JP7217875B2 - Preventive and/or therapeutic agent for blood cancer - Google Patents

Description

Application of Article 30, Paragraph 2 of the Patent Law 1. Published by the publication of conference abstracts (Journal of the Japan Endocrine Society) Society name: The 91st Annual Meeting of the Japan Endocrine Society Publisher: Japan Endocrine Society Secretariat Publication date: April 1, 2018 2. Publication by academic conference presentation Name of conference: The 91st Annual Meeting of the Japan Endocrine Society Publication date: April 26, 2018 Date: April 26-28, 2018

TECHNICAL FIELD The present invention relates to an anticancer agent containing an SGLT2 inhibitor.

Cancer is said to be a disease that afflicts one in two Japanese people every year. Therefore, there is a need for cancer treatment, management and prevention.
In addition, various anticancer agents have been marketed and developed, but they are still under development. That is, current anticancer agents have problems such as difficulty in complete cure, individual differences in efficacy, and large side effects, and further development or improvement of anticancer agents is desired.

Cancer cells create an environment that is advantageous for their growth and metastasis, and acquire traits that allow them to survive even in poor environments such as low nutrition and low oxygen. The Warburg effect, proposed more than 90 years ago, is a phenomenon in which cancer cells rely more on glycolytic adenosine triphosphate (ATP) production than on mitochondrial oxidative phosphorylation, even in an aerobic environment. , the details of the molecular mechanism have not yet been fully elucidated.
In cancer cells, combined with a decline in mitochondrial function, sugar metabolism is shifted to the glycolytic system, which is less efficient for energy production, so sugar uptake is more active than in normal cells. FDG-PET examination using this characteristic is widely used for cancer localization diagnosis and metastasis diagnosis (Non-Patent Document 1). It has also been reported that PET examination is useful for stratifying the malignancy of adult T-cell leukemia (hereinafter also referred to as "ATL"), which is an extremely intractable blood cancer (Non-Patent Document 2). .

Adult T-cell leukemia (ATL) is caused by infection with HTLV-1, a type of virus. In the world, the southwestern part of Japan (Kyushu and Okinawa regions) has the highest number of HTLV-1 infected people, and there are about 1.1 million people infected with HTLV-1 in Japan, mainly in the coastal areas of southwestern Japan. 0.6 to 0.7 people. Due to the long incubation period from infection to onset, the lifetime probability of developing HTLV-1 infection is about 5%. prognosis is poor due to

The development of anti-cancer drugs that target the glycolysis system in cancer cells has been attempted so far. Although some drugs are being used (Non-Patent Document 1), they have not yet been applied to clinical practice.

On the other hand, sodium-glucose cotransporter 2 (hereinafter also referred to as “SGLT2”) inhibitors are antidiabetic agents that inhibit glucose reabsorption from the proximal renal tubules and promote urinary glucose excretion, thereby reducing blood sugar levels. Correct the value. SGLT2 inhibitors are therapeutic drugs that have fewer side effects and have different effects from drugs that act on insulin secretion.
These SGLT2 inhibitors are known to have antitumor effects against certain types of cancer (Non-Patent Document 3), and dapagliflozin, canagliflozin, and tofogliflozin have been used in cancer model mice and various Its action on cancer cells has been reported (Non-Patent Documents 4 to 8).

For example, Non-Patent Document 4 reports that dapagliflozin and canagliflozin have anticancer effects in pancreatic cancer and prostate cancer transplantation model mice.
Non-Patent Document 5 reports that dapagliflozin has anticancer effects in human renal cancer cell lines and renal cancer cell line-derived mouse xenograft models.
Non-Patent Document 6 reports that canagliflozin has an anticancer effect in a human liver cancer cell line and a human liver cancer cell line-derived mouse xenograft model.
Non-Patent Document 7 reports that canagliflozin has an anticancer effect on human prostate cancer cell lines, lung cancer cell lines, liver cancer cell lines, and breast cancer cell lines. In addition, although its activity is weak, anticancer activity in colon cancer cell lines and ovarian cancer cell lines, and anticancer activity in mouse xenograft models derived from human prostate cancer cell lines have also been reported.
Non-Patent Document 8 reports that tofogliflozin has a preventive effect on the development of liver cancer in diabetic mice.
Although these documents report that SGLT2 inhibitors act on various cancers, there is no report on blood cancer.

In addition, regarding the combined use of an SGLT2 inhibitor and an anticancer agent, Non-Patent Document 4 reports that combined use of canagliflozin and gemcitabine has an effect on pancreatic cancer.
Non-Patent Document 7 reports the effects of combined use of canagliflozin and docetaxel or radiotherapy on prostate cancer.
Patent Document 1 reports the effects of combined use of canagliflozin and docetaxel, cisplatin, or radiotherapy on prostate cancer and lung cancer.
Patent document 2 reports combination therapy with tapagliflozin and glufosfamide.
However, in these documents, there is no report on the combined use of luseogliflozin and an anticancer drug, and there is no report on combination therapy for blood cancer.

International Publication No. 2016/134486 Pamphlet JP 2015-514756 A

Oncogene, 2006, 25, 4633-4646. Nakachi S et al. Hematology 22:536, 2017 Pharmacol. Ther., 2017, 170, 148-165. Proc. Natl. Acad. Sci. USA, 2015, 112, E4111-E4119. Med. Sci. Monit., 2017, 23, 3737-3745. International Journal of Cancer, 2018, 142, 1712-1722. Molecular Metabolism, 2016, 5, 1048-1056. ObaraK, et al. Oncotarget.2017i8:58353

An object of the present invention is to provide a new drug that has anti-blood cancer activity and few side effects.

As a result of intensive studies to solve the above problems, the present inventors have focused on the fact that cancer strongly requires glucose uptake for growth, and have obtained the following findings. , found to be useful in the treatment of hematologic cancers.
(1) High expression of the SGLT2 gene in blood cancer cells.
(2) Luseogliflozin, an SGLT2 inhibitor, suppresses glucose uptake in blood cancer cells.
(3) Luseogliflozin, an SGLT2 inhibitor, suppresses ATP production in blood cancer cells.
(4) luseogliflozin, which is an SGLT2 inhibitor, exerts an effect of suppressing cell proliferation in blood cancer cells;

Furthermore, the inventors have found that the combined use of luseogliflozin and an anticancer drug enhances the effect of suppressing proliferation of blood cancer cells, and have completed the present invention.

That is, the present invention
(1) a preventive and/or therapeutic agent for blood cancer, characterized by containing luseogliflozin;
(2) The preventive and/or therapeutic agent for blood cancer according to (1), which is characterized by a combination of luseogliflozin and an anticancer agent;
(3) the preventive and/or therapeutic agent for blood cancer according to (1) or (2), wherein the blood cancer is selected from the group consisting of malignant lymphoma, leukemia and multiple myeloma;
(4) The preventive and/or therapeutic agent for blood cancer according to any one of (1) to (3), wherein the blood cancer is adult T-cell leukemia;
is.

Luseogliflozin can suppress the proliferation of hematological cancer cells, and by using it for the prevention and/or treatment of hematological cancer, it is possible to expand the range of cancer treatment options. In addition, such a drug can be used as a safe therapeutic drug with few side effects, and can be a drug having a new action different from conventional anti-blood cancer therapeutic drugs.
Furthermore, it has been suggested that the combined use of luseogliflozin with conventionally used anticancer agents has a therapeutic enhancing effect, and is extremely useful in that it can provide a new treatment method. .

1 is a graph showing the effect of glucose concentration in a culture medium on growth of ATL cell line (MT-2) in Test Example 1. FIG. 4 is a graph showing SGLT2 mRNA expression in Test Example 2. FIG. 2 is a graph showing the effect of luseogliflozin on glucose uptake in Test Example 3. FIG. 2 is a graph showing the effect of luseogliflozin on ATP production in Test Example 4. FIG. 10 is a graph showing the cell viability of ATL cell lines with luseogliflozin at 50 μM or 100 μM in Test Example 5 over time. 10 is a graph showing the cell viability of ATL cell lines when cultured for 48 hours at each etoposide concentration and luseogliflozin concentration in Test Example 6. FIG. 10 is a graph showing the effects of 0.2 μM etoposide, 50 μM luseogliflozin, and the combined use of 0.2 μM etoposide and 50 μM luseogliflozin on the cell viability of ATL cell lines in Test Example 6. FIG.

Luseogliflozin in the present invention is a kind of SGLT2 inhibitor. The luseogliflozin used in the present invention is not particularly limited as long as it can exert an SGLT2 inhibitory effect, and various chemical forms can be used. For example, luseogliflozin itself or a hydrate thereof can be used.
SGLT2 inhibitors inhibit sodium-glucose exchange in sodium-glucose cotransporter 2 (SGLT-2) and excrete excess blood glucose in the urine. It is a drug that lowers blood sugar levels in the body and suppresses increases in blood glucose concentration. As a result, SGLT2 inhibitors can reduce the burden on exhausted pancreatic islets of Langerhans β-cells by lowering blood glucose levels and restore their secretory capacity. In addition, SGLT2 inhibitors improve insulin resistance by improving glucose toxicity by improving blood sugar. Luseogliflozin is marketed as a non-insulin dependent diabetes drug.

The anticancer agent in the present invention is a drug intended to suppress the growth of cancer, and includes chemotherapeutic agents, molecular target therapeutic agents, hormone therapy agents, immunotherapeutic agents, etc., which are intended to suppress cancer. It includes drugs with any mechanism of action.
Chemotherapeutic agents include various types such as alkylating agents, platinum compounds, antimetabolites, topoisomerase inhibitors, microtubule inhibitors, and antibiotics, but are not particularly limited in the present invention.
Since the present invention is intended for blood cancer, the anticancer agent is preferably a drug that is effective against blood cancer. Purine, methotrexate, epirubicin hydrochloride, etoposide, vincristine, interferon α, imatinib, dasatinib, nilotinib, ofatumumab, retinoin, revlimid, mogalizumab, etc., but from the perspective of targeting leukemia and malignant lymphoma where cancer cells proliferate rapidly Cyclophosphamide, cytarabine, epirubicin hydrochloride, etoposide, and vincristine, which are cytotoxic anticancer agents that inhibit DNA synthesis, are preferred from the standpoint of high frequency of concomitant use and a certain degree of antitumor effect even when used alone. Etoposide is preferred because it is the best in

The blood cancer in the present invention includes leukemia, malignant lymphoma and multiple myeloma, which are said to be the three major cancers of blood.
Leukemia is classified into acute and chronic according to its course, and into myeloid and lymphatic according to the origin of cancer cells. In acute cases, the symptoms progress rapidly, and early treatment is necessary because the symptoms appear suddenly. Myeloid cells become malignant due to the occurrence of genetic mutations in myeloblasts differentiated from myeloid stem cells. Lymphoid cells become malignant at an immature stage. As a result, cancerous cells (leukemia cells) proliferate indefinitely and develop symptoms.
Acute lymphocytic leukemia (ALL) is an acute and lymphocytic leukemia among the above-mentioned leukemias, and is roughly divided into B cell lineage and T cell lineage according to the type of cancerous cells. Acute lymphoblastic leukemia cells (ALL cells) grow primarily in the bone marrow, but also in the blood. Symptoms are systemic, and symptoms associated with decreased hematopoietic function due to proliferation of leukemia cells in the bone marrow include anemia, bleeding, and infection. Symptoms associated with the infiltration of leukemia cells into organs include liver and Swelling of organs such as the spleen, lumbago, joint pain, headache, nausea/vomiting, lymphadenopathy, etc. Since ALL cells are particularly prone to infiltrate the central nervous system, attention should be paid to headache, nausea, and vomiting. Lymphoblastic lymphoma is one in which leukemia cells proliferate mainly in lymphoid tissues such as lymph nodes.
Malignant lymphoma is classified into more than 30 types according to the morphology and properties of cancer cells, but it is broadly divided into Hodgkin's lymphoma and non-Hodgkin's lymphoma. is also included.
Multiple myeloma develops when plasma cells differentiated from B cells, a type of white blood cell, among the blood cells produced in the bone marrow become cancerous and become myeloma cells.

