Leukemia cell growth inhibitor containing FKI-7550A substance and FKI-7550B substance as active ingredients

Abstract

To provide a novel leukemia cell-growth inhibitor.SOLUTION: There is provided, a FKI-7550A substance represented by a following formula, or a salt thereof, or a hydrate or solvate thereof, produced by the filamentous fungus Fusarium sp. FKI-7550 strain.SELECTED DRAWING: None

Description

Technical Field

[0001] The present invention relates to the field of leukemia cell growth inhibitors.

Background Art

[0002] Acute lymphoblastic leukemia (ALL) is an aggressive leukemia characterized by the presence of excessive lymphoblasts or lymphocytes in the bone marrow and peripheral blood. It can spread to other organs in addition to lymph nodes, spleen, liver, and central nervous system. ALL often progresses rapidly if left untreated. The causes of acute lymphoblastic leukemia are said to include a high incidence rate among atomic bomb survivors, association with exposure to ionizing radiation and benzene, and infection by Epstein-Barr virus or c-type RNA virus. However, in most cases, the exact cause is unknown.

[0003] The incidence rate of ALL is said to be high in Europe and North America and slightly lower in Asia and Africa. In Japan, the mortality rate for leukemia as a whole is 4 to 5 per 100,000 people per year, and the number of ALL patients is high among children and the elderly over 60 years old. It is said that 3.8 out of 100,000 children under 15 years old are affected. That is, approximately 800 to 900 children develop the disease each year.

[0004] The treatment of ALL is broadly classified into surgery, radiation therapy, and chemotherapy. However, in ALL, leukemia cells exist throughout the body by riding on the blood flow, so chemotherapy that administers anticancer drugs systemically by oral administration or intravenous drip is the first choice. The treatment process is divided into induction therapy, which is the first treatment, consolidation therapy that follows, and maintenance therapy that continues the treatment thereafter. However, for types with a high risk of recurrence, bone marrow transplantation therapy is considered according to the situation.

[0005] The remission induction therapy is carried out with the aim of sufficiently reducing leukemia cells so that the proportion of leukemia cells in the bone marrow is 5% or less. In addition, it aims for a remission state in which the peripheral blood and bone marrow normalize, the hematopoiesis of normal white blood cells, red blood cells, and platelets recovers, no leukemia cells are found in the blood flowing in the body, and white blood cells and platelets recover. The therapeutic drugs (anticancer drugs) used for ALL are adrenal cortical steroids (prednisolone) and vincristine, to which daunorubicin, asparaginase, and cyclophosphamide are added.

[0006] The consolidation therapy is a therapy that further reduces the leukemia cells that still remain after the remission induction and prevents recurrence. It is a treatment with strong anticancer drugs, and in addition to the above-mentioned anticancer drugs, methotrexate, cytarabine, etc. are used.

[0007] If complete remission persists at the end of the remission induction therapy and the consolidation therapy, maintenance therapy is performed for 1 to 2 years thereafter. It is said that the recurrence rate increases if this maintenance therapy is not carried out. Usually, mercaptopurine and methotrexate are used as anticancer drugs.

[0008] However, many of these anticancer drugs cause necrosis not only of cancer cells but also of normal cells and normal tissues. It is difficult to optimally set the dosage and usage period, etc. of such anticancer drugs that cause necrosis of cells and tissues, and furthermore, their side effects are a great burden on cancer patients.

[0009] In recent years, in the field of anticancer drug research, research on selective leukemia cell growth inhibitors has been actively conducted. If leukemia cells can be specifically growth-inhibited, it will be an anticancer drug with few side effects without damaging the functions that play important roles in the formation of other tissues and organs and the maintenance and defense of the body's homeostasis. In order to specifically inhibit the growth of leukemia cells, it is necessary to show growth inhibition specifically for leukemia cells without inhibiting the growth of solid cancer cell lines. Therefore, the investigation of the inducer substances that specifically cause growth inhibition of leukemia cells is very important for the prevention and treatment of leukemia.

[0010] So far, substances such as tolypoalbin, F-14329 (Non-Patent Document 1), PKS-NRPS metabolites (Non-Patent Document 2), and Epicoccarine have been reported, but no relationship between these substances and leukemia cells has been reported at all.

Prior Art Documents

Non-Patent Documents

[0011]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Summary of the Invention

Problems to be Solved by the Invention

[0012] ALL is one type of leukemia, which is called blood cancer. Cancer cells including leukemia stem cells develop through multi-step gene abnormalities, and drugs that specifically suppress the growth of leukemia cells are expected to cause the cancerous leukemia cells to undergo cell death. Therefore, an object of the present invention is to provide such a novel leukemia cell growth inhibitor.

Means for Solving the Problems

[0013] Therefore, the present inventors searched for substances that specifically exhibit growth inhibitory activity against human acute promyelocytic leukemia-derived cell lines, human T-cell leukemia-derived cell lines, and human acute monocytic leukemia-derived cell lines, targeting the metabolites produced by microorganisms. As a result, the present inventors found that a substance that specifically exhibits growth inhibitory activity against human leukemia cells is produced in the culture broth of the filamentous fungus Fusarium sp. FKI-7550 strain newly isolated from soil. Subsequently, a substance that specifically exhibits growth inhibitory activity against human acute promyelocytic leukemia-derived cell lines, human T-cell leukemia-derived cell lines, and human acute monocytic leukemia-derived cell lines was purified and isolated from the culture. As a result, the present inventors succeeded in obtaining the FKI-7550A substance and the FKI-7550B substance, which are novel substances not reported so far, and completed the present invention.

[0014] The present invention has been completed based on such findings, and relates to the FKI-7550A substance and the FKI-7550B substance represented by the following formula, or a salt thereof, or a hydrate or solvate thereof.

[0015]

Chemical formula

[0016]

Chemical formula

[0017] As used herein, "salt" refers to a salt formed by the combination of a compound of the present invention with an inorganic or organic base or acid, and is a salt that is acceptable for administration into the body as a medicine. Such salts are described, for example, in Berge et al., J. Pharm. Sci. 66: 1-19 (1977). Examples of salts include, when an acidic group is present, alkali metal and alkaline earth metal salts such as lithium, sodium, potassium, magnesium, calcium; salts of amines such as ammonia, methylamine, dimethylamine, trimethylamine, dicyclohexylamine, tris(hydroxymethyl)aminomethane, N,N-bis(hydroxyethyl)piperazine, 2-amino-2-methyl-1-propanol, ethanolamine, N-methylglucamine, L-glucamine; or salts with basic amino acids such as lysine, δ-hydroxylysine, arginine. When a basic group is present, salts of mineral acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid; salts with organic acids such as methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, acetic acid, propionate, tartaric acid, fumaric acid, maleic acid, malic acid, oxalic acid, succinic acid, citric acid, benzoic acid, mandelic acid, cinnamic acid, lactic acid, glycolic acid, glucuronic acid, ascorbic acid, nicotinic acid, salicylic acid; or salts with acidic amino acids such as aspartic acid, glutamic acid, etc. can be mentioned.

[0018] Furthermore, hydrates or solvates of compound (I) and hydrates or solvates of pharmacologically acceptable salts of compound (I) are also included in the compounds of the present invention. Also, as used herein, "compound (I)" includes, unless it is clearly inappropriate, pharmacologically acceptable salts, hydrates and solvates of compound (I), even when not explicitly stated, and hydrates or solvates of pharmacologically acceptable salts of compound (I).

[0019] In addition, the compound represented by formula (I) of the present invention or a salt thereof may be crystalline or amorphous. Further, since the compound of the present invention has an asymmetric carbon, optical isomers exist. As the compound of the present invention, it may be any of dextrorotatory (+) or levorotatory (-) compounds, or a mixture of these isomers in any ratio such as a racemate. Further, the compound of the present invention includes any tautomer or geometric isomer (for example, E-form, Z-form, etc.) unless otherwise specified.

Effects of the Invention

[0020] Since the FKI-7550A substance and FKI-7550B substance of the present invention can induce growth inhibition against human acute promyelocytic leukemia-derived cell lines, human T-cell leukemia-derived cell lines, and human acute monocytic leukemia-derived cell lines, they become novel leukemia cell growth inhibitors. In addition, since the Fusarium sp. FKI-7550 strain of the present invention produces the FKI-7550A substance and the KI-7550B substance, it can be used for the production of the FKI-7550A substance and the FKI-7550B substance.

Modes for Carrying Out the Invention

[0021] In one aspect, the present invention relates to a method for producing an FKI-7550A substance and / or an FKI-7550B substance, which includes culturing a microorganism belonging to the filamentous fungi having the ability to produce the FKI-7550A substance and / or the FKI-7550B substance in a medium, accumulating the FKI-7550A substance and / or the FKI-7550B substance in the culture, and isolating the FKI-7550A substance and / or the FKI-7550B substance by separating, extracting, and purifying them from the culture.

[0022] As used herein, the "microorganism belonging to the filamentous fungi having the ability to produce the FKI-7550A substance and / or the FKI-7550B substance" is a bacterium belonging to the filamentous fungi and is not particularly limited as long as it is a microorganism having the ability to produce the FKI-7550A substance and / or the FKI-7550B substance. Preferably, it is a microorganism belonging to the genus Fusarium having the ability to produce the FKI-7550A substance and / or the FKI-7550B substance. Any of the FKI-7550A substance and / or FKI-7550B substance-producing filamentous fungi, including the Fusarium sp. FKI-7550 strain and its mutant strains, can be used. As used herein, the "mutant strain" refers to a strain having mycological properties or genes different from those of the Fusarium sp. FKI-7550 strain due to artificial or mutagenic stimuli in nature. Such mutant strains include not only strains derived from the Fusarium sp. FKI-7550 strain but also the original strains from which the Fusarium sp. FKI-7550 strain was derived. As used herein, the mutant strain does not depend on the presence or absence of actual traces of derivation. For example, strains having genes with high homology (e.g., 90% or more, 95% or more, etc.) to the Fusarium sp. FKI-7550 strain gene (e.g., the 28S rRNA gene) are also included in the mutant strains. Also, such mutant strains are not limited as to whether they are artificially produced or collected from nature as long as they maintain the ability to produce the FKI-7550A substance and / or the FKI-7550B substance.

[0023] Whether the microorganism is a "microorganism belonging to a filamentous fungus having the ability to produce FKI-7550A substance and / or FKI-7550B substance" can be determined, for example, by the following method. 1 mL of the test microorganism cultured in a liquid medium is inoculated into a 500 mL Erlenmeyer flask containing 100 mL of a liquid medium (pH 7.0) consisting of 2.4% starch, 0.1% glucose, 0.3% peptone, 0.3% skipjack extract, 0.5% yeast extract, and 0.4% calcium hydrogen carbonate. After shaking culture at 27°C for 3 days, the obtained seed culture solution is inoculated with 1 mL into a 500 mL Erlenmeyer flask containing 100 mL of a liquid medium (pH 7.0) consisting of a liquid medium composed of 3.3% nutrient broth, 3.3% soy bean meal, 2.2% glycerol, 2.2% available starch, and 2.2% calcium hydrogen carbonate. If the FKI-7550A substance and / or FKI-7550B substance is present in the culture obtained by shaking culture at 27°C for 7 days, it can be determined that the microorganism belongs to a filamentous fungus having the ability to produce the FKI-7550A substance and / or FKI-7550B substance. In the present specification, as the microorganism belonging to a filamentous fungus having the ability to produce the FKI-7550A substance and / or FKI-7550B substance, preferably, it is Fusarium sp. FKI-7550 strain.

[0024] The "medium" for culturing a microorganism belonging to the genus filamentous fungus having the ability to produce FKI-7550A substance and / or FKI-7550B substance can contain, as a nutrient source, those that can be used as a nutrient source for filamentous fungi. For example, rice, commercially available peptone, meat extract, corn steep liquor, cottonseed meal, peanut meal, soybean meal, yeast extract, NZ-amine, casein hydrate, sodium nitrate, ammonium nitrate, ammonium sulfate and other nitrogen sources, glycerin, starch, glucose, galactose, mannose and other carbohydrates, or carbon sources such as fats, and inorganic salts such as sodium chloride, phosphates, calcium carbonate, magnesium sulfate, etc. can be used alone or in combination. In addition, trace metal salts and animal, plant, and mineral oils as antifoaming agents can be added to the medium as needed. These can be any substances that are useful for the production of FKI-7550A substance and / or FKI-7550B substance using the production bacterium, and all known culture materials for filamentous fungi can be used. Trace metal salts and animal, plant, and mineral oils as antifoaming agents can also be added to the medium as needed. These additives can be any substances that are useful for the production of FKI-7550A substance and / or FKI-7550B substance using the production bacterium, and all known culture materials such as filamentous fungi can be used.

[0025] The culture temperature can be applied within the range in which the production bacterium can grow and produce FKI-7550A substance and / or FKI-7550B substance. For example, the culture of a microorganism belonging to the genus filamentous fungus having the ability to produce FKI-7550A substance and / or FKI-7550B substance can be carried out by shaking culture for several days to 2 weeks within the temperature range (for example, 10°C to 40°C, preferably 25 to 30°C) in which the production bacterium can grow and produce FKI-7550A substance and / or FKI-7550B substance. The culture conditions can be appropriately selected according to the properties of the production bacterium used while referring to the description in this specification.

[0026] The FKI-7550A substance and / or the FKI-7550B substance can be extracted from the culture solution with a water-immiscible organic solvent such as ethyl acetate. In addition to the said extraction method, known methods used for collecting fat-soluble substances, for example, adsorption chromatography, partition chromatography, gel filtration chromatography, scraping from thin-layer chromatography, countercurrent distribution chromatography, high-performance liquid chromatography, etc. can be appropriately combined or repeated to purify until it becomes pure.

[0027] In another aspect, the present invention further relates to a novel filamentous fungus, Fusarium sp. FKI-7550 strain (NITE AP-2944) (NITE P-03741) In this specification, the Fusarium sp. FKI-7550 strain is a microorganism newly isolated by the present inventors from the soil in Tokushima City, Tokushima Prefecture, and Fusarium sp. FKI-7550 (NITE AP-2944) (NITE P-03741) is the strain deposited on March 27, 2019, at the Patent Microorganisms Depositary, National Institute of Technology and Evaluation (2-5-8 Kazusa Kamashima, Kisarazu City, Chiba Prefecture). The Fusarium sp. FKI-7550 strain of the present invention has the mycological properties described in Example 1 of this specification.

[0028] In yet another aspect, the present invention relates to a pharmaceutical composition containing the FKI-7550A substance and / or the FKI-7550B substance as an active ingredient. Preferably, the pharmaceutical composition of the present invention is a leukemia cell growth inhibitor, a leukemia therapeutic agent, or a leukemia preventive agent. Specifically, since the FKI-7550A substance and / or the FKI-7550B substance of the present invention can induce growth inhibition against cell lines derived from human acute promyelocytic leukemia, human T-cell leukemia, and human acute monocytic leukemia, it can be used as a growth inhibitor of human acute promyelocytic leukemia cells, human T-cell leukemia cells, or human acute monocytic leukemia cells, or as a therapeutic agent or preventive agent for human acute promyelocytic leukemia, human T-cell leukemia, or human acute monocytic leukemia.

[0029] The pharmaceutical composition of the present invention can be used in an oral administration form or a parenteral administration form such as an injection or a drip infusion. When administering this compound to mammals or the like, it may be orally administered as tablets, powders, granules, syrups, etc., or parenterally administered as injections or drip infusions. The dosage varies depending on the degree of symptoms, age, type of disease, etc., but usually 50 mg to 500 mg per day for adults is administered in 1 to several divided doses per day.

[0030] The pharmaceutical composition of the present invention can be formulated by a conventional method using a normal pharmaceutically acceptable carrier. When preparing a solid preparation for oral use, after adding an excipient to the main drug and further, if necessary, a binder, a disintegrant, a lubricant, etc., it is made into a solvent, granules, powders, capsules, etc. by a conventional method. When preparing an injection, a pH adjuster, a buffer, a stabilizer, a solubilizer, etc. can be added to the main drug as necessary, and it can be made into a subcutaneous or intravenous injection by a conventional method.

Examples

[0031] Examples of the present invention are given below to specifically explain the present invention, but the present invention is not limited thereto. The documents cited throughout this specification are incorporated herein by reference in their entirety.

[0032] (Example 1) Mycological Characteristics of Fusarium sp. Strain FKI-7550 The inventors newly isolated Fusarium sp. strain FKI-7550 from the soil in Tokushima City, Tokushima Prefecture. The mycological characteristics of Fusarium sp. strain FKI-7550 were as follows.

[0033] 1. Morphological Characteristics This strain grew well on malt extract agar medium and synthetic nutrient agar medium, and conidia grew well on various agar media. When observing the colonies grown on synthetic nutrient agar medium under a microscope, the hyphae were colorless, 2.1 - 4.2 (-6.2) μm in width, septate, and thin-walled. The conidia showed dimorphism on the medium, forming microconidia and macroconidia. The microconidia were formed in a pseudocapitate manner on short phialides arising from septate hyphae. The size of the phialides was (10.4 -) 18.7 - 37.4 × 2.1 - 3.1 (-4.2) μm. The conidia were oblong or elliptical, with 0 - 1 septum, and the size was 6.2 - 12.5 (-17.7) × (2.1 -) 4.0 - 4.2 μm. The macroconidia were formed in a clustered manner on phialides branched 2 - 3 times on the conidiophores. The size of the conidiophores was 0 - 6.2 (-10.5) × 2.1 - 4.2 μm, and the size of the phialides was 4.2 - 6.2 (-14.6) × 2.1 - 3.1 (-4.2) μm. The conidia were crescent-shaped, with 3 - 4 septa, and the size was (31.2 -) 43.7 - 56.2 (-72.8) × 3.1 - 4.2 (-6.2) μm. The top was slightly curved like a sickle, the base was heel-shaped, and the size of the base was 1.0 - 3.1 × 1.0 - 2.1 μm. The chlamydospores were formed laterally on the hyphae or in a chain at the tips of their branches, colorless, thick-walled, subglobose to broadly elliptical, and the size was (6.2 -) 10.4 - 16.6 × (10.4 -) 16.6 - 20.8 μm.

[0034] 2. Culture Characteristics Table 1 shows the macroscopic observation results when cultured on various agar media at 25°C for 7 days.

[0035]

Table 1

[0036] 3. ITS Gene Analysis The sequence of 562 bases of the ITS gene was determined, and as a result of homology search using the data of filamentous fungi registered and published in the DNA database and phylogenetic analysis by the neighbor-joining method, it was reasonable to classify this strain into the genus Fusarium.

[0037] 4. Conclusion Based on the morphological characteristics and cultural properties of the above FKI-7550 strain, as a result of attempting to compare with known bacterial species, and from the analysis results of the ITS gene, this strain is judged to be a bacterial species belonging to the genus Fusarium. This strain was deposited at the Patent Biological Deposit Center of the National Institute of Technology and Evaluation on May 9, 2019, as Fusarium sp. FKI-7550 (NITE AP-2944). (NITE P-03741) 。

[0038] (Example 2) Acquisition of FKI-7550A Substance and FKI-7550B Substance 100 pieces were placed in a 500 mL Erlenmeyer flask containing 100 mL of a liquid medium (pH 7.0) consisting of 2.4% starch, 0.1% glucose, 0.3% peptone (manufactured by Kyokuto Pharmaceutical Industry Co., Ltd.), 0.3% skipjack extract (manufactured by Kyokuto Pharmaceutical Industry Co., Ltd.), 0.5% yeast extract (manufactured by Oriental Yeast Co., Ltd.), and 0.4% calcium hydrogen carbonate, and inoculated with 1 mL each of Fusarium sp. FKI-7550 strain (NITE AP-2944) cultured in the liquid medium, and shake-cultured at 27°C for 3 days. (NITE P-03741) The obtained seed culture solution was inoculated into 60 pieces in a 500 mL Erlenmeyer flask containing 100 mL of a liquid medium (pH 7.0) consisting of 3.3% nutrient broth (Difco), 3.3% soy flour (Tokyo Preservation Food Co., Ltd.), 2.2% glycerol, 2.2% available starch, and 2.2% calcium hydrogen carbonate, 1 mL each, and shake-cultured at 27°C for 7 days.

[0039] To each of 60 500-mL Erlenmeyer flasks after the completion of cultivation, 100 mL of ethanol was added and the mixture was vigorously stirred for 1 hour. Next, the ethanol in the extract was distilled off under reduced pressure. To the resulting aqueous solution, 3 L of ethyl acetate was added and the mixture was well stirred, after which the ethyl acetate layer was recovered. Using an evaporator, it was concentrated to dryness to obtain crude product 1. Next, this was dissolved in a small amount of methanol, and using silica gel (manufactured by MERCK) open column chromatography, stepwise elution was carried out with a chloroform-methanol solvent system (100:0, 100:1, 50:1, 10:1, 1:1, 0:100), and 600 mg of a 10:1 fraction (crude product 2) containing the FKI-7550A substance and the FKI-7550B substance was obtained.

[0040] Crude product 2 was dissolved in a small amount of methanol and injected into an octadecylsilyl column (Chromatorex C8 SPS100-5HE, φ20×250 mm, flow rate 10.0 mL / min, detection at 210 nm) by high performance liquid chromatography, and elution was carried out with acetonitrile-water (70:30) containing 0.1% trifluoroacetic acid. The active fraction was collected and concentrated under reduced pressure to obtain 70.7 mg and 11.5 mg of the FKI-7550A substance and the FKI-7550B substance, respectively, as pale yellow solids.

[0041] As a result of measuring the physicochemical properties of the obtained FKI-7550A substance, the following were obtained. (1) Appearance: Pale yellow solid (2) Molecular weight: 455 (3) Molecular formula: C27H37O4N [M+H] by high speed atom collision ionization + Theoretical value (m / z) 456.2750, measured value (m / z) 456.2738 (4) Specific rotation: [α]D27 = -15 (c = 0.1, methanol) (5) Ultraviolet absorption maximum λmax nm (in methanol): 243.0 (49,322), 275 (14,970) (ε in parentheses) (6) Infrared absorption maxima vmax(ART): It has maximum absorption at 3394, 2958, 2925, 2870, 1770, 1715, 1601, 1438, 1318, 1238, 1074, 1040, 963 cm-1. (7) Proton nuclear magnetic resonance spectrum: The chemical shifts (ppm) and spin coupling constants (Hz) in deuterated dimethyl sulfoxide are shown in Table 2. (8) Carbon nuclear magnetic resonance spectrum: The chemical shifts (ppm) in deuterated dimethyl sulfoxide are shown in Table 2. (9) Solubility in solvents: Freely soluble in methanol, dimethyl sulfoxide, acetone, and chloroform. Sparingly soluble in water.

[0042]

Table 2

[0043] As a result of measuring the physicochemical properties of the obtained FKI-7550B substance, the following were obtained. (1) Appearance: Pale yellow solid (2) Molecular weight: 487 (3) Molecular formula: C28H41O6N [M-H] by high-speed atom collision ionization - Theoretical value (m / z) 486.2857, measured value (m / z) 486.2856 (4) Specific rotation: [α]D25 = -140 (c = 0.1, methanol) (5) Ultraviolet absorption maximum λmax nm (in methanol): 243.0 (52,791), 281 (15,633) (ε in parentheses) (6) Infrared absorption maxima vmax(ART): It has maximum absorption at 3324, 2958, 2927, 1721, 1655, 1608, 1440, 1375, 1208, 1084, 963 cm-1. (7) Proton nuclear magnetic resonance spectrum: The chemical shifts (ppm) and spin coupling constants (Hz) in deuterated dimethyl sulfoxide are shown in Table 2. (8) Carbon nuclear magnetic resonance spectrum: The chemical shifts (ppm) in deuterated dimethyl sulfoxide are shown in Table 2. (9) Solubility in solvents: Freely soluble in methanol, dimethyl sulfoxide, acetone, and chloroform. Sparingly soluble in water.

[0044]

Table 3

[0045] As described above, various physicochemical properties of the FKI-7550A substance and the FKI-7550B substance have been described in detail. However, no compounds that match such properties have been reported so far, and it was determined that the FKI-7550A substance and the FKI-7550B substance are novel substances.

[0046] (Example 3) In vitro leukemia cell growth inhibitory activity of the FKI-7550A substance and the FKI-7550B substance As cells, the HL-60 human acute promyelocytic leukemia-derived cell line, the Jurkat human T-cell leukemia-derived cell line, and the THP-1 human acute monocytic leukemia-derived cell line were used. Cells maintained and subcultured in RPMI-1640 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS) and antibiotics were used. Each cell was suspended in RPMI-1640 medium containing 10% FBS to adjust the suspension to 3×10 6 cells / mL, 100 μL was added to a 96-well plate and mixed, and then cultured at 37 °C in a 5% CO2 - 95% air environment for 1 day. Thereafter, 1 μL of an aqueous methanol solution of this compound was added to each well, mixed, and then cultured in the aforementioned gas environment for 2 days. The presence or absence of growth of each cell was colorimetrically quantified by the WST-8 method. The 50% cell growth inhibitory concentration (IC50 value) of this compound was determined from the compound concentration-action curve. The results are shown in Table 3.

[0047]

Table 4

[0048] The FKI-7550A substance and FKI-7550B substance of the present invention showed the activity of suppressing the proliferation of leukemia-derived cells. Therefore, it was shown that the FKI-7550A substance and FKI-7550B substance of the present invention can be used as agents that induce the suppression of the proliferation of human acute promyelocytic leukemia cells, human T-cell leukemia cells, and human acute monocytic leukemia cells.

Claims

1. The FKJ-7550A substance or FKJ-7550B substance represented by the following formula, or a salt thereof, or a hydrate or solvate thereof. 【Chemical Formula 1】 【Chemical Formula 2】

2. A method for producing the FKJ-7550A substance or FKJ-7550B substance, which comprises culturing a microorganism belonging to the genus filamentous fungus having the ability to produce the FKJ-7550A substance or FKJ-7550B substance according to Claim 1 in a medium, accumulating the FKJ-7550A substance or FKJ-7550B substance in the culture, and collecting the FKJ-7550A substance or FKJ-7550B substance from the culture, The method, wherein the microorganism belonging to the genus filamentous fungus having the ability to produce the FKJ-7550A substance or FKJ-7550B substance is Fusarium sp. FKJ-7550 strain (Accession No.: NITE P-03741).

3. A pharmaceutical composition containing, as an active ingredient, the FKJ-7550A substance or FKJ-7550B substance according to Claim 1, or a salt thereof, or a hydrate or solvate thereof.

4. The pharmaceutical composition according to Claim 3, which is an inhibitor of leukemia cell growth, a therapeutic agent or a prophylactic agent for leukemia.

5. The pharmaceutical composition according to Claim 4, which is an inhibitor of the growth of human acute promyelocytic leukemia cells, human T-cell leukemia cells, or human acute monocytic leukemia cells, or a therapeutic agent or a prophylactic agent for human acute promyelocytic leukemia, human T-cell leukemia, or human acute monocytic leukemia.

Images