Pharmaceutical composition for treatment of lymphoma

Abstract

To provide a pharmaceutical composition for the treatment of lymphoma.SOLUTION: A pharmaceutical composition comprising brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof is used for treating lymphoma. The lymphoma may be EBV-positive lymphoma. The lymphoma may be MYC-positive lymphoma. The pharmaceutical composition may also be applied in combination with a chemotherapeutic agent.SELECTED DRAWING: Figure 1A

Description

[Technical field]

[0001] The technical field of the invention relates to the treatment of lymphoma. [Background technology]

[0002] Lymphoma is a type of blood cancer, known as a disease caused by lymphocytes becoming cancerous. Non-Patent Document 1 describes that non-Hodgkin's lymphoma is the most common hematological malignancy in the world, accounting for approximately 3% of cancer diagnoses and deaths. Non-Patent Document 2 describes that the incidence of Hodgkin's lymphoma has been increasing over the past decade, especially among women, young people, and people from Asian countries.

[0003] There have been several reports in recent years regarding methods for treating lymphoma. Non-Patent Document 3 describes the possibility of treatment targeting CD19. Non-Patent Document 4 describes the possibility of treatment targeting EZH2. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] "Epidemiology of Non-Hodgkin's Lymphoma" Thandra et al., Med Sci (Basel). 2021 Jan 30;9(1):5. [Non-Patent Document 2] "Incidence, mortality, risk factors, and trends for Hodgkin lymphoma: a global data analysis" Huang et al., J Hematol Oncol. 2022 May 11;15(1):57. [Non-Patent Document 3] "Targeting CD19 for diffuse large B cell lymphoma in the era of CARs: Other modes of transportation" Blood Rev. 2022 Aug 17;101002. [Non-Patent Document 4] "Taking the EZ way: Targeting enhancer of zeste homolog 2 in B-cell lymphomas" Morschhauser et al., Blood Rev. 2022 Jul 9;100988. Summary of the Invention [Problem to be solved by the invention]

[0005] Although a variety of findings have been accumulated to date, many aspects of the mechanisms of lymphoma remain unknown, and conventional treatment approaches alone have not been sufficient. [Means for solving the problem]

[0006] Meanwhile, the present inventors, as a result of extensive research, have discovered that brincidofovir (hereinafter, referred to as BCV) exhibits excellent effects in inhibiting the proliferation of lymphoma cells.

[0007] That is, according to one aspect of the present invention, there is provided a pharmaceutical composition for treating lymphoma, comprising BCV, a pharma- ceutical acceptable salt thereof, or a solvate thereof, which can be used to treat lymphoma. [Brief description of the drawings]

[0008] [Figure 1A-C] Figures 1A-C show the results of investigating the effect of BCV treatment on lymphoma cell proliferation, with Figures 1A and 1B showing the results using EBV-positive cells, and Figure 1C showing the results using EBV-negative cells. [Diagram 2]FIG. 2 shows the results of investigating the effect of treatment with BCV or various antiviral drugs on the proliferation of EBV-positive NK / T lymphoma cells. [Figure 3A-D] Figures 3A-D show the results of administering BCV to NSG mice transplanted with lymphoma cells. Figure 3A shows the results of examining tumor size on days 1, 5, 8, 12, and 15. Figure 3B shows the results of examining body weight on days 1, 5, 8, 12, and 15. Figure 3C shows the results of measuring tumor weight. The values ​​in the bar graph indicate vehicle administration and BCV administration from the left. Figure 3D shows photographs of tumors taken from each individual. [Figure 4] FIG. 4 shows the results of examining the effect of BCV treatment on the gene expression of EBNA1 and LMP1 in lymphoma cells. [Figure 5A-C] Figure 5A shows the results of investigating the effect of BCV treatment on the gene expression of Myc in lymphoma cells. The values ​​of the bar graphs in Figures 5A-C indicate, from the left, DMSO, BCV 0.1 μg / mL (0.17 μM), and BCV 1 μg / mL (1.7 μM). Figures 5B and C show the results of investigating the effect of BCV treatment on the expression of genes in the STING pathway in lymphoma cells. [Figure 6] Figure 6 shows the results of investigating the effect of BCV treatment on immunogenic cell death. The values ​​in the bar graph indicate, from the left, DMSO, BCV 0.1 μg / mL (0.17 μM), and BCV 1 μg / mL (1.7 μM). [Figure 7A-C] Figures 7A-C show the results of investigating the effect of BCV and various anticancer drug treatments, or BCV treatment and radiation exposure on lymphoma cell proliferation. Figure 7A shows the results of etoposide or gemcitabine treatment. Figure 7B shows the results of BCV, BCV and etoposide, or BCV and gemcitabine treatment. Figure 7C shows the results of BCV treatment and radiation exposure. [Figure 8] FIG. 8 shows the results of investigating the effect of BCV treatment on the proliferation of MYC-amplified lymphoma (EBV positive or negative). [Figure 9] FIG. 9 shows the results of investigating the sensitivity of 11 lymphoma cell lines to the effect of BCV treatment on their proliferation. [Figure 10A-B]Figures 10A and 10B show the results of gene expression analysis. Figure 10A shows a set of genes that showed increased expression in BCV-susceptible cell lines. Figure 10B shows an enrichment plot of MYC TARGETS V2. [Figure 11A-B] Figures 11A and 11B show the results of investigating the prognosis of a cohort of human patients: Figure 11A shows progression-free survival, and Figure 11B shows overall survival. [Figure 12] FIG. 12 shows the results of examining genes that showed increased expression in the poor prognosis group of a human patient cohort. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0009] Hereinafter, the embodiments of the present invention will be described in detail. Note that, in order to avoid repetition and complexity, the description of similar contents will be omitted as appropriate.

[0010] (1) Treatment method According to one embodiment of the present invention, there is provided a method for treating lymphoma, comprising administering to a subject BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof, which provides a novel therapeutic approach to the treatment of lymphoma.

[0011] The above-mentioned lymphoma may be MYC-positive lymphoma. In this case, BCV, its pharma- ceutically acceptable salt, or their solvates can provide a particularly excellent therapeutic effect. For example, in MYC-positive lymphoma, a high growth suppression effect can be obtained not only against EBV-positive lymphoma, but also against EBV-negative lymphoma.

[0012] In the above-mentioned treatment method, the subject may be, for example, a MYC-positive subject.In this case, BCV, its pharma- ceutically acceptable salt, or their solvates can provide a particularly excellent therapeutic effect.For example, in the MYC-positive subject, a high growth suppression effect can be obtained not only for EBV-positive lymphoma, but also for subjects suffering from EBV-negative lymphoma.

[0013] The above-mentioned treatment method may, for example, include a step of identifying a MYC-positive subject. In this case, a particularly excellent therapeutic effect of BCV, its pharma- ceutically acceptable salt, or a solvate thereof can be obtained. For example, in a MYC-positive subject, a high growth suppression effect can be obtained not only for EBV-positive lymphoma, but also for subjects suffering from EBV-negative lymphoma.

[0014] In the above-mentioned treatment method, the subject may be, for example, a subject that is specified by the expression level of MYC. In this case, BCV, its pharma- ceutically acceptable salt, or a solvate thereof can provide a particularly excellent therapeutic effect. For example, by identifying and treating a subject with a high expression level of MYC, a higher lymphoma proliferation suppression effect can be obtained than by treating a subject with a low expression level of MYC.

[0015] The above-mentioned treatment method may include, for example, a step of identifying a subject with high MYC expression as a treatment subject. In this case, a particularly excellent therapeutic effect can be obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, treating a subject with high MYC expression can obtain a higher lymphoma proliferation suppression effect than treating a subject without high MYC expression.

[0016] The above-mentioned treatment method may include, for example, a step of detecting the expression level of MYC in a subject's specimen. In this case, a particularly excellent therapeutic effect can be obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, a high therapeutic effect can be obtained by treating a subject provided with a specimen having a high expression level of MYC. For example, when the expression level of MYC in a subject's specimen is high, the subject may be identified as a subject to be administered with BCV or a subject to be administered with a low dose. On the other hand, for example, when the expression level of MYC in a subject's specimen is low, the subject may be identified as a subject to not be administered with BCV or a subject to be administered with a high dose of BCV.

[0017] The above-mentioned therapeutic method may include, for example, administering a higher dose of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject provided with a sample having a high MYC expression level than to a subject provided with a sample having a low MYC expression level, in which case a particularly excellent therapeutic effect of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof can be obtained.

[0018] The above-mentioned treatment method may, for example, comprise a step of administering brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject, and the subject may be a subject whose MYC expression level in a sample from the subject has been shown to be higher than that in a sample previously confirmed not to have high MYC expression, by detecting the expression level of MYC in the sample from the subject. In this case, a particularly excellent therapeutic effect of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof can be obtained.

[0019] The above-mentioned treatment method may include, for example, a step of administering brincidofovir, a pharma- ceutical acceptable salt, or a solvate thereof to a subject in which the expression level of MYC in the subject's specimen is shown to be higher than that in a MYC-negative specimen by detecting the expression level of MYC in the subject's specimen, in which case a particularly excellent therapeutic effect of BCV, a pharma- ceutical acceptable salt, or a solvate thereof can be obtained.

[0020] The lymphoma may be EBV-positive lymphoma. In this case, BCV, its pharma- ceutically acceptable salt, or a solvate thereof can provide a particularly excellent therapeutic effect. For example, in this EBV-positive lymphoma, a higher growth suppression effect can be obtained than in EBV-negative lymphoma.

[0021] In the above-mentioned treatment method, the subject may be, for example, an EBV-positive subject. In this case, a particularly excellent therapeutic effect is obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, a higher growth suppression effect is obtained in an EBV-positive subject than in a subject suffering from EBV-negative lymphoma.

[0022] The above-mentioned treatment method may include, for example, a step of identifying an EBV-positive subject. In this case, a particularly excellent therapeutic effect of BCV, its pharma- ceutically acceptable salt, or a solvate thereof can be obtained. For example, a higher growth inhibitory effect can be obtained in an EBV-positive subject than in a subject suffering from EBV-negative lymphoma.

[0023] In the above-mentioned treatment method, the subject may be, for example, a subject identified by the expression level of EBV. In this case, a particularly excellent therapeutic effect can be obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, when a subject with a high expression level of EBV is identified and treated, a higher lymphoma proliferation suppression effect can be obtained than when a subject with a low expression level of EBV is treated.

[0024] The above-mentioned treatment method may include, for example, a step of identifying a subject with high EBV expression as a treatment subject. In this case, a particularly excellent therapeutic effect can be obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, treating a subject with high EBV expression can obtain a higher lymphoma proliferation suppression effect than treating a subject without high EBV expression.

[0025] The above-mentioned treatment method may include, for example, a step of detecting the expression level of EBV in a subject's specimen. In this case, a particularly excellent therapeutic effect is obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, a high therapeutic effect is obtained by treating a subject provided with a specimen having a high expression level of EBV. For example, when the expression level of EBV in a subject's specimen is high, the subject may be identified as a subject to be administered with BCV or a subject to be administered with a low dose. On the other hand, for example, when the expression level of EBV in a subject's specimen is low, the subject may be identified as a subject not to be administered with BCV or a subject to be administered with a high dose of BCV.

[0026] The above-mentioned therapeutic method may include, for example, administering a higher dose of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject providing a specimen with a high EBV expression level than to a subject providing a specimen with a low EBV expression level, in which case a particularly excellent therapeutic effect of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof can be obtained.

[0027] The above-mentioned treatment method may, for example, comprise a step of administering brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject, and the subject may be a subject whose EBV expression level in a specimen from the subject has been shown to be higher than that in a specimen previously confirmed not to have high EBV expression, by detecting the expression level of EBV in the specimen from the subject. In this case, a particularly excellent therapeutic effect of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof can be obtained.

[0028] The above-mentioned therapeutic method may include, for example, a step of administering brincidofovir, a pharma- ceutical acceptable salt, or a solvate thereof to a subject in which the expression level of EBV in the subject's specimen has been shown to be higher than that in an EBV-negative specimen by detecting the expression level of EBV in the subject's specimen, in which case a particularly excellent therapeutic effect of BCV, a pharma- ceutical acceptable salt, or a solvate thereof can be obtained.

[0029] The above-mentioned treatment method may include, for example, a step of identifying a MYC-positive and EBV-positive subject as a treatment subject, a step of identifying a subject using the expression levels of MYC and EBV as an index, or a step of identifying a subject with high expression of MYC and EBV as a treatment subject. In this case, a particularly excellent therapeutic effect is obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. The above-mentioned lymphoma may be MYC-positive and EBV-positive lymphoma. In this case, a particularly excellent therapeutic effect is obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. The above-mentioned treatment method may include, for example, a step of identifying a MYC-positive and EBV-negative subject. The above-mentioned lymphoma may be MYC-positive and EBV-negative lymphoma.

[0030] The lymphoma may be, for example, EBV-positive NK / T lymphoma, MYC-positive EBV-positive NK / T lymphoma, MYC-positive Burkitt's lymphoma, MYC-positive DLBCL, MYC-positive double-hit DLBCL, or MYC-positive triple-hit DLBCL, in which case a particularly excellent therapeutic effect is obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof.

[0031] The above-mentioned treatment method may include a step of administering a chemotherapeutic agent to the subject. In this case, a particularly excellent therapeutic effect is obtained by the combination. For example, this combination treatment provides a synergistic effect by BCV and the chemotherapeutic agent. The synergistic effect includes, for example, an effect that exceeds the effect obtained by adding the lymphoma growth inhibitory effect obtained by BCV treatment alone and the lymphoma growth inhibitory effect obtained by chemotherapy treatment alone. A particularly excellent therapeutic effect is obtained by using an antimetabolite or a topoisomerase inhibitor in combination with BCV. In addition, a particularly excellent therapeutic effect is obtained by using an immune checkpoint inhibitor in combination with BCV. The chemotherapeutic agent may be administered to the subject before, simultaneously with, or after administration of BCV. The term "simultaneous with administration" includes the same period, and may be substantially simultaneous in consideration of normal treatment procedures. The term "simultaneous with administration" includes the case where BCV and the chemotherapeutic agent are administered as a combination drug.

[0032] The above-mentioned treatment method may include a step of irradiating the subject. The irradiation may include, for example, gamma irradiation. The irradiation may be administered to the subject before, simultaneously with, or after the administration of BCV.

[0033] The above-mentioned therapeutic method may, for example, decrease MYC expression in lymphoma cells of a subject by administering BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof to the subject, and this therapeutic method may improve the prognosis of human lymphoma patients due to decreased MYC expression.

[0034] The above-mentioned therapeutic methods may, for example, induce immunogenic cell death by administering to a subject BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof.

[0035] In the above-mentioned treatment method, the subject (including the patient) includes humans or non-human mammals (e.g., one or more of mice, guinea pigs, hamsters, rats, mice, rabbits, pigs, sheep, goats, cows, horses, cats, dogs, marmosets, monkeys, or chimpanzees). The patient may be a patient diagnosed with lymphoma (e.g., MYC-positive or EBV-positive lymphoma) or a patient in need of lymphoma treatment. The patient may be, for example, a patient administered a chemotherapeutic agent, a patient undergoing treatment with a chemotherapeutic agent, a patient administered BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof, or a patient undergoing treatment with BCV, a pharma-ceutically acceptable salt thereof, or a solvate thereof. In the treatment with BCV, a pharma-ceutically acceptable salt thereof, or a solvate thereof, it is preferable to treat patients with NK / T-cell lymphoma. In this case, excellent therapeutic effects on NK / T-cell lymphoma, effects of minimizing side effects, effects of reducing MYC expression, or effects of improving prognosis are particularly obtained. In this case, it is preferable that the NK / T cell lymphoma is EBV positive or highly expresses EBV. In this case, a particularly excellent therapeutic effect on the NK / T cell lymphoma can be obtained. In addition, it is preferable that the NK / T cell lymphoma is MYC positive or highly expresses MYC. In this case, a particularly excellent therapeutic effect on the NK / T cell lymphoma can be obtained.

[0036] The above-mentioned treatment method may include, for example, (i) a step of identifying a MYC-positive or EBV-positive subject as a subject for lymphoma treatment, (ii) a step of identifying a subject with MYC-positive lymphoma or EBV-positive lymphoma as a subject for lymphoma treatment, (iii) a step of detecting MYC or EBV in the subject, (iv) a step of detecting the presence or absence of MYC-positive or EBV-positive in the subject, (v) a step of collecting a subject's sample (e.g., lymphoma cells, lymph node tissue, or a blood sample (e.g., plasma, serum, or whole blood)), (vi) a step of detecting MYC or EBV in the collected sample, or (vii) a step of identifying a subject who provided a sample in which MYC or EBV was detected as a subject for treatment. The adoption of one or more of these steps is useful for administering treatment to a population that is likely to respond to treatment. When two or more of these steps are adopted, the order is arbitrary and can be determined according to the desired treatment method. These steps may be performed before or after the step of administering BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof, or a chemotherapeutic agent to the subject. Detection may be based on gene dosage, mRNA dosage, protein dosage, or gene translocation. Detection may be by testing.

[0037] The above-mentioned treatment methods may include, for example, a step of identifying a subject having lymphoma as a subject for lymphoma treatment, a step of identifying a subject who is positive for a lymphoma marker as a subject for lymphoma treatment, a step of administering a therapeutically effective amount of BCV or a chemotherapeutic agent to the subject, a step of inhibiting the proliferation of lymphoma cells in the subject, a step of reducing a lymphoma marker in the subject, a step of reducing expression of MYC in the lymphoma of the subject, or a step of inhibiting swelling of lymph nodes in the subject. Additionally, the above treatment methods or subjects may or may not include, for example, any of the following: identifying a subject having a 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 transplant or allogeneic transplant (e.g., hematopoietic cell transplant); identifying a subject in need of an immunosuppressant; identifying an immunosuppressed subject; a method for treating virus-induced tumors in an immunosuppressed subject; or a method for preventing or treating a virus (e.g., CMV, AdV, BKV, or VaV) infection.

[0038] (2) Pharmaceutical Composition According to one embodiment of the present invention, there is provided a pharmaceutical composition for treating lymphoma, comprising BCV, a pharma- ceutically acceptable salt, or a solvate thereof, which can be used to treat lymphoma through a novel therapeutic approach.

[0039] The above-mentioned lymphoma may be MYC-positive lymphoma. In this case, BCV, its pharma- ceutically acceptable salt, or their solvates can provide a particularly excellent therapeutic effect. For example, in MYC-positive lymphoma, a high growth suppression effect can be obtained not only against EBV-positive lymphoma, but also against EBV-negative lymphoma.

[0040] The lymphoma may be EBV-positive lymphoma. In this case, BCV, its pharma- ceutically acceptable salt, or a solvate thereof can provide a particularly excellent therapeutic effect. For example, in EBV-positive lymphoma, a higher growth suppression effect can be obtained than in EBV-negative lymphoma.

[0041] In the above treatment, the subject may be, for example, a subject identified using the expression level of EBV or MYC as an index. In this case, a particularly excellent therapeutic effect can be obtained by BCV, its pharma- ceutically acceptable salt, or a solvate thereof. For example, by identifying and treating a subject with a high expression level of EBV or MYC, a higher lymphoma proliferation suppression effect can be obtained than by treating a subject with a low expression level of EBV or MYC.

[0042] The above treatment may include, for example, detecting the expression level of EBV or MYC in a subject's sample. In this case, a particularly excellent therapeutic effect can be obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. For example, a high therapeutic effect can be obtained by treating a subject who has provided a sample with a high expression level of EBV or MYC.

[0043] The above treatment may, for example, comprise administering brincidofovir, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject, and the subject may be a subject whose EBV or MYC expression level in a specimen from the subject has been shown to be higher than that in a specimen previously confirmed not to have high EBV or MYC expression. In this case, a particularly excellent therapeutic effect of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof can be obtained.

[0044] The lymphoma may be MYC-positive and EBV-positive lymphoma, in which case BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof can provide a particularly excellent therapeutic effect.

[0045] The lymphoma may be, for example, EBV-positive NK / T lymphoma, MYC-positive EBV-positive NK / T lymphoma, MYC-positive Burkitt's lymphoma, MYC-positive DLBCL, MYC-positive double-hit DLBCL, or MYC-positive triple-hit DLBCL, in which case a particularly excellent therapeutic effect is obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof.

[0046] The above pharmaceutical composition may be used in a combination treatment of BCV, its pharma- ceutically acceptable salt, or a solvate thereof, and a chemotherapeutic agent. In this case, a particularly excellent therapeutic effect is obtained by the combination. For example, this combination treatment can provide a synergistic effect by BCV and a chemotherapeutic agent. The synergistic effect includes, for example, an effect that exceeds the effect obtained by adding together the lymphoma growth suppression effect obtained by BCV treatment alone and the lymphoma growth suppression effect obtained by chemotherapeutic agent treatment alone.

[0047] The pharmaceutical compositions include those containing BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof for use in treating lymphoma. In another aspect, there is provided a use of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof for the manufacture of a pharmaceutical composition for treating lymphoma.

[0048] According to one embodiment of the present invention, there is provided a pharmaceutical composition for treating lymphoma, comprising BCV, a pharma- ceutically acceptable salt or solvate thereof, or a chemotherapeutic agent, for use in combination therapy with BCV, a pharma- ceutically acceptable salt or solvate thereof, or a chemotherapeutic agent, which provides a particularly excellent therapeutic effect of the combination, as described above.

[0049] The pharmaceutical composition includes a pharmaceutical composition for use in the treatment method of (1) above. This treatment method may include at least one of the steps of (1) above (e.g., steps (i) to (vii)).

[0050] (3) Inhibition method According to one embodiment of the present invention, there is provided a method for inhibiting MYC, comprising a step of administering BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject. The proliferation of MYC-positive lymphoma cells is suppressed by inhibiting MYC. Thus, by using this inhibition method, the proliferation of MYC-positive lymphoma cells can be more effectively suppressed. This method may include at least one of the steps (1) above (e.g., steps (i) to (vii)). This method includes a method for treating a disease by inhibiting MYC. The disease includes, for example, a disease caused by MYC. The disease includes, for example, lymphoma (e.g., MYC-positive lymphoma).

[0051] According to one embodiment of the present invention, a composition for inhibiting MYC is provided, comprising BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. The proliferation of MYC-positive lymphoma cells is suppressed by inhibiting MYC. Therefore, the use of this composition can more effectively inhibit the proliferation of MYC-positive lymphoma cells. This composition includes a composition for use in the above-mentioned inhibition method.

[0052] The above-mentioned compositions include compositions comprising BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof for use in inhibiting MYC. In another aspect, there is provided a use of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof for the manufacture of a composition for inhibiting MYC.

[0053] According to one embodiment of the present invention, there is provided a method for inhibiting MYC comprising contacting MYC with BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof, including, for example, in vitro, ex vivo, or in vivo inhibition.

[0054] According to one embodiment of the present invention, there is provided a method for inhibiting EBNA1 or LMP1, comprising a step of administering BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof to a subject. By using this inhibition method, the proliferation of EBV-positive lymphoma cells can be more effectively suppressed. This method may comprise at least one of any of the steps (1) above (e.g., steps (i) to (vii)). This method includes a method for treating a disease by inhibiting EBNA1 or LMP1. The disease includes, for example, a disease caused by EBV. The disease includes, for example, lymphoma (e.g., EBV-positive lymphoma). From another aspect, there is provided a composition for inhibiting EBNA1 or LMP1, comprising BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. From another aspect, there is provided a use of BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof for the manufacture of a composition for inhibiting EBNA1 or LMP1. In another aspect, there is provided a method for inhibiting EBNA1 or LMP1, the method comprising the step of contacting EBNA1 or LMP1 with BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. Inhibition includes, for example, in vitro, ex vivo, or in vivo inhibition.

[0055] (4) Diagnostic method According to one embodiment of the present invention, a companion diagnostic method is provided, which comprises detecting the presence or absence of MYC positivity in a subject. This method includes a diagnostic method for evaluating the efficacy of treatment with BCV, its pharma- ceutically acceptable salt, or a solvate thereof before administration. According to this diagnostic method, treatment can be administered to a population that is likely to respond to treatment. Here, the subject includes a patient suffering from lymphoma or the lymphoma of the patient.

[0056] According to one embodiment of the present invention, a companion diagnostic method is provided, which comprises a step of detecting the presence or absence of EBV positivity in a subject. This method includes a diagnostic method for evaluating the efficacy of treatment with BCV, its pharma- ceutically acceptable salt, or a solvate thereof before administration. This diagnostic method allows treatment to be administered to a population that is likely to respond to treatment. Here, the subject includes a patient suffering from lymphoma or a lymphoma of the patient.

[0057] According to one embodiment of the present invention, a companion diagnostic method is provided, which comprises detecting the presence or absence of MYC positivity and EBV positivity in a subject. This method includes a diagnostic method for evaluating the effectiveness of treatment with BCV, its pharma- ceutically acceptable salt, or a solvate thereof before administration. According to this diagnostic method, treatment can be administered to a population that is likely to respond to treatment. In the diagnostic method, the subject includes a patient suffering from lymphoma or the lymphoma of the patient.

[0058] According to one embodiment of the present invention, a companion diagnostic method is provided, which comprises detecting EBV or MYC expression in a subject. The method includes a diagnostic method for evaluating the efficacy of treatment with BCV, its pharma- ceutically acceptable salt, or a solvate thereof before administration. According to this diagnostic method, treatment can be administered to a population that is likely to respond to treatment. Here, the subject includes a patient suffering from lymphoma or the lymphoma of the patient.

[0059] According to one embodiment of the present invention, a method for diagnosing whether a subject has lymphoma that is highly sensitive to BCV, its pharmaceutically acceptable salt, or a solvate thereof is provided, comprising a step of examining the expression level of EBV or MYC in a specimen derived from a lymphoma subject. This method can be used to diagnose whether a subject has lymphoma that is highly sensitive to BCV using the expression level of EBV or MYC as an index. Therefore, by using this method, more appropriate BCV treatment can be performed. For example, if a subject is diagnosed as having a lymphoma that is highly sensitive, treatment may be performed such that BCV is administered or at a low dose. On the other hand, for example, if a subject is diagnosed as not having a lymphoma that is highly sensitive, treatment may be performed such that BCV is not administered or at a high dose. This diagnosis may be a companion diagnosis for treatment using BCV, its pharmaceutically acceptable salt, or a solvate thereof.

[0060] From another perspective, according to one embodiment of the present invention, there is provided a kit or composition for any of the above diagnostic methods or companion diagnostic methods. The kit or composition includes a means for measuring, for example, the gene amount, mRNA amount, protein amount, or gene translocation of MYC or EBV. The measuring means may include, for example, a primer (for example, a primer capable of amplifying at least a part of the MYC or EBV gene (for example, a primer capable of binding to the MYC or EBV gene, or an upstream or downstream site thereof)) or a probe (a probe capable of detecting translocation of the MYC gene (for example, a FISH probe)).

[0061] (5) Other methods According to one embodiment of the present invention, there is provided a method for identifying a subject to be treated with BCV, its pharma- ceutically acceptable salt, or a solvate thereof, comprising a step of detecting MYC or EBV in a subject. According to this method, treatment can be administered to a population with high therapeutic efficacy. For example, if a subject's expression level of EBV or MYC is high and it is predicted that the subject is highly sensitive, the subject may be identified as a subject to be administered BCV or a subject to be administered a low dose of BCV. On the other hand, for example, if a subject's expression level of EBV or MYC is low and it is predicted that the subject is less sensitive, the subject may be identified as a subject to not be administered BCV or a subject to be administered a high dose of BCV. Here, the subject includes a patient suffering from lymphoma or a specimen from the patient.

[0062] According to one embodiment of the present invention, there is provided a method for improving the prognosis of a subject in lymphoma treatment, comprising administering BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof to the subject. BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof has the effect of reducing MYC expression, and therefore may improve the prognosis of a subject in lymphoma treatment. The improvement of prognosis includes, for example, improving the prognosis. The improvement of prognosis includes, for example, extending progression-free survival or survival time. The improvement of prognosis includes, for example, a state in which the prognosis is improved compared to that of a patient administered a treatment method that does not reduce MYC expression. The prognosis includes, for example, a change in the patient's symptoms as the disease progresses.

[0063] According to one embodiment of the present invention, there is provided a method for predicting or evaluating the prognosis of a malignant tumor, comprising a step of using the expression level of MYC in a lymphoma subject as an index. By using this method, the prognosis of a patient can be predicted or evaluated. For example, when the expression level of MYC in a specimen of a lymphoma subject is high, the prognosis may be predicted or evaluated as poor (poor prognosis). For example, when the expression level of MYC in a specimen of a lymphoma subject is low, the prognosis may be predicted or evaluated as good (good prognosis).

[0064] The above embodiments (1) to (5) will be described in more detail below. The following detailed embodiments or explanations of each term can be applied to any of the above embodiments (1) to (5).

[0065] In the above (1) to (5), BCV (brincidofovir) includes a compound having a structure represented by the following formula. BCV can also be represented by the IUPAC name of [(2S)-1-(4-amino-2-oxopyrimidin-1-yl)-3-hydroxypropan-2-yl]oxymethyl-(3-hexadecoxypropoxy)phosphinic acid or ({[(2S)-1-(4-amino-2-oxo-1,2-dihydropyrimidin-1-yl)-3-hydroxypropan-2-yl]oxy}methyl)[3-(hexadecyloxy)propoxy]phosphinic acid. BCV includes a compound represented by the CAS registry number of 444805-28-1. In this specification, BCV is an abbreviation of brincidofovir, and they have the same meaning. [ka]

[0066] In the above (1) to (5), 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 or NK / T cell lymphoma. B cell lymphoma includes, for example, 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. NK / T cell lymphomas include, for example, 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). Hodgkin lymphomas include, for example, classical Hodgkin lymphoma, or nodular lymphocyte-predominant Hodgkin lymphoma. For details of 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, or Voltin et al., Cancers (Basel). 2020 Mar 5;12(3):601. The therapeutic effect or onset of lymphoma may be diagnosed by examining swollen lymph nodes with CT, PET, or MRI, histological examination of lymph nodes or tumor masses, or by examining tumor markers for malignant lymphoma in blood (e.g., sIL2-R). Lymphoma may be diagnosed, for example, by carrying out the method described in Nirmal (supra) or Voltin et al. (supra). The therapeutic effect of lymphoma may be evaluated, for example, by observing changes in lymphoma 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 index, or may be determined from image data. The therapeutic effect of lymphoma may be evaluated, for example, by observing a reduction in the amount of tumor mass after drug administration.In this case, the therapeutic effect may be judged to be present when the tumor mass is significantly decreased compared to before administration of the drug or when the negative control is administered. The therapeutic effect against lymphoma may be measured, for example, using the amount of lymphoma marker in the patient or a patient-derived sample as an index. In this case, the therapeutic effect may be judged to be present when the marker amount is significantly decreased compared to before administration of the drug or when the negative control is administered. The tumor mass or marker amount after administration of the drug may be decreased to 0.9, 0.7, 0.5, 0.3, or 0.1 times or less compared to before administration or when the negative control is administered.

[0067] In the above (1) to (5), the lymphoma includes MYC-positive, MYC-negative, EBV-positive, or EBV-negative lymphoma. By targeting MYC-positive or EBV-positive lymphoma, a high therapeutic effect can be obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof. By targeting MYC-positive lymphoma, a high therapeutic effect can be obtained by BCV, a pharma- ceutically acceptable salt thereof, or a solvate thereof for lymphoma that is either EBV-positive or EBV-negative or both.

[0068] In the above (1) to (5), positive includes positive gene expression. Positive may also include increased gene expression compared to a healthy person or a person previously confirmed to be negative. In the above (1) to (5), the expression level of MYC or EBV in the subject or the subject's lymphoma, or the positive / negative state of MYC or EBV can be detected, for example, by collecting a subject's sample (e.g., lymphoma cells, plasma, whole blood, or tissue) and measuring the gene expression of MYC or EBV in the sample. The gene expression can be measured, for example, by RT-PCR, DNA chip, or immunostaining. In this case, the sample collected from the subject may be compared with a sample collected from a healthy person or a person previously confirmed to be negative, and if the gene expression of MYC or EBV is significantly enhanced in the sample collected from the subject, it may be determined that the subject is positive for MYC or EBV. The gene expression of EBV may be indicated by the expression of an EBV-derived transcription product (e.g., EBER or EBNA1, etc.). In addition, the expression level of MYC or EBV in a subject or a subject's lymphoma, or the positive / negative state of MYC or EBV can be detected, for example, by collecting lymph nodes from the subject 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 person or a person previously confirmed to be negative, and the sample collected from the subject may be judged as positive for MYC or EBV if the staining intensity of MYC or EBV is significantly increased. The above-mentioned enhancement of gene expression or staining intensity 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 within a range of any two of these values. In addition, in the detection of the positive / negative state of MYC or EBV in a subject or a subject's lymphoma, the sample may be judged as positive for MYC or EBV if the proportion of cells in the sample immunostained for MYC or EBV is equal to or greater than a certain value. This ratio may be, for example, 10, 20, 30, 40, 50, 60, 70, 80, or 90% or more, or 100%, or may be within a range between any two of these values. When the expression level of EBV or MYC is used as an index, a high expression level of EBV or MYC in a subject specimen may include, for example, being higher than the expression level of EBV or MYC in a comparative subject.The height may be, for example, 1.5, 2, 3, 4, 5, 10, 20, or 50 times or more, or within a range of any two of these values. The comparison subject may be a sample previously confirmed not to have high expression of EBV or MYC, or an EBV-negative or MYC-negative sample. Also, for example, a high expression level of EBV or MYC in a subject sample may include a relatively high expression level in a group of samples of lymphoma subjects. The group of samples may include, for example, samples of general patients with lymphoma. The evaluation of high expression or positivity may be performed based on the technical common knowledge of a person skilled in the art, for example, by using any one of the methods shown in this paragraph. In addition, the expression level of MYC in the subject or the subject's lymphoma, or the positive / negative state of MYC can be detected, for example, by detecting MYC translocation by fluorescent 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). In addition, the expression level of EBV in the subject or the subject's lymphoma, or the positive / negative state of EBV can be detected, 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 LLC)), 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.)). Detection of the expression level of MYC or EBV in a subject or in a subject's lymphoma, or MYC or EBV positivity / negativity, may be performed, for example, in vitro, ex vivo or in vivo.

[0069] In the above (1) to (5), lymphomas include, for example, double-hit or triple-hit lymphomas. Double-hit lymphomas include, for example, lymphomas with gene rearrangements of MYC and BCL2. Triple-hit lymphomas include, for example, lymphomas with gene rearrangements of MYC, BCL2, and BCL6. Detection of double-hit or triple-hit lymphomas can be performed, for example, by detecting translocations by the FISH method (see, for example, Salam et al. (supra) or Epperla et al. (supra)). Lymphomas include, for example, high-grade lymphomas.

[0070] In the above (1) to (5), MYC includes a protein known as a transcriptional regulator. In the present specification, MYC, Myc, and c-Myc can be used interchangeably or with substantially the same intent. The primary accession number of MYC described in UniProt is, for example, P01106. EBV is an abbreviation for Epstein-Barr Virus. EBV is known as a virus having 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).

[0071] In the above (1) to (5), the chemotherapeutic agent is not particularly limited and includes, for example, an anticancer agent. The anticancer agent includes, for example, a microtubule inhibitor, a DNA synthesis inhibitor, a growth factor inhibitor, a tyrosine kinase inhibitor, a cytotoxic substance, an immune checkpoint inhibitor, or other anticancer agent. The microtubule inhibitor includes, for example, a vinca alkaloid drug or a taxane drug. The vinca alkaloid drug includes, for example, vincristine, vinblastine, vindesine, vinorelbine, or eribulin. The taxane drug includes, for example, paclitaxel or docetaxel. The DNA synthesis inhibitor includes, for example, an antimetabolite, a topoisomerase inhibitor, a platinum agent, an antitumor antibiotic, or an alkylating agent. The antimetabolite includes, for example, pemetrexed, 5-fluorouracil, S-1, gemcitabine, or capecitabine. Topoisomerase inhibitors include, for example, irinotecan, nogitecan, etoposide, or zobuzoxacin. Platinum agents include, for example, cisplatin, oxaliplatin, nedaplatin, or carboplatin. Antitumor antibiotics include, for example, anthracyclines (e.g., doxorubicin, liposomal doxorubicin, daunorubicin, epirubicin, idarubicin, aclarubicin, amrubicin, mitoxatrone, or pirarubicin), mitomycin C, actinomycin D, bleomycin, beplomycin, or zinostatin stimer. Alkylating agents include, for example, bendamustine, cyclophosphamide, dacarbacine, or ifosfamide. Growth factor inhibitors include, for example, inhibitors of EGF, VEGF, FGF, or IGF. Examples of growth factor inhibitors include bevacizumab, cetuximab, and panitumumab. Examples of tyrosine kinase inhibitors include gefitinib or erlotinib. Examples of cytotoxic substances include saporin, emtansine, deruxtecan, or vedotin. Examples of immune checkpoint inhibitors include drugs that bind to immune checkpoint molecules or their ligands to inhibit the transmission of immunosuppressive signals, thereby releasing the inhibition of T cell activation by immune checkpoint molecules.Immune checkpoint inhibitors include, for example, anti-CTLA-4 antibodies (e.g., ipilimumab), anti-PD-1 antibodies (e.g., nivolumab or pembrolizumab), or anti-PD-L1 antibodies (e.g., atezolizumab or avelumab). Other anticancer drugs include, for example, L-asparaginase. Forms of chemotherapeutic agents include, for example, low molecular weight compounds or high molecular weight compounds. Chemotherapeutic agents include salts of any one or more of the compounds shown herein (including the salts shown below).

[0072] In the above (1) to (5), 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."). The salt includes, 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. The 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. The 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 that have reasonable benefits for pharmaceutical use. In one embodiment of the present invention, one form of the compound or its salt includes the form of their solvates. In the above (1) to (5), the solvate includes the form of the compound formed by the solute and the solvent (see, for example, Healy et al., Adv Drug Deliv Rev. 2017 Aug 1;117:25-46.). The solvate is not particularly limited, but includes, for example, a hydrate (e.g., a monohydrate, a dihydrate, a trihydrate, etc.) or an organic solvent solvate (e.g., a solvate with an alcohol (e.g., methanol, ethanol, propanol, etc.), acetone, dimethylformamide, or ethyl acetate, etc.).Solvents include those that are capable of substantially maintaining the biological activity of the solute after forming the solvate. Solvates include pharma- ceutically acceptable solvates.

[0073] In the above (1) to (5), the treatment includes exerting a symptom-improving effect, a suppressing effect, a recurrence-suppressing effect, or a preventive effect on the patient's disease or one or more symptoms associated with the disease. The treatment also includes, for example, treatment of suppressing the proliferation of lymphoma cells in the patient, reducing lymphoma markers, suppressing swelling of lymph nodes, and suppressing recurrence of lymph nodes. In the above (1) to (5), the pharmaceutical composition may be produced by, for example, mixing an active ingredient with one or more pharma-ceutically acceptable carriers and using any method known in the technical field of pharmaceuticals. The pharmaceutical composition may be used in any form as long as it is used for treatment, and may be an active ingredient alone or a mixture of an active ingredient and any ingredient. The shape 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 pharmacy-effective amount. The effective amount may be, for example, an amount sufficient for pharmacy-based stability or delivery of the active ingredient. For example, a buffer solution is effective for stabilizing the active ingredient in a vial. The pharmaceutical composition may also contain a stabilizer (e.g., mannitol), a buffer (e.g., arginine), or a pH adjuster (NaOH). The dosage, administration interval, administration method, and administration route are not particularly limited and can be appropriately selected depending on the age and weight of the patient, symptoms, target organ, and the like. The pharmaceutical composition preferably contains a therapeutically effective amount, or an effective amount of an active ingredient that exerts a desired effect. In one embodiment of the present invention, the therapeutically effective amount includes an amount necessary for clinically observing improvement of symptoms in the patient. In one embodiment of the present invention, pharma- ceutical acceptable includes a state suitable for use in accordance with a reasonable benefit / risk ratio within the scope of reasonable medical judgment. There are no particular limitations on the components other than BCV in the pharmaceutical composition as long as they do not impair the effects of the present invention, and they can be appropriately selected according to the purpose. In the above (1) to (5), the treatment may not include, for example, co-administration with probenecid, uricosuric agents, or drugs that suppress side effects, if specifically specified. Side effects include, for example, diarrhea or nephrotoxicity.

[0074] In the above (1) to (5), the route of administration of BCV, its pharma- ceutically acceptable salt, or solvate thereof, or its pharmaceutical composition to a subject is preferably one that is effective for the treatment, and may be, for example, oral, intravenous, subcutaneous, intramuscular, or intraperitoneal. The administration form is preferably one that is effective for the treatment, and may be, for example, a solid preparation (e.g., tablet), a liquid preparation (e.g., oral suspension), or an injection (e.g., intravenous injection).

[0075] In the above (1) to (5), the dosage, administration interval, and administration method of BCV, its pharma- ceutically acceptable salt, or solvate thereof, or its pharmaceutical composition to a subject can be appropriately selected depending on the age, body weight, symptoms, target organ, and the like of the patient. The dosage 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 1 to 4 weeks. More specifically, it includes intravenous injection of 20 to 80 mg / day. It may also include intravenous injection of 20 to 80 mg / day twice a week. In addition, during combined treatment of BCV and a chemotherapeutic agent (e.g., gemcitabine, etoposide, or an immune checkpoint inhibitor), the dosage of BCV may include intravenous injection of 10 to 40 mg or 10 to 20 mg / day. It may also include intravenous injection of 10 to 40 mg or 10 to 20 mg / day twice a week. The dosage for humans is preferably 10-80 mg / day, twice a week, and the administration route is preferably intravenous injection. In this case, it is possible to minimize side effects while showing excellent therapeutic effects on lymphoma. The range of 10-80 mg shown in this paragraph may be, for example, 10, 20, 30, 40, 50, 60, 70, or 80 mg, or may be within the range of any two of them.

[0076] In one embodiment of the present invention, "significantly" may mean, for example, a state where a statistically significant difference is evaluated using a Student's t-test (one-tailed or two-tailed) and p<0.05 or p<0.01, or a state where a substantial difference is generated.

[0077] All publications cited herein are incorporated by reference in their entirety. In this specification, "or" is used when "at least one or more" of the items listed in the text can be employed. The same applies to "or." When it is stated herein that "within a range of two values," the range includes the two values ​​themselves. In this specification, "A to B" includes A and B. In this specification, "having" includes suffering from a disease when it relates to a disease.

[0078] 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 be adopted. Furthermore, the present invention may be adopted in combination with each of the configurations or features described in the above embodiments, or independently. EXAMPLES

[0079] The present invention will be further explained below with reference to examples, but is not limited to these.

[0080] Example 1: Inhibition of lymphoma cell proliferation Cell viability was measured using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol: lymphoma cells were cultured at 2 × 10 3Cells were seeded in 96-well plates at a concentration of 100 mM, and each well was added with various concentrations of drug (BCV). At each time point, Promega CellTiter-Glo® 2.0 Cell Viability Assay reagent was added to the wells, and after 10 min of incubation at room temperature, the absorbance at 450 nm was measured using a Tecan M200 Infinite 96-well plate reader and IControl software 1.6 (Tecan, Männedorf, Switzerland). Cell viability was evaluated as a percentage of control absorbance. Growth inhibition effects were analyzed by generating dose-response curves with plots of the percentage of viable cells against drug concentrations, and their IC50s were estimated using GraphPad Prism version 8.0.2 (GraphPad Software). All reactions were performed in triplicate. In addition, the EBV-positive / negative status of cells was confirmed by the presence or absence of detection of EBV-derived transcripts (EBER, EBNA1, etc.).

[0081] The results are shown in Figure 1. BCV suppressed the proliferation of NK / T lymphoma cells. Regarding this cell proliferation inhibitory effect, EBV-positive NK / T lymphoma cells (KAI-3, NK-S1) showed significantly lower IC50 values ​​than EBV-negative NK / T lymphoma cells (KHYG-1).

[0082] Example 2: Comparison with antiviral drugs The experimental procedure was the same as in Example 1, except that the drugs used were BCV, acyclovir, ganciclovir, adefovir, foscarnet, and penciclovir.

[0083] The results are shown in Figure 2. BCV showed a significantly superior cell proliferation inhibitory effect against EBV-positive NK / T lymphoma cells (NK-S1) compared with other antiviral drugs (acyclovir, ganciclovir, adefovir, foscarnet, and penciclovir).

[0084] Example 3: In vivo drug treatment For in vivo drug treatment with BCV, 6-week-old female NSG mice were treated with 0.5x10 6The mice were inoculated with NK-S1 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 2000 mm 3 Tumor measurements were recorded twice weekly until tumor mass reached 0.01 mg / kg / day. Mice were euthanized according to IACUC guidelines. Tumor sizes in 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.

[0085] The results are shown in Figures 3A to 3D. The administration of BCV significantly suppressed the growth of EBV-positive NK / T lymphoma. BCV was shown to have a high therapeutic effect in vivo. Furthermore, the animals were in good health and no diarrhea was observed. This suggests that side effect management can be omitted or simplified.

[0086] Example 4: Reduction of EBNA1 and LMP1 Expression NK-S1 and KAI-3 cell lines were treated with 0.1 and 1 μg / mL BCV for 72 hours, and then whole cell lysates were incubated with 4-15% Mini-PROTEAN TM TGX Stain-Free TMThe proteins were separated by SDS-PAGE using Protein Gel (Bio-Rad Laboratories, Hercules, CA, USA) and transferred to a 0.2 μm PVDF membrane (Bio-Rad Laboratories, Hercules, CA, USA). After blocking, the membrane was gently shaken overnight at 4°C in a solution containing 5% nonfat dry milk (Bio-Rad Laboratories, Hercules, CA, USA) or 5% bovine serum albumin (Sigma-Aldrich, Darmstadt, Germany) TBST solution (50 mM Tris / HCl pH 7.4, 150 mM NaCl, 0.1% Tween-20) and primary antibodies (anti-EBNA1 antibody (Santa Cruz), anti-LMP1 antibody (Dako)). Exposure to the appropriate HRP-conjugated 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). ChemiDoc TM XRS+ (Image Lab TM Imaging was performed using a system equipped with software (Bio-Rad Laboratories, Hercules, CA, USA).

[0087] The results are shown in Figure 4. BCV treatment reduced the expression levels of EBNA1 and LMP1 in lymphoma cells.

[0088] Example 5: Reduction of Myc Expression Whole transcript sequencing was performed on NK-S1 and KAI-3 cell lines treated with BCV. Gene set enrichment analysis (GSEA) was performed using the Molecular Signatures Database (MSigDB) Hallmark gene set. 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. Quantitative PCR was performed using Maxima SYBR Green / ROX qPCR Master Mix (Thermo Scientific, USA). The ΔCt of the mRNA expression of a gene was taken as the difference from the Ct value of the internal control GAPDH, and ΔΔCt was calculated by subtracting the ΔCt value of the treated group from the ΔCt value of the untreated control group. Expression ratios were calculated using the formula 2 -ΔΔCt was used to calculate.

[0089] The results are shown in Figure 5A-C. BCV significantly reduced Myc expression and the expression of genes controlled by MYC, while increasing the expression of genes in the STING pathway.

[0090] Example 6: Induction of immunogenic cell death NK-S1 and KAI-3 cell lines were treated with BCV (0.1ug / mL or 1ug / mL) or DMSO alone for 72 hours. For calreticulin staining, cells were incubated with calreticulin antibody (1:100, 30 minutes at room temperature) (#ab92516, Abcam), washed and resuspended in PBS before analysis and analyzed on a cell analyzer (BD LSR Fortessa, BD Biosciences, San Jose, CA, USA) and data were analyzed using FlowJo version 10.8.0 (BD Biosciences, San Jose, CA, USA). Extracellular levels of HMGB1 upon BCV treatment were measured using the Lumit HMGB1 immunoassay (Promega, Madison, WI, USA).

[0091] The results are shown in Figure 6. Typical phenotypes reflecting immunogenic cell death, such as an increase in calreticulin-positive cells and release of HMGB1, were clearly elevated by BCV treatment.

[0092] Example 7: Combined Treatment NK-S1 cell lines were treated with selected doses of etoposide, gemcitabine, or gamma irradiation with or without BCV (0.1 μg / mL). Cell viability was quantified using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol. For the calculation of Combination Index (CI), Chou-Talalay median effect analysis was used to determine synergy, additivity, or antagonism; i.e., CI values ​​<1 or >1 indicate synergy or antagonism, respectively. CI values ​​close to 1 (i.e., between 0.9 and 1.1) were considered additive.

[0093] The results are shown in Figures 7A to 7C. A significant enhancement in efficacy was observed by combined treatment of BCV with the drugs etoposide or gemcitabine compared to treatment with each drug alone. The effect was found to be synergistic based on the combination index value.

[0094] Example 8: Inhibition of proliferation of MYC-positive lymphoma cells Various B-cell lymphoma cell lines were treated with various concentrations of BCV for 96 hours. Cell viability was quantified using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol. The presence or absence of EBV and molecular information were referenced from public databases. The cell lines shown in Figure 8 have been reported to be classified into the following types: Raji and Daudi: Burkitt's lymphoma (BL) cell lines, VAL and DB: diffuse large B-cell lymphoma (DLBCL) cell lines, BJAB: BL or DLBCL cell lines. All are B lymphomas with MYC translocation or amplification. Raji, Daudi, and VAL are EBV-positive, while BJAB and DB are EBV-negative cell lines. VAL is a triple-hit lymphoma, and DB is a double-hit lymphoma.

[0095] The results are shown in Figure 8. BCV exhibited a clear cell proliferation inhibitory effect on MYC-positive lymphoma cells. At this time, it showed a high proliferation inhibitory effect on both EBV-positive and -negative cells. In Example 5, BCV reduced Myc expression, and in Example 8, BCV inhibited the proliferation of MYC-positive lymphoma cells. These results demonstrated that BCV is particularly suitable for the treatment of MYC-positive lymphoma.

[0096] Example 9: Inhibition of proliferation of 11 types of lymphoma cells The effect of BCV on cell viability was examined for 11 NK / T lymphoma cell lines (KAI-3, NK-S1, NK-92, KHYG-1, NK-YS, MEC-04, SNK-1, SNK-6, YT, HANK-1, or SNT-8). Cell viability was quantified using the Promega CellTiter-Glo® 2.0 Cell Viability Assay (Promega, Madison, WI, USA) according to the manufacturer's protocol. Briefly, cells were diluted at 2 × 10 in 100 μL medium. 3Cells were seeded in 96-well plates at a concentration of 0.01 mg / mL and drugs were added at each concentration. After each time point, Promega CellTiter-Glo® 2.0 Cell Viability Assay reagent was added to the wells and incubated at room temperature for 10 min before measuring the absorbance at 480 nm 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 inhibition effects were analyzed by generating dose-response curves as plots of percentage of viable cells against drug concentration, and their IC50s were estimated using GraphPad Prism version 8.0.2 (GraphPad Software). All reactions were performed in triplicate.

[0097] The results are shown in Figure 9. Four cell lines (KAI-3, NK-S1, NK-92, and KHYG-1) showed lower IC50 values ​​and higher susceptibility to BCV than the other cell lines.

[0098] Example 10: Whole-transcriptome sequencing analysis of 11 types of lymphoma cells Whole transcriptome sequencing was performed on the same 11 NK / T lymphoma cell lines as in Example 9. Gene set enrichment analysis (GSEA) was performed using the Molecular Signatures Database (MSigDB) Hallmark gene set to compare the top four BCV-susceptible cell lines (KAI-3, NK-S1, NK-92, and KHYG-1) with the other seven cell lines. Gene sets were considered to be significantly altered if the False Discovery Rate (FDR) q value of the normalized enrichment score (NES) was less than 0.05.

[0099] The results are shown in Figures 10A and 10B. In the figures, MYC TARGETS V2, which showed a significant increase, is a group of genes regulated by MYC. As a result of the analysis, a group of genes that were characteristically highly expressed in the four sensitive lines (KAI-3, NK-S1, NK-92, KHYG-1) compared to the other seven lines was found, and among them, the high expression was remarkable in the group of genes regulated by MYC. It was also shown that the higher the expression of MYC, the higher the antitumor effect of BCV.

[0100] Example 11: Impact on prognosis The progression-free survival and overall survival were investigated for a cohort of human NK / T-cell lymphoma patients (n=36). Furthermore, based on the results of genetic analysis, the patient cohort was divided into group A (n=19) and group B (n=17). The enrolled patients were those who had not received BCV treatment. The results are shown in Figures 11A and 11B. Comparing group A and group B, group A showed a good prognosis, while group B showed a tendency toward a poor prognosis. Statistical analysis was performed using MedCalc for Windows, version 18.2.1 (MedCalc Software).

[0101] Furthermore, based on the results obtained from whole-transcriptome sequencing in the above human patient cohort (n=36), we performed gene set enrichment analysis (GSEA) using the Molecular Signatures Database (MSigDB) Hallmark gene sets to compare the good and bad prognosis groups. Gene sets were considered to be significantly altered if the False Discovery Rate (FDR) q value of the normalized enrichment score (NES) was less than 0.05.

[0102] The results are shown in FIG. 12. In the figure, MYC TARGETS V1 and MYC TARGETS V2, which showed a significant increase, are gene groups regulated by MYC. As a result of the analysis, the gene group regulated by MYC was significantly more highly expressed in the poor prognosis group than in the good prognosis group. Here, as described in Example 5, BCV significantly reduces Myc expression and also significantly reduces the expression of the gene group controlled by MYC. Therefore, it is considered that administration of BCV has the effect of improving prognosis.

[0103] 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 the treatment of malignant lymphoma, comprising blinatumomab, a pharmaceutically acceptable salt thereof, or a solvate thereof.

2. The pharmaceutical composition according to claim 1, comprising blinatumomab.

3. The pharmaceutical composition according to claim 1, wherein the malignant lymphoma is NK / T cell lymphoma or B cell lymphoma.

4. The pharmaceutical composition according to claim 1, wherein the malignant lymphoma is NK / T cell lymphoma.

5. The pharmaceutical composition according to claim 1, which reduces the MYC expression of the malignant lymphoma cells to be treated.

6. The pharmaceutical composition according to claim 1, wherein the treatment includes identifying EBV-positive subjects as treatment targets.

7. The pharmaceutical composition according to claim 1, wherein the malignant lymphoma is EBV-positive lymphoma.

8. The pharmaceutical composition according to claim 1, wherein the treatment includes detecting the expression level of EBV in a subject's sample.

9. The pharmaceutical composition according to claim 1, wherein the treatment includes identifying MYC-positive subjects as treatment targets.

10. The pharmaceutical composition according to claim 1, wherein the malignant lymphoma is MYC-positive lymphoma.

11. The pharmaceutical composition according to claim 1, wherein the treatment includes detecting the expression level of MYC in a subject's sample.

12. The pharmaceutical composition according to claim 1, wherein the treatment includes identifying EBV-positive and MYC-positive subjects as treatment targets.

13. The pharmaceutical composition according to claim 1, wherein the malignant lymphoma is EBV-positive and MYC-positive.

14. The pharmaceutical composition according to claim 1, wherein the treatment includes detecting the expression levels of EBV and MYC in a sample of a subject.

15. The pharmaceutical composition according to any one of claims 1 to 14, which improves prognosis.

16. The pharmaceutical composition according to claim 1, which suppresses the proliferation of malignant lymphoma cells.

17. The pharmaceutical composition according to claim 1, for use in combination therapy with brincidofovir, a pharmaceutically acceptable salt thereof, or a solvate thereof and a chemotherapeutic agent.

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