JPWO2018056470A1 - Epithelial-mesenchymal transition induction cell inhibitor - Google Patents

Abstract

An object of the present invention is to provide a novel compound which selectively damages cells which have been mesenchymalized by epithelial-mesenchymal transition.
The compound etc. which are represented by following formula (I).
[Chemical formula 1]

[In the Formula, R ¹ represents a C1-6 alkyl group, and R ² represents a C1-6 alkoxycarbonyl group, a carboxy group, or a C1-6 alkyl group substituted with a hydroxyl group. ]

Description

Cross reference

  The present application claims priority from Japanese Patent Application No. 2016-186970 filed on September 26, 2016 to the Japanese Patent Office. The contents of Japanese Patent No. 2016-186970 for which the present application claims priority are all incorporated by reference in their entirety. Also, all documents cited throughout the present application are incorporated herein by reference in their entirety.

  The present invention relates to a novel nanaomycin compound, a method for producing the same, a pharmaceutical composition containing nanaomycin as an active ingredient, and an Epithelial Mesenchymal Transition (EMT) -induced cell inhibitor.

  Epithelial cells are known to transform into mesenchymal cells by various stimuli. This transformation is called epithelial-mesenchymal transition, and is considered to be involved in wound healing and fibrosis in normal cells.

  Nanaomycins A and B are compounds found as new antibiotics from Streptomyces OS-3966, and their antibacterial activity on Mycoplasma gallisepticum KP-13 has been reported (Non-patent Document 1).

Haruo Tanaka et al., The Journal of Antibiotics (1975) Vol. XXVIII; 11: pp. 860-867.

  An object of the present invention is to provide a novel compound which selectively damages cells which have been mesenchymalized by epithelial-mesenchymal transition.

  The present inventors searched for various compounds and conducted intensive studies on their biological activities, and as a result, the novel substance nanaomycin and its derivatives were selected against the cells which were mesenchymalized by epithelial-mesenchymal transition. Found to be injured.

The present invention has been made based on such findings, and accordingly, the present invention relates to the following inventions.
(1) A compound represented by the following formula (I) or an ester derivative thereof or a salt, hydrate or solvate thereof (hereinafter referred to as “compound represented by the above-mentioned formula (I) of the present invention, etc. Sometimes

[Wherein, R ¹ represents a C1-6 alkyl group, R ² represents a C1-6 alkoxycarbonyl group, a carboxy group, or a C1-6 alkyl group substituted with a hydroxyl group, and R ³ represents The group represented by either is shown. ]
(2) In the compound according to (1), a microorganism belonging to an actinomycete having the ability to produce a compound wherein R ¹ is a methyl group and R ² is a carboxy group is cultured in a medium, and The method for producing a compound according to claim 1, wherein the compound is accumulated and the compound is collected from the culture, wherein R ¹ is a methyl group and R ² is a carboxy group.
(3) In the compound described in (1), a microorganism belonging to an actinomycete having the ability to produce a compound wherein R ¹ is a methyl group and R ² is a carboxy group is Streptomyces sp. 3. The process according to claim 2, which is K15-0591 (reception number NITE ABP-02304).
(4) Streptomyces sp. K15-0591 (reception number NITE ABP-02304).
(5) A pharmaceutical composition comprising the compound according to (1) as an active ingredient.
(6) An agent for damaging cells in which epithelial-mesenchymal transition has been induced, comprising the compound according to (1) as an active ingredient.

  Therefore, in one aspect, the present invention provides a novel compound nanaomycin or an ester derivative thereof represented by the formula (I), which is isolated from a culture solution of Streptomyces sp. Strain K15-0591. These salts or hydrates are provided.

In the compound represented by the formula (I) of the present invention, R ¹ is a linear or branched C1-6 alkyl group. In the present specification, the “C1-6 alkyl group” means a linear or branched saturated hydrocarbon group having 1 to 6 carbon atoms, and examples thereof include a methyl group, an ethyl group and an n-propyl group, Examples thereof include i-propyl group, n-butyl group, sec-butyl group, t-butyl group, isobutyl group, pentyl group, isopentyl group, 2,3-dimethylpropyl group, hexyl group, cyclohexyl group and the like, preferably And C.sub.1-5 alkyl group, more preferably methyl group, ethyl group, n-propyl group, i-propyl group, n-butyl group, sec-butyl group, t-butyl group, isobutyl group, pentyl group, It is an isopentyl group or a 2,3-dimethylpropyl group. More preferably, it is a C1-3 alkyl group, for example, a methyl group, an ethyl group, an n-propyl group, and an i-propyl group, and most preferably a methyl group, an ethyl group or a propyl group.

In the compound represented by the formula (I) of the present invention, R ² is a C1-6 alkoxycarbonyl group, a carboxy group (COOH group), or a C1-6 alkyl group substituted with a hydroxyl group. In the present specification, the term "C1-6 alkoxycarbonyl group" refers to a (C1-6 alkyl group) -OC (= O)-group, and the alkyl group portion may be linear. It may be branched. The term "C1-6 alkoxycarbonyl group" means that the number of carbon atoms in the alkyl group portion is 1 to 6. As the C1-6 alkoxycarbonyl group, for example, methoxycarbonyl group, ethoxycarbonyl group, 1-propyloxycarbonyl group, 2-propyloxycarbonyl group, 2-methyl-1-propyloxycarbonyl group, 2-methyl-2- Propyloxycarbonyl group, 2,2-dimethyl-1-propyloxycarbonyl group, 1-butyloxycarbonyl group, 2-butyloxycarbonyl group, 2-methyl-1-butyloxycarbonyl group, 3-methyl-1-butyl Oxycarbonyl group, 2-methyl-2-butyloxycarbonyl group, 3-methyl-2-butyloxycarbonyl group, 1-pentyloxycarbonyl group, 2-pentyloxycarbonyl group, 3-pentyloxycarbonyl group, 2-methyl -1-pentyloxycarbonyl group, 3- Tyl-1-pentyloxycarbonyl group, 2-methyl-2-pentyloxycarbonyl group, 3-methyl-2-pentyloxycarbonyl group, 1-hexyloxycarbonyl group, 2-hexyloxycarbonyl group, 3-hexyloxycarbonyl group Groups and the like. The C1-6 alkoxy group is preferably a C1-5 alkoxy group, and more preferably a methoxy group, an ethoxy group, an n-propyloxycarbonyl group, an i-propyloxycarbonyl group, an n-butyloxycarbonyl group, sec- A butyloxycarbonyl group, a t-butyloxycarbonyl group, an isobutyloxycarbonyl group, a pentyloxycarbonyl group, an isopentyloxycarbonyl group, and a 2,3-dimethylpropyloxycarbonyl group, and more preferably a C1-3 alkoxycarbonyl group It is a group (a methoxycarbonyl group, an ethoxycarbonyl group and a propyloxycarbonyl group), and still more preferably a methoxycarbonyl group or an ethoxycarbonyl group.

  In the present specification, a C1-6 alkyl group substituted by a hydroxyl group means the above C1-6 alkyl group substituted by one hydroxyl group, and examples thereof include a hydroxymethyl group, a 1-hydroxyethyl group, 2 -Hydroxyethyl group, 1-hydroxypropyl group, 2-hydroxypropyl group, 1-hydroxybutyl group, 2-hydroxybutyl group, 3-hydroxybutyl group, 1-hydroxypentyl group, 2-hydroxypentyl group, 3-hydroxy group Examples thereof include pentyl group, 4-hydroxypentyl group, 1-hydroxyhexyl group, 2-hydroxyhexyl group, 3-hydroxyhexyl group, 4-hydroxyhexyl group, 5-hydroxyhexyl group, and hydroxycyclohexyl group, with preference given to , A C 1-3 alkyl group substituted with one hydroxyl group, for example, Hydroxymethyl group, 1-hydroxyethyl group, a 2-hydroxyethyl group.

  The compounds represented by the formula (I) of the present invention include nanaomycin H represented by the following formula.

  In another aspect, the present invention relates to culturing a microorganism belonging to an actinomycete having the ability to produce nanaomycin H represented by the above formula in a culture medium to accumulate nanaomycin H represented by the above formula in the culture. The present invention relates to a method for producing nanaomycin H represented by the above formula, which comprises collecting nanaomycin H represented by the above formula from the culture.

In the present specification, naomycin H is a compound having the following physical properties:
(1) Properties: White powder or pale yellow powder (2) Molecular weight: 804
(3) Molecular formula: C ₃₃ H ₄₄ N ₂ O ₁₉ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 805.2307, actual value (m / z) 805.2337
(5) Specific rotation: [α] _D ^25.7 = -7.675 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 231 (40 200), 270 (16643, sh), 353 (13346)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3412 and 1637 cm ^-1 (8) ¹ H NMR (in heavy methanol) δ ppm: 1.752 (3 H, d, 8.0) , 1.907 (3H, s), 2.053 (1 H, d, 12.0), 2.278 (1 H, dd, 9.5, 12.0), 2.433 (1 H, dd, 8. 0, 12.5), 2.612 (2H, d, 6.0), 2.808 (1H, dd, 5.0, 12.5), 3.151 (1H, t, 9.5), 3.309 (1 H, dd, 9.5, 10.0), 3.34 (1 H, dd, 2.5, 9.5), 3.43 (1 H, dd, 3.0, 10.0) , 3.592 (1 H, dd, 9.5, 10.0), 3.645 (1 H, dd, 7.0, 12.0), 3.719 (1 H, dd, 9.5, 0.0), 3.772 (1H, dd, 9.5, 10.0), 3.819 (1H, ddd, 2.5, 7.0, 10.0), 3.85 (1H, dd) , 2.5, 12.0), 3.85 (1 H, dd, 3.5, 10.0), 4. 18 (1 H, dd, 2.5. 3.0), 4.22 (1 H, 1 H, dd) dd, 8.0, 15.0), 4.44 (1 H, dd, 5.0, 9.0), 4.7 (1 H, ddd, 6.0, 6.0, 9.5), 5 .042 (1 H, d, 3.5), 7. 242 (1 H, dd, 1.0, 8.5), 7.57 (1 H, dd, 1.0, 8.0), 7.698 ( 1H, dd, 8.0, 8.5)
(9) ¹³ C NMR (in heavy methanol) δ ppm: 16.2, 22.6, 31.5, 33.6, 41.5, 54.2, 55.6, 60.1, 62.8, 64.6, 72, 37.2, 87.2, 73.2, 74.0, 74.1, 74.4, 75.5, 76.4, 77.5, 80.4, 100. 1, 116.0, 120.0, 124.6, 134.7, 137.9, 162.8, 172.0, 173.6, 175.6, 190.6, 200.5
(10) Solubility: Easily soluble in water, ethanol and methanol, poorly soluble in acetone and hexane.

  The compounds represented by the formula (I) of the present invention include nanaomycin I represented by the following formula.

  In another aspect, the present invention relates to culturing a microorganism belonging to an actinomycete having the ability to produce nanaomycin I represented by the above formula in a culture medium to accumulate nanaomycin I represented by the above formula in the culture. The present invention relates to a method for producing nanaomycin I represented by the above formula, which comprises collecting nanaomycin I represented by the above formula from the culture.

As used herein, nanaomycin I is a compound having the following physical properties:
(1) Properties: light yellow powder or light yellow amorphous solid (2) molecular weight: 481
(3) Molecular formula: C ₂₁ H ₂₃ NO ₁₀ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 482.1115, actual value (m / z) 482.1129
(5) Specific rotation: [α] _D ²⁶ = -142.7 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 204 (15392), 231 (15969), 267 (sh), 353 (5050)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3451, 1646, 1527 cm ^-1 (8) ¹ H NMR (in heavy dimethyl sulfoxide) δ ppm: 1.61 (3H, d, 7 2), 1.72 (1H, s), 1.82 (1H, d, 13.5), 2.13 (1H, dd, 13.5, 12.0), 2.33 (1H, dd) , 12.6, 8.4), 2. 39 (1 H, dd, 15.3, 9.0), 2.54 (1 H, dd, 15.3, 3.6), 2.64 (1 H, dd, 12.6, 4.8), 4.13 (1 H, q, 7.2), 4. 19 (1 H, m), 4.50 (1 H, m), 7. 28 (1 H, d, 8.1), 7.45 (1 H, d, 7.6), 7.72 (1 H, dd, 8.1, 7.6)
(9) ¹³ C NMR (in heavy dimethyl sulfoxide) δ ppm: 15.5, 22.2, 29.9, 31.9, 40.3, 51.3, 58.4, 62.8, 72.6 , 75.9, 114.4, 118.6, 123.6, 132.9, 137.0, 160.3, 169.2, 171.3, 172.1, 189.2, 198.7
(10) Solubility: Easily soluble in DMSO and methanol. Poorly soluble in hexane, ethyl acetate and chloroform.

  The compounds represented by the formula (I) of the present invention include nanaomycin J represented by the following formula.

  In another aspect, the present invention relates to culturing a microorganism belonging to an actinomycete having the ability to produce nanaomycin J represented by the above formula in a culture medium to accumulate nanaomycin J represented by the above formula in the culture. The present invention relates to a method for producing nanaomycin J represented by the above formula, which comprises collecting nanaomycin J represented by the above formula from the culture.

As used herein, nanaomycin J is a compound having the following physical properties:
(1) Properties: light yellow powder or light yellow amorphous solid (2) molecular weight: 643
(3) Molecular formula: C ₂₇ H ₃₄ N ₂ O ₁₄ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 643.1803, found value (m / z) 643.1833
(5) Specific rotation: [α] _D ²⁶ = -125.5 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 204 (240 75), 232 (18746), 267 (sh), 353 (5778)
(7) Infrared absorption maximum ν _max (KBr tablet): 3463, 1643, 1527 cm ⁻¹ having maximum absorption (8) ¹ H NMR (in heavy methanol) δ ppm: 1.76 (3 H, d, 7. 2), 1.87 (1 H, s), 2.06 (1 H, d, 14.1), 2.27 (1 H, dd, 14.1, 4.8), 2.40 (1 H, dd, 12.6, 9.0), 2.61 (1 H, d, 6.0), 2.77 (1 H, dd, 12.6, 5.1), 3.37 (1 H, dd, 9.0) , 9.0), 3.68 (1 H, dd, 10.0, 9.0), 3.69 (1 H, dd, 12.0, 6.0), 3. 76 (1 H, dd, 10. 0, 3.0), 3.79 (1H, dd, 12.0, 3.0), 3.79 (1H, ddd, 9.0, 6.0, 3.0), 4.21 (1H , Q, 7. ), 4.50 (1 H, dd, 9.0, 5.1), 4.69 (1 H, m), 5.03 (1 H, d, 3.0), 7.24 (1 H, d, 8) .4, 0.9), 7.56 (1 H, d, 7.5, 0.9), 7.69 (1 H, dd, 8.4, 7.5)
(9) ¹³ C NMR (in heavy methanol) δ ppm: 16.1, 22.4, 31.3, 33.8, 41.6, 53.7, 56.1, 60.1, 62.8, 64.6, 72.4, 72.6, 73.2, 75.5, 77.5, 92.4, 116.0, 119.9, 124.6, 134.6, 137.9, 162. 6, 172.1, 173.2, 175.6, 190.7, 200.5
(10) Solubility: Easily soluble in DMSO and methanol. Poorly soluble in hexane, ethyl acetate and chloroform.

  Examples of the compound represented by the formula (I) of the present invention include nanaomycin K represented by the following formula.

  In another aspect, the present invention relates to culturing a microorganism belonging to an actinomycete having the ability to produce nanaomycin K represented by the above formula in a culture medium to accumulate nanaomycin K represented by the above formula in the culture. The present invention relates to a method for producing nanaomycin K represented by the above formula, which comprises collecting nanaomycin K represented by the above formula from the culture.

As used herein, nanaomycin K is a compound having the following physical properties:
(1) Properties: orange powder or orange amorphous solid (2) molecular weight: 547
(3) Molecular formula: C ₂₅ H ₂₉ N ₃ O ₉ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 548.1697, found value (m / z) 548.1709
(5) Specific rotation: [α] _D ²⁶ = −200.1 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 204 (18488), 235 (26967), 356 (5907)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3432, 1677, 1527 cm ^-1 (8) ¹ H NMR (in heavy methanol) δ ppm: 1.87 (3 H, br d), 1.94 (1 H, d, 4.9), 2.27 (1 H, dd, 12.0, 12.0), 2.57 (1 H, d, 14.9, 8.0), 2.70 (1H, dd, 14.9, 4.0), 3.10-3.28 (2H, m), 3.25 (9H, s), 3.95 (1 H, brd), 4.28 ( 1 H, q, 6.9), 4.86 (1 H, overlap), 7.03 (1 H, s), 7.24 (1 H, d, 8.2), 7.41 (1 H, d, 6. 9), 7.66 (1 H, br t)
(9) ¹³ C NMR (in heavy methanol) δ ppm: 16.2, 27.4, 32.1, 41.8, 52.9, 63.5, 64.5, 75.4, 77.0, 79.4, 116.1, 120.5, 122.5, 122.7, 134.8, 138.1, 138.6, 163.0, 171.0, 175.2, 190. 7,200.3
(10) Solubility: Easily soