days.

6. Statistical analysis JMP Start Statistics version 14 (Statistical Discovery Software; SAS Institute, Cary, NC, USA) was used for statistical analysis.

7. Results Boxplots of CXCL12 scores in the group (CS group) where paclitaxel therapy was effective and conversion surgery was applicable, and the group (-group) where paclitaxel therapy was not effective and conversion surgery was not applied. show. All patients in the CS group had a CXCL12 score below 6. On the other hand, the score of CXCL12 tended to be higher in the-group than in the CS group (P = 0.048).

From this result, it was shown that the paclitaxel-based compound is effective for pancreatic tumor cells of patients with a CXCL12 score of less than 6. It was also shown that conversion surgery can be applied to these patients.

In addition, the group in which the expression of CXCR4 was scored instead of CXCL12 and paclitaxel therapy was remarkably effective and conversion surgery could be applied (CS group), and the group in which paclitaxel therapy did not show any effect and conversion surgery was excluded. (-Group) Scores were compared. The results are shown in FIG. Patients in the CS group had a CXCR4 score of 10 or higher. However, in the significance test between the two groups, P = 0.423, and no superiority was observed.

101 Processing unit 10 Evaluation device

Claims (10)

1. Including the step of detecting CXCL12 in a region of interest containing pancreatic tumor cells in a tissue collected from a subject.
   A method for detecting an evaluation marker that reflects the effectiveness of a composition for treating cancer in patients with pancreatic tumors having peritoneal dissemination.
   A method for detecting the evaluation marker, wherein the composition for treating cancer contains a compound represented by the following general formula (I).
   

   

   (here,
   R ¹ has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent.
   ^R2 has (2-1) a linear or branched alkyl group of C1-4 on the phenyl ring, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, and the like. A phenylcarbonyl group which may have at least one selected from the group consisting of a carboxy group and a cyano group as a substituent, or an alkenylcarbonyl group having (2-2) an alkenyl moiety having 2 to 6 carbon atoms. Indicates an amino group that may have.
   R ³ , R ⁴ and R ⁵ represent the same or different hydrogen atom or hydroxyl group.
   R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ represent the same or different linear alkyl groups of C1-3.
   R ¹² has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent. ).
2. Further comprising the step of presenting a label indicating that the composition for treating cancer is effective when the score indicating the abundance of CXCL12 in the region of interest is equal to or less than the reference value.
   The score is based on the ratio of (1) the staining intensity of CXCL12 protein in the pancreatic tumor cells and (2) the number of pancreatic tumor cells positive for CXCL12 protein staining when the number of pancreatic tumor cells is 100%. The method for detecting the evaluation marker according to claim 1, which is obtained in the above-mentioned method.
3. The method for detecting an evaluation marker according to claim 1 or 2, further comprising a step of presenting a label indicating that the conversation surgery is applicable when the score in the area of interest falls below the reference value.
4. The compound represented by the general formula (I) is paclitaxel.
   The method for detecting an evaluation marker according to any one of claims 1 to 3.
5. A composition for treating cancer for use in patients with pancreatic tumors having peritoneal dissemination, which comprises a compound represented by the following general formula (I).
   The cancer therapeutic composition is used for administration to the subject having a CXCL12 score of less than or equal to a reference value in an area of interest containing pancreatic tumor cells in a tissue collected from the subject.
   The score is obtained based on the ratio of (1) the staining intensity of CXCL12 in the pancreatic tumor cells and (2) the number of pancreatic tumor cells positive for CXCL12 staining when the number of pancreatic tumor cells is 100%. The composition for treating cancer:
   

   

   (here,
   R ¹ has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent.
   ^R2 has (2-1) a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C1-4, a halogen atom, an amino group, a hydroxyl group, and a nitro group on the phenyl ring. A phenylcarbonyl group which may have at least one selected from the group consisting of a carboxy group and a cyano group as a substituent, or an alkenylcarbonyl group having (2-2) an alkenyl moiety having 2 to 6 carbon atoms. Indicates an amino group that may have.
   R ³ , R ⁴ and R ⁵ represent the same or different hydrogen atom or hydroxyl group.
   R ⁶ , R ⁷ , R ⁸ , R ⁹ , R ¹⁰ and R ¹¹ represent the same or different linear alkyl groups of C1-3.
   R ¹² has a linear or branched alkyl group of C1-4, a linear or branched alkoxy group of C-4, a halogen atom, an amino group, a hydroxyl group, a nitro group, a carboxy group and a cyano group on the phenyl ring. Indicates a phenyl group which may have at least one selected from the group consisting of as a substituent. ).
6. The composition for cancer treatment according to claim 6, wherein the compound represented by the general formula (I) is paclitaxel.
7. A method of using CXCL12 in pancreatic tumor cells in a tissue collected from a subject as a marker for evaluating the efficacy of the cancer therapeutic composition according to claim 5 or 6, wherein the subject is said to have the same. The above method comprises peritoneal dissemination of a pancreatic tumor.
8. An evaluation marker comprising CXCL12 for evaluating the efficacy of the cancer therapeutic composition according to claim 5 or 6 for a subject having peritoneal dissemination of a pancreatic tumor.
9. A detection reagent for detecting CXCL12 in pancreatic tumor cells in a tissue collected from a subject as a marker for evaluating the efficacy of the cancer therapeutic composition according to claim 5 or 6, wherein the detection reagent is used. The reagent contains an antibody for detecting CXCL12 and contains
   Here, the subject has a peritoneal dissemination of a pancreatic tumor.
   The detection reagent.
10. A device for evaluating the effectiveness of a composition for treating cancer, which comprises a processing unit.
    The processing unit
    Obtain a score of CXCL12 in the area of interest containing pancreatic tumor cells in the tissue collected from the subject.
    Compare the obtained score with the standard value and
    When the obtained score falls below the reference value, a label indicating that the cancer therapeutic composition according to claim 5 or 6 is effective is output, and / or the obtained score is the reference value. When the value falls below, a label indicating that conversion surgery is applicable is output.
    Here, the subject has a peritoneal dissemination of a pancreatic tumor.
    The evaluation device.

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