Adult T-cell leukemia (ATL) is a type of virus in which HTLV-1 (human T-lymphotropic virus type-I) infects T cells, which are white blood cells, and transforms the infected T cells into cancerous cells (ATL It develops when cells (cells) proliferate indefinitely. HTLV-1-infected T cells can infect others through direct contact with normal lymphocytes, but the disease affects only a minority of infected individuals and has an incubation period of approximately 30-50 years.
ATL is classified into malignant lymphoma of lymphocytic leukemia or non-Hodgkin's lymphoma among the three major cancers. Since ATL cells proliferate not only in the blood but also in the lymph nodes, swelling of the lymph nodes is often observed, but the spread of the lesions is systemic. Disease types are classified into acute type, lymphoma type, chronic type, and smoldering type, but are not particularly limited in the present invention.

From the viewpoint that the hematological cancer in the present invention is intractable and a new therapeutic strategy is desired, it is preferably lymphocytic cancer, more preferably adult T-cell leukemia or acute lymphocytic leukemia, and still more preferably. is adult T-cell leukemia.
In addition, the prophylactic and/or therapeutic agent for blood cancer of the present invention can also function as a growth inhibitor for blood cancer cells, an agent for reducing ATP production in blood cancer cells, and a sugar uptake inhibitor for blood cancer cells. .
The preventive and/or therapeutic agent for hematological cancer has a feature of exerting an effect on the cancer cells of hematological cancers, particularly adult T-cell leukemia or acute lymphoblastic leukemia. Effective in cancer cells in leukemia. Cancer cells (ATL cells) in adult T-cell leukemia include cells obtained from peripheral blood, bone marrow, and ATL cell lines of ATL patients. An ATL cell line is a subcultured ATL cell, and stably has the characteristics of an ATL cell. Cancer cells in acute lymphoblastic leukemia (ALL cells) include cells obtained from peripheral blood of ALL patients and ALL cell lines. ALL cell lines are cells obtained by subculturing ALL cells, and stably have the characteristics of ALL cells.

In the present invention, the prevention and/or treatment of cancer means suppressing the proliferation of cancer cells, reducing the dose of cancer therapeutic agents and therapeutic adjuvants, suppressing and stabilizing various symptoms caused by cancer. Improving the quality of life of cancer patients, including all medical treatments related to cancer, such as the suppression of cancer onset, recurrence, and progression, the reduction of side effects in cancer treatment, and the prolongation of survival of cancer patients. Also includes
Suppression of cancer development refers to suppressing the onset of cancer in advance when it develops due to external factors such as viral infection. Suppression of cancer recurrence refers to patients whose cancer cells or lesions have disappeared due to treatment or resection. Suppressing the recurrence of cancer in advance and suppressing the progression of cancer mean delaying or reversing the progression of cancer.

The prophylactic and/or therapeutic agent according to the present invention is generally administered systemically or locally, orally or parenterally. The dose varies depending on age, body weight, symptoms, therapeutic effect, administration method, treatment time, etc., and the dose varies depending on various conditions.

The "preventive and/or therapeutic agent characterized by a combination" according to the present invention is a single formulation (combination formulation) of luseogliflozin and an anticancer agent, which are active ingredients, or two types obtained by separately formulating It can be a formulation of
The above preparations can be made into tablets, granules, powders, capsules, emulsions, suspensions, syrups, etc., or injections such as sterile solutions, suspensions, etc., according to commonly practiced means. When these active ingredients are formulated separately to form two formulations, the individual formulations can be administered simultaneously or separately.

When the prophylactic and / or therapeutic agent according to the present invention is made into two or more formulations different for each active ingredient, it is likely to be administered at the same time or at extremely short intervals (continuously). It is preferable to state in the package inserts and sales pamphlets of the medicines that are used in combination that they are used together. It is also preferable to prepare a kit mainly composed of a combination of luseogliflozin and an anticancer drug.

