JP2015071566A - Pharmaceutical composition of FGF4 gene amplified tumor - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a pharmaceutical composition of FGF4 gene amplification tumor in one or more embodiments.SOLUTION: In one aspect, the pharmaceutical composition for improvement of an FGF 4 (fibroblast growth factor 4) gene amplification tumor, progression inhibition, and/or treatment comprises at least one selected from the group consisting of anti-FGF4 antibody, FGF receptor inhibitor, FGF 4 siRNA, and FGF 4 shRNA as an active ingredient.

Description

  The present disclosure relates to a pharmaceutical composition of an FGF4 gene-amplified tumor, a method for predicting the effect of the pharmaceutical composition, a kit used therefor, use of the pharmaceutical composition, and a method for improving, suppressing progression and / or treating FGF4 gene-amplified tumor. .

  Patent Document 1 predicts whether therapeutic administration of an epidermal growth factor receptor (EGFR) inhibitor is effective or ineffective is an increase in EGFR gene copy number, tyrosine kinase receptor type receptor (HER2) copy. Disclose what is possible by detecting an increase in number.

  Patent Document 2 discloses that the prediction of the therapeutic effect of sorafenib, which is a multi-target kinase inhibitor and a cancer molecular target therapeutic agent, becomes possible by detecting an increase in the copy number of the FGF3 gene.

  Patent Document 3 discloses that an increase in the copy number of the α-actinin-4 gene can indicate the presence of cancer.

JP2013-5800A JP 2012-249633 A JP 2009-100777 A

  In one or more embodiments, the present disclosure provides a pharmaceutical composition of an FGF4 gene amplified tumor. Moreover, this indication provides the effect prediction method of this pharmaceutical composition in one or some embodiment.

  In one or a plurality of embodiments, the present disclosure provides a pharmaceutical composition for improving, suppressing progression, and / or treating an FGF4 gene-amplified tumor, comprising an anti-FGF4 antibody, an FGF receptor inhibitor, an FGF4 siRNA, and The present invention relates to a pharmaceutical composition comprising as an active ingredient at least one selected from the group consisting of FGF4shRNA.

  In one or a plurality of other embodiments, the present disclosure detects the gene amplification level of the FGF4 gene in the target tumor tissue sample or tumor cell sample, and the target FGF4 gene is amplified. If the effectiveness of therapeutic administration of the pharmaceutical composition according to the present invention is predicted, the subject is classified, or the therapeutic administration of the pharmaceutical composition according to the present disclosure is effective when the subject FGF4 gene is not amplified. It relates to a method comprising classifying said object if gender is not predicted.

  In one or a plurality of other embodiments of the present disclosure, the present disclosure detects the gene amplification level of the FGF4 gene in the tumor tissue sample or tumor cell sample of the subject, and the FGF4 protein when the subject FGF4 gene is amplified. The present invention relates to a method for improving, suppressing the progression of and / or treating FGF4 gene-amplified tumor, comprising administering to the subject a pharmaceutical composition comprising a substance that suppresses the expression of or inhibits the function thereof as an active ingredient.

FIG. 1 is a graph showing an example of the results of measuring the FGF4 gene copy number of 194 esophageal cancer specimens. The horizontal axis is the sample sample and is arranged in order of copy number. The vertical axis represents the measured copy number. FIG. 2 shows an example of the results of measuring the tumor size of a cell line (WiDr / FGF4) over time when FGF4 gene is introduced into colon cancer cell line WiDr and FGF4 protein is overexpressed. It is a graph. As a control, an EGFP gene was introduced (WiDr / EGFP). FIG. 3 is a graph showing an example of the results of measuring the tumor size of a cell line (A549 / FGF4) over time when the FGF4 gene was introduced into the lung cancer cell line A549 and the FGF4 protein was overexpressed. It is. As a control, an EGFP gene was introduced (A549 / EGFP). FIG. 4 shows the results of measuring the tumor size of the cell line (KYSE-270 / FGF4) over time when the FGF4 gene was introduced into the esophageal cancer cell line KYSE-270 and the FGF4 protein was overexpressed. It is a graph which shows an example. As a control, an EGFP gene was introduced (KYSE-270 / EGFP). FIG. 5 is a graph showing an example of the results of confirming the effect of anti-FGF4 antibody on FGF4 overexpressing esophageal cancer cell line KYSE-270 / FGF4 transplanted into mice. As a control, goat polyclonal IgG was used instead of anti-FGF4 antibody. In the left diagram of FIG. 5, the vertical axis represents the tumor size (mm ³ ), and the horizontal axis represents the culture time (days). In the right diagram of FIG. 5, the vertical axis represents the mouse body weight (g), and the horizontal axis represents the culture time (days). FIG. 6 is a graph showing an example of the results of confirming the effect of an FGFR inhibitor on the FGF4-overexpressing lung cancer cell line A549 / FGF4 transplanted into mice. The control is an example in which no FGFR inhibitor is administered. The vertical axis represents the tumor size (mm ³ ), and the horizontal axis represents the culture time (days).

  According to the present disclosure, some tumors and cancers have an amplified copy number of the FGF4 gene, and suppress or inhibit the function of the FGF4 protein against such an FGF4 gene-amplified tumor. Based on the finding that a pharmaceutical composition containing a substance as an active ingredient has an antitumor effect. The present disclosure is also based on the finding that, in one or more embodiments, amplification of the FGF4 gene copy number occurs in about 40% of esophageal cancers.

  Therefore, in one aspect, the present disclosure is a pharmaceutical composition for improving, suppressing progression and / or treating FGF4 gene-amplified tumor, and contains a substance that suppresses the expression of FGF4 protein or inhibits its function as an active ingredient. The present invention relates to a pharmaceutical composition.

[Target tumor]
In one or a plurality of embodiments, the tumor that is the target of the pharmaceutical composition according to the present disclosure is a tumor in which the copy number of FGF4 (Fibroblast growth factor 4) gene is amplified (hereinafter also referred to as “FGF4 gene amplified tumor”). It is. FGF4 gene amplified tumors can be present in various cancers. For example, about 40% of esophageal cancers are FGF4 gene amplified tumors. In addition, for example, there is a report that FGF4 gene amplification is observed in tumor samples of about 10% of bladder cancer and about 35% of head and neck squamous cell carcinoma. In one or a plurality of embodiments, the pharmaceutical composition according to the present disclosure can exert an antitumor effect on an FGF4 gene-amplified tumor. In one or a plurality of embodiments, the pharmaceutical composition according to the present disclosure can be used for improvement, suppression of progression, and / or treatment of an individual having an FGF4 gene-amplified tumor. In one or more embodiments of the present disclosure, the FGF4 gene amplified tumor is esophageal cancer.

[Detect number of copies]
Therefore, the effectiveness of the pharmaceutical composition according to the present disclosure can be determined by examining the presence or absence of amplification of the copy number of the FGF4 gene in the subject tumor. Therefore, in another aspect, the present disclosure detects the gene amplification level of the FGF4 gene in the target tumor tissue sample or tumor cell sample, and the pharmaceutical composition according to the present disclosure when the target FGF4 gene is amplified Classifying the subject as the efficacy of therapeutic administration of the product is predicted, or the efficacy of therapeutic administration of the pharmaceutical composition according to the present disclosure is not expected when the subject's FGF4 gene is not amplified And a method including classifying the object (hereinafter, also referred to as “classification method according to the present disclosure”). Examples of the classification method according to the present disclosure include humans and mammals other than humans in one or a plurality of embodiments.

  For the presence or absence of amplification of the copy number of the FGF4 gene, in one or more embodiments, a conventionally performed method or a method developed in the future can be used. In one or a plurality of embodiments, the presence or absence of amplification of the copy number of the FGF4 gene can be performed by fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, or real-time quantitative PCR. In one or a plurality of embodiments of the present disclosure, the fact that the copy number of the FGF4 gene is increased means that the copy number of the FGF4 gene of the target tumor is larger than the copy number of the control gene of normal cells.

Amplification of gene copy number by fluorescence in situ hybridization (FISH) can be detected / measured in one or more embodiments as follows.
In the FISH method, a fragment DNA (probe) of a specific gene is hybridized with a chromosome or nuclear DNA using a fluorescent substance, and the increase / decrease (deletion or amplification) of the gene and the position (mapping) in a staining pair are examined. It is. Therefore, cells can be stained with a probe specific to the sequence of the target gene, and the increase or decrease in copy number can be measured according to the number of signals. Also, “https://www.yodosha.co.jp/jikkenigaku/keyword/234.html” and “Hepatology. 2013 Apr; 57 (4): 1407-15. FGF3 / FGF4 amplification and multiple lung metastases in responders to sorafenib in hepatocellular carcinoma. ”

In one or a plurality of embodiments, amplification of gene copy number by a next-generation sequencer can be detected / measured as follows.
Unlike the conventional Sanger method, the next-generation sequencer can process tens of millions to hundreds of millions of DNA fragments in parallel. Therefore, it is possible to estimate the increase or decrease in the copy number of the target gene by sequencing the DNA from the tumor sample and the control sample and comparing the coverage of the target gene. Also, “PLoS One. 2013; 8 (5): e64271. Targeted Sequencing of Cancer-Related Genes in Colorectal Cancer Using Next-Generation Sequencing” and “Clin Cancer Res. 2013 May 15; 19 (10): 2668-76. Relapsed classic E-cadherin (CDH1) -mutated invasive lobular breast cancer shows a high frequency of HER2 (ERBB2) gene mutations. ”

In one or a plurality of embodiments, amplification of gene copy number by a DNA chip can be detected / measured as follows.
That is, CGH (comparative genomic hybridization) analysis is performed using a slide glass (DNA chip) in which a large number of DNA fragments are arrayed. Tumor samples and control samples labeled with different fluorescent dyes are hybridized with DNA fragments on the DNA chip. The number of copies of tumor DNA in the region corresponding to the arrayed DNA fragments is quantified based on the intensity ratio of the fluorescent signal derived from each sample. Also, “http://www.cghtmd.jp/CGHDatabase/about/about_high.jsp” and “Hepatology. 2013 Apr; 57 (4): 1407-15. FGF3 / FGF4 amplification and multiple lung metastases in responders to sorafenib in hepatocellular carcinoma. "

Amplification of gene copy number by real-time quantitative PCR can be detected / measured as follows.
That is, using a sample DNA as a template, a target gene site is amplified by a PCR reaction using primers specific to the target gene. The amplification reaction is detected in real time, the number of PCR cycles for amplification up to a certain amount is compared between the control sample and the tumor sample, and the copy number is calculated according to the difference. Also, “http://tools.invitrogen.com/content/sfs/manuals/cms_062368.pdf” and “Hepatology. 2013 Apr; 57 (4): 1407-15. FGF3 / FGF4 amplification and multiple lung metastases in responders to sorafenib in hepatocellular carcinoma. ”

[Active ingredients]
The active ingredient of the pharmaceutical composition according to the present disclosure is a substance that suppresses the expression of FGF4 protein or a substance that inhibits the function. In one or a plurality of embodiments, the active ingredient of the pharmaceutical composition according to the present disclosure includes at least one selected from the group consisting of an anti-FGF4 antibody, an FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA.

  In view of improving the antitumor effect of the pharmaceutical composition, the anti-FGF4 antibody has neutralizing activity or high neutralizing activity in one or more embodiments. The anti-FGF4 antibody is a monoclonal antibody in one or more embodiments. Further, in one or a plurality of embodiments, the anti-FGF4 antibody may be in the form of an antibody fragment or may be modified with a sugar chain.

