JP2015013825A - Novel compound quinofuracins, method for producing same, and application thereof, and novel microorganisms - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a novel compound having an excellent anti-cancer effect, to provide a method for producing the novel compound, to provide a novel microorganism which is a production fungus of the novel compound, and to provide a compound-containing composition and an anticancer drug using the novel compound.SOLUTION: Provided are a compound represented by any of the structural formula (A)-(E) and a method for producing the same; a microorganism having an ability to produce the compound represented by any of the (A)-(E); and a compound-containing composition and an anticancer drug containing the compound.

Description

  The present invention relates to a novel compound and a method for producing the same, a novel microorganism having the ability to produce the novel compound, a composition containing the novel compound, and an anticancer agent.

  Cancer has the property of allowing unlimited cell growth as a result of the escape of cell death from normal cells that should cause cell death due to DNA damage or the like. The cancer is a disease that accounts for about 30% of Japanese deaths. With the elucidation of the remarkable mechanism of canceration of cells in recent years, molecular targeted therapeutic drugs targeting cancer cell-specific genes have been vigorously developed both in Japan and overseas.

  Up to now, attention has been focused on Murine double minute 2 (hereinafter sometimes referred to as “MDM2”), which is known to bind to p53, a transcription factor involved in cell cycle control and apoptosis induction. And inhibitors that inhibit the binding to p53 have been proposed (see, for example, Non-Patent Documents 1 and 2).

  However, because cancer has diversity such as organ-specific properties and the presence or absence of metastasis, treatment methods suitable for individual cancers are necessary, and cancer therapeutic agents also have specificity and diversity. It has been demanded. Currently used cancer therapeutics are insufficient to deal with all cancers, and there is a strong demand for the development of new cancer therapeutics.

Yosup Rew et al, Structure-Based Design of Novell Inhibitors of the MDM2-p53 Interaction, J. MoI. Med. Chem. , 55, 4936-4594 (2012) Lyubomir T.W. Vassilev et al, In Vivo Activation of the p53 Pathway by Small-Molecular Agents of MDM2, SCIENCE, 303, 844-848 (2004)

  An object of the present invention is to solve the above-described problems and achieve the following objects. That is, the present invention relates to a novel compound having an excellent anticancer activity, a method for producing the novel compound, a novel microorganism that is a bacterium producing the novel compound, and a compound-containing composition using the novel compound, And it aims at providing an anticancer agent.

Means for solving the problems are as follows. That is,
<1> A compound represented by any one of the following structural formulas (A) to (E).
<2> A method for producing the compound according to <1>,
A culture step for culturing a microorganism belonging to the genus Staphylotricum and having the ability to produce the compound according to <1>above;
And a collecting step of collecting the compound according to <1> from the culture obtained in the culturing step.
<3> A microorganism belonging to the genus Staphylotrichum and having the ability to produce the compound according to <1>.
<4> A compound-containing composition comprising the compound according to <1>.
<5> An anticancer agent comprising the compound according to <1>.

  According to the present invention, it is possible to solve the conventional problems and achieve the object, and to provide a novel compound having excellent anticancer activity, a method for producing the novel compound, and a bacterium producing the novel compound. A novel microorganism, a compound-containing composition and an anticancer agent using the novel compound can be provided.

FIG. 1 is a chart of a proton nuclear magnetic resonance spectrum of the compound represented by the structural formula (A). Horizontal axis: ppm unit. FIG. 2 is a chart of carbon-13 nuclear magnetic resonance spectrum of the compound represented by the structural formula (A). Horizontal axis: ppm unit. FIG. 3 is a chart of a proton nuclear magnetic resonance spectrum of the compound represented by the structural formula (B). Horizontal axis: ppm unit. FIG. 4 is a chart of the carbon-13 nuclear magnetic resonance spectrum of the compound represented by the structural formula (B). Horizontal axis: ppm unit. FIG. 5 is a chart of a proton nuclear magnetic resonance spectrum of the compound represented by the structural formula (C). Horizontal axis: ppm unit. FIG. 6 is a chart of carbon-13 nuclear magnetic resonance spectrum of the compound represented by the structural formula (C). Horizontal axis: ppm unit. FIG. 7 is a chart of a proton nuclear magnetic resonance spectrum of the compound represented by the structural formula (D). Horizontal axis: ppm unit. FIG. 8 is a chart of carbon-13 nuclear magnetic resonance spectrum of the compound represented by the structural formula (D). Horizontal axis: ppm unit. FIG. 9 is a chart of a proton nuclear magnetic resonance spectrum of the compound represented by the structural formula (E). Horizontal axis: ppm unit. FIG. 10 is a chart of a carbon-13 nuclear magnetic resonance spectrum of the compound represented by the structural formula (E). Horizontal axis: ppm unit.

(New compound)
The compound of the present invention is a compound represented by any one of the following structural formulas (A) to (E) (hereinafter sometimes referred to as “Quinofuracin A to E”). It is a novel compound found.

  Among the compounds represented by the structural formulas (A) to (E), the compound represented by any one of the structural formulas (A), (B), (C), and (E) is preferable. A compound represented by any one of A) and (B) is more preferable. The preferred compound is advantageous in that it can further suppress the growth of glioblastoma cells.

<Physicochemical properties of compound (quinofuracin A) represented by structural formula (A)>
The physicochemical properties of the quinofuracin A are as follows.
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₆ H ₃₀ O ₁₂ and molecular weight is 534.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 557.1614 (M + Na) ⁺ and the calculated value is m / z 557.1629 (as C ₂₆ H ₃₀ O ₁₂ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.38), 262 (4.16), 295 (4.27), 314 (4.23), 469 (3.78)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 211 (4.63), 231 (4.29), 265 (4.10), 324 (4.60), 398 (3. 79), 546 (3.82)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 224 (4.40), 253 (4.14, sh), 264 (4.18), 293 (4.40), 320 ( 3.92), 453 (3.90)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.15.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 10.8 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 620, 770, 1070, 1170, 1210, 1260, 1400, 1470, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.

<Physicochemical properties of compound (quinofuracin B) represented by structural formula (B)>
The physicochemical properties of the quinofuracin B are as follows.
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₈ H ₃₂ O ₁₃ , and the molecular weight is 576.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 599.1724 (M + Na) ⁺ and the calculated value is m / z 599.1735 (as C ₂₈ H ₃₂ O ₁₃ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.37), 263 (4.11), 294 (4.23), 314 (4.09), 460 (3.75)
(Ii) When measured with methanol-NaOH solution λ _max nm (log ε): 211 (4.67), 231 (4.29), 264 (4.10), 325 (4.55), 397 (3. 74), 545 (3.78)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 224 (4.40), 253 (4.10, sh), 264 (4.13), 293 (4.35), 320 ( 3.88, sh), 453 (3.93)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.39.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 15.2 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 630, 780, 1070, 1170, 1210, 1260, 1380, 1450, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.

<Physicochemical properties of compound (quinofuracin C) represented by structural formula (C)>
The physicochemical properties of quinofuracin C are as follows.
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₆ H ₃₀ O ₁₂ and molecular weight is 534.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 557.1616 (M + Na) ⁺ and the calculated value is m / z 557.1629 (as C ₂₆ H ₃₀ O ₁₂ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.42), 261 (4.22), 290 (4.25), 314 (4.16), 455 (3.85)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 212 (4.63), 237 (4.42), 258 (4.19), 290 (4.17, sh), 314 ( 4.34), 527 (3.91)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 225 (4.44), 253 (4.16, sh), 264 (4.23, sh), 288 (4.35), 319 (3.94), 448 (3.94)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.16.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 15.4 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 650, 770, 1080, 1170, 1210, 1280, 1400, 1470, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.

<Physicochemical properties of compound (quinofuracin D) represented by structural formula (D)>
The physicochemical properties of the quinofuracin D are as follows.
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₆ H ₂₈ O ₁₃ , and the molecular weight is 548.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 571.1422 (M + Na) ⁺ and the calculated value is m / z 571.1422 (as C ₂₆ H ₂₈ O ₁₃ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.39), 263 (4.14), 290 (4.30), 314 (4.12), 460 (3.82)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 211 (4.64), 232 (4.29), 263 (4.10), 324 (4.56), 397 (3. 73), 542 (3.84)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 223 (4.41), 253 (4.09, sh), 265 (4.14), 293 (4.39), 321 ( 3.86), 453 (3.91)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.11.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 4.2 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 620, 770, 1090, 1160, 1210, 1260, 1400, 1470, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.

<Physicochemical properties of compound (quinofuracin E) represented by structural formula (E)>
The physicochemical properties of quinofuracin E are as follows.
(1) Appearance: Yellowish orange powder.
(2) Molecular formula: It is represented by C ₂₄ H ₂₂ O ₁₂ and its molecular weight is 502.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental values are m / z 503, 1173 (M + H) ⁺ and the calculated value is m / z 503.1184 (as C ₂₄ H ₂₃ O ₁₂ ).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 222 (4.41), 251 (4.10, sh), 266 (4.20, sh), 287 (4.35), 317 ( 3.84), 442 (3.89)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 212 (4.61), 240 (4.26, sh), 263 (4.32), 311 (4.03), 512 ( 3.84)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 223 (4.40), 251 (4.12, sh), 266 (4.21, sh), 287 (4.36), 318 (3.83), 443 (3.90)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.25.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 6.0 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 650, 770, 950, 1070, 1160, 1190, 1250, 1320, 1390, 1470, 1630, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.

  Whether or not the compound has a structure represented by any one of the structural formulas (A) to (E) can be confirmed by various analytical methods selected as appropriate, for example, the mass spectrometry method, Examples include ultraviolet spectroscopy, infrared spectroscopy, proton nuclear magnetic resonance spectroscopy, and carbon-13 nuclear magnetic resonance spectroscopy. The measurement values obtained by the respective analytical methods may have some errors, but those skilled in the art will have a compound represented by any one of the structural formulas (A) to (E). It can be easily identified.

The compound represented by any one of the structural formulas (A) to (E) may be in the form of a salt.
The salt is not particularly limited as long as it is a pharmacologically acceptable salt, and can be appropriately selected according to the purpose. Examples thereof include organic salts such as acetates and citrates, hydrochlorides and carbonates. Examples include salt.
The compound represented by any one of the structural formulas (A) to (E) may be a tautomer thereof.

  There is no restriction | limiting in particular as a manufacturing method of the compound represented by either of the said structural formula (A) to (E), Although it can select suitably according to the objective, It manufactures with the manufacturing method of this invention mentioned later. It is preferable to do.

<Application>
The compound represented by any one of the structural formulas (A) to (E) is a highly safe compound having excellent anticancer activity as shown in Test Examples described later. Therefore, the compound represented by any one of the structural formulas (A) to (E) can be suitably used as an active ingredient such as a compound-containing composition of the present invention and an anticancer agent described later.

(Method for producing compound)
The method for producing the compound of the present invention includes at least a culture step and a collection step, and further includes other steps as necessary.

<Culture process>
The culturing step is a step of culturing a microorganism that belongs to the genus Staphylotrichum and has the ability to produce a compound represented by any one of the structural formulas (A) to (E).

The microorganism belongs to the genus Staphylotrichum and has the ability to produce the above-described compound of the present invention, that is, the compound represented by any one of the structural formulas (A) to (E). If there is no particular limitation, it can be appropriately selected according to the purpose. For example, Staphylotricum boninense PF1444 strain (accession number NITE BP-1450, details of the microorganism of the present invention described later) ). Other strains capable of producing the compound represented by any one of the structural formulas (A) to (E) can also be isolated from the natural world by conventional methods. Incidentally, the Staphylococcus tri Kum Boninense (Staphylotrichum boninense) including PF1444 strain, a producing strain of the compound represented by any one of structural formulas (A) (E), be subjected to irradiation or other mutagenesis Thus, it is also possible to increase the productivity of the compound represented by any one of the structural formulas (A) to (E). Furthermore, it is possible to produce a compound represented by any one of the structural formulas (A) to (E) by genetic engineering techniques.

  The method for analyzing that the microorganism has the ability to produce the compound represented by any one of the structural formulas (A) to (E) is not particularly limited and can be appropriately selected depending on the purpose. For example, the above structural formula (A) can be obtained by a method for analyzing the anticancer activity of a component of a culture of the microorganism, preferably a culture supernatant after liquid culture or a solid medium after solid culture, and various analytical methods. ) To (E), and the like.

  In the culture, a producing bacterium producing a compound represented by any one of the structural formulas (A) to (E) (hereinafter sometimes simply referred to as “compound producing bacterium”) is used as a nutrient medium (hereinafter simply referred to as “ Medium (which may be referred to as “medium”) and culturing at a temperature suitable for the production of the compound represented by any of the structural formulas (A) to (E).

The nutrient medium is not particularly limited and can be appropriately selected according to the purpose. For example, a known medium conventionally used for culturing ascomycetes can be used, and a liquid medium can be used. It may be a solid (agar, brown rice, etc.) medium.
The nutrient source added to the nutrient medium is not particularly limited and may be appropriately selected depending on the intended purpose. For example, commercially available rice, brown rice, soybean flour, wheat germ, pressed wheat, peptone, cottonseed meal Nitrogen sources such as yeast extract, meat extract, corn steep liquor, ammonium sulfate, sodium nitrate, urea, etc .; carbon sources such as tomato paste, glycerin, starch, glucose, galactose, dextrin, bacto-soyton; Can be mentioned.
Furthermore, inorganic salts such as sodium chloride and calcium carbonate can be added to the medium for use, and in addition, a trace amount of metal salt can be added to the medium for use.
Any of these materials may be used as long as they are useful for the production of the compound represented by any one of the structural formulas (A) to (E) by the compound-producing bacteria, and all known culture materials may be used. it can.

  There is no restriction | limiting in particular as seed | species culture | cultivation liquid for production of the compound represented by either of said structural formula (A) to (E), According to the objective, it can select suitably, A liquid culture medium, a flat plate A growth product obtained by culturing the above-described compound-producing bacteria on a medium such as a medium, a slant medium, or a half slant medium can be used.

The culture method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include shaking culture, static culture, and tank culture.
The temperature of the culture is not particularly limited as long as the compound-producing bacteria are not substantially inhibited from being grown and can produce the compound represented by any one of the structural formulas (A) to (E). There is no restriction | limiting, Although it can select suitably according to the production microbe to be used, 20 to 35 degreeC is preferable.
The pH of the culture solution is within a range in which the compound represented by any one of the structural formulas (A) to (E) can be produced without substantially inhibiting the growth of the compound-producing bacteria. There is no restriction | limiting in particular, According to the production microbe to be used, it can select suitably.
There is no restriction | limiting in particular as the period of the said culture | cultivation, According to accumulation | storage of the compound represented by either of the said structural formula (A) to (E), it can select suitably.

<Collection process>
The collecting step is a step of collecting the compound represented by any one of the structural formulas (A) to (E) from the culture obtained in the culturing step.
Since the compound represented by any one of the structural formulas (A) to (E) has the physicochemical properties described above, it can be collected from the culture according to the properties.

The culture is not particularly limited as long as it contains a compound obtained by the culture step and represented by any one of the structural formulas (A) to (E), and is appropriately selected depending on the purpose. Examples include bacterial cells, culture supernatant after liquid culture, solid medium after solid culture, and mixtures thereof.
When the cells are used as the culture, the compound represented by the structural formula (A) is obtained from the cells by an extraction method using an appropriate organic solvent or an elution method by disrupting the cells. It may be extracted and subjected to separation and / or purification.

  The collection method is not particularly limited, and a method used for collecting a metabolite produced by a microorganism can be appropriately selected. For example, a solvent extraction method, a difference in adsorption affinity for various adsorbents can be selected. Examples thereof include a method used and a chromatographic method. The compounds represented by any one of the structural formulas (A) to (E) that have been separated and / or purified can be collected by using these methods alone or in combination as appropriate, and repeatedly in some cases. .

  There is no restriction | limiting in particular as a solvent used for the said solvent extraction method, According to the objective, it can select suitably, For example, ethanol, methanol, acetone, butanol, acetonitrile etc. are mentioned.

  There is no restriction | limiting in particular as said adsorbent, According to the objective, it can select suitably from well-known adsorbents, For example, a polystyrene type adsorption resin etc. are mentioned.

The chromatographic method is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include a thin layer chromatographic method, a high-performance liquid chromatograph for fractionation using a normal phase or reverse phase column (preparation). HPLC) method and the like.
There is no restriction | limiting in particular as a support | carrier used for the said chromatographic method, According to the objective, it can select suitably, For example, ion exchange resin, gel filtration, a silica gel, an alumina, activated carbon etc. are mentioned.
Specific examples of commercially available carriers used in the chromatographic method include ion exchange resins such as Amberlite (registered trademark) CG50 (manufactured by Sigma Aldrich); Toyopearl (registered trademark) HW-40F (manufactured by Tosoh Corporation) Gel filtration such as Sephadex (registered trademark) LH-20 (manufactured by GE Healthcare); Silica gel such as CAPCELL PAK UG120 (manufactured by Shiseido Co., Ltd.);

  The method for eluting the compound represented by any of the structural formulas (A) to (E) from the adsorbent or the carrier in the chromatographic method is not particularly limited, and the type of the adsorbent and the carrier It can select suitably according to a property etc. For example, in the case of a polystyrene-based adsorption resin, a method of elution using water-containing alcohol, water-containing acetone or the like as an elution solvent can be mentioned.

As described above, the compound represented by any one of the structural formulas (A) to (E) can be produced.
The degree of purification of the compounds represented by structural formulas (A) to (E) in the collecting step is not particularly limited, and may be a crude product or a purified product. It may be.

<Other processes>
There is no restriction | limiting in particular as said other process, According to the objective, it can select suitably, A washing | cleaning process, a refinement | purification process, etc. are mentioned.
The washing step is a step of washing the culture obtained in the culturing step or the compound represented by any one of the structural formulas (A) to (E) obtained in the collecting step. There is no restriction | limiting in particular as the said washing | cleaning method, A well-known method can be selected suitably and can be performed.
The purification step is a step of purifying the compound represented by any one of the structural formulas (A) to (E) obtained in the collecting step. There is no restriction | limiting in particular as the said purification method, A well-known method can be selected suitably and can be performed, for example, the method similar to the method described in the said collection process is mentioned.

(Microorganism)
The microorganism of the present invention belongs to the genus Staphylotricum and has the ability to produce the above-described compound of the present invention, that is, the compound represented by any one of the structural formulas (A) to (E). The microorganism has the ability to produce the compound represented by any one of the structural formulas (A) to (E). For this reason, in the above-described method for producing the compound of the present invention, the structural formula (A) To (E) are not particularly limited as long as they are microorganisms that can be used as a microorganism producing the compound represented by any one of (E), and can be appropriately selected according to the purpose.

  Among such microorganisms, in particular, it is preferable to use a microorganism that is an ascomycete isolated from soil sampled in Fukuoka Prefecture and is assigned a strain number of PF1444 strain. The mycological properties of the PF1444 strain are as follows.

1. Morphological properties Using potato dextrose agar medium, 2% malt agar medium, oatmeal agar medium, and LCA medium (Miura medium), PF1444 strain was cultured at 25 ° C. for 2 weeks to 7 weeks, macroscopically and finely Visual morphological observation was performed.
As a result of colony properties and morphology observation, the size is about 65 mm to 70 mm, white to yellow, velvety to wooly colonies are formed (cultured for 2 weeks), and from the tip of the handle upright from the vegetative mycelium A branched conidial pattern was formed, and T-shaped to L-shaped peduncle cells were observed at the pattern base. From the conidia-forming cells at the tip of the conidial pattern, it was observed that allelo-type spherical, subspherical, pear-shaped, thick wall, colorless to light brown, 1-cell conidia were formed.

2.28S rDNA-D1 / D2 gene analysis The base sequence (603 bp) of the 28S rDNA-D1 / D2 gene was determined and compared with the data of known strains registered in the international base sequence database BLASTA. As a result, the base sequence of the 28S rDNA-D1 / D2 gene of the PF1444 strain, as shown below, is the homology with the base sequence of the 28S rDNA-D1 / D2 gene of a plurality of Staphylotrichum bonins , which is a kind of Ascomycota. It showed 100% homology. That is, Staphylotrichum boninense JCM17910 strain (AB625570, 100%), Staphylotrichum boninense JCM17909 strain (AB6256969, 100%), and Staphylotrichum boninense J1001782 T In the non Staphylotrichum boninense, Staphylotrichum coccosporum NBRC33272 strain (AB625574,99.1%), Staphylotrichum coccosporum CBS364.58T strain (AB625572,98.9%), Staphylotrichum coccosporum JCM17911 strain (AB625571,98.9%), Staphylotrichum coccosporum NBRC31817 shares (AB625573,98.8%), Humicola fuscoatra NBRC9530 shares (AB625576,98.4%), Chaetomium sphaerale MUCL40089 strain (AF286407, Is 8.4%). In the parentheses, the accession number of the international base sequence database and the base sequence homology rate are shown in this order.

2. ITS-5.8S rDNA gene analysis The base sequence (599 bp) of the ITS-5.8S rDNA gene was determined and compared with the data of known strains registered in the international base sequence database BLASTA. As a result, as shown below, the base sequence of the ITS-5.8S rDNA gene of the PF1444 strain is a kind of ascomycetes, and the homology with the base sequences of the ITS-5.8S rDNA genes of a plurality of Staphylotrichum bonins. High homology of 99.5% -100% was shown. That, Staphylotrichum boninense JCM17909 strain (AB625581,100%), Staphylotrichum boninense JCM17908T strain (AB625580,99.5%), a Staphylotrichum boninense JCM17910 strain (AB625582,99.5%). In addition, except for Staphylotrichum boninense , Fungal. sp ARIZ B362 strain (FJ612994, 100%), Fungal. sp 229c4c Ltd. (EU563611,100%), Staphylotrichum coccosporum NBRC31817 shares (AB625585,96.1%), Staphylotrichum coccosporum NBRC33272 shares (AB625586,96.1%), Staphylotrichum coccosporum CBS364.58T Ltd. (AB625584,95.6%) It is. In the parentheses, the accession number of the international base sequence database and the base sequence homology rate are shown in this order.

From the above results, it is considered that the PF1444 strain belongs to the genus Staphylotrichum . Therefore, the PF1444 strain was designated as Staphylotricum boninense PF1444 strain.
The PF1444 strain was filed with NITE BP-1450 by filing a deposit application with the Patent Microorganisms Deposit Center (National Institute of Technology and Evaluation, 25-2-8, Kazusa-Kamashita, Kisarazu City, Chiba Prefecture 292-0818). (Contract date: November 6, 2012).

  As seen in other bacteria, the properties of the PF1444 strain are likely to change. Even if it is an artificial mutant strain that can be obtained by mutation treatment, etc.), a zygote, a genetic recombinant, etc., it has the ability to produce a compound represented by any of the structural formulas (A) to (E) Is included in the microorganism of the present invention.

(Compound-containing composition, anticancer agent)
<Compound-containing composition>
The compound-containing composition of the present invention contains at least the compound represented by any one of the structural formulas (A) to (E), and further contains other components as necessary.
The compounds represented by any one of the structural formulas (A) to (E) may be used alone or in combination of two or more.

  There is no restriction | limiting in particular as content of the compound represented by either of said structural formula (A) to (E) in the said compound containing composition, According to the objective, it can select suitably. The compound-containing composition may be the compound itself represented by any one of the structural formulas (A) to (E).

-Other ingredients-
There is no restriction | limiting in particular as said other component, According to the objective, it can select suitably from the carriers accept | permitted pharmacologically, For example, an additive, an adjuvant, water, etc. are mentioned. These may be used alone or in combination of two or more.

  The additive or the adjuvant is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include bactericides, preservatives, binders, thickeners, fixing agents, binders, and coloring agents. , Stabilizers, pH adjusters, buffers, isotonic agents, solvents, antioxidants, UV inhibitors, crystal precipitation inhibitors, antifoaming agents, physical property improvers, preservatives, and the like.

  There is no restriction | limiting in particular as said disinfectant, According to the objective, it can select suitably, For example, cationic surfactants, such as benzalkonium chloride, benzethonium chloride, and cetylpyridinium chloride, etc. are mentioned.

  There is no restriction | limiting in particular as said preservative, According to the objective, it can select suitably, For example, paraoxybenzoic acid esters, chlorobutanol, cresol, etc. are mentioned.

  The binder, thickener and fixing agent are not particularly limited and may be appropriately selected depending on the intended purpose. For example, starch, dextrin, cellulose, methylcellulose, ethylcellulose, carboxymethylcellulose, hydroxyethylcellulose, hydroxypropyl Cellulose, hydroxypropylmethylcellulose, carboxymethyl starch, pullulan, sodium alginate, ammonium alginate, propylene glycol alginate, guar gum, locust bean gum, gum arabic, xanthan gum, gelatin, casein, polyvinyl alcohol, polyethylene oxide, polyethylene glycol, ethylene propylene Block polymer, sodium polyacrylate, polyvinylpyrrolidone, etc.

  The binder is not particularly limited and may be appropriately selected depending on the intended purpose. For example, water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, hydroxypropylcellulose, hydroxy Examples include propyl starch, methyl cellulose, ethyl cellulose, shellac, calcium phosphate, and polyvinyl pyrrolidone.

  The colorant is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include titanium oxide and iron oxide.

  The stabilizer is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include tragacanth, gum arabic, gelatin, sodium pyrosulfite, ethylenediaminetetraacetic acid (EDTA), thioglycolic acid, and thiolactic acid. Is mentioned.

  There is no restriction | limiting in particular as said pH adjuster or said buffering agent, According to the objective, it can select suitably, For example, sodium citrate, sodium acetate, sodium phosphate etc. are mentioned.

  The isotonizing agent is not particularly limited and may be appropriately selected depending on the intended purpose. Examples thereof include sodium chloride and glucose.

  There is no restriction | limiting in particular as content of the said other component in the said compound containing composition, For the purpose in the range which does not impair the effect of the compound represented by either of the said structural formula (A) to (E). It can be appropriately selected depending on the case.

-Use-
Since the compound-containing composition contains a compound represented by any one of the structural formulas (A) to (E), the compound-containing composition has excellent anticancer activity and high safety. It can be suitably used as a composition, an anticancer agent and the like.
In addition, the said compound containing composition may be used independently and may be used in combination with the pharmaceutical which uses another component as an active ingredient. Moreover, the said compound containing composition may be used in the state mix | blended in the pharmaceutical which uses another component as an active ingredient.

<Anticancer agent>
The anticancer agent of the present invention contains at least the compound represented by any one of the structural formulas (A) to (E), and further contains other components as necessary.
The compounds represented by any one of the structural formulas (A) to (E) may be used alone or in combination of two or more.

  There is no restriction | limiting in particular as content of the compound represented by either of said structural formula (A) to (E) in the said anticancer agent, According to the objective, it can select suitably. The anticancer agent may be the compound itself represented by any one of the structural formulas (A) to (E).

-Other ingredients-
There is no restriction | limiting in particular as said other component, According to the objective, it can select suitably, For example, the thing similar to the other component described with the said compound containing composition is mentioned. These may be used alone or in combination of two or more.

  There is no restriction | limiting in particular as content of the said other component in the said anticancer agent, For the objective in the range which does not impair the effect of the compound represented by either of the said structural formula (A) to (E). It can be appropriately selected depending on the case.

-Use-
Since the anticancer agent contains a compound represented by any one of the structural formulas (A) to (E), it has excellent anticancer activity, high safety, and prevention of various cancers. It can be suitably used as an agent or therapeutic agent. Among these, it can be particularly suitably used for glioblastoma.
In addition, the said anticancer agent may be used independently and may be used in combination with the pharmaceutical which uses another component as an active ingredient. Moreover, the said anticancer agent may be used in the state mix | blended in the pharmaceutical which uses another component as an active ingredient.
Further, as shown in the test examples described later, the compound represented by any one of the structural formulas (A) to (E) of the present invention is a specific cancer cell depending on the expression level of the wild-type p53 protein. Has the effect of inhibiting the growth of

<Dosage form>
There is no restriction | limiting in particular as a dosage form of the said compound containing composition and an anticancer agent, According to the objective, it can select suitably, For example, a solid agent, a semi-solid agent, a liquid agent etc. are mentioned. These compound-containing compositions and anticancer agents in these dosage forms can be produced according to conventional methods.

-Solid agent-
The solid preparation is not particularly limited and may be appropriately selected depending on the intended purpose. However, when used as an internal preparation, for example, a tablet, chewable tablet, effervescent tablet, orally disintegrating tablet, troche, drop Agents, hard capsules, soft capsules, granules, powders, pills, dry syrups, soaking agents and the like.
When the said solid agent is used as an external preparation, a suppository, a poultice, a plaster agent etc. are mentioned, for example.

-Semi-solid agent-
There is no restriction | limiting in particular as said semi-solid preparation, Although it can select suitably according to the objective, When using as an internal preparation, a licking agent, a chewing gum agent, a whipping agent, a jelly agent etc. are mentioned, for example.
When the semi-solid preparation is used as an external preparation, examples thereof include an ointment, a cream, a mousse, an inhaler, and a nazar gel.

-Liquid-
There is no restriction | limiting in particular as said liquid agent, Although it can select suitably according to the objective, When used as an internal preparation, a syrup agent, a drink agent, a suspension agent, an alcoholic agent etc. are mentioned, for example.
When the said liquid agent is used as an external preparation, a liquid agent, eye drops, an aerosol agent, a spray agent etc. are mentioned, for example.

<Administration>
There is no restriction | limiting in particular as an administration method, dosage amount, administration time, and administration object of the said compound containing composition and an anticancer agent, According to the objective, it can select suitably.
Examples of the administration method include a local administration method, an enteral administration method, a parenteral administration method and the like.
The dose is not particularly limited, and is appropriately selected in consideration of various factors such as the age, weight, constitution, symptom, and presence / absence of administration of a drug or drug containing other ingredients as active ingredients. be able to.
The animal species to be administered is not particularly limited and can be appropriately selected according to the purpose. For example, human, monkey, pig, cow, sheep, goat, dog, cat, mouse, rat, bird, etc. Among them, among these, it can be suitably used for humans.

  The present invention will be specifically described below with reference to production examples and test examples of the present invention, but the present invention is not limited to these production examples and test examples. In the following production examples and test examples, “%” represents “% by mass” unless otherwise specified.

(Production Example 1)
<Production of compound represented by any one of structural formulas (A) to (E)>
-Preparation of seed culture medium-
TS medium (2.0% soluble starch (manufactured by Wako Pure Chemical Industries, Ltd.), 1.0% glucose (manufactured by Wako Pure Chemical Industries, Ltd.), 0.5% polypeptone (manufactured by Wako Pure Chemical Industries, Ltd.) 0.6% wheat germ (Nisshin Pharma Co., Ltd.), 0.3% yeast extract (Wako Pure Chemical Industries, Ltd.), 0.2% soybean meal (Showa Sangyo Co., Ltd.), 0.2% CaCO ₃ (manufactured by Wako Pure Chemical Industries, Ltd., pH 7.0 (adjusted with NaOH)) was added and sterilized at 121 ° C. for 20 minutes. Next, Staphylotricum boninense PF1444 strain was inoculated and cultured on a rotary shaker (220 rpm) at 25 ° C. for 3 days to 6 days, and this culture solution was used as a seed culture solution.

-Culture process-
In a 500 mL Erlenmeyer flask, a mixture of 80 g of brown rice (Akitakomachi) dipped in water overnight and drained for 1 hour the following morning and 2 g of oatmeal (Quaker Oats) was sterilized at 121 ° C for 20 minutes, The medium used for the following culture (hereinafter sometimes referred to as “production medium”) was used.
The production medium was inoculated with 3 mL of the seed culture solution and statically cultured at 25 ° C. for 14 days. The culture was performed using 25 500 mL Erlenmeyer flasks.

-Sampling process-
After the culture, 160 mL of acetone water (67% by volume) was dispensed into each Erlenmeyer flask and extracted to obtain a sample preparation solution.
The sample preparation solution was concentrated to dryness under reduced pressure. To this, 2 L of deionized water was added and suspended, and eluted with an equal amount of ethyl acetate. The obtained elution fraction was concentrated to dryness under reduced pressure to obtain 5 g of an extract. Next, the extract was subjected to column chromatography on 250 g of silica gel (Wakogel C-200, manufactured by Wako Pure Chemical Industries, Ltd.), and concentration gradient elution of chloroform and methanol was performed. The resulting elution fraction was concentrated to dryness to obtain a crude product.

-Purification step-
The obtained crude product is dissolved in methanol and subjected to high performance liquid chromatography (column: CAPCELL PAK UG120, particle size 5 μm, inner diameter 30 mm, length 250 mm, manufactured by Shiseido Co., Ltd.), and contains 0.1% by volume acetic acid. Elution was performed with 30% by volume and 40% by volume acetonitrile aqueous solution.
The elution fraction obtained was concentrated to dryness, 30 mg of red powder quinofuracin A, 5 mg of red powder quinofuracin B, 10 mg of red powder quinofuracin C, 60 mg of red powder quinofuracin D, quinofuracin E of yellow orange powder. 25 mg of each was isolated and purified.

-Physicochemical properties of compound (quinofuracin A) represented by structural formula (A)-
The physicochemical properties of the obtained quinofuracin A are as follows, and from these, it was confirmed that the quinofuracin A is a novel compound having a structure represented by the following structural formula (A). .
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₆ H ₃₀ O ₁₂ and molecular weight is 534.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 557.1614 (M + Na) ⁺ and the calculated value is m / z 557.1629 (as C ₂₆ H ₃₀ O ₁₂ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.38), 262 (4.16), 295 (4.27), 314 (4.23), 469 (3.78)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 211 (4.63), 231 (4.29), 265 (4.10), 324 (4.60), 398 (3. 79), 546 (3.82)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 224 (4.40), 253 (4.14, sh), 264 (4.18), 293 (4.40), 320 ( 3.92), 453 (3.90)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.15.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 10.8 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 620, 770, 1070, 1170, 1210, 1260, 1400, 1470, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.

-Physicochemical properties of the compound represented by the structural formula (B) (quinofuracin B)-
The physicochemical properties of the obtained quinofuracin B are as follows, and from these, it was confirmed that the quinofuracin B is a novel compound having a structure represented by the following structural formula (B). .
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₈ H ₃₂ O ₁₃ , and the molecular weight is 576.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 599.1724 (M + Na) ⁺ and the calculated value is m / z 599.1735 (as C ₂₈ H ₃₂ O ₁₃ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.37), 263 (4.11), 294 (4.23), 314 (4.09), 460 (3.75)
(Ii) When measured with methanol-NaOH solution λ _max nm (log ε): 211 (4.67), 231 (4.29), 264 (4.10), 325 (4.55), 397 (3. 74), 545 (3.78)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 224 (4.40), 253 (4.10, sh), 264 (4.13), 293 (4.35), 320 ( 3.88, sh), 453 (3.93)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.39.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 15.2 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 630, 780, 1070, 1170, 1210, 1260, 1380, 1450, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.

-Physicochemical properties of the compound represented by the structural formula (C) (quinofuracin C)-
The physicochemical properties of the obtained quinofuracin C are as follows, and from these, it was confirmed that the quinofuracin C is a novel compound having a structure represented by the following structural formula (C). .
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₆ H ₃₀ O ₁₂ and molecular weight is 534.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 557.1616 (M + Na) ⁺ and the calculated value is m / z 557.1629 (as C ₂₆ H ₃₀ O ₁₂ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.42), 261 (4.22), 290 (4.25), 314 (4.16), 455 (3.85)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 212 (4.63), 237 (4.42), 258 (4.19), 290 (4.17, sh), 314 ( 4.34), 527 (3.91)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 225 (4.44), 253 (4.16, sh), 264 (4.23, sh), 288 (4.35), 319 (3.94), 448 (3.94)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.16.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 15.4 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 650, 770, 1080, 1170, 1210, 1280, 1400, 1470, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.

-Physicochemical properties of compound (quinofuracin D) represented by structural formula (D)-
The physicochemical properties of the obtained quinofuracin D are as follows, and from these, it was confirmed that the quinofuracin D is a novel compound having a structure represented by the following structural formula (D). .
(1) Appearance: Red powder.
(2) Molecular formula: represented by C ₂₆ H ₂₈ O ₁₃ , and the molecular weight is 548.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental value is m / z 571.1422 (M + Na) ⁺ and the calculated value is m / z 571.1422 (as C ₂₆ H ₂₈ O ₁₃ Na).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 223 (4.39), 263 (4.14), 290 (4.30), 314 (4.12), 460 (3.82)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 211 (4.64), 232 (4.29), 263 (4.10), 324 (4.56), 397 (3. 73), 542 (3.84)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 223 (4.41), 253 (4.09, sh), 265 (4.14), 293 (4.39), 321 ( 3.86), 453 (3.91)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.11.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 4.2 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 620, 770, 1090, 1160, 1210, 1260, 1400, 1470, 1620, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, CD ₃ OD / TMS):
The measurement results are as shown in FIG.

-Physicochemical properties of compound (quinofuracin E) represented by structural formula (E)-
The physicochemical properties of the obtained quinofuracin E are as follows. From these, it was confirmed that the quinofuracin E is a novel compound having a structure represented by the following structural formula (E). .
(1) Appearance: Yellowish orange powder.
(2) Molecular formula: It is represented by C ₂₄ H ₂₂ O ₁₂ and its molecular weight is 502.
(3) High resolution mass spectrometry (HRESIMS: positive ion mode):
The experimental values are m / z 503, 1173 (M + H) ⁺ and the calculated value is m / z 503.1184 (as C ₂₄ H ₂₃ O ₁₂ ).
(4) Ultraviolet absorption spectrum:
The main peaks are as shown below.
(I) When measured with a methanol solution λ _max nm (log ε): 222 (4.41), 251 (4.10, sh), 266 (4.20, sh), 287 (4.35), 317 ( 3.84), 442 (3.89)
(Ii) When measured with a methanol-NaOH solution λ _max nm (log ε): 212 (4.61), 240 (4.26, sh), 263 (4.32), 311 (4.03), 512 ( 3.84)
(Iii) When measured with a methanol-HCl solution λ _max nm (log ε): 223 (4.40), 251 (4.12, sh), 266 (4.21, sh), 287 (4.36), 318 (3.83), 443 (3.90)
(5) Thin layer chromatography:
The Rf value measured by developing with a developing solvent [chloroform: methanol: acetic acid (10: 1: 0.03, volume ratio)] in a thin layer chromatography of a TLC plate (TLC Silica gel 60F ₂₅₄ , manufactured by Merck) 0.25.
(6) High performance liquid chromatography:
As the HPLC column, CAPCELLPAK UG120 (particle diameter 5 μm, inner diameter 4.6 mm, length 250 mm, manufactured by Shiseido Co., Ltd.) was used, and as the solvent, 40 volume% acetonitrile aqueous solution containing 0.1 volume% acetic acid was used. The holding time when the flow rate is 1 mL / min is 6.0 minutes.
(7) Infrared absorption spectrum:
The main peaks of the infrared absorption spectrum (KBr tablet method) are as shown below.
ν _max cm ⁻¹ : 650, 770, 950, 1070, 1160, 1190, 1250, 1320, 1390, 1470, 1630, 2930, 3430
(8) Proton nuclear magnetic resonance spectrum (600 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.
(9) Carbon-13 nuclear magnetic resonance spectrum (150 MHz, Pyridine- d ₅ / TMS):
The measurement results are as shown in FIG.

(Test Example 1)
For each of the quinofuracins A to E, the cancer cell proliferation inhibitory action was examined by the following method.

-Examination using LNZTA3 cells-
As the cancer cells, human glioblastoma cell line LNZTA3 cells (obtained from ATCC) that can regulate the expression level of human wild-type p53 protein by the concentration of tetracycline were used.
Using the LNZTA3 cells, 1 μg / mL or 0.004 μg / mL tetracycline containing 1% by volume FBS (manufactured by Sigma Aldrich) and 1% by volume Opti-MEM (manufactured by Life Technologies Japan). 1 × 10 ⁵ cells / mL were seeded at 100 μL per well of a 96-well plate and cultured at 37 ° C. in an atmosphere of 5% CO ₂ for 24 hours to completely adhere the cells.
The LNZTA3 cells have a low expression level of human wild-type p53 protein when the content of tetracycline in the medium is 1 μg / mL, and a high expression level of human wild-type p53 protein at 0.004 μg / mL.

In the 96-well plate to which the LNZTA3 cells are adhered, the final concentrations of quinofuracins A to E are 0.79 μg / mL, 1.57 μg / mL, 3.13 μg / mL, 6.25 μg / mL, 12 The solution was added to 5 μg / mL, 25 μg / mL, 50 μg / mL, or 100 μg / mL, and cultured for 3 days at 37 ° C. in a 5% CO ₂ atmosphere.
As a comparative control, the 96-well plate to which the LNZTA3 cells were adhered was cultured for 3 days at 37 ° C. in a 5% CO ₂ atmosphere without adding quinofuracin A to E.
After culturing for 3 days, cell proliferation of LNZTA3 cells was measured using the MTT method. The cell growth inhibition rate (%) was calculated by the following formula (1), where “A” was the absorbance of the system to which any of quinofuracin A to E was added, and “B” was the absorbance of the comparative control system.
Cell growth inhibition rate (%) = {(B−A) / B} × 100 (1)
Next, from the cell growth inhibition rate, the concentration (IC ₅₀ value) at which cell growth of LNZTA3 cells was inhibited by 50% was determined. The results are shown in Table 1.

As is clear from the results in Table 1, all of quinofuracins A to E show a cancer cell growth inhibitory effect, and also show a specific cell growth inhibitory effect depending on the expression level of wild-type p53 protein. all right.
Moreover, it was found that among quinofuracins A to E, kinofuracin A, B, C, and D have a high cell growth inhibitory effect, and in particular, quinofuracin A and B have a high cell growth inhibitory effect.

Examples of the aspect of the present invention include the following.
<1> A compound represented by any one of the following structural formulas (A) to (E).
<2> A method for producing the compound according to <1>,
A culture step for culturing a microorganism belonging to the genus Staphylotricum and having the ability to produce the compound according to <1>above;
And a collecting step of collecting the compound according to <1> from the culture obtained in the culturing step.
<3> belongs to Staphylococcus tri Kum (Staphylotrichum) genus, the microorganism having the ability to produce a compound according to <1> is, Accession No. NITE BP-1450 of Staphylococcus tri Kum Boninense (Staphylotrichum boninense) in PF1444 strain A method for producing the compound according to <2>.
<4> A microorganism belonging to the genus Staphylotricum and having the ability to produce the compound according to <1>.
<5> The microorganism according to the above <4>, which is Staphylotrichum boninense PF1444 strain having an accession number of NITE BP-1450.
<6> A compound-containing composition comprising the compound according to <1>.
<7> An anticancer agent comprising the compound according to <1>.

  NITE BP-1450

  Since the compound represented by any one of the structural formulas (A) to (E) of the present invention is a highly safe compound having an excellent anticancer activity, a pharmaceutical composition, an anticancer agent, It can be suitably used as an active ingredient.

Claims (7)

A compound represented by any one of the following structural formulas (A) to (E):
It is a manufacturing method of the compound of Claim 1, Comprising:
A culture step of culturing a microorganism belonging to the genus Staphylotricum and having the ability to produce the compound of claim 1;
A method for producing a compound comprising the step of collecting the compound according to claim 1 from the culture obtained in the culturing step. The microorganism belonging to the genus Staphylotrichum and capable of producing the compound according to claim 1 is Staphylotrichum boninense PF1444 strain having the accession number NITE BP-1450. A method for producing the compound described in 1. A microorganism belonging to the genus Staphylotricum and having the ability to produce the compound according to claim 1. The microorganism according to claim 4, which is Staphylotrichum boninense PF1444 strain having an accession number of NITE BP-1450.   A compound-containing composition comprising the compound according to claim 1.   An anticancer agent comprising the compound according to claim 1.