The dose of luseogliflozin of the present invention varies depending on the subject of administration, administration method, etc. For example, in the case of oral administration, luseogliflozin is 0.1 to 50 mg per day per patient (60 kg), preferably It is preferable to administer 0.5 to 5 mg, more preferably 0.5 to 2.5 mg.
On the other hand, the daily dose of anticancer drugs combined with luseogliflozin is explained by giving an example. Although 200 mg can be administered, it is preferable to administer 10 to 100 mg because of side effects such as bone marrow suppression, and more preferably 25 to 50 mg because malignant lymphoma is often taken orally for 3 consecutive weeks for one course. . In the case of etoposide (injection), a daily dose of 1 to 150 mg can be administered per patient's body surface area (1 m ² ). Since it is often used in combination with other anticancer agents, it is preferable to administer 10 to 60 mg.
This amount is the amount indicated in the package insert for doctors or an amount less than the dose, but the reason why even a small amount can be expected to be effective is that multiple drugs can be used in combination and that it is time-dependent, not dose-dependent. This is because it is effective for
Therefore, anticancer agents other than etoposide can be expected to be effective even in the amount indicated in the package insert for doctors or in an amount smaller than the dose, as in the case of etoposide.
As a result, the present invention is able to exert its effect at a dose lower than the dose normally used as an anticancer drug, and is expected to be effective even at a dose lower than that instructed by a doctor's package insert, for example, 25 to 50 mg of etoposide. can.

The compounding ratio of luseogliflozin of the present invention and the anticancer agent varies depending on the type of drug, the subject of administration, the administration method, etc. A combination of 3.0×10 ^-5 to 8.0×10 ^-3 parts by weight of an anticancer agent per part by weight can provide a superior anti-blood cancer effect compared to administration of individual drugs. It is possible.
In particular, when the anticancer agent is etoposide, it is preferable to combine 22.0×10 ⁻³ to 4.0×10 ⁻³ parts by mass of etoposide with 1 part by mass of luseogliflozin. As a result, a sufficient effect can be obtained with a smaller amount than when each drug is administered alone. In addition, it is possible to produce a drug with few side effects.

The prophylactic and/or therapeutic agent according to the present invention is used together with other drugs (including other SGLT inhibitors other than luseogliflozin, or anticancer agents) for the prevention and/or treatment of blood cancer. They may be used in combination, or may be used in combination with other treatments (radiation therapy, etc.).
In this case, the timing of administration and the timing of treatment of the pharmaceutical of the present invention and other agents are not limited, and they may be administered to the subject at the same time or at different times. Furthermore, the prophylactic and/or therapeutic agent according to the present invention and other agents may be administered as different formulations, or as a single formulation containing all active ingredients.
The dosage of other drugs can be appropriately selected based on clinically used dosages. Moreover, the compounding ratio of the prophylactic and/or therapeutic agent according to the present invention and other agents can be appropriately selected depending on the subject of administration, administration route, target disease, symptom, combination, and the like.
Similarly, when used in combination with other treatments, the timing of administration and the timing of treatment are not limited, and may be performed simultaneously or with a time lag.

EXAMPLES The present invention will be described in more detail below with reference to Examples and the like, but the present invention is not limited to these Examples and the like.

<<Test Example 1. Evaluation of cell proliferation ability by difference in medium glucose concentration in ATL cell lines>>
The aim of this study was to investigate whether changes in cell growth ability were observed by changing the glucose concentration in the medium using ATL cell lines.

<Experimental method>
1. ATL cell line MT-2 (provided by Dr. Takuya Fukushima, Department of Hematology, Faculty of Health Sciences, University of the Ryukyus) was cultured in two types of media with different glucose concentrations (medium: RPMI1640, RPMI1640 Hybri-Max, culture conditions : 37° C., 5% CO ₂ T25-flask, cell number: 3.9×10 ² ). Each glucose concentration was adjusted to 11 mM for normal concentration and 25 mM for high concentration, and cultured for 72 hours.
2. Cell counts (WST-8 method, Cell counting kit-8, Dojindo Laboratories, CK04) were performed 24 hours, 48 hours, and 72 hours after culture. Cell count was performed by measuring absorbance at 450 nm using a microplate reader (TEKAN, Spark (registered trademark)).

<Experimental results>
1. The results are shown in FIG.
(1) The number of cells increased over time in both normal and high glucose concentrations.
(2) In addition, at all time points, the number of cells increased significantly at high concentration compared to normal concentration, and the difference was significant.
2. These results indicated that the proliferation of ATL cells was promoted under high glucose concentration.

<<Test Example 2. Evaluation of SGLT2 mRNA expression in ATL cell lines, etc.>>
Using ATL cell lines and peripheral blood of ATL patients as samples, RT-PCR was used to examine the expression of SGLT2mRNA.

<Experimental method>
1. Each cell (MT-2, ATL acute type and B-ALL cell number: 1-6×10 ⁶ , PBML was isolated from 40 ml of peripheral blood using Ficoll. Human colon cancer RNA was purchased from OriGene Technologies and used. RNA was extracted from the cells), and nucleic acid amplification was performed by RT-PCR (using Applied Biosystems StepOne Plus, 95°C, 10 minutes → 40 cycles (95°C, 15 seconds → 60°C, 1 minute)).
2. SGLT2mRNA was calculated by dividing the amplified nucleic acid of β-actin as an endogenous control in each cell.

<Experimental results>
1. The results are shown in FIG. 2 (positive control is set to 1, RQ=relative quantification).
In FIG. 2, MT-2 is an ATL cell line, PBML is peripheral blood-derived healthy human mononuclear cells (negative control), human colon cancer is human colon cancer cells (positive control), and ATL acute type is peripheral blood ATL cells. , B-ALL indicates acute lymphocytic leukemia cells (peripheral blood).
(1) No increase in SGLT2 mRNA expression was observed in the negative control PBML. A clear increase in the expression level of SGLT2mRNA was observed in human colon cancer, a positive control (expression was observed in human colon cancer as a positive control, and expression was low in PBML, a negative control).
(2) On the other hand, in MT-2, ATL acute type, and B-ALL, SGLT2 mRNA was clearly increased (expressed in ATL acute type, B-ALL, and MT-2 compared to the negative control). .).
2. From these results, the following was considered.
(1) In ATL samples, etc., it is speculated that SGLT2 promotes glucose uptake into cells.
(2) Combined with the results of Test Example 1, it can be considered that in ATL, cell proliferation is more active due to promotion of glucose uptake into cells.

<<Test Example 3. Effects of SGLT2 inhibitors on glucose uptake in ATL cell lines>>
The purpose of this study was to investigate the effects of luseogliflozin, an SGLT2 inhibitor, on glucose uptake using ATL cell lines.

<Experimental method>
1. In the presence of luseogliflozin at various concentrations, the ATL cell line MT-2 was cultured in two types of media with different glucose concentrations under the same conditions as in Test Example 1.
2. At each time point, intracellular glucose uptake was measured using a glucose uptake kit (Glucose Uptake-Glo Assay, Promega, J1341).

<Experimental results>
1. The results are shown in FIG.
(1) Luseogliflozin reduced glucose uptake in a concentration-dependent manner at both normal and high glucose concentrations.
(2) Among these conditions, high glucose concentration (high) and luseogliflozin 100 μM significantly decreased.
2. From the results of Test Examples 3 and 1, luseogliflozin was used to reduce glucose uptake in ATL cells, thus presuming suppression of ATL cell proliferation.

<<Test Example 4. Effects of SGLT2 inhibitors on ATP production in ATL cell lines >>
The purpose of this study was to examine the effects of luseogliflozin, an SGLT2 inhibitor, on intracellular ATP production in ATL cell lines.

<Experimental method>
1. In the presence of luseogliflozin at each concentration, ATL cell line MT-2 was cultured under the same conditions as in Test Example 1 except for the medium adjusted to a glucose concentration of 11 mM under the same conditions as in Test Example 1.
2. Forty-eight hours after the culture, the intracellular ATP concentration was measured using an ATP measurement kit (CellTiter-Glo (registered trademark) 2.0 Assay, Promega, G9241) using a luminescence method.

<Experimental results>
1. The results are shown in FIG.
(1) In the presence of luseogliflozin, both 50 μM and 100 μM, the amount of ATP was significantly decreased compared to the control.
(2) In addition, the decrease in ATP decreased in a luseogliflozin concentration-dependent manner.
2. These results confirmed that addition of luseogliflozin reduced intracellular ATP production in ATL cells.

<<Test Example 5. Cell growth inhibitory effect by SGLT2 inhibitor >>
The purpose of this study was to investigate the effects of luseogliflozin, an SGLT2 inhibitor, on cell proliferation in ATL cell lines.

<Experimental method>
1. In the presence of luseogliflozin at each concentration, ATL cell line MT-2 was cultured under the same conditions as in Test Example 1 except for the medium adjusted to a glucose concentration of 11 mM under the same conditions as in Test Example 1.
2. Cell counting (WST-8 method, Cell counting kit-8, Dojindo Laboratories, CK04) at each time point (24 hours, 48 hours, 72 hours) was performed in the same manner as in Test Example 1.

<Experimental results>
1. FIG. 5 is a diagram showing the cell viability at each time point at luseogliflozin concentrations of 50 μM and 100 μM, in comparison with that in the absence of luseogliflozin.
(1) In the absence of luseogliflozin, the cell viability was nearly 100% in all of the studies.
(2) On the other hand, in the presence of luseogliflozin, cell viability was significantly reduced at all time points compared to that in the absence of luseogliflozin, and the reduction was concentration-dependent. was targeted.
(3) In addition, in the presence of luseogliflozin, cell viability decreased over time.

2. From these results, it was confirmed that the addition of luseogliflozin inhibited the proliferation of ATL cells.
The following mechanism is conceivable for this proliferation suppression.
(1) Increased SGLT2 mRNA expression in ATL cells.
(2) Suppression of glucose uptake by luseogliflozin.
(3) decrease in intracellular ATP production by luseogliflozin;
(4) Suppression of ATL cell proliferation by luseogliflozin.

<<Test Example 6. Investigation of anticancer drug effect in ATL cell line when SGLT2 inhibitor is used in combination >>
Using etoposide as an anticancer agent and luseogliflozin as an SGLT2 inhibitor, we investigated the effects of anticancer agents on ATL cell lines.

<Experimental method>
1. In the presence of various concentrations of etoposide and luseogliflozin, using only the medium adjusted to a glucose concentration of 11 mM under the same conditions as in Test Example 1, other conditions were the same as in Test Example 1 (however, the number of cells was 5.4 × 10 ⁴ ). ) ATL cell line MT-2 was cultured.
2. 48 hours after culturing, cell count (WST-8 method, Cell counting kit-8, Dojindo Laboratories, CK04) was performed in the same manner as in Test Example 1 (however, the number of cells was 5.4×10 ⁴ ). .

<Experimental results>
1. FIG. 6 shows the results of cell viability after 48 hours of culture in the presence of etoposide or luseogliflozin at each concentration.
(1) The cell viability of ATL cells decreased in a dose-dependent manner with etoposide, confirming that etoposide exhibits anticancer effects.
(2) At low etoposide concentrations of 0.1 to 0.2 μM, luseogliflozin at 30 μM and 50 μM combined with etoposide showed a lower cell viability than etoposide alone. In particular, at an etoposide concentration of 0.2 μM, both 30 μM and 50 μM combined with luseogliflozin showed a significant decrease in cell viability compared to etoposide alone.

2. FIG. 7 is a diagram showing the results of cell viability 48 hours after culturing with 0.2 μM etoposide, 50 μM luseogliflozin, and a combination thereof, in comparison with the control.
(1) At 0.2 μM etoposide, no decrease in cell viability was observed compared to the control.
(2) On the other hand, luseogliflozin 50 μM significantly decreased the cell viability compared to the control.
(3) Furthermore, when etoposide 0.2 μM, luseogliflozin 50 μM, or both of them were used in combination, the cell viability was decreased compared to when each was used alone, and both of them were significantly decreased.

3. From these results, etoposide 0.1-0.2 μM was effective in combination with luseogliflozin.
4. When combined with SGLT2 inhibitors, low doses of topoisomerase 2 inhibitors, which are indicated for malignant lymphoma and acute leukemia, suppressed proliferation of ATL cells.

INDUSTRIAL APPLICABILITY According to the present invention, it is possible to provide a preventive and/or therapeutic agent for blood cancer that is safer and has fewer side effects, and can be used as a useful agent for appropriate treatment, extension of survival period, improvement of quality of life, and the like.

Claims (4)

A prophylactic and/or therapeutic agent for blood cancer, characterized by containing luseogliflozin. 2. The preventive and/or therapeutic agent for blood cancer according to claim 1, which is a combination of luseogliflozin and an anticancer agent. 3. The preventive and/or therapeutic agent for blood cancer according to claim 1 or 2, wherein the blood cancer is selected from the group consisting of malignant lymphoma, leukemia and multiple myeloma. The preventive and/or therapeutic agent for blood cancer according to any one of claims 1 to 3, wherein the blood cancer is adult T-cell leukemia.

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