  The FGF receptor is a receptor for the FGF4 protein. Therefore, the FGF receptor inhibitor can be used as an active ingredient of the pharmaceutical composition according to the present disclosure. As the FGF receptor inhibitor, conventionally known and future developed ones can be used.

  FGF4 gene siRNA (small interfering RNA) and shRNA (small hairpin) can also be used as an active ingredient of the pharmaceutical composition according to the present disclosure. A person skilled in the art can design and manufacture appropriate FGF4 gene siRNA and shRNA according to standard methods.

  In one or more embodiments of the present disclosure, the “pharmaceutical composition” can be made into a dosage form suitable for an administration form by applying a well-known formulation technique. Examples of the dosage form include, but are not limited to, oral administration in a dosage form such as a tablet, capsule, granule, powder, pill, troche, syrup, and liquid. Alternatively, parenteral administration in dosage forms such as injections, liquids, aerosols, suppositories, patches, lotions, liniments, ointments, eye drops and the like can be mentioned. These preparations can be produced by known methods using additives such as, but not limited to, excipients, lubricants, binders, disintegrants, stabilizers, flavoring agents, and diluents.

  In one or a plurality of embodiments, the pharmaceutical composition according to the present disclosure does not contain other active ingredients having a therapeutic effect, or further contains one or more active ingredients.

  Examples of the excipient include, but are not limited to, starch such as starch, potato starch, and corn starch, lactose, crystalline cellulose, and calcium hydrogen phosphate. Examples of the coating agent include, but are not limited to, ethyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, shellac, talc, carnauba wax, paraffin, and the like. Examples of the binder include, but are not limited to, polyvinyl pyrrolidone, macrogol and the same compound as the excipient. Examples of the disintegrant include, but are not limited to, compounds similar to the excipients and chemically modified starch / celluloses such as croscarmellose sodium, sodium carboxymethyl starch, and crosslinked polyvinylpyrrolidone. . Examples of the stabilizer include, but are not limited to, paraoxybenzoates such as methylparaben and propylparaben; alcohols such as chlorobutanol, benzyl alcohol, and phenylethyl alcohol; benzalkonium chloride; phenol, cresol Mention may be made of such phenols; thimerosal; dehydroacetic acid; and sorbic acid. Examples of the flavoring agent include, but are not limited to, sweeteners, acidulants, and fragrances that are commonly used.

  In the production of the liquid agent, the solvent is not limited to these, but ethanol, phenol, chlorocresol, purified water, distilled water and the like can be used, and a surfactant or an emulsifier can also be used as necessary. . Examples of the surfactant or emulsifier include, but are not limited to, polysorbate 80, polyoxyl 40 stearate, lauromacrogol, and the like.

  The method of using the pharmaceutical composition according to the present disclosure may vary depending on symptoms, age, administration method, and the like. The method of use is not limited to these, but intermittently or continuously, orally, transdermally, submucosally, subcutaneously, intramuscularly, vascularly, so that the in vivo concentration of the active ingredient is between 100 nM and 1 mM. It can be administered internally, intracerebrally, or intraperitoneally. As a non-limiting embodiment, in the case of oral administration, the lower limit is 0.01 mg (preferably 0.1 mg) and the upper limit is 2000 mg as converted into the active ingredient per day for a subject (adult if human). (Preferably 500 mg, more preferably 100 mg) may be divided into 1 or several times and administered according to symptoms. As a non-limiting embodiment, in the case of intravenous administration, the lower limit is 0.001 mg (preferably 0.01 mg) and the upper limit is 500 mg (preferably 50 mg) per day for a subject (adult if human). Is divided into one or several times and administered according to symptoms.

[Method and use]
In one aspect, the present disclosure relates to at least one use selected from the group consisting of an anti-FGF4 antibody, an FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA for producing a pharmaceutical composition according to the present disclosure. In addition, in one aspect, the present disclosure relates to a method for improving, suppressing progression, and / or treating an FGF4 gene-amplified tumor, comprising administering a pharmaceutical composition according to the present disclosure to a subject in need. Further, in one aspect, the present disclosure detects the gene amplification level of the FGF4 gene in the target tumor tissue sample or tumor cell sample, and suppresses the expression of the FGF4 protein when the target FGF4 gene is amplified. Alternatively, the present invention relates to a method for improving, suppressing progression and / or treating an FGF4 gene-amplified tumor, comprising administering to the subject a pharmaceutical composition containing a substance that inhibits function as an active ingredient. Furthermore, the present disclosure provides at least one selected from the group consisting of an anti-FGF4 antibody, an FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA in the above-described improvement, suppression of progression, and / or treatment of an FGF4 gene amplified tumor. Use, or use of a pharmaceutical composition according to the present disclosure.

[kit]
In another aspect, the present disclosure provides a reagent for detecting the FGF4 gene amplification level by a method selected from the group consisting of fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, and real-time quantitative PCR, And a reagent for detecting the FGF4 gene amplification level by a method selected from the group consisting of fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, and real-time quantitative PCR, and a nucleotide that hybridizes to the FGF4 gene Relates to a kit comprising:

The present disclosure further relates to one or more of the following embodiments.
[1] A pharmaceutical composition for improvement, suppression of progression and / or treatment of FGF4 gene-amplified tumor, at least selected from the group consisting of anti-FGF4 antibody, FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA A pharmaceutical composition containing one as an active ingredient.
[2] The pharmaceutical composition according to [1], wherein the FGF4 gene-amplified tumor is esophageal cancer.
[3] The pharmaceutical composition according to [1] or [2], wherein the anti-FGF4 antibody is a neutralizing antibody having specificity for FGF4.
[4] detecting the gene amplification level of the FGF4 gene in the tumor tissue sample or tumor cell sample of the subject; and
Classifying the subject when the efficacy of therapeutic administration of the pharmaceutical composition according to any one of [1] to [3] is predicted when the subject's FGF4 gene is amplified, or Categorizing the subject as not predicting the effectiveness of therapeutic administration of the pharmaceutical composition of any of [1] to [3] when the FGF4 gene is not amplified.
[5] The method according to [4], wherein the FGF4 gene amplification level is detected by a method selected from the group consisting of fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, and real-time quantitative PCR.
[6] The method according to [4] or [5], wherein the tumor tissue or tumor cell is collected from the subject's esophagus.
[7] A reagent for detecting the FGF4 gene amplification level and the FGF4 gene are hybridized by a method selected from the group consisting of fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, and real-time quantitative PCR. A kit for use in the method according to any one of [4] to [6], comprising nucleotides.
[8] Use of at least one selected from the group consisting of an anti-FGF4 antibody, an FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA for producing the pharmaceutical composition according to any one of [1] to [3] .
[9] A method for improving, suppressing progression and / or treating an FGF4 gene-amplified tumor, comprising administering the pharmaceutical composition according to any one of [1] to [3] to a subject in need.
[10] Use of at least one selected from the group consisting of an anti-FGF4 antibody, an FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA in the improvement, suppression of progression, and / or treatment of the FGF4 gene amplified tumor of [9] .
[11] Detecting the gene amplification level of the FGF4 gene in the target tumor tissue sample or tumor cell sample, and effectively inhibiting the function or inhibiting the function of the FGF4 protein when the target FGF4 gene is amplified A method for improving, suppressing progression and / or treating FGF4 gene-amplified tumor, comprising administering a pharmaceutical composition as a component to the subject.
[12] Use of the pharmaceutical composition according to any one of [1] to [3] in the improvement, suppression of progression, and / or treatment of the FGF4 gene-amplified tumor according to [11].

  Hereinafter, although it demonstrates based on an Example and a comparative example, this indication is not limited to this.

[Analysis of FGF4 gene copy number in surgical specimen of esophageal cancer]
Copy number analysis of the FGF4 gene was performed in 194 esophageal cancer surgical specimens.
(Sample)
A specimen of esophageal cancer (ESCC: esophageal squamous cell carcinoma) which was a frozen specimen excised at surgery or a FFPE (formalin fixed paraffin embedded) specimen was used as a test sample.
(Copy number assay)
DNA was extracted from esophageal cancer specimens and copy number assays were performed using TaqMan Copy Number Assays (Applied Biosystems, Foster City, Calif.). The ID of the primer used is HS01235235_cn (Forward: 5′-GAG CAG CAA GGG CAA GCTCTA -3 ′, Reverse: 5′-ACC TTC ATG GTG GGCGAC A -3 ′). In addition, TERT locus was used as an internal control (Forward: 5'-GCA TCG AGGCTG AGA AC-3 ', Reverse: 5'-ATG GTG GTGAAG ACG CCA GT-3'). In addition, Human Genomic DNA (Clontech) was used as a normal control. The result is shown in FIG.

  FIG. 1 is a graph showing an example of the results of measuring the FGF4 gene copy number of 194 esophageal cancer specimens. The horizontal axis is the sample sample and is arranged in order of copy number. The vertical axis represents the measured copy number. As shown in the figure, the copy number of the FGF4 gene was 5 or more in about 40% of the esophageal cancer specimens. In addition, when the same esophageal cancer specimen was analyzed by CGH (comparative genomic hybridization) analysis method and FISH, amplification of FGF4 gene was recognized in the same tendency (data not shown). The CGH method was performed using the trade name: Oncoscan, and the FISH used a probe obtained from a human genomic clone library (GSP Laboratory, Kawasaki, Japan).

[Preparation of FGF4 overexpression strain]
The FGF4 gene was introduced into the colon cancer cell line WiDr, lung cancer cell line A549, and esophageal cancer cell line KYSE-270, respectively, and cell lines overexpressing FGF4 protein were prepared. The produced FGF4 overexpression strains are referred to as WiDr / FGF4, A549 / FGF4, and KYSE-270 / FGF4, respectively.
On the other hand, cell lines WiDr / EGFP, A549 / EGFP, and KYSE-270 / EGFP in which EGFP gene was introduced and EGFP protein was overexpressed were prepared as these controls.
(Evaluation of tumorigenicity)
The prepared FGF4 overexpression strain was examined for tumorigenicity. Specifically, about 5 × 10 ⁶ cells are transplanted subcutaneously into mice, and 6, 10, 11, 13, 15, 18, 21 days after culture, or 4, 7, 11, 14, 18 On the 21st day, tumor size was measured. Tumor size was measured with calipers. The results are shown in FIGS.

  FIG. 2 is a graph showing an example of the results of measuring the tumor size of a cell line (WiDr / FGF4) over time when FGF4 gene is introduced into a colon cancer cell line WiDr and overexpressed in culture. . FIG. 3 is a graph showing an example of the results of measuring the tumor size of the cell line (A549 / FGF4) over time when the FGF4 gene was introduced into the lung cancer cell line A549 and overexpressed. FIG. 4 shows an example of the results of measuring the tumor size of a cell line (KYSE-270 / FGF4) over time when the FGF4 gene was introduced into the esophageal cancer cell line KYSE-270 and overexpressed. It is a graph to show. In any case, significant enhancement of tumor-forming ability was observed with respect to the control into which the EGFP gene was introduced. Moreover, these results show that the FGF4 gene can exert a function (oncogenic activity) as an oncogene.

[Effect of anti-FGF4 antibody on FGF4-overexpressing tumor in vivo]
The effect of anti-FGF4 antibody on FGF4 overexpressing tumors was confirmed in vivo.
As the anti-FGF4 antibody, a commercially available product (trade name: AB-235-NA) was used. The FGF4 neutralizing ability of this antibody was IC ₅₀ = 10 to 20 μg / ml (data not shown).
An FGF4-overexpressing esophageal cancer cell line KYSE-270 / FGF4 transplanted tumor-bearing mouse model was prepared. Specifically, a mouse model was prepared by transplanting KYSE-270 / FGF4 cells subcutaneously into nude mice. Mice were divided into 2 groups of 4 mice, goat polyclonal IgG was administered to the control group, and anti-FGF4 antibody was administered intraperitoneally to the other group (5 mg / kg, 2 times / week, 2 weeks). . Tumor diameter and body weight were measured every few days.

FIG. 5 is a graph showing an example of the results of confirming the effect of the anti-FGF4 antibody on the tumor of KYSE-270 / FGF4 transplanted into mice. In the left diagram of FIG. 5, the vertical axis represents the tumor size (mm ³ ), and the horizontal axis represents the culture time (days). In the right diagram of FIG. 5, the vertical axis represents the mouse body weight (g), and the horizontal axis represents the culture time (days). As shown in the left diagram of FIG. 5, significant antitumor activity (T / C = 0.7) was observed for the anti-FGF4 antibody. In addition, as shown in the right diagram of FIG. 5, even when the anti-FGF4 antibody was administered, the weight of the mouse was not reduced.

[Effect of FGFR inhibitor on FGF4-overexpressing tumor in vivo]
The effect of FGFR inhibitors on FGF4 overexpressing tumors was confirmed in vivo.
A commercially available product (trade name: PD173074) was used as the FGFR inhibitor. The PD173074 inhibits the FGFR signal pathway (data not shown).
A tumor-bearing mouse model in which the FGF4 overexpressing lung cancer cell line A549 / FGF4 was transplanted was prepared. This tumor-bearing mouse model was intraperitoneally administered with the FGFR inhibitor at 50 mg / kg, 7 times / week for 1 week. As a control, DMSO was used instead of the FGFR inhibitor. The result is shown in FIG.

FIG. 6 is a graph showing an example of the results of confirming the effect of an FGFR inhibitor on the FGF4-overexpressing lung cancer cell line A549 / FGF4 transplanted into mice. The vertical axis represents the tumor size (mm ³ ), and the horizontal axis represents the culture time (days). As shown in the figure, the FGFR inhibitor showed significant antitumor activity.

Claims (8)

  A pharmaceutical composition for FGF4 (Fibroblast growth factor 4) gene amplification tumor improvement, progression inhibition and / or treatment, selected from the group consisting of anti-FGF4 antibody, FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA A pharmaceutical composition containing at least one of the above as an active ingredient.   The pharmaceutical composition according to claim 1, wherein the FGF4 gene-amplified tumor is esophageal cancer.   The pharmaceutical composition according to claim 1 or 2, wherein the anti-FGF4 antibody is a neutralizing antibody having specificity for FGF4. Detecting the level of gene amplification of the FGF4 gene in a tumor tissue sample or tumor cell sample of interest; and
When the subject FGF4 gene is amplified, the subject is classified as being effective for the therapeutic administration of the pharmaceutical composition according to any one of claims 1 to 3, or the subject FGF4 gene A method comprising classifying said subject as not predicting the effectiveness of therapeutic administration of a pharmaceutical composition according to any of claims 1 to 3 when is not amplified.   The method according to claim 4, wherein the detection of the FGF4 gene amplification level is performed by a method selected from the group consisting of fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, and real-time quantitative PCR.   The method according to claim 4 or 5, wherein the tumor tissue or tumor cell is collected from a subject's esophagus.   A reagent for detecting the FGF4 gene amplification level by a method selected from the group consisting of fluorescence in situ hybridization (FISH), next-generation sequencer, DNA chip, and real-time quantitative PCR, and a nucleotide that hybridizes to the FGF4 gene A kit for use in the method according to any one of claims 4 to 6.   Use of at least one selected from the group consisting of an anti-FGF4 antibody, an FGF receptor inhibitor, FGF4 siRNA, and FGF4 shRNA for producing the pharmaceutical composition according to any one of claims 1 to 3.

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