Treatment of malignant tumor

Abstract

Provided is a method relating to the treatment of a malignant tumor. The present invention uses a pharmaceutical composition for the treatment of a malignant tumor, the pharmaceutical composition containing brincidofovir, a pharmaceutically acceptable salt of brincidofovir, or a solvate of brincidofovir or the pharmaceutically acceptable salt, wherein a target for the treatment has a malignant tumor with reduced expression of TLE1. The present invention may use a pharmaceutical composition for the treatment of a malignant tumor, the pharmaceutical composition containing brincidofovir, a pharmaceutically acceptable salt of brincidofovir, or a solvate of brincidofovir or the pharmaceutically acceptable salt, wherein a target for the treatment is specified by using the expression level of TLE1 as an indicator. The present invention may use a method for predicting the sensitivity to brincidofovir, a pharmaceutically acceptable salt of brincidofovir, or a solvate of brincidofovir or the pharmaceutically acceptable salt in a target having a malignant tumor, the method including a step for examining the expression level of TLE1 in a sample derived from the target.

Description

Treatment of malignant tumors

[0001] The technical field of the present invention relates to the treatment of malignant tumors.

[0002] Brincidofovir (BCV) was approved by the U.S. Food and Drug Administration (FDA) in 2021 as a treatment for smallpox. Non-Patent Document 1 describes a benefit-risk assessment of BCV in smallpox treatment. Non-Patent Document 2 also describes that monkeypox epidemics have begun in May 2022, and that the use of BCV is recommended for the management of confirmed cases.

[0003] "Benefit-risk assessment for brincidofovir for the treatment of smallpox: US Food and Drug Administration's Evaluation" Chan-Tack et al., Antiviral Res. 2021 Nov;195:105182. "Monkeypox: A Mini-Review on the Globally Emerging Orthopoxvirus" Evans et al., Int J Environ Res Public Health. 2022 Nov 25;19(23):15684.

[0004] Many unknowns remain regarding the therapeutic effectiveness and methods of using BCV for malignant tumors.

[0005] Meanwhile, as described in the Examples below, the present inventors have discovered that BCV susceptibility in malignant tumors is associated with the expression level of TLE1. This finding has been shown to enable BCV treatment to be performed with improved therapeutic efficacy or an increased probability of therapeutic efficacy.

[0006] One aspect of the present invention provides a pharmaceutical composition for treating a malignant tumor, the pharmaceutical composition comprising brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, wherein the subject of the treatment has a malignant tumor with low TLE1 expression. Using this pharmaceutical composition, appropriate treatment with brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof can be carried out.

[0007] One aspect of the present invention provides a pharmaceutical composition for treating malignant tumors, comprising brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, wherein the subject of the treatment is a subject identified based on the expression level of TLE1. Use of this pharmaceutical composition enables appropriate treatment with brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0008] According to one aspect of the present invention, there is provided a composition for use in predicting sensitivity to brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof in a subject with a malignant tumor, the composition comprising a reagent for testing TLE1 expression levels. The use of this composition enables the prediction of sensitivity.

[0009] According to one aspect of the present invention, there is provided a composition for use in determining whether a subject with a malignant tumor should be administered brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, the composition comprising a reagent for testing TLE1 expression levels. The composition can be used to determine whether a subject is suitable for administration of brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0010] According to one aspect of the present invention, there is provided a composition for use in diagnosing whether a subject has a malignant tumor highly susceptible to brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, the composition comprising a reagent for testing TLE1 expression levels. Use of this composition enables diagnosis of whether a subject has a highly susceptible malignant tumor.

[0011] According to one aspect of the present invention, there is provided a method for predicting sensitivity to brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof in a subject with a malignant tumor, the method comprising examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor.

[0012] According to one aspect of the present invention, there is provided a method for determining whether a subject with a malignant tumor should be administered brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor. By using this method, it is possible to determine whether the subject is suitable for administration of brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0013] According to one aspect of the present invention, there is provided a method for diagnosing whether a subject has a malignant tumor highly susceptible to brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor. Using this method, it is possible to diagnose whether a subject has a highly susceptible malignant tumor.

[0014] According to one aspect of the present invention, there is provided a composition for use in predicting the prognosis of a malignant tumor, the composition comprising a reagent for testing TLE1 expression levels. The composition can be used to predict prognosis.

[0015] According to one aspect of the present invention, there is provided a method for predicting the prognosis of NK / T lymphoma or B-cell lymphoma using the expression level of TLE1 in a sample from a malignant tumor subject as an index. This method can be used to predict the prognosis.

[0016] According to one aspect of the present invention, there is provided a composition for use in selecting a treatment method for a malignant tumor, the composition comprising a reagent for testing TLE1 expression levels, which can be used to select an appropriate treatment method.

[0017] According to one aspect of the present invention, there is provided a method for selecting a treatment method for a malignant tumor using the expression level of TLE1 in a subject sample as an index, which allows for the selection of an appropriate treatment method.

[0018] According to one aspect of the present invention, there is provided a pharmaceutical composition for treating a malignant tumor, the pharmaceutical composition comprising brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof, wherein the malignant tumor is T-cell lymphoma. The pharmaceutical composition can be used to perform an appropriate treatment with brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0019] The following figures show the effects of BCV on the viability of NK / T-cell lymphoma cell lines. Figure 2A shows the results of administering BCV to NSG mice transplanted with NK / T-cell lymphoma cells (NK-S1). Figure 2A shows tumor size on days 1, 5, 8, 12, and 15. Figure 2B shows body weight on days 1, 5, 8, 12, and 15. Figure 2C shows tumor weight measurements. The bar graph values ​​indicate vehicle administration and BCV administration, from left to right. Figure 2D shows a photograph of tumors from each individual. The results of whole-transcriptome sequencing of 11 NK / T-cell lymphoma cell lines are shown. The plot in Figure 3 shows the relative relationship between the top four cell lines and the remaining seven cell lines. Figure 3 is divided into six sections by dotted lines. The circles in the upper left section, including TLE1, FAM89A, and AK4, indicate genes with elevated expression in the remaining seven cell lines (i.e., genes with decreased expression in the top four cell lines). The circles in the upper right section, including ZNF441 and ZNF141, indicate genes with elevated expression in the top four cell lines. The other four sections contain relatively weak genes with expression differences of less than 2-fold between the top four cell lines and the remaining seven cell lines, or genes with a correlation significance of 0.01 or greater between the top four cell lines and the remaining seven cell lines. Throughout this specification and figures, NKTL is synonymous with NK / T-cell lymphoma. Figure 6A shows the results of an investigation into TLE1 protein expression levels. Figure 6B shows the results of gene mutation analysis. Figure 7A shows the results of gene set expression analysis. Figure 6A shows the gene set with elevated expression in the top four cell lines. Figure 6B shows an enrichment plot for MYC TARGETS V2. Figure 7A shows the results of an investigation into the impact of TLE1 expression levels on prognosis. Figure 7A shows the results of an investigation into progression-free survival. Figure 7B shows the results of an investigation into overall survival. Figure 7B shows the results of expression analysis of a gene set from a cohort of NK / T-cell lymphoma patients. Figure 7C shows the results of an investigation into the effect of BCV on the survival rate of T-cell lymphoma cell lines. Figure 7D shows the results of administering BCV to NSG mice transplanted with T-cell lymphoma cells (Z4739). Figure 10A shows the results of an investigation into tumor size on days 4, 7, 11, 18, and 21. Figure 10B shows the results of an investigation into body weight on days 1, 4, 7, 11, 18, and 21. Figure 10C shows the results of tumor weight measurements.The values ​​in the bar graph indicate, from the left, vehicle administration and BCV administration. Figure 10D shows photographs of tumors collected from each individual. The results of whole-transcriptome sequencing analysis of (i) the top four cell lines with respect to susceptibility to BCV as described in Example 1, (ii) the remaining seven cell lines, and (iii) the 11 cell lines that tended to be highly susceptible to BCV as described in Example 9 are shown. The results of investigating the effect of BCV on gene expression in T-cell lymphoma cell lines are shown. Figure 12A shows the results of investigating normalized enrichment scores. Figure 12B shows an enrichment plot for MYC TARGETS V2 and V1. Figure 12C shows the results of investigating gene expression profiles. The results of investigating the effect of BCV on gene expression in T-cell lymphoma cell lines (Western blotting results) are shown. The results of investigating the effect of BCV on the viability of B-cell lymphoma cell lines are shown. The results of administering BCV to NSG mice transplanted with B-cell lymphoma cells (OCI-LY19) are shown. Figure 15A shows the results of tumor size on days 1, 5, 6, and 7. Figure 15B shows the results of body weight on days 1, 5, 6, and 7. Figure 15C shows the results of tumor weight measurements. The bar graph values ​​indicate, from left to right, vehicle administration and BCV administration. Figure 15D shows a photograph of tumors taken from each individual. The results of whole-transcriptome sequencing analysis of 19 B-cell lymphoma cell lines are shown. The plot in Figure 16 shows the relative relationship between the top nine cell lines and the remaining 10 cell lines. Figure 16 is divided into six sections by dotted lines. Circles in the upper left section indicate genes with decreased expression in the top nine cell lines, while circles in the upper right section, including TLE1, CERS6, etc., indicate genes with increased expression in the top nine cell lines. The other four compartments contain genes with a two-fold or less difference in expression between the top nine cell lines and the remaining 10 cell lines, or genes with a correlation significance of 0.01 or more between the top nine cell lines and the remaining 10 cell lines. The following shows the results of investigating the impact of TLE1 expression levels on prognosis. Figure 17A shows the results of investigating overall survival based on GSE11318 (Lenz et al., Proc Natl Acad Sci USA. 2008 Sep 9;105(36):13520-5.).Figure 17B shows the results of examining overall survival time based on GSE10846 (Lenz et al., N Engl J Med. 2008 Nov 27;359(22):2313-23.).

[0020] Hereinafter, embodiments of the present invention will be described in detail, with the same contents omitted as appropriate to avoid repetition.

[0021] (1) Method According to one embodiment of the present invention, a method for treating a malignant tumor is provided, comprising administering brincidofovir (BCV), a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject with a malignant tumor. The subject may be a subject with a malignant tumor with low TLE1 expression or a subject identified using TLE1 expression level as an indicator. In the Examples described below, BCV susceptibility in malignant tumors was associated with TLE1 expression level. For example, it has been shown that NK / T-cell lymphoma or T-cell lymphoma with low TLE1 expression level has high BCV susceptibility. Using this method, BCV can be administered to subjects with low TLE1 expression who are expected to have high BCV susceptibility. In such cases, improved therapeutic effects or a higher probability of therapeutic efficacy can be expected compared to administration to unspecified subjects. Furthermore, using this method, susceptibility can be predicted using TLE1 expression level as an indicator, allowing for more appropriate BCV treatment. For example, if a subject has a low TLE1 expression level and is predicted to have high susceptibility, treatment may be performed by administering BCV or administering it at a low dose. Furthermore, for example, if a subject has a high TLE1 expression level and is predicted to have low sensitivity, treatment may be performed by not administering BCV or by administering it at a high dose. Furthermore, as described in the Examples below, patients with NK / T-cell lymphoma with low TLE1 expression who have not been treated with BCV had a particularly poor prognosis. Therefore, BCV can be used to treat malignant tumors with poor prognosis. Furthermore, the Examples below showed that B-cell lymphomas with high TLE1 expression levels are highly susceptible to BCV. Therefore, using this method, BCV can be administered to subjects who are expected to have high TLE1 expression and high BCV sensitivity. In such cases, improved therapeutic effects or a higher probability of therapeutic effects can be expected compared to administration to unspecified subjects. Furthermore, using this method, even for malignant tumors with high TLE1 expression and high BCV sensitivity, sensitivity can be predicted using the TLE1 expression level as an indicator, allowing for more appropriate BCV treatment. For example, if a subject has a high TLE1 expression level and is predicted to have high sensitivity, treatment may be performed by administering BCV or by administering it at a low dose.Furthermore, for example, if the subject's TLE1 expression level is low and sensitivity is predicted to be low, treatment may be performed by not administering BCV or by administering it at a high dose. Furthermore, as described in the Examples below, patients with BCV-untreated B-cell lymphomas with high TLE1 expression had a particularly poor prognosis. Therefore, BCV can be used to treat malignant tumors with poor prognosis.

[0022] (2) Method According to one embodiment of the present invention, there is provided a method for treating a malignant tumor with a poor prognosis, the method comprising the step of administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject having a malignant tumor, the subject being identified using the expression level of TLE1 as an index. This method can be used to treat a malignant tumor with a poor prognosis.

[0023] (3) Methods According to one embodiment of the present invention, there is provided a method for treating a malignant tumor, the method comprising the step of administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject with a malignant tumor (including a subject having a malignant tumor) that expresses TLE1 underexpression. By using this method to administer to a subject with a malignant tumor that expresses TLE1 underexpression and is highly susceptible to BCV, a particularly high therapeutic effect of BCV can be achieved. By using this method to treat a subject predicted to be highly susceptible to BCV, the probability of achieving a therapeutic effect is increased. According to one embodiment of the present invention, there is provided a method for treating a malignant tumor, the method comprising the step of administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject with a malignant tumor that expresses TLE1 overexpression. By using this method to treat a subject predicted to be highly susceptible to BCV, the probability of achieving a therapeutic effect is increased.

[0024] (4) Methods According to one embodiment of the present invention, there is provided a method for inhibiting the proliferation of malignant tumor cells, the method comprising the step of contacting BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof with malignant tumor cells that express a low level of TLE1 (malignant tumor cells with low TLE1 expression). By using this method, a particularly strong effect of inhibiting the proliferation of malignant tumor cells can be achieved by contacting BCV with malignant tumor cells that express a low level of TLE1 and are highly sensitive to BCV. According to one embodiment of the present invention, there is provided a method for inhibiting the proliferation of malignant tumor cells, the method comprising the step of contacting BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof with malignant tumor cells that express a high level of TLE1 (malignant tumor cells with high TLE1 expression). By using this method, a particularly strong effect of inhibiting the proliferation of malignant tumor cells can be achieved by contacting BCV with malignant tumor cells that express a high level of TLE1 and are highly sensitive to BCV.

[0025] (5) Methods According to one embodiment of the present invention, a method for predicting susceptibility to BCV, its pharmaceutically acceptable salt, or a solvate thereof in a subject with a malignant tumor is provided, comprising a step of examining the expression level of TLE1 in a sample derived from the subject. This method can be used to predict or identify subjects with high or low susceptibility to BCV using the expression level of TLE1 as an indicator. Therefore, this method can be used to implement more appropriate BCV treatment. For example, if a subject's TLE1 expression level is low and the subject is predicted to be highly susceptible, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject's TLE1 expression level is high and the subject is predicted to be highly susceptible, treatment may be performed by not administering BCV or administering it at a high dose. For example, if a subject's TLE1 expression level is high and the subject is predicted to be highly susceptible, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject's TLE1 expression level is low and the subject is predicted to be highly susceptible, treatment may be performed by not administering BCV or administering it at a high dose. From another perspective, one embodiment of the present invention provides a method for predicting susceptibility to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof in a malignant tumor subject, the method comprising a step of examining the expression level of TLE1 in malignant tumor cells derived from the malignant tumor subject.

[0026] (6) Methods According to one embodiment of the present invention, there is provided a method for determining a subject with a malignant tumor to be administered with BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, comprising a step of examining the expression level of TLE1 in a sample derived from the subject. This method allows for more appropriate BCV treatment. For example, if a subject's TLE1 expression level is low and predicted to be highly sensitive, the subject may be selected as a subject for administration of BCV or a low dose of BCV. For example, if a subject's TLE1 expression level is high and predicted to be highly sensitive, the subject may be selected as a subject for administration of BCV or a low dose of BCV. For example, if a subject's TLE1 expression level is high and predicted to be highly sensitive, the subject may be selected as a subject for administration of BCV or a low dose of BCV. For example, if a subject's TLE1 expression level is low and predicted to be highly sensitive, the subject may be selected as a subject for administration of BCV. The subject to be administered may also be a subject to be administered with BCV. From another perspective, according to one embodiment of the present invention, there is provided a method for selecting a malignant tumor subject suitable for treatment with BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the malignant tumor subject.

[0027] (7) Methods According to one embodiment of the present invention, a method for diagnosing whether a subject has a malignant tumor highly susceptible to BCV, its pharmaceutically acceptable salt, or a solvate thereof is provided, comprising a step of examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor. This method can be used to diagnose whether a subject has a malignant tumor highly susceptible to BCV using the expression level of TLE1 as an indicator. Therefore, use of this method enables more appropriate BCV treatment to be implemented. For example, if a subject has a low TLE1 expression level and is diagnosed as having a malignant tumor highly susceptible to BCV, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject has a high TLE1 expression level and is diagnosed as not having a malignant tumor highly susceptible to BCV, treatment may be performed by not administering BCV or administering it at a high dose. For example, if a subject has a high TLE1 expression level and is diagnosed as having a malignant tumor highly susceptible to BCV, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject is diagnosed as having a low TLE1 expression level and not having a highly susceptible malignant tumor, treatment may be performed by not administering BCV or administering it at a high dose. This diagnosis may be a companion diagnosis for treatment with BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof.

[0028] (8) Methods According to one embodiment of the present invention, there is provided a method for detecting the expression level of TLE1 in malignant tumor cells of a subject with a malignant tumor sensitive to BCV, its pharmaceutically acceptable salt, or a solvate thereof, the method comprising: (a) obtaining a sample from the subject (wherein the sample may be malignant tumor cells, a malignant tumor cell lysate, or a solution containing nucleic acids or proteins contained in malignant tumor cells); or (b) contacting the sample with a TLE1 detection reagent (e.g., containing a nucleic acid or peptide as described below) and then detecting the TLE1 expression level in the sample. This method allows the detection of the expression level of TLE1 in malignant tumor cells of a subject with a BCV-susceptible malignant tumor. For example, detecting low TLE1 expression levels in malignant tumor cells with low TLE1 expression and high BCV susceptibility allows treatment to be performed while anticipating the effective therapeutic effect of BCV. For example, detecting high TLE1 expression levels in malignant tumor cells with high TLE1 expression and high BCV susceptibility allows treatment to be performed while anticipating the effective therapeutic effect of BCV. For example, detecting a low expression level of TLE1 in malignant tumor cells with low TLE1 expression and high BCV susceptibility can be used as an indicator for BCV dose setting.For example, detecting a high expression level of TLE1 in malignant tumor cells with high TLE1 expression and high BCV susceptibility can be used as an indicator for BCV dose setting.

[0029] (9) Methods According to one embodiment of the present invention, there is provided a method for detecting an indicator for identifying a subject with a malignant tumor who is likely to be susceptible to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor. This method can be used to identify a subject with a malignant tumor who is likely to be susceptible to BCV using the expression level of TLE1 as an indicator. Therefore, use of this method allows for more appropriate BCV treatment.

[0030] (10) Method According to one embodiment of the present invention, there is provided a method for detecting an indicator for identifying a malignant tumor subject who is likely to respond or react to BCV, its pharmaceutically acceptable salt, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the malignant tumor subject. This method can be used to identify a malignant tumor subject who is likely to respond or react to BCV using the expression level of TLE1 as an indicator. Therefore, use of this method allows for more appropriate BCV treatment to be implemented.

[0031] (11) Methods According to one embodiment of the present invention, there is provided a method for detecting low levels of TLE1 expression in malignant tumor cells of a subject to be administered BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, or in a subject for whom administration of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof is being considered. This method includes, for example, (a) obtaining a sample from a subject (wherein the sample may be malignant tumor cells, a malignant tumor cell lysate, or a solution containing nucleic acids or proteins contained in malignant tumor cells), or (b) contacting the sample with a TLE1 detection reagent (e.g., containing a nucleic acid or peptide described below) and then detecting a low level of TLE1 expression in the sample. According to this method, for example, by detecting a low level of TLE1 expression in malignant tumor cells that underexpress TLE1 and are highly susceptible to BCV, it is possible to predict that BCV will have an effective therapeutic effect and perform treatment. According to one embodiment of the present invention, there is provided a method for detecting high levels of TLE1 expression in malignant tumor cells of a subject to be administered BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, or in a subject for whom administration of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof is being considered. This method includes, for example, (a) obtaining a sample from a subject (wherein the sample may be malignant tumor cells, a malignant tumor cell lysate, or a solution containing nucleic acids or proteins contained in malignant tumor cells), or (b) detecting a high level of TLE1 expression in the sample by contacting the sample with a TLE1 detection reagent (e.g., containing a nucleic acid or peptide described below). According to this method, for example, by detecting a high level of TLE1 expression in malignant tumor cells that highly express TLE1 and are highly susceptible to BCV, it is possible to predict that BCV will have an effective therapeutic effect and perform treatment.

[0032] (12) Method According to one embodiment of the present invention, there is provided a method for predicting the response of a subject with a malignant tumor to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the subject. This method can be used to identify subjects who are likely to respond to BCV using the expression level of TLE1 as an indicator. Therefore, use of this method allows for more appropriate BCV treatment.

[0033] (13) Method According to one embodiment of the present invention, there is provided a method for selecting a subject with a malignant tumor who is expected to benefit from treatment with BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor. This method can be used to select subjects who are expected to benefit from treatment with BCV using the expression level of TLE1 as an indicator. Therefore, use of this method allows for more appropriate BCV treatment.

[0034] (14) Method According to one embodiment of the present invention, there is provided a method for obtaining an indicator for determining whether to perform a treatment method using BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from a subject with a malignant tumor. Use of this method allows for more appropriate BCV treatment to be performed.

[0035] (15) Methods According to one embodiment of the present invention, a method for examining the expression level of TLE1 in a sample from a subject having or suspected of developing a malignant tumor is provided. This method may include, for example, measuring the expression level of TLE1 in a sample from the subject, or identifying a subject who provided a TLE1-low expression sample as a recipient of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof. By using this method, a subject with a malignant tumor that expresses TLE1 at a low level and is highly susceptible to BCV can be identified, thereby enabling more appropriate BCV treatment. This method may also include measuring the expression level of TLE1 in a sample from the subject, or identifying a subject who provided a TLE1-high expression sample or a highly expressing sample as a recipient of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof. By using this method, a subject with a malignant tumor that expresses TLE1 at a high level and is highly susceptible to BCV can be identified, thereby enabling more appropriate BCV treatment.

[0036] (16) Methods According to one embodiment of the present invention, there is provided a method for selecting a therapeutic method or an active ingredient for use in a therapeutic method, the method comprising a step of using the expression level of TLE1 as an index. This method may comprise, for example, a step of examining the expression level of TLE1 in a sample derived from a malignant tumor subject. This method may comprise, for example, selecting BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof for use in a therapeutic treatment when the subject is predicted to have low TLE1 expression and high susceptibility to BCV. This method may comprise, for example, adjusting or selecting the dosage of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof using the expression level of TLE1 as an index. The therapeutic method may comprise a step of administering brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject. This method may comprise, for example, selecting BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof for use in a therapeutic treatment when the subject is predicted to have high TLE1 expression and high susceptibility to BCV. This method may comprise, for example, a companion diagnostic method for selecting a therapeutic method or an active ingredient for use in a therapeutic treatment when the subject is predicted to have high TLE1 expression and high susceptibility to BCV.

[0037] (17) Method According to one embodiment of the present invention, there is provided a method for predicting BCV susceptibility in tumor cell proliferation, comprising a step of examining the expression level of TLE1 in a sample derived from a subject with a malignant tumor. By using this method, tumor cell proliferation can be more effectively suppressed.

[0038] (18) Method According to one embodiment of the present invention, there is provided a method for treating a malignant tumor, comprising the step of administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject having a malignant tumor. This method can be used to treat malignant tumors. The treatment may be carried out by administering a therapeutically effective amount of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to the subject. The subject may be a subject having lymphoma. The lymphoma may be NK / T-cell lymphoma, T-cell lymphoma, or B-cell lymphoma. In the Examples described below, it has been shown that the antitumor effect of BCV is particularly strong against T-cell lymphoma. Therefore, T-cell lymphoma is a particularly suitable target for the treatment of BCV.

[0039] (19) Method According to one embodiment of the present invention, there is provided a method for predicting drug sensitivity or determining a target for drug administration, the method comprising a step of examining the expression level of TLE1 in a sample derived from a subject with a malignant tumor. Use of this method enables more effective treatment of malignant tumors.

[0040] (20) Method According to one embodiment of the present invention, there is provided a method for predicting the prognosis of a malignant tumor, comprising a step of using the expression level of TLE1 in a subject with a malignant tumor as an index. This method can be used to predict the prognosis of a patient. For example, a poor prognosis may be predicted when the expression level of TLE1 in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) is low. For example, a good prognosis may be predicted when the expression level of TLE1 in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) is high. This prediction may further include predicting the prognosis based on the expression level of MYC in the subject with a malignant tumor. For example, a poor prognosis may be assessed when the expression level of TLE1 in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) is low and the subject is MYC-positive. For example, if the expression level of TLE1 in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) is high and the sample is MYC-negative, the prognosis may be evaluated as good. For example, if the malignant tumor is a B-cell lymphoma, a poor prognosis (poor prognosis) may be predicted if the expression level of TLE1 in the sample is high, and a good prognosis (good prognosis) may be predicted if the expression level of TLE1 in the sample is low. This prognosis may also be predicted based on the expression level of MYC in the malignant tumor subject. For example, a poor prognosis may be evaluated if the expression level of TLE1 in a sample from a B-cell lymphoma subject is high and MYC-positive. For example, a good prognosis may be evaluated if the expression level of TLE1 in a sample from a B-cell lymphoma subject is low and MYC-negative.

[0041] (21) Method According to one embodiment of the present invention, the methods described in (1) to (20) above may use ZNF441 as an indicator or test subject instead of or in addition to using TLE1 as an indicator or test subject. In this case, "when the expression level of TLE1 is low" described in (1) to (20) above can be interpreted as "when the expression level of ZNF441 is high," "the expression level of TLE1 in the subject is low" can be interpreted as "the expression level of ZNF441 in the subject," "the expression level of TLE1 in the subject is high" can be interpreted as "the expression level of ZNF441 in the subject," "the expression level of TLE1 in the subject is high" can be interpreted as "the expression level of ZNF441 in the subject," "low expression of TLE1" can be interpreted as "high expression of ZNF441," "the expression level of TLE1" can be interpreted as "the expression level of ZNF441," "low expression amount of TLE1" can be interpreted as "high expression amount of ZNF441," and "high expression amount of TLE1" can be interpreted as "low expression amount of ZNF441." When ZNF441 is used as an indicator or test subject, the target disease may be NK / T-cell lymphoma. Furthermore, according to one embodiment of the present invention, the methods described in (1) to (20) above may use, as an indicator, one or more of the proteins or genes named in Figure 3, instead of or in addition to using TLE1 or ZNF441 as an indicator or test subject. In this case, the same embodiments as for TLE1 (e.g., the above (1) to (20)) may be adopted for FAM89A and AK4 in Figure 3 (in this case, TLE1 may be read as FAM89A or AK4), and the same embodiments as for ZNF441 may be adopted for proteins or genes other than FAM89A and AK4 (in this case, ZNF441 may be read as the name of each protein or gene).

[0042] (22) Method According to one embodiment of the present invention, the methods described in (1) to (20) above may use ceramide synthase 6 (CERS6) as an indicator or test subject instead of or in addition to using TLE1 as an indicator or test subject. In this case, "when the expression level of TLE1 is low" in (1) to (20) above can be interpreted as "when the expression level of CERS6 is high," "the expression level of TLE1 in the subject is low" can be interpreted as "the expression level of CERS6 in the subject is high," "the expression level of TLE1 in the subject is high" can be interpreted as "the expression level of CERS6 in the subject," "the expression level of TLE1 in the subject is high" can be interpreted as "the expression level of CERS6 in the subject," "low expression of TLE1" can be interpreted as "high expression of CERS6," "the expression level of TLE1" can be interpreted as "the expression level of CERS6," "low expression amount of TLE1" can be interpreted as "high expression amount of CERS6," and "high expression amount of TLE1" can be interpreted as "low expression amount of CERS6." The target disease when CERS6 is used as an indicator or test subject may be B-cell lymphoma. Furthermore, according to one embodiment of the present invention, the methods described in (1) to (20) above may use, as an indicator, one or more of the proteins or genes named in Figure 16, instead of or in addition to using TLE1 or CERS6 as an indicator or test subject. The embodiments of these proteins or genes may be the same as those of TLE1 or CERS6 (e.g., (1) to (21) above) (in this case, TLE1 or CERS6 may be read as the name of each protein or gene).

[0043] (23) Methods According to one embodiment of the present invention, a method for treating a malignant tumor is provided, comprising administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject with a malignant tumor, the subject being identified using ZNF441 expression level as an indicator. As described in the Examples below, BCV susceptibility in malignant tumors was associated with ZNF441 expression level. For example, it has been shown that a high ZNF441 expression level is associated with a high inhibitory effect of BCV and high susceptibility. Therefore, this method can be used to predict susceptibility using ZNF441 expression level as an indicator, allowing for more appropriate BCV treatment. For example, if a subject's ZNF441 expression level is high and susceptibility is predicted, treatment may be performed by administering BCV or at a low dose. On the other hand, if a subject's ZNF441 expression level is low and susceptibility is predicted, treatment may be performed by not administering BCV or by administering a high dose.

[0044] (24) Methods According to one embodiment of the present invention, a method for treating a malignant tumor is provided, comprising administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject with a malignant tumor, the subject being identified using CERS6 expression level as an indicator. As described in the Examples below, BCV susceptibility in malignant tumors was associated with CERS6 expression level. For example, it has been shown that a high CERS6 expression level is associated with a high inhibitory effect of BCV and high susceptibility. Therefore, this method can be used to predict susceptibility using CERS6 expression level as an indicator, allowing for more appropriate BCV treatment. For example, if a subject's CERS6 expression level is high and susceptibility is predicted, treatment may be performed by administering BCV or at a low dose. On the other hand, if a subject's CERS6 expression level is low and susceptibility is predicted, treatment may be performed by not administering BCV or by administering a high dose of BCV.

[0045] (25) Methods According to one embodiment of the present invention, there is provided a method for predicting susceptibility to BCV, its pharmaceutically acceptable salt, or a solvate thereof in a malignant tumor subject, the method comprising a step of examining the expression level of ZNF441 in a sample derived from the malignant tumor subject. This method can be used to predict or identify subjects with high or low susceptibility to BCV using the expression level of ZNF441 as an indicator. Therefore, this method can be used to implement more appropriate BCV treatment. For example, if a subject's ZNF441 expression level is high and high susceptibility is predicted, treatment may be implemented by administering BCV or administering it at a low dose. On the other hand, if a subject's ZNF441 expression level is low and low susceptibility is predicted, treatment may be implemented by not administering BCV or administering it at a high dose. From another perspective, according to one embodiment of the present invention, there is provided a method for predicting susceptibility to BCV, its pharmaceutically acceptable salt, or a solvate thereof in a malignant tumor subject, the method comprising a step of examining the expression level of ZNF441 in malignant tumor cells derived from the malignant tumor subject.

[0046] (26) Methods According to one embodiment of the present invention, there is provided a method for predicting susceptibility to BCV, its pharmaceutically acceptable salt, or a solvate thereof in a subject with a malignant tumor, comprising a step of examining the expression level of CERS6 in a sample derived from the subject. This method can be used to predict or identify subjects with high or low susceptibility to BCV using the expression level of CERS6 as an indicator. Therefore, this method can be used to implement more appropriate BCV treatment. For example, if a subject's CERS6 expression level is high and thus highly susceptible, treatment may be implemented by administering BCV or a low dose of BCV. On the other hand, if a subject's CERS6 expression level is low and thus highly susceptible, treatment may be implemented by not administering BCV or administering a high dose of BCV. From another perspective, according to one embodiment of the present invention, there is provided a method for predicting susceptibility to BCV, its pharmaceutically acceptable salt, or a solvate thereof in a subject with a malignant tumor, comprising a step of examining the expression level of CERS6 in malignant tumor cells derived from the subject.

[0047] (27) Methods According to one embodiment of the present invention, a method for detecting the expression level of ZNF441 in malignant tumor cells of a subject with a malignant tumor susceptible to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof is provided. This method may include, for example, (a) obtaining a sample from the subject (wherein the sample may be malignant tumor cells, a malignant tumor cell lysate, or a solution containing nucleic acids or proteins contained in malignant tumor cells), or (b) contacting the sample with a ZNF441 detection reagent (e.g., containing a nucleic acid or peptide described below) and then detecting the ZNF441 expression level in the sample. This method allows the detection of the ZNF441 expression level in malignant tumor cells of a subject with a BCV-susceptible malignant tumor. For example, by detecting a high expression level of ZNF441 in malignant tumor cells that are highly ZNF441-expressing and highly susceptible to BCV, treatment can be performed while predicting that BCV will have an effective therapeutic effect. For example, detecting a high expression level of ZNF441 in malignant tumor cells that highly express ZNF441 and are highly susceptible to BCV can be used as an indicator for setting the dose of BCV.

[0048] (28) Methods According to one embodiment of the present invention, a method for detecting the expression level of CERS6 in malignant tumor cells of a subject with a malignant tumor sensitive to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof is provided. This method may include, for example, (a) obtaining a sample from the subject (wherein the sample may be malignant tumor cells, a malignant tumor cell lysate, or a solution containing nucleic acids or proteins contained in malignant tumor cells), or (b) contacting the sample with a CERS6 detection reagent (e.g., containing a nucleic acid or peptide described below) and then detecting the CERS6 expression level in the sample. This method allows the detection of the expression level of CERS6 in malignant tumor cells of a subject with a BCV-susceptible malignant tumor. For example, detecting a high expression level of CERS6 in malignant tumor cells that are highly CERS6-expressing and highly susceptible to BCV allows treatment with the expectation that BCV will have an effective therapeutic effect. For example, detecting a high expression level of CERS6 in malignant tumor cells that are highly CERS6-expressing and highly susceptible to BCV can be used as an indicator for BCV dose adjustment.

[0049] (29) Methods The methods of the embodiments of the present invention (e.g., (1) to (28) above) include, for example, (i) testing whether a subject has a malignant tumor, (ii) identifying a subject with a malignant tumor, (iii) identifying a subject who is positive for a malignant tumor marker, (iv) recovering, obtaining, or preparing a subject sample (e.g., malignant tumor cells or cell lysates thereof), (v) harvesting malignant tumor cells from the subject, (vi) extracting nucleic acid (e.g., genomic DNA or total RNA) from the subject sample, and (vii) determining the TLE1 expression level in the subject or the subject sample. (viii) performing the method of interest using the TLE1 expression level of the subject or a sample from the subject as an index; (ix) identifying the subject as a target for treatment with BCV using the TLE1 expression level of the subject or a sample from the subject as an index; (x) determining the malignant tumor subject to be administered BCV using the TLE1 expression level of the subject or a sample from the subject as an index; (xi) predicting the subject's susceptibility to BCV using the TLE1 expression level of the subject or a sample from the subject as an index; (xii) predicting the subject's susceptibility to BCV using the TLE1 expression level of the subject or a sample from the subject as an index. (xiii) identifying a subject as a candidate for treatment with BCV when the TLE1 expression level in the subject or in a sample from the subject is low, or identifying a subject as a candidate for treatment with BCV when the TLE1 expression level in the subject or in a sample from the subject is high; (xiv) identifying a subject with a malignant tumor with low TLE1 expression as a candidate for treatment, or identifying a subject with a malignant tumor with high TLE1 expression as a candidate for treatment; (xv) identifying a subject with a malignant tumor with low TLE1 expression as a candidate for treatment with low-dose BC (xvi) identifying a subject with a malignant tumor with high TLE1 expression as a subject for treatment with high dose BCV, or identifying a subject with a malignant tumor with low TLE1 expression as a subject for treatment with low dose BCV; (xvii) administering BCV when the subject or a sample from the subject has a low level of TLE1 expression, or administering BCV when the subject or a sample from the subject has a high level of TLE1 expression;(xviii) administering a low dose of BCV when the TLE1 expression level in the subject or a sample from the subject is low, or administering a low dose of BCV when the TLE1 expression level in the subject or a sample from the subject is high; (xix) not administering BCV when the TLE1 expression level in the subject or a sample from the subject is high, or not administering BCV when the TLE1 expression level in the subject or a sample from the subject is low; (xx) administering a high dose of BCV when the TLE1 expression level in the subject or a sample from the subject is high, or administering a high dose of BCV when the TLE1 expression level in the subject or a sample from the subject is low; (xxi) indicating that the subject is (or is likely to be) susceptible to BCV when the TLE1 expression level in the subject or a sample from the subject is low, or indicating that the subject is (or is likely to be) susceptible to BCV when the TLE1 expression level in the subject or a sample from the subject is high; (xxii) indicating that the subject is (or is likely to be) not susceptible to BCV when the TLE1 expression level in the subject or a sample from the subject is high; (xxiii) examining the ZNF441 or CERS6 expression level in the subject or a sample from the subject, (xxiv) performing the method of interest using the ZNF441 or CERS6 expression level in the subject or a sample from the subject as an index, (xxv) identifying the subject as a candidate for treatment with BCV using the ZNF441 or CERS6 expression level in the subject or a sample from the subject as an index, and (xxvi) examining the ZNF441 or CERS6 expression level in the subject or a sample from the subject as an index. (xxvii) a step of predicting the subject's susceptibility to BCV using the ZNF441 or CERS6 expression level in the subject or a sample from the subject as an index; (xxviii) a step of diagnosing whether the subject has a malignant tumor that is highly susceptible to BCV using the ZNF441 or CERS6 expression level in the subject or a sample from the subject as an index; (xxix) a step of identifying the subject as a candidate for treatment with BCV if the ZNF441 or CERS6 expression level in the subject or a sample from the subject is high;(xxx) identifying a subject having a malignant tumor with high ZNF441 or CERS6 expression as a treatment target, (xxxi) identifying a subject having a malignant tumor with high ZNF441 or CERS6 expression as a treatment target for low dose BCV, (xxxii) identifying a subject having a malignant tumor with low ZNF441 or CERS6 expression as a treatment target for high dose BCV, (xxxiii) administering BCV when the ZNF441 or CERS6 expression level in the subject or a sample from the subject is high, (xxxiv) administering BCV at a low dose when the ZNF441 or CERS6 expression level in the subject or a sample from the subject is high, (xxxv) not administering BCV when the ZNF441 or CERS6 expression level in the subject or a sample from the subject is low, (xxxvi) administering BCV at a high dose when the ZNF441 or CERS6 expression level in the subject or a sample from the subject is low, (xxxvii) administering BCV at a high dose when the ZNF441 or CERS6 expression level in the subject or a sample from the subject is high (xxxviii) indicating that the subject is (or is likely to be) susceptible to BCV if the ZNF441 or CERS6 expression level in the subject or a sample from the subject is low, (xxxix) testing the presence or absence of MYC positivity in the subject or a sample from the subject, (xxxx) identifying the subject as a candidate for BCV treatment using the presence or absence of MYC positivity in the subject or a sample from the subject as an indicator, (xxxxi) administering BCV to the subject if the subject or a sample from the subject is MYC positive, (xxxxii) predicting the prognosis of malignant tumors using the MYC expression level in the subject or a sample from the subject as an indicator, (xxxxiii) administering a therapeutically effective amount of BCV to the subject, (xxxxiv) suppressing the proliferation of malignant tumor cells in the subject, (xxxxv) reducing a malignant tumor marker in the subject, or (xxxxvi) reducing the size of a malignant tumor in the subject. The methods of the embodiments of the present invention (e.g., the above (1) to (28)) can employ one or more of the steps (i) to (xxxxvi), which is useful for more appropriate BCV treatment (e.g., for administering treatment to a population that is likely to respond to treatment). When two or more steps are employed, the order of the steps can be arbitrary.The order can be determined depending on the desired treatment method. Any one or more of these steps (i) to (xxxxvi) may be performed before or after the step of administering BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject. Furthermore, methods of embodiments of the present invention (e.g., the above-mentioned steps (1) to (28)) may or may not include, for example, the following, where specified: identifying a subject with cytomegalovirus (CMV), adenovirus (AdV), BK virus (BKV), or variola virus (VaV) infection; identifying a CMV, AdV, BKV, or VaV-seropositive subject; identifying a subject in need of prevention or treatment of CMV, AdV, BKV, or VaV infection; identifying a subject after allogeneic transplantation or allogeneic transplantation (e.g., hematopoietic cell transplantation); identifying a subject in need of an immunosuppressant; identifying an immunosuppressed subject; a method for treating virus-induced tumors in immunosuppressed subjects; or a method for preventing or treating virus (e.g., CMV, AdV, BKV, or VaV) infection. ,

[0050] (30) Composition According to one embodiment of the present invention, a pharmaceutical composition for treating malignant tumors is provided, comprising BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof. The subject of treatment may be a subject with a malignant tumor with low TLE1 expression or a subject identified using the TLE1 expression level as an indicator. In the Examples described below, BCV susceptibility in malignant tumors was associated with the TLE1 expression level. For example, it has been shown that NK / T cell lymphoma or T cell lymphoma with low TLE1 expression levels exhibits high susceptibility. Using this pharmaceutical composition, BCV can be administered to subjects with low TLE1 expression who are expected to be highly susceptible to BCV. In such cases, improved therapeutic effects or a higher probability of therapeutic effects can be expected compared to administration to unspecified subjects. Furthermore, this pharmaceutical composition can be used to predict susceptibility using the TLE1 expression level as an indicator, allowing for more appropriate BCV treatment. For example, if a subject has a low TLE1 expression level and is predicted to be highly susceptible, treatment may be performed by administering BCV or administering it at a low dose. Furthermore, for example, if a subject has a high TLE1 expression level and is predicted to have low susceptibility, treatment may be performed by not administering BCV or by administering it at a high dose. Furthermore, as described in the Examples below, patients with NK / T-cell lymphoma with low TLE1 expression who have not been treated with BCV have a particularly poor prognosis. Therefore, this pharmaceutical composition can be used to treat malignant tumors with poor prognosis. Furthermore, the Examples below show that B-cell lymphomas with high TLE1 expression levels are highly susceptible. Therefore, this pharmaceutical composition can be used to administer BCV to subjects who are predicted to have high TLE1 expression and high BCV susceptibility. In such cases, improved therapeutic effects or a higher probability of therapeutic effects can be expected compared to administration to unspecified subjects. Furthermore, the use of the pharmaceutical composition can predict susceptibility using the TLE1 expression level as an indicator, allowing for more appropriate BCV treatment. For example, if a subject has a high TLE1 expression level and is predicted to have high susceptibility, treatment may be performed by administering BCV either alone or at a low dose.Furthermore, for example, if the subject's TLE1 expression level is low and sensitivity is predicted to be low, treatment may be performed by not administering BCV or by administering it at a high dose. Furthermore, as described in the Examples below, patients with BCV-untreated B-cell lymphomas with high TLE1 expression had a particularly poor prognosis. Therefore, this pharmaceutical composition can be used to treat malignant tumors with poor prognosis.

[0051] (31) Composition According to one embodiment of the present invention, a composition for use in predicting susceptibility to BCV, its pharmaceutically acceptable salt, or a solvate thereof in a malignant tumor subject is provided, the composition comprising a reagent for testing TLE1 expression levels. This composition can be used to predict or identify subjects with high or low susceptibility to BCV using the TLE1 expression level as an indicator. Therefore, use of this composition enables more appropriate BCV treatment. For example, if a subject's TLE1 expression level is low and high susceptibility is predicted, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject's TLE1 expression level is high and susceptibility is predicted, treatment may be performed by not administering BCV or administering it at a high dose. For example, if a subject's TLE1 expression level is high and susceptibility is predicted, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject's TLE1 expression level is high and susceptibility is predicted, treatment may be performed by administering BCV or administering it at a low dose. For example, if a subject's TLE1 expression level is low and susceptibility is predicted, treatment may be performed by not administering BCV or administering it at a high dose.

[0052] (32) Composition According to one embodiment of the present invention, a composition for use in determining which malignant tumor subject should receive BCV, its pharmaceutically acceptable salt, or a solvate thereof is provided, the composition comprising a reagent for testing TLE1 expression levels. Use of this composition enables more appropriate BCV treatment. For example, if a subject's TLE1 expression level is low and predicted to be highly sensitive, the subject may be selected to receive BCV or a low dose of BCV. If a subject's TLE1 expression level is high and predicted to be highly sensitive, the subject may be selected to receive BCV or a low dose of BCV. If a subject's TLE1 expression level is low and predicted to be highly sensitive, the subject may be selected to receive BCV or a low dose of BCV. If a subject's TLE1 expression level is low and predicted to be low sensitive, the subject may be selected to receive BCV. The subject to be administered may also be the subject to be administered. From another perspective, one embodiment of the present invention provides a composition for use in selecting a subject with a malignant tumor suitable for treatment with BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the composition comprising a reagent for testing TLE1 expression levels.

[0053] (33) Compositions According to one embodiment of the present invention, a composition for use in diagnosing whether a subject has a malignant tumor highly susceptible to brincidofovir, its pharmaceutically acceptable salt, or a solvate thereof is provided, the composition comprising a reagent for testing TLE1 expression levels. This composition can be used to diagnose whether a subject has a malignant tumor highly susceptible to BCV using the TLE1 expression level as an indicator. Therefore, use of this composition enables more appropriate BCV treatment. For example, if a subject is diagnosed as having a highly susceptible malignant tumor, treatment may be performed by administering BCV or a low dose of BCV. On the other hand, for example, if a subject is diagnosed as not having a highly susceptible malignant tumor, treatment may be performed by not administering BCV or administering a high dose of BCV. This diagnosis may also be a companion diagnostic for treatment with BCV, its pharmaceutically acceptable salt, or a solvate thereof.

[0054] (34) Compositions According to one embodiment of the present invention, there is provided a composition for use in detecting an indicator for identifying a subject with a malignant tumor who is likely to be sensitive to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the composition comprising a reagent for testing TLE1 expression levels. From another aspect, according to one embodiment of the present invention, there is provided a composition for use in detecting an indicator for identifying a subject with a malignant tumor who is likely to respond or react to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, the composition comprising a reagent for testing TLE1 expression levels. From another aspect, according to one embodiment of the present invention, there is provided a composition for use in detecting the expression level of TLE1 in a sample from a subject who has a malignant tumor or is suspected of developing a malignant tumor, the composition comprising a reagent for testing TLE1 expression levels. Use of any of these methods enables more appropriate BCV treatment to be implemented.

[0055] (35) Composition According to one embodiment of the present invention, there is provided a pharmaceutical composition for treating malignant tumors, comprising BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof. This pharmaceutical composition can be used to treat malignant tumors. The pharmaceutical composition may comprise BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof as an active ingredient. The malignant tumor may be lymphoma. The lymphoma may be NK / T-cell lymphoma, T-cell lymphoma, or B-cell lymphoma. The Examples described below demonstrate that the antitumor effect of BCV is particularly strong against T-cell lymphoma. Therefore, T-cell lymphoma is a particularly suitable target for the treatment of BCV.

[0056] (36) Composition According to one embodiment of the present invention, there is provided a composition for use in predicting the prognosis of a malignant tumor, the composition comprising a reagent for testing TLE1 expression levels. This composition can be used to predict the prognosis of a patient. This composition may be used to predict the prognosis based on the expression level of TLE1 in a subject with a malignant tumor. For example, a low TLE1 expression level in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) may be evaluated as a poor prognosis. For example, a high TLE1 expression level in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) may be evaluated as a good prognosis. This prediction may further include a prognosis prediction based on the expression level of MYC in the subject with a malignant tumor. For example, a low TLE1 expression level in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) and a positive MYC level may be evaluated as a poor prognosis. For example, if the expression level of TLE1 in a sample from a subject with a malignant tumor (e.g., NK / T-cell lymphoma or T-cell lymphoma) is high and the sample is MYC-negative, the prognosis may be evaluated as good. The expression level of MYC may be tested using a MYC test reagent. In the case of a malignant tumor such as a B-cell lymphoma, for example, if the expression level of TLE1 in the subject's sample is high, the prognosis may be predicted as poor (poor prognosis), and if the expression level of TLE1 in the subject's sample is low, the prognosis may be predicted as good (good prognosis). This prediction may further be based on the expression level of MYC in the subject with a malignant tumor. For example, if the expression level of TLE1 in a sample from a B-cell lymphoma subject is high and the sample is MYC-positive, the prognosis may be evaluated as poor. For example, if the expression level of TLE1 in a sample from a B-cell lymphoma subject is low and the sample is MYC-negative, the prognosis may be evaluated as good.

[0057] (37) Composition According to one embodiment of the present invention, there is provided a composition for use in the methods described in (1) to (29) above, comprising a reagent for testing the expression level of TLE1, ZNF441, CERS6, or a protein or gene described in Figure 3 or 16.

[0058] (38) Composition According to one embodiment of the present invention, there is provided a composition comprising BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof, for use in the methods described in (1) to (29) above.

[0059] (39) Kit According to one embodiment of the present invention, there is provided a kit for use in the methods described in (1) to (29) above, comprising a reagent for testing the expression level of TLE1, ZNF441, CERS6, or a protein or gene described in Figure 3 or 16. The kit may also include, for example, an instruction manual, a buffer solution, a container, or packaging.

[0060] (40) Use According to one embodiment of the present invention, there is provided use of TLE1, ZNF441, CERS6, or a protein or gene described in Figure 3 or 16 as a biomarker for use in the methods described in (1) to (29) above.

[0061] (41) Use According to one embodiment of the present invention, there is provided a use of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof for producing the composition according to any one of (30) to (38) above.

[0062] In embodiments of the present invention (e.g., (1) to (41) above), brincidofovir (BCV) includes a compound having the structure shown in the following formula: BCV can also be referred to by its IUPAC name, [(2S)-1-(4-amino-2-oxopyrimidin-1-yl)-3-hydroxypropan-2-yl]oxymethyl-(3-hexadecoxypropoxy)phosphinic acid. BCV includes the compound represented by the CAS registry number 444805-28-1. As used herein, BCV is an abbreviation for brincidofovir, and the two terms have the same meaning.

[0063] In embodiments of the present invention (e.g., (1) to (41) above), TLE1 includes a protein designated as Transducin-like enhancer of split 1, TLE family member 1, or transcriptional corepressor. Details of TLE1, such as its amino acid sequence, can be found on websites such as NCBI or UniProt. At the time of filing, the UniProt primary accession number for TLE1 was Q04724, its HGNC ID was HGNC:11837, and its NCBI Gene ID was 7088 (updated on 9-Apr-2023). In the Examples described below, TLE1 corresponding to Q04724 was detected. TLE1 may be derived from, for example, human, monkey, mouse, rat, rabbit, dog, or cat.

[0064] In embodiments of the present invention (e.g., (1) to (41) above), ZNF441 includes a protein designated Zinc finger protein 441. Details of ZNF441, such as its amino acid sequence, can be found on websites such as NCBI or UniProt. At the time of filing, the UniProt primary accession number for ZNF441 was Q8N8Z8, its HGNC ID was HGNC:20875, and its NCBI Gene ID was 126068. In the Examples described below, ZNF441 corresponding to Q8N8Z8 was detected. ZNF441 may be derived from, for example, humans, monkeys, mice, rats, rabbits, dogs, or cats. In embodiments of the present invention (e.g., (1) to (41) above), CERS6 includes a protein designated Ceramide synthase 6, LASS6. Details of CERS6, such as its amino acid sequence, can be found on websites such as NCBI or UniProt. At the time of filing the present application, the UniProt Primary accession number for CERS6 is Q6ZMG9, the HGNC ID is HGNC:23826, and the NCBI Gene ID is 253782. In the Examples described below, a CERS6 corresponding to Q6ZMG9 was detected. CERS6 may be CERS6 derived from, for example, a human, monkey, mouse, rat, rabbit, dog, or cat.

[0065] In embodiments of the present invention (e.g., (1) to (41) above), malignant tumors include, for example, tumors that develop as a result of mutation of normal cells. Malignant tumors can arise in any organ or tissue throughout the body. Examples of such malignant tumors include one or more selected from the group consisting of hematological malignancies (e.g., lymphoma, leukemia, or multiple myeloma), lung cancer, esophageal cancer, gastric cancer, liver cancer, pancreatic cancer, kidney cancer, adrenal cancer, biliary tract cancer, breast cancer, colon cancer, small intestine cancer, ovarian cancer, uterine cancer, bladder cancer, prostate cancer, ureteral cancer, renal pelvis cancer, ureteral cancer, penile cancer, testicular cancer, brain tumors (e.g., glioma), central nervous system cancer, peripheral nervous system cancer, head and neck cancer, skin cancer (e.g., melanoma), thyroid cancer, salivary gland cancer, carcinoma, and sarcoma. The hematological malignant tumor may be a blood cancer. The above-mentioned leukemias include, for example, myeloid leukemia (e.g., acute myeloid leukemia or chronic myeloid leukemia) and lymphocytic leukemia (e.g., acute lymphocytic leukemia or chronic lymphocytic leukemia). The therapeutic effect on malignant tumors may be evaluated, for example, by observing changes in malignant tumor cell proliferation over time after drug administration. A state in which cell proliferation is inhibited includes a state in which the proliferation rate of the test cells is significantly reduced compared to before drug treatment. The proliferation rate may be measured, for example, using absorbance as an indicator, or may be determined from image data. The therapeutic effect on malignant tumors may be evaluated, for example, by observing a decrease in tumor size or tumor mass after drug administration. In this case, a significant decrease in tumor size or tumor mass compared to before drug administration or after administration of a negative control may be considered to be therapeutically effective. The therapeutic effect on malignant tumors may be measured, for example, using the amount of a malignant tumor marker in a patient or a patient-derived sample as an indicator. In this case, a significant decrease in the amount of the marker compared to before drug administration or after administration of a negative control may be considered to be therapeutically effective. The tumor mass or marker amount after drug administration may be reduced to 0.9, 0.7, 0.5, 0.3, or 0.1 times or less compared to before administration or after administration of a negative control. The malignant tumor may be a malignant tumor with low TLE1 expression. A malignant tumor with low TLE1 expression may include a TLE1-negative malignant tumor. The malignant tumor may be a malignant tumor with high TLE1 expression. A malignant tumor with high TLE1 expression may include a TLE1-positive malignant tumor.The malignant tumor may be a malignant tumor with high expression of a protein or gene other than ZNF441, CERS6, FAM89A and AK4 in Figure 3, or a protein or gene listed in Figure 16. This high-expression malignant tumor may include a positive malignant tumor.

[0066] In the embodiments of the present invention (e.g., (1) to (41) above), when the malignant tumor is a blood cancer, the blood cancer may include, for example, lymphoma, leukemia, or multiple myeloma. Leukemia may include, for example, acute myeloid leukemia, acute lymphocytic leukemia, chronic myeloid leukemia, or chronic lymphocytic leukemia. When the malignant tumor is lymphoma, the lymphoma includes malignant lymphoma. Malignant lymphoma includes, for example, diseases caused by canceration of lymphocytes. Malignant lymphoma includes, for example, non-Hodgkin's lymphoma or Hodgkin's lymphoma. Non-Hodgkin's lymphoma includes, for example, B-cell lymphoma, NK / T-cell lymphoma, or T-cell lymphoma. Examples of B-cell lymphomas include B-cell lymphomas classified into one or more of the following: follicular lymphoma, MALT lymphoma, lymphoplasmacytic lymphoma, mantle cell lymphoma, diffuse large B-cell lymphoma (DLBCL), Burkitt's lymphoma, primary effusion lymphoma, or chronic lymphocytic leukemia / small lymphocytic lymphoma. Examples of NK / T-cell lymphomas include NK / T-cell lymphomas classified into one or more of the following: peripheral T-cell lymphoma, angioimmunoblastic T-cell lymphoma, anaplastic large cell lymphoma, adult T-cell leukemia / lymphoma, extranodal NK / T-cell lymphoma, nasal type, and cutaneous lymphoma (e.g., mycosis fungoides). T-cell lymphomas include T-cell lymphomas classified as one or more of peripheral T-cell lymphoma (PTCL), cutaneous T-cell lymphoma, anaplastic large cell lymphoma, enteropathy-associated T-cell lymphoma, or lymphoblastic lymphoma. Non-Hodgkin's lymphomas include, for example, peripheral T-cell lymphomas. Peripheral T-cell lymphomas include, for example, peripheral T-cell lymphomas classified as one or more of nodal peripheral T-cell lymphoma or extranodal peripheral T-cell lymphoma. Nodal peripheral T-cell lymphomas include, for example, nodal peripheral T-cell lymphomas classified as one or more of PTCL-NOS, PTCL with a follicular helper T-cell phenotype, or ALK-positive anaplastic large cell lymphoma. Extranodal peripheral T-cell lymphoma includes, for example, extranodal peripheral T-cell lymphoma classified as extranodal NK / T-cell lymphoma, nasal type. From the viewpoint of improving the therapeutic effect or increasing the probability of achieving the therapeutic effect, the T-cell lymphoma is preferably peripheral T-cell lymphoma.Peripheral T-cell lymphoma is generally a difficult disease to treat, but in the examples described below, BCV was also shown to be effective in treating peripheral T-cell lymphoma cell lines. Hodgkin's lymphoma includes, for example, classical Hodgkin's lymphoma and nodular lymphocyte-predominant Hodgkin's lymphoma. For details on lymphoma, see, for example, Nirmal, J Oral Maxillofac Pathol. 2020 May-Aug;24(2):195-199; Singh et al., J Family Med Prim Care. 2020 Apr;9(4):1834-1840; Voltin et al., Cancers (Basel). 2020 Mar 5;12(3):601; or Pu et al., Front Immunol. 2022;13:1008695. From the viewpoint of improving the therapeutic effect or increasing the probability of achieving a therapeutic effect, the lymphoma is preferably NK / T-cell lymphoma, T-cell lymphoma, or B-cell lymphoma. The therapeutic effect or onset of lymphoma may be diagnosed by examining lymph node swelling using CT, PET, or MRI, by histological examination of lymph nodes or tumor masses, or by testing for tumor markers for malignant lymphoma (e.g., sIL2-R) in the blood. Lymphoma may be diagnosed, for example, by the methods described in Nirmal (supra), Voltin et al. (supra), or Pu et al. (supra).

[0067] In embodiments of the present invention (e.g., (1) to (41) above), malignant tumors with low TLE1 expression and high BCV susceptibility include NK / T cell lymphomas or T cell lymphomas with low TLE1 expression. Furthermore, malignant tumors with high TLE1 expression and high BCV susceptibility include B cell lymphomas with high TLE1 expression. Furthermore, malignant tumors with high ZNF441 expression and high BCV susceptibility include NK / T cell lymphomas with high ZNF441 expression. Furthermore, malignant tumors with high CERS6 expression and high BCV susceptibility include B cell lymphomas with high CERS6 expression.

[0068] In embodiments of the present invention (e.g., (1) to (41) above), the expression level of a protein (e.g., TLE1) may be tested by methods such as PCR (polymerase chain reaction), antigen-antibody reaction, immunohistochemical staining, or DNA methylation analysis. PCR includes, for example, qPCR. Antigen-antibody reaction includes, for example, Western blotting. DNA methylation analysis includes, for example, analysis of DNA methylation upstream (e.g., promoter) of a gene (e.g., TLE1 gene). Known methods can be used to test the expression level of TLE1, and reference may be made to the method described in Xu et al., Mol Cell Biochem. 2018 Jan;438(1-2):85-96. When the expression level of a protein (e.g., TLE1) is low in a subject sample (e.g., malignant tumor cells or their cell lysate), the level of the low expression may be significantly reduced compared to a sample from a healthy individual (e.g., normal cells or their cell lysate). This degree of underexpression may be, for example, 10, 5, 3, 1, 0.1, or 0.01% or less, or 0%, or may be within a range between any two of these values. This underexpression may be, for example, 10 to 0, 5 to 0, 1 to 0, or 0.1 to 0%. This degree of underexpression may be less than or equal to the mean value minus 1 standard deviation of a group of samples from healthy individuals. This value may be, for example, -1, -1.5, -2, -2.5, or -3 SD or less, or may be within a range between any two of these values. When the expression level of a protein (e.g., TLE1) in a subject's sample is low, the degree of underexpression may be less than or equal to -1 SD of the mean value of a group of samples from malignant tumors of the same type as the malignant tumor present in the subject but obtained from patients other than the subject. This value may be, for example, -1, -1.5, -2, -2.5, or -3 SD or less, or may be within a range between any two of these values. The group of malignant tumor samples obtained from patients other than the subject may be, for example, a group of samples previously confirmed to have a high expression level of a protein (e.g., TLE1) or a group of samples previously confirmed to have low BCV susceptibility. For example, if the malignant tumor of the subject is lymphoma, the malignant tumor of the same type as the malignant tumor of the subject is lymphoma.When the expression level of a protein (e.g., TLE1) in a subject sample is low, the degree of underexpression may be comparable to that of a sample previously confirmed to have low expression levels of the protein (e.g., TLE1) or a sample previously confirmed to have high BCV susceptibility. Examples of such samples include KAI-3, NK-S1, NK-92, KHYG-1, FE-PD, Z4739, SUP-M2, SUDHL-1, DEL, L-82, SMZ-1, SR-786, K-299, HH, or KI-JK cell lines. This comparable degree may refer to a difference in expression level of, for example, ±30, 20, 10, 5, 3, 2, 1, or 0.1% or less, or 0%, or within a range between any two of these values. This value may be, for example, ±10-0, 5-0, 1-0, or 0.1-0%. When the expression level of a protein (e.g., TLE1) in a subject sample is low, the degree of underexpression may be significantly lower than that of a sample with a high expression level of that protein, a sample previously confirmed to have a high expression level of that protein, a sample with a non-low expression level of that protein, or a sample previously confirmed to have low BCV susceptibility. Such samples may be, for example, SNK-1, SNK-6, YT, HANK-1, or SNT-8 cell lines. This significantly lower level may be, for example, 10%, 5%, 3%, 1%, 0.1%, 0.01%, or less, or 0%, or may be within a range of any two of these values. This value may be, for example, 10-0, 5-0, 1-0, or 0.1-0%. The expression level may be evaluated, for example, by measuring the amount of mRNA or protein. Considering the impact on protein function, it is preferable to evaluate the expression level by measuring the amount of protein. Evaluating the expression level may include, for example, comparing the expression level of the subject with a predetermined threshold, or evaluating the expression level as low when the expression level of the subject is lower than the threshold. Evaluating the expression level may include a step of evaluating the expression level as low when the expression amount in the subject is significantly reduced compared to the control value.In embodiments of the present invention (e.g., (1) to (41) above or this paragraph), low TLE1 expression may be TLE1 negative, and high TLE1 expression may be TLE1 positive or TLE1 overexpression. Thus, the above embodiments regarding low or high TLE1 expression are applicable to TLE1 negative or positive embodiments, respectively. Low TLE1 expression may be detected, for example, in a sample from a subject with NK / T-cell lymphoma or T-cell lymphoma. Examination of protein expression levels may be performed, for example, in vitro, ex vivo, or in vivo.

[0069] In embodiments of the present invention (e.g., (1) to (41) above), when the expression level of a protein (e.g., TLE1, ZNF441, or CERS6) is high in a subject sample (e.g., malignant tumor cells or their cell lysates), the level may be significantly increased compared to a sample from a healthy subject (e.g., normal cells or their cell lysates). This level of high expression may be, for example, 1.3, 1.5, 2, 3, 4, 5, 10, 20, 30, 40, 50, 100, or 500 times higher, or within a range of any two of these values. This level of high expression may be, for example, 1.3 to 500, 2 to 500, 5 to 500, or 50 to 500 times higher. This level of high expression may be equal to or greater than the mean value of a group of healthy subject samples plus one standard deviation. This value may be, for example, +1, +1.5, +2, +2.5, or +3 SD or higher, or within a range of any two of these values. When the expression level of a protein (e.g., TLE1, ZNF441, or CERS6) is high in a subject's sample, the degree of high expression may be at least 1 SD above the mean value of a group of samples of malignant tumors of the same type as the malignant tumor in the subject but obtained from patients other than the subject. This value may be, for example, +1, +1.5, +2, +2.5, or +3 SD or more, or may be within a range of any two of these values. The group of malignant tumor samples obtained from patients other than the subject may be, for example, a group of samples previously confirmed to have low expression levels of a protein (e.g., TLE1, ZNF441, or CERS6) or a group of samples previously confirmed to have low BCV susceptibility. For example, if the subject's malignant tumor is lymphoma, the malignant tumor of the same type as the malignant tumor in the subject is lymphoma. When the expression level of a protein (e.g., TLE1, ZNF441, or CERS6) is high in a subject sample, the degree of high expression may be comparable to that of a sample previously confirmed to have a high expression level of the protein (e.g., TLE1, ZNF441, or CERS6) or a sample previously confirmed to have high BCV susceptibility, such as a cell line such as SUDHL-5, DB, BJAB, OCI-LY19, L-428, OCI-LY18, OCI-LY3, WSU-NHL, or RL.This degree of similarity may be, for example, a difference in expression level of 30, 20, 10, 5, 3, 2, 1, or 0.1% or less, or 0%, or may be within a range of any two of these values. This value may be, for example, 10-0, 5-0, 1-0, or 0.1-0%. When the expression level of a protein (e.g., TLE1, ZNF441, or CERS6) in a subject sample is high, the degree of high expression may be significantly higher than a sample with low expression level of that protein, a sample previously confirmed to have low expression level of that protein, a sample without high expression level of that protein, or a sample previously confirmed to have low BCV susceptibility. Such samples may be, for example, SC-1 or K231 cell lines. This significantly higher degree may be, for example, 1.3-, 1.5-, 2-, 3-, 4-, 5-, 10-, 20-, 30-, 40-, 50-, 100-, or 500-fold or more, or may be within a range of any two of these values. This value may be, for example, 1.3 to 500, 2 to 500, 5 to 500, or 50 to 500 times. The expression level may be evaluated, for example, by measuring the amount of mRNA or protein. Considering the impact on protein function, it is preferable to evaluate the expression level by measuring the amount of protein. Evaluating the expression level may include, for example, comparing the expression level of a target with a predetermined threshold, or evaluating the expression level as high when the expression level of the target is higher than the threshold. Evaluating the expression level may include evaluating the expression level as high when the expression level of a target is significantly increased compared to the control value. In embodiments of the present invention (e.g., (1) to (41) above or this paragraph), high expression of TLE1, ZNF441, or CERS6 may refer to positive or overexpression of TLE1, ZNF441, or CERS6. Accordingly, the above-described embodiments regarding high or low expression of TLE1, ZNF441, or CERS6 are applicable to positive or negative embodiments of TLE1, ZNF441, or CERS6, respectively. High expression of TLE1 or CERS6 may be detected in a sample from a subject with, for example, B-cell lymphoma. High expression of ZNF441 may be detected in a sample from a subject with, for example, NK / T-cell lymphoma.Examination of protein expression levels may be carried out, for example, in vitro, ex vivo or in vivo.

[0070] In embodiments of the present invention (e.g., (1) to (41) above), the malignant tumor may include a MYC-positive, MYC-negative, EBV-positive, or EBV-negative malignant tumor. As described above, the MYC-positive or -negative nature may be used to predict prognosis. Positive includes positive gene expression. Positive may also include increased gene expression in a subject's sample compared to gene expression in a sample from a healthy individual or an individual previously confirmed to be negative. Detection of MYC or EBV positivity / negativity (or expression level) in a subject or a subject's malignant tumor may be performed, for example, by collecting a subject's sample (e.g., malignant tumor cells, plasma, whole blood, or tissue) and measuring gene expression of MYC (or a gene whose expression is upregulated in association with MYC) or EBV in the sample. Gene expression can be measured, for example, by qPCR, DNA chip, or immunostaining. In this case, a sample collected from a subject may be compared with a sample from a healthy individual (e.g., normal cells) or a sample previously confirmed to be negative, and if MYC gene expression or expression of a gene whose expression is upregulated in association with MYC is significantly increased in the sample collected from the subject, the sample may be determined to be MYC-positive. Alternatively, a sample collected from a subject may be compared with a sample from a healthy individual (e.g., normal cells) or a sample previously confirmed to be negative, and if EBV gene expression is significantly increased in the sample collected from the subject, the sample may be determined to be EBV-positive. EBV gene expression may be measured using the expression of EBV-derived transcription products (e.g., EBER or EBNA1). Detection of MYC or EBV positivity / negative in a subject or a subject's lymphoma may be performed, for example, by collecting the subject's lymph nodes and immunostaining them for MYC or EBV. In this case, a sample collected from the subject may be compared with a sample collected from a healthy individual or an individual previously confirmed to be negative, and if the staining intensity of MYC or EBV in the sample collected from the subject is significantly increased, the sample may be determined to be positive for MYC or EBV. The increase in gene expression or staining intensity mentioned above includes, for example, an increase of 1.5, 2, 3, 4, 5, 10, 20, or 50 times or more compared to the comparison subject, or an increase within a range of any two of these values.This multiple may be, for example, 1.5 to 50, 1.5 to 10, 1.5 to 5, 2 to 50, 3 to 50, or 5 to 50. Furthermore, in detecting MYC or EBV positivity / negativity in a subject or a subject's lymphoma, for example, a certain percentage of cells in a sample immunostained for MYC or EBV may be determined to be MYC or EBV positive. This percentage may be, for example, 10, 20, 30, 40, 50, 60, 70, 80, or 90% or more, or 100%, or may be within a range of any two of these values. This percentage may be, for example, 10 to 100, 10 to 70, 10 to 50, 20 to 100, 40 to 100, or 60 to 100%. Detection of MYC positivity / negativity in a subject or a subject's lymphoma can be performed, for example, by detecting MYC translocations using fluorescence in situ hybridization (FISH) (see, for example, Salam et al., J Cancer 2020; 11(1):190-198 or Epperla et al., Cancer. 2017 Nov 15;123(22):4411-4418). Detection of EBV positivity / negativity in a subject or a subject's lymphoma can be performed, for example, by quantifying EBV DNA in plasma or whole blood using a commercially available DNA quantification kit (e.g., Accugene m-EBV (Abbott Japan G.K.)), or by detecting IgM antibodies against EBV capsid antigens in serum using a commercially available antibody detection kit (e.g., BioPlex EBV IgM Kit (Bio-Rad Laboratories, Inc.)). MYC positive / negative detection may be performed, for example, in vitro, ex vivo or in vivo.

[0071] In embodiments of the present invention (e.g., (1) to (41) above), MYC includes a protein known as a transcriptional regulator. For example, the primary accession number for MYC in UniProt at the time of filing this application is P01106. EBV is an abbreviation for Epstein-Barr virus. EBV is known as a virus with double-stranded DNA (see, for example, Rivailler et al., J. Virol. 2002 Dec; 76(23): 12055-12068 or Correia et al., J. Virol. 2018 Nov 15; 92(22): e01132-18). Targeting MYC-positive or EBV-positive lymphomas can provide a high therapeutic effect with BCV, its pharmaceutically acceptable salt, or a solvate thereof.

[0072] In the methods of the embodiments of the present invention (e.g., (1) to (29) above), the step of identifying a subject may include a step of identifying a subject as a treatment subject. The step of identifying a subject as a treatment subject may include a step of identifying a subject as a drug administration subject. The step of identifying a subject may include a step of selecting a subject or a step of identifying a subject.

[0073] In the methods of the embodiments of the present invention (e.g., the above (1) to (29)), the step of identifying a subject with a malignant tumor may be performed using a malignant tumor biomarker in the subject's body fluid (e.g., blood) as an indicator, or by diagnostic imaging (e.g., CT) or tissue findings. This identification step may also be performed using an increase in the amount of the biomarker in the subject compared to the amount of the malignant tumor biomarker in a control subject (e.g., a healthy individual) as an indicator.

[0074] In the embodiments of the present invention (e.g., (1) to (29) above), the method may include a method of assisting. Assistance may include assistance in the investigation or determination of a treatment target, treatment method, or diagnostic method. The method may be performed, for example, in vitro, ex vivo, or in vivo, depending on the purpose.

[0075] In embodiments of the present invention (e.g., (1) to (41) above), in predicting BCV susceptibility, a subject may be predicted to be susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is low. In predicting BCV susceptibility, a subject may be predicted to be not susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is high. In this case, the subject's disease may be a malignant tumor with low TLE1 expression and high BCV susceptibility. In embodiments of the present invention (e.g., (1) to (41) above), in predicting BCV susceptibility, a subject may be predicted to be susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is high. In predicting BCV susceptibility, a subject may be predicted to be not susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is low. In this case, the subject's disease may be a malignant tumor with high TLE1 expression and high BCV susceptibility.

[0076] In embodiments of the present invention (e.g., (1) to (41) above), when determining a subject with a malignant tumor to be administered BCV, the subject may be determined to be a subject to be administered BCV if the TLE1 expression level in the subject or a subject's sample is low. When determining a subject with a malignant tumor to be administered BCV, the subject may be determined to be a subject to be administered a high dose of BCV if the TLE1 expression level in the subject or a subject's sample is high. In this case, the malignant tumor may be a malignant tumor with low TLE1 expression and high BCV sensitivity. In embodiments of the present invention (e.g., (1) to (41) above), when determining a subject with a malignant tumor to be administered BCV, the subject may be determined to be a subject to be administered BCV if the TLE1 expression level in the subject or a subject's sample is high. When determining a subject with a malignant tumor to be administered BCV, the subject may be determined to be a subject to be administered a high dose of BCV if the TLE1 expression level in the subject or a subject's sample is low. In this case, the malignant tumor may be a malignant tumor with high TLE1 expression and high BCV sensitivity.

[0077] In embodiments of the present invention (e.g., (1) to (41) above), in diagnosing whether a subject has a malignant tumor highly susceptible to BCV, a subject may be diagnosed as having a malignant tumor highly susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is low. In diagnosing whether a subject has a malignant tumor highly susceptible to BCV, a subject may be diagnosed as not having a malignant tumor highly susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is high. In this case, the malignant tumor may be a malignant tumor with low TLE1 expression and high BCV susceptibility. In embodiments of the present invention (e.g., (1) to (41) above), in diagnosing whether a subject has a malignant tumor highly susceptible to BCV, a subject may be diagnosed as having a malignant tumor highly susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is high. In diagnosing whether a subject has a malignant tumor highly susceptible to BCV, a subject may be diagnosed as not having a malignant tumor highly susceptible to BCV if the TLE1 expression level in the subject or a subject's sample is low. In this case, the malignant tumor may be a malignant tumor that highly expresses TLE1 and is highly susceptible to BCV.

[0078] In embodiments of the present invention (e.g., (1) to (41) above), the test includes, for example, detection or measurement. In embodiments of the present invention (e.g., (1) to (41) above), the test reagent may be, for example, a nucleic acid (e.g., a DNA strand or an RNA strand), a peptide (e.g., an oligopeptide or a polypeptide (e.g., a protein)), or a composition containing them. The nucleic acid may be, for example, a primer or a probe. The polypeptide may be, for example, an antibody. The nucleic acid may be, for example, a nucleic acid capable of binding to an upstream or downstream region of DNA (e.g., the TLE1 gene) encoding a protein (e.g., TLE1), or a nucleic acid capable of amplifying the entire length or a portion of DNA encoding a protein (e.g., TLE1). The peptide or antibody may be, for example, a peptide or antibody capable of binding to TLE1. An oligopeptide may refer to a peptide having 1 to 20 amino acids. A polypeptide may refer to a peptide having 21 or more amino acids. The antibody may be, for example, a human, mouse, rabbit, rat, hamster, or guinea pig antibody. The antibody may be a chimeric antibody of any two or more of the above organisms. The antibody may be a monoclonal antibody. The testing reagent may be substituted for the detection means. In embodiments of the present invention (e.g., (1) to (41) above), the testing step may be performed by, for example, PCR, antigen-antibody reaction, immunohistochemical staining, or DNA methylation analysis of the promoter region of a gene (e.g., the TLE1 gene). The testing may be performed using, for example, the amount of nucleic acid (e.g., gene or mRNA), the amount of protein, DNA methylation, or gene translocation as an indicator. DNA methylation analysis may be performed using a means for detecting methylation or demethylation of the promoter region of a gene (e.g., reagents used in methylation-specific PCR, demethylation-specific PCR, or pyrosequencing). DNA methylation analysis may be performed using a primer (e.g., a primer capable of amplifying at least a portion of the promoter region (e.g., a primer capable of binding to an upstream or downstream site of the promoter region)) or an antibody. The primer may be a methylation- or demethylation-specific primer. The antibody may be an antibody specific to the methylated or unmethylated promoter region of a gene or a partial region thereof.The above-mentioned test reagents may include those used in these detection steps. In embodiments of the present invention (e.g., (1) to (41) above), the step of examining the expression level may be replaced with, for example, a step of examining expression or a step of examining the presence of a positive / negative result.

[0079] In embodiments of the present invention (e.g., (1) to (41) above), when the expression level of TLE1 is used as an index, the index may be low expression of TLE1. For example, when identifying a patient with a malignant tumor who is a target for BCV treatment, low expression of TLE1 may be used as an index. The expression level may be examined qualitatively or quantitatively. In this case, the malignant tumor may be a malignant tumor with low TLE1 expression and high BCV susceptibility. In embodiments of the present invention (e.g., (1) to (41) above), when the expression level of TLE1 is used as an index, high expression of TLE1 may be used as an index. For example, when identifying a patient with a malignant tumor who is a target for BCV treatment, high expression of TLE1 may be used as an index. The expression level may be examined qualitatively or quantitatively. In this case, the malignant tumor may be a malignant tumor with high TLE1 expression and high BCV susceptibility.

[0080] In embodiments of the present invention (e.g., (1) to (41) above), the sample may include, for example, a biological sample. The sample may include, for example, cells (e.g., malignant tumor cells), cell lysates, blood specimens (e.g., plasma, serum, or whole blood), and tissues. The sample may include, for example, a specimen. Alternatively, the sample may include malignant tumor cells from a malignant tumor subject.

[0081] In embodiments of the present invention (e.g., (1) to (41) above), a more appropriate BCV treatment may be a treatment that provides a greater therapeutic effect than a BCV treatment that does not use the TLE1 expression level as an indicator or that does not take BCV susceptibility into consideration. A more appropriate BCV treatment may be a treatment administered to a population with a higher therapeutic effect, a treatment administered to subjects more likely to demonstrate therapeutic benefit, or a treatment that implements a more appropriate treatment regimen. Demonstration of a therapeutic effect may include a significant reduction in tumor size compared to before administration of an active ingredient or after administration of a non-active ingredient (e.g., a placebo).

[0082] In embodiments of the present invention (e.g., (1) to (41) above), treatment includes exerting an effect that brings about a beneficial result in a patient with respect to a patient's disease (e.g., a malignant tumor (e.g., lymphoma)) or one or more symptoms associated with the disease (e.g., improvement, alleviation, palliation, cure, remission, or suppression (e.g., suppression of onset (e.g., prevention), suppression of progression, or suppression of recurrence). Treatment may be carried out by administering a therapeutically effective amount of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof to a subject. In embodiments of the present invention (e.g., (1) to (41) above), the pharmaceutical composition may be produced by, for example, mixing an active ingredient with one or more pharmaceutically acceptable carriers using any method known in the technical field of pharmaceuticals. The pharmaceutical composition may be in any form used as long as it is used for treatment, and may contain the active ingredient alone, or may be a mixture of the active ingredient and any ingredient. The form of the carrier is not particularly limited, and may be, for example, a solid or liquid (e.g., a buffer solution). The content of the carrier may be, for example, a pharmaceutically effective amount. The effective amount may be, for example, a pharmaceutically effective amount of the active ingredient. The amount may be sufficient for effective stability or delivery. For example, a buffer is effective for stabilizing the active ingredient in the vial. The pharmaceutical composition may also contain a stabilizer (e.g., mannitol), a buffering agent (e.g., arginine), or a pH adjuster (e.g., NaOH). The dosage, administration interval, administration method, and administration route are not particularly limited and can be selected appropriately depending on the patient's age, weight, symptoms, target organ, etc. Furthermore, the pharmaceutical composition preferably contains a therapeutically effective amount, or an effective amount of the active ingredient that exerts the desired effect. In one embodiment of the present invention, a therapeutically effective amount includes the amount necessary to clinically observe improvement in symptoms in the patient. In one embodiment of the present invention, pharmaceutically acceptable includes a state that is suitable for use within the scope of sound medical judgment and commensurate with a reasonable benefit / risk ratio. The components other than BCV in the pharmaceutical composition are not particularly limited as long as they do not impair the effects of the present invention and can be selected appropriately depending on the purpose. The therapeutic effect of malignant tumors may be determined to be successful if the patient's malignant tumor cells are significantly reduced after administration of the therapeutic agent.

[0083] In embodiments of the present invention (e.g., (1) to (41) above), the subject (including a patient) may be, for example, a subject diagnosed with a malignant tumor, a subject suspected of developing a malignant tumor, or a subject in need of treatment for a malignant tumor. The subject includes a population of subjects. The subject may be, for example, a subject predicted to be highly sensitive to BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof. The subject may be, for example, a subject with a malignant tumor with a low expression level of TLE1 (a malignant tumor subject with low TLE1 expression). The subject may be, for example, a subject with a malignant tumor with a high expression level of TLE1 (a malignant tumor subject with high TLE1 expression). The malignant tumor subject includes a subject with a malignant tumor. The subject may be, for example, a subject in which a malignant tumor biomarker is increased compared to a comparison subject (e.g., a healthy subject), or a subject in which a malignant tumor is confirmed by imaging diagnosis or histological findings. The subject may be, for example, a subject who has previously experienced a malignant tumor. The subject may be, for example, a subject diagnosed with a malignant tumor with low TLE1 expression. The subject may be, for example, a subject diagnosed with a malignant tumor with high TLE1 expression. The subject includes a human or a non-human mammal (e.g., one or more of a mouse, guinea pig, hamster, rat, mouse, rabbit, pig, sheep, goat, cow, horse, cat, dog, marmoset, monkey, or chimpanzee). The subject includes an adult human patient.

[0084] In embodiments of the present invention (e.g., (1) to (41) above), the route of administration of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof (or a pharmaceutical composition containing them) to a subject (e.g., a human subject) is preferably one that is effective for the treatment, and may be, for example, intravenous, oral, subcutaneous, intramuscular, or intraperitoneal. The dosage form is preferably one that is effective for the treatment, and may be, for example, a solid formulation (e.g., a tablet), a liquid formulation (e.g., an oral solution), or an injection (e.g., an intravenous injection).

[0085] In embodiments of the present invention (e.g., (1) to (41) above), the dose, administration interval, and administration method of BCV, its pharmaceutically acceptable salt, or solvate thereof (or a pharmaceutical composition containing them) to a subject (e.g., a human subject) can be appropriately selected depending on the patient's age, body weight, symptoms, target organ, etc. The dose may be, for example, 0.01 to 200 mg / kg body weight per administration. The administration interval may be, for example, once or twice every 1 to 28 days or every 1 to 4 weeks. More specifically, this includes intravenous injection of 20 to 80 mg / day. It may also include twice-weekly intravenous injection of 20 to 80 mg / body / day. Furthermore, during combination therapy with BCV and a chemotherapeutic agent (e.g., gemcitabine, etoposide, or an immune checkpoint inhibitor), the BCV dose may include intravenous injection of 10 to 40 mg or 10 to 20 mg / body / day. It may also include twice-weekly intravenous injection of 10 to 40 mg or 10 to 20 mg / body / day. The range of 10 to 80 mg / body / day set forth herein may be, for example, 10, 20, 30, 40, 50, 60, 70, or 80 mg / body / day, or may be within any two ranges thereof. In embodiments of the present invention (e.g., (1) to (41) above), a low dose of BCV, a pharmaceutically acceptable salt thereof, or a solvate thereof may include, for example, 5 to 80, 10 to 40, or 10 to 60 mg / body / day. A high dose may include, for example, 90 to 200, 100 to 180, or 120 to 160 mg / body / day.

[0086] In embodiments of the present invention (e.g., (1) to (41) above), the salt is not particularly limited and includes, for example, inorganic salts or organic salts (see, for example, Bharate et al., Drug Discov Today. 2021 Feb;26(2):384-398 or Berge et al., J Pharm Sci. 1977 Jan;66(1):1-19). Salts include, for example, metal salts, ammonium salts, salts with organic bases, salts with inorganic acids, salts with organic acids, salts with basic or acidic amino acids, and the like. Metal salts include, for example, alkali metal salts (sodium salts, potassium salts, etc.), alkaline earth metal salts (calcium salts, magnesium salts, barium salts, etc.), aluminum salts, and the like. Salts with organic bases include, for example, salts with trimethylamine, triethylamine, pyridine, picoline, 2,6-lutidine, ethanolamine, diethanolamine, triethanolamine, cyclohexylamine, dicyclohexylamine, N,N'-dibenzylethylenediamine, and the like. Salts with inorganic acids include, for example, salts with hydrochloric acid, hydrobromic acid, nitric acid, sulfuric acid, phosphoric acid, etc. Salts with organic acids include, for example, salts with formic acid, acetic acid, trifluoroacetic acid, phthalic acid, fumaric acid, mesylic acid, tosylic acid, oxalic acid, tartaric acid, maleic acid, citric acid, succinic acid, malic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc. Salts with basic amino acids include, for example, salts with arginine, lysine, ornithine, etc. Salts with acidic amino acids include, for example, salts with aspartic acid, glutamic acid, etc. Salts include pharmaceutically acceptable salts. In one embodiment of the present invention, pharmaceutically acceptable includes forms having reasonable benefits for pharmaceutical use. In one embodiment of the present invention, a form of the compound or a salt thereof includes a solvate form thereof. In embodiments of the present invention (e.g., the above (1) to (41)), solvates include forms of compounds formed by a solute and a solvent (see, for example, Healy et al., Adv Drug Deliv Rev. 2017 Aug 1;117:25-46).Solvates include, but are not limited to, hydrates (e.g., monohydrates, dihydrates, trihydrates, etc.) or organic solvent solvates (e.g., solvates with alcohols (e.g., methanol, ethanol, propanol, etc.), acetone, dimethylformamide, or ethyl acetate, etc.). Solvents include those that can substantially maintain the physiological activity of the solute after forming the solvate. Solvates include pharmaceutically acceptable solvates.

[0087] In one embodiment of the present invention, "significantly" may mean, for example, that unless otherwise specified, a statistically significant difference is evaluated using a Student's t-test (one-tailed or two-tailed), where p<0.05 or p<0.01, or that a substantial difference is present.

[0088] All publications cited herein are incorporated by reference in their entirety. As used herein, "or" is used when "at least one or more" of the items listed in the text can be employed. The same applies to "alternative." As used herein, a phrase in the form "at least one or more of A, B, and C" means that any of (A), (B), (C), (A and B), (A and C), (B and C), or (A, B, and C) can be employed. When "within a range of two values" is stated herein, the range also includes the two values ​​themselves. As used herein, "A to B" includes A and B as well as values ​​between A and B. As used herein, "a," "an," and "the" may indicate that one or more elements or steps may be present, unless the context clearly dictates otherwise. As used herein, "having" in relation to a disease includes being affected. As used herein, "the above (1) to (41)" includes reference to any one or more of (1), (2), (3), (4), (5), (6), (7), (8), (9), (10), (11), (12), (13), (14), (15), (16), (17), (18), (19), (20), (22), (22), (23), (24), (25), (26), (27), (28), (29), (30), (31), (32), (33), (34), (35), (36), (37), (38), (39), (40), or (41).

[0089] Although the embodiments of the present invention have been described above, these are merely examples of forms that may be included in the present invention, and the present invention is not limited to these, and various configurations other than those described above may also be adopted. Furthermore, the present invention may be adopted by combining or independently adopting each of the configurations or features described in the above embodiments.

[0090] The present invention will be further explained below with reference to examples, but is not limited to these.

[0091] Example 1: Effect of BCV on the viability of malignant tumor cells The effect of BCV on cell viability was investigated for 11 different NK / T cell lymphoma cell lines. Cell viability was quantified using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol. Cells were cultured at 2 × 10 in 100 μL of medium. 3 Cells were seeded into 96-well plates at a concentration of 0.01 mg / mL and drug (BCV) was added at various concentrations. After each time point, Promega CellTiter-Glo® 2.0 Cell Viability Assay reagent was added to the wells. After 10 minutes of incubation at room temperature, absorbance at 480 nm was measured using a Tecan M200 Infinite 96-well plate reader and iConrol software 1.6 (Tecan, Männedorf, Switzerland). Cell viability was calculated as a percentage of control absorbance. Growth inhibitory effects were analyzed by generating dose-response curves as a plot of the percentage of viable cells against drug concentration, and their IC50 values ​​were estimated using GraphPad Prism version 8.0.2 (GraphPad Software). All reactions were performed in triplicate.

[0092] The results are shown in Figure 1. The addition of BCV reduced the survival rate of NK / T cell lymphoma cells. The susceptibility to BCV varied among cell lines.

[0093] Example 2: In vivo test 0.5 x 10 6NK-S1 cells were inoculated into 6-week-old female NSG mice, and BCV or a control vehicle was intraperitoneally administered twice a week at a dose of 40 mg / kg. The tumor size in the control group was approximately 2000 mm. 3 Tumor measurements were recorded twice weekly until tumor size reached 100 mg / kg. Mice were euthanized according to IACUC guidelines. Tumor sizes in the experimental and control groups (n = 8 per group) were averaged at each time point (days 1, 5, 8, 12, and 15) and statistically compared. Signs of toxicity, including diarrhea and weight loss, were monitored throughout the experiment.

[0094] The results are shown in Figures 2A-D. Administration of BCV significantly suppressed lymphoma growth, and the animals were in good health with no diarrhea observed.

[0095] Example 3 Whole Transcriptome Sequencing Analysis Whole transcriptome sequencing was performed on the same 11 NK / T cell lymphoma cell lines as in Example 1. Differentially expressed genes between the top four cell lines (KAI-3, NK-S1, NK-92, and KHYG-1) for BCV susceptibility and the remaining seven cell lines were identified using DESeq2 (Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014, 15(12):550) and displayed as a volcano plot.

[0096] The results are shown in Figure 3. TLE1 was found to be the gene with the lowest expression level in the top four cell lines. This result indicates that TLE1 can be used as an indicator for predicting susceptibility to BCV. Furthermore, this result indicates that administering BCV to patients with NK / T-cell lymphoma with low TLE1 expression can be expected to improve therapeutic efficacy or the probability of therapeutic efficacy compared to administering BCV to unspecified patients.

[0097] At the same time, ZNF441 was found to be a gene whose expression was upregulated and strongly associated with BCV susceptibility, suggesting that it may be an indicator of susceptibility similar to TLE1. The other gene groups shown in the figure may also serve as complementary indicators for predicting susceptibility.

[0098] Example 4: Analysis of TLE1 expression levels Cell lysates from each of the same 11 NK / T cell lymphoma cell lines as in Example 1 were analyzed using 4 to 15% Mini-PROTEAN TM TGX Stain-Free TM Components separated by SDS-PAGE using Protein Gels (Bio-Rad Laboratories, Hercules, CA, USA) were adsorbed onto a 0.2 μm PVDF membrane (Bio-Rad Laboratories, Hercules, CA, USA). After blocking for 1 hour in TBST solution (50 mM Tris / HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) containing 5% nonfat dry milk (Bio-Rad Laboratories, Hercules, CA, USA) or 5% bovine serum albumin (Sigma-Aldrich, Darmstadt, Germany), the membrane was exposed to primary antibody against TLE1 (Abcam) overnight at 4°C with gentle shaking. Further exposure to HRP-conjugated anti-rabbit or anti-mouse antibodies (Cytiva, Washington, DC, USA) was performed for 1 h, and finally chemiluminescence detection was performed using the SuperSignal Substrate Western Blotting Kit (Thermo Fisher Scientific, MA, USA) and analyzed by Image Lab. TM ChemiDoc using software TM Imaging was performed on an XRS+ system (Bio-Rad Laboratories, Hercules, CA, USA).

[0099] The results are shown in Figure 4. The expression levels of TLE1 in the top four cell lines in terms of susceptibility to BCV were clearly lower than those in the remaining cell lines.

[0100] Example 5: Gene Mutation Analysis Somatic gene mutation information for the same 11 NK / T-cell lymphoma cell lines as in Example 1 was obtained by whole-genome sequencing followed by variant calling using an in-house bioinformatics pipeline. Whole-genome sequencing was performed on the Illumina HiSeq X platform as paired-end 150-bp reads.

[0101] The results are shown in Figure 5. No clear association was found between the 25 somatic gene mutations examined and BCV susceptibility.

[0102] Example 6: Gene Set Expression Analysis Whole transcriptome sequencing was performed on the same 11 NK / T cell lymphoma cell lines as in Example 1. Gene set enrichment analysis (GSEA) was performed using the Molecular Signatures Database (MSigDB) Hallmark gene set to compare the top four cell lines for susceptibility to BCV. Gene sets were considered significantly altered if the False Discovery Rate (FDR) q value of the normalized enrichment score (NES) was less than 0.05.

[0103] The results are shown in Figure 6A-B. Various gene sets, including the MYC target genes, showed significantly increased expression in the top four cell lines.

[0104] Example 7: Prognostic Impact of TLE1 Expression Levels. The association between TLE1 gene expression levels obtained from whole-transcriptome sequencing and progression-free survival and overall survival was examined in a cohort of NK / T-cell lymphoma patients (n = 36). High TLE1 expression was defined as TLE1 gene expression above the median level. Kaplan-Meier analysis was performed to evaluate TLE1 expression as a predictor of survival, revealing a hazard ratio (HR) of 3.44 for progression-free survival with a 95% confidence interval (CI) of 1.23-10.49. Statistical analysis was performed using MedCalc for Windows, version 18.2.1 (MedCalc Software).

[0105] The results are shown in Figures 7A-B. It was revealed that low levels of TLE1 expression were associated with a poor prognosis in patients with NK / T-cell lymphoma.

[0106] Example 8: Gene Set Expression Analysis Using the results obtained from whole transcriptome sequencing in the same NK / T lymphoma patient cohort (n=36) as in Example 7, gene set enrichment analysis (GSEA) was performed using the Molecular Signatures Database (MSigDB) Hallmark gene set to compare TLE1 low-expressing and TLE1 high-expressing patient samples. Gene sets were considered significantly altered if the False Discovery Rate (FDR) q value of the normalized enrichment score (NES) was less than 0.05.

[0107] The results are shown in Figure 8. Similar to the results obtained using NK / T cell lymphoma cell lines, various gene sets, including the MYC target gene group, showed significantly increased expression in patients with low TLE1 expression.

[0108] Example 9: Effect of BCV on the viability of malignant tumor cells The effect of BCV on the viability of 11 different T-cell lymphoma cell lines was investigated. Cell viability was quantified using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol. Cells were cultured at 2 × 10 in 100 μL of medium. 3Cells were seeded into 96-well plates at a concentration of 0.01% and drugs were added at various concentrations. After each time point, Promega CellTiter-Glo® 2.0 Cell Viability Assay reagent was added to the wells, and after 10 minutes of incubation at room temperature, absorbance at 480 nm was measured using a Tecan M200 Infinite 96-well plate reader and iConrol software 1.6 (Tecan, Männedorf, Switzerland). Cell viability was calculated as a percentage of control absorbance. Growth inhibitory effects were analyzed by generating dose-response curves as plots of the percentage of viable cells against drug concentration, and their IC50 values ​​were estimated using GraphPad Prism version 8.0.2 (GraphPad Software). All reactions were performed in triplicate.

[0109] The results are shown in Figure 9. The addition of BCV reduced the viability of T-cell lymphoma cells. Unexpectedly, all cell lines tended to be highly sensitive to BCV. This suggests that T-cell lymphoma is a particularly suitable target for BCV therapy compared to other malignant tumors.

[0110] Example 10: In vivo test 0.5 x 10 6 Six-week-old female NSG mice were inoculated with Z4739 cells and intraperitoneally administered BCV or control vehicle at a dose of 40 mg / kg twice a week. The tumor size in the control group was approximately 2500 mm. 3 Tumor measurements were recorded at each time point (days 4, 7, 11, 18, and 21) until tumor size reached 0.05%. Mice were euthanized according to IACUC guidelines. Tumor size and weight in the experimental and control groups (n = 5 per group) were averaged on day 21 and statistically compared. Signs of toxicity, including diarrhea and weight loss, were monitored throughout the experiment.

[0111] The results are shown in Figures 10A–D. A significant increase in lymphoma was observed in the vehicle-treated group. In contrast, lymphoma growth was significantly suppressed in the BCV-treated group. Furthermore, the animals in the BCV-treated group remained in good health, with no significant weight loss or diarrhea (Figure 10B). The tumor volume in the BCV-treated group remained consistently low between days 0 and 21, and the minimal increase in tumor volume was particularly unexpected (Figure 10A). These results suggest that T-cell lymphomas are particularly suitable targets for BCV therapy compared with other malignancies.

[0112] Example 11: Whole-transcriptome sequencing analysis We performed whole-transcriptome sequencing on (i) the four cell lines (KAI-3, NK-S1, NK-92, and KHYG-1) that were ranked highly susceptible to BCV as described in Example 1, (ii) the remaining seven cell lines (NK-YS, MEC-04, SNK-1, SNK-6, YT, HANK-1, and SNT-8), and (iii) the eleven cell lines (FE-PD, Z4739, SUP-M2, SUDHL-1, DEL, L-82, SMZ-1, SR-786, K-299, HH, and KI-JK) that showed a tendency toward high susceptibility to BCV as described in Example 9. TLE1 gene expression was evaluated from the obtained data.

[0113] The results are shown in Figure 11. The above four cell line groups and the above 11 cell line groups had significantly lower TLE1 gene expression than the above seven cell line group (p = 0.001302, Kruskal-Wallis test). These results indicate that TLE1 can be used as an indicator for predicting susceptibility to BCV. These results also indicate that administering BCV to patients with NK / T-cell lymphoma or T-cell lymphoma with low TLE1 expression can be expected to improve therapeutic efficacy or the probability of achieving therapeutic efficacy compared to administering BCV to unspecified patients.

[0114] Example 12: Effect of BCV on gene expression in malignant tumor cells. The effect of BCV on gene expression in T-cell lymphoma cell lines was investigated. Whole-transcriptome sequencing of Z4739 cell lines treated with 0.1 and 1 μg / mL BCV was performed, and gene set enrichment analysis (GSEA) was performed using the Hallmark gene set from the Molecular Signatures Database (MSigDB). Gene sets were considered significantly enriched if the false discovery rate (FDR) q value of the normalized enrichment score (NES) was less than 0.05. Individual gene expression profiles (IFNB1, IFNY, CCL5, CXCL10, and MYC) obtained from whole-transcriptome sequencing results were expressed as the number of transcripts detected per million reads.

[0115] The results are shown in Figure 12A-C. BCV administration reduced MYC expression, MYC-related gene expression, and other oncogenic genes, including those related to mTOR. Meanwhile, the expression of immune activation-related genes, such as INFγ, increased. This suggests the utility of BCV in tumors with overexpressed oncogenic genes or suppressed cancer immunity.

[0116] Example 13: Effect of BCV on gene expression in malignant tumor cells (Western blotting results) Lysates of Z4739 cells treated with 0.1 and 1 μg / mL of BCV were analyzed by 4 to 15% Mini-PROTEAN TM TGX Stain-Free TMComponents separated by SDS-PAGE using Protein Gels (Bio-Rad Laboratories, Hercules, CA, USA) were adsorbed onto a 0.2 μm PVDF membrane (Bio-Rad Laboratories, Hercules, CA, USA). After blocking for 1 h in TBST solution (50 mM Tris / HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) containing 5% nonfat dry milk (Bio-Rad Laboratories, Hercules, CA, USA) or 5% bovine serum albumin (Sigma-Aldrich, Darmstadt, Germany), the membrane was exposed to the respective primary antibodies overnight at 4°C with gentle shaking. Further exposure to HRP-conjugated anti-rabbit or anti-mouse antibodies (Cytiva, Washington, DC, USA) was performed for 1 h. Finally, chemiluminescence detection was performed using the SuperSignal Substrate Western Blotting Kit (Thermo Fisher Scientific, MA, USA) and analyzed by Image Lab. TM ChemiDoc using software TM Imaging was performed on an XRS+ system (Bio-Rad Laboratories, Hercules, CA, USA).

[0117] The results are shown in Figure 13. BCV treatment resulted in a decrease in MYC protein and an increase in DNA damage (phospho-H2AX), as well as increases in apoptosis markers (cleaved PARP, cleaved caspase-3), cell cycle-related markers (phospho-CHK1 Ser317 / Ser345), and PDL1, suggesting a possible mechanism for the antitumor effect of BCV.

[0118] Example 14: Effect of BCV on the Viability of Malignant Tumor Cells The effect of BCV on cell viability was examined for 19 different B-cell lymphoma cell lines and a CML cell line (K-562) as a control. Cell viability was quantified using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol. Cells were cultured at 2 × 10 in 100 μL of medium. 3 Cells were seeded into 96-well plates at a concentration of 0.01 mg / mL and drug (BCV) was added at various concentrations. After each time point, Promega CellTiter-Glo® 2.0 Cell Viability Assay reagent was added to the wells. After 10 minutes of incubation at room temperature, absorbance at 480 nm was measured using a Tecan M200 Infinite 96-well plate reader and iConrol software 1.6 (Tecan, Männedorf, Switzerland). Cell viability was calculated as a percentage of control absorbance. Growth inhibitory effects were analyzed by generating dose-response curves as a plot of the percentage of viable cells against drug concentration, and their IC50 values ​​were estimated using GraphPad Prism version 8.0.2 (GraphPad Software). All reactions were performed in triplicate.

[0119] The results are shown in Figure 14. The addition of BCV reduced the viability of B cell lymphoma cells. The susceptibility to BCV varied among cell lines.

[0120] Example 15: In vivo test 0.5 x 10 6 Six-week-old female NSG mice were inoculated with OCI-LY19 cells and intraperitoneally administered BCV or a control vehicle at a dose of 40 mg / kg twice a week. The tumor size in the control group was approximately 2500 mm. 3Tumor measurements were recorded at each time point (days 1, 5, 6, and 7) until tumor size reached 0.05%. Mice were euthanized according to IACUC guidelines. Tumor size and weight in the experimental and control groups (n = 5 per group) were averaged on day 7 and statistically compared. Signs of toxicity, including diarrhea and weight loss, were monitored throughout the experiment.

[0121] The results are shown in Figures 15A to 15D. Administration of BCV significantly suppressed lymphoma growth, and the animals were in good health with no weight loss or diarrhea observed.

[0122] Example 16 Whole Transcriptome Sequencing Analysis Whole transcriptome sequencing was performed on the same 19 B-cell lymphoma cell lines as in Example 9. Differentially expressed genes between the top nine cell lines (SUDHL5, DB, BJAB, OCI-LY19, L-428, OCI-LY18, OCI-LY3, WSU-NHL, and RL) for susceptibility to BCV and the remaining 10 cell lines were identified using DESeq2 (Love MI, Huber W, Anders S. Moderated estimation of fold change and dispersion for RNA-seq data with DESeq2. Genome Biol. 2014, 15(12):550) and displayed as a volcano plot.

[0123] The results are shown in Figure 16. TLE1 was found to be one of the genes with elevated expression in the top nine cell lines. These results indicate that TLE1 can be used as an indicator for predicting susceptibility to BCV. These results also indicate that administering BCV to patients with B-cell lymphomas with high TLE1 expression can be expected to improve therapeutic efficacy or the probability of therapeutic efficacy compared to administering BCV to unspecified patients.

[0124] At the same time, CERS6 was found to be the gene whose expression was most strongly correlated with susceptibility, suggesting that it may be an indicator for predicting susceptibility, similar to TLE1. The other gene groups shown in the figure may also serve as complementary indicators for predicting susceptibility.

[0125] Example 17: Patient Cohort Analysis. TLE1 expression levels were analyzed in previously published genetic analysis data for B-cell lymphoma patient cohorts. The association between TLE1 gene expression and overall survival was examined using gene expression data from two previously published DLBCL patient cohorts: a) GSE11318 and b) GSE10846. High TLE1 expression was defined as TLE1 gene expression above the median level. Kaplan-Meier analysis was performed to evaluate TLE1 expression as a predictor of survival. The results were a) hazard ratio (HR) of 2.77 (95% CI: 1.78-4.30) for GSE11318 and b) HR of 1.94 (95% CI: 1.35-2.80) for GSE10846. Statistical analysis was performed using MedCalc for Windows, version 18.2.1 (MedCalc Software).

[0126] The results are shown in Figures 17A-B. When TLE1 expression was divided into low and high levels, high expression was associated with a poor prognosis. This finding was consistent across two separate cohort analyses, suggesting that it is a common feature of B-cell lymphoma.

[0127] The above examples have revealed that BCV susceptibility in malignant tumors is associated with the expression level of TLE1. Specifically, it has been shown that NK / T cell lymphomas and T cell lymphomas with low TLE1 expression levels have high BCV susceptibility. It has also been shown that B cell lymphomas with high TLE1 expression levels have high BCV susceptibility. Therefore, it is believed that more appropriate BCV treatment can be implemented by predicting susceptibility using the TLE1 expression level as an indicator. For example, a patient's BCV susceptibility can be examined using the TLE1 expression level as an indicator, and BCV can be administered to patients predicted to be highly susceptible. This can be expected to improve the therapeutic effect or increase the probability of therapeutic efficacy compared to administration to unspecified patients. Furthermore, for example, when BCV is administered to subjects with low TLE1 expression and predicted to be highly susceptible to BCV, it can be expected to improve the therapeutic effect or increase the probability of therapeutic efficacy compared to administration to unspecified patients. Furthermore, it has been shown that patients with BCV-naive NK / T lymphomas with low TLE1 expression or B cell lymphomas with high TLE1 expression have a poor prognosis. Therefore, BCV therapy is considered to be an effective treatment for malignant tumors with poor prognosis. Furthermore, the antitumor effect of BCV was shown to be particularly strong against T-cell lymphoma. Therefore, it became clear that T-cell lymphoma is particularly suitable as a target for BCV treatment.

[0128] The present invention has been described above based on the embodiments. However, these embodiments are merely illustrative, and it will be understood by those skilled in the art that various modifications are possible and that such modifications are also within the scope of the present invention.

Claims

1. A pharmaceutical composition for treating a malignant tumor, comprising brincidofovir, a pharma- ceutical acceptable salt thereof, or a solvate thereof, wherein the subject of said treatment has a malignant tumor with low expression of TLE1.

2. The pharmaceutical composition according to claim 1, wherein the malignant tumor is a malignant tumor that has low expression of TLE1 and is highly sensitive to brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof.

3. The pharmaceutical composition according to claim 1 or 2, which inhibits the proliferation of malignant tumor cells.

4. The pharmaceutical composition described in any one of claims 1 to 3, wherein the malignant tumor is a blood cancer with low expression of TLE1.

5. The pharmaceutical composition described in any one of claims 1 to 4, wherein the malignant tumor is a lymphoma with low expression of TLE1.

6. The pharmaceutical composition described in any one of claims 1 to 5, wherein the malignant tumor is a NK / T cell lymphoma with low expression of TLE1, or a T cell lymphoma with low expression of TLE1.

7. A pharmaceutical composition according to any one of claims 1 to 6, comprising brincidofovir.

8. A pharmaceutical composition for treating a malignant tumor, comprising brincidofovir, a pharma- ceutical acceptable salt thereof, or a solvate thereof, wherein the subject of the treatment is a subject identified based on the expression level of TLE1.

9. The pharmaceutical composition of claim 8, comprising brincidofovir.

10. The pharmaceutical composition described in claim 8 or 9, wherein the subject is predicted to be highly sensitive to brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof, based on the expression level of TLE1.

11. The pharmaceutical composition according to any one of claims 8 to 10, wherein the subject is identified using the expression level of TLE1 in the subject's malignant tumor cells as an index.

12. A pharmaceutical composition according to any one of claims 8 to 11, wherein the treatment includes a step of examining the TLE1 expression level of the subject.

13. The pharmaceutical composition described in any one of claims 8 to 12, wherein the treatment includes a step of identifying a subject having high sensitivity to brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof using the expression level of TLE1 as an index.

14. The pharmaceutical composition described in any one of claims 8 to 13, wherein the subject has been diagnosed with a malignant tumor with low expression of TLE1.

15. The pharmaceutical composition described in any one of claims 8 to 13, wherein the subject has been diagnosed with a malignant tumor with high expression of TLE1.

16. The pharmaceutical composition according to any one of claims 8 to 15, wherein the malignant tumor is a MYC-positive malignant tumor.

17. The pharmaceutical composition according to any one of claims 8 to 16, wherein the malignant tumor is a blood cancer.

18. The pharmaceutical composition according to any one of claims 8 to 17, wherein the malignant tumor is lymphoma.

19. The pharmaceutical composition of any one of claims 8 to 18, wherein the malignant tumor is NK / T cell lymphoma, B cell lymphoma, or T cell lymphoma.

20. The pharmaceutical composition of claim 14, wherein the malignant tumor is NK / T cell lymphoma.

21. The pharmaceutical composition of claim 14, wherein the malignant tumor is a T-cell lymphoma.

22. The pharmaceutical composition of claim 15, wherein the malignant tumor is a B-cell lymphoma.

23. A composition for use in predicting sensitivity to brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof in a subject with a malignant tumor, the composition comprising a reagent for testing TLE1 expression levels.

24. A composition for use in determining whether a subject has a malignant tumor to which brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof is to be administered, the composition comprising a reagent for testing TLE1 expression levels.

25. A composition for use in diagnosing whether a subject has a malignant tumor highly sensitive to brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof, the composition comprising a reagent for testing TLE1 expression levels.

26. A method for predicting sensitivity to brincidofovir, a pharmacologic acceptable salt thereof, or a solvate thereof in a subject with a malignant tumor, the method comprising a step of examining the expression level of TLE1 in a sample derived from the subject with a malignant tumor.

27. A method for determining which malignant tumor subject to administer brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from the malignant tumor subject.

28. A method for diagnosing whether a subject has a malignant tumor that is highly sensitive to brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof, the method comprising a step of examining the expression level of TLE1 in a sample derived from a subject with a malignant tumor.

29. A kit comprising a reagent for testing TLE1 expression levels, for use in the method according to any one of claims 26 to 28.

30. A composition for use in predicting the prognosis of NK / T cell lymphoma, comprising a reagent for testing TLE1 expression levels.

31. A method for predicting the prognosis of NK / T-cell lymphoma using the expression level of TLE1 in a sample from a subject with NK / T-cell lymphoma as an index.

32. The method of claim 31, wherein low expression of TLE1 predicts a poor prognosis, or high expression predicts a good prognosis.

33. The method according to claim 31 or 32, further comprising predicting the prognosis of NK / T cell lymphoma using the expression level of MYC in the sample as an index.

34. A composition for use in selecting a treatment method for NK / T cell lymphoma, comprising a reagent for testing TLE1 expression levels.

35. A method for selecting a treatment method for NK / T-cell lymphoma using the expression level of TLE1 in a subject sample as an index.

36. A composition for use in selecting a treatment method for B-cell lymphoma, the treatment method comprising a step of administering brincidofovir, a pharmaceutical acceptable salt thereof, or a solvate thereof to a subject, the composition comprising a reagent for testing TLE1 expression levels.

37. A method for selecting a treatment method for B-cell lymphoma using the expression level of TLE1 in a subject's sample as an index, the treatment method comprising a step of administering brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject.

38. A pharmaceutical composition for treating a malignant tumor, comprising brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof, wherein the malignant tumor is a T-cell lymphoma.

39. The pharmaceutical composition of claim 38, wherein the malignant tumor has reduced MYC expression due to the treatment.

40. The pharmaceutical composition of claim 38 or 39, wherein the malignant tumor has increased expression of INFβ1, INFγ, CCL5, CXCL10, or CD274 as a result of the treatment.

41. A pharmaceutical composition according to any one of claims 38 to 40, which inhibits the proliferation of the T-cell lymphoma.

42. The pharmaceutical composition of any one of claims 38 to 41, wherein the malignant tumor is peripheral T-cell lymphoma.

43. A pharmaceutical composition according to any one of claims 38 to 42, comprising brincidofovir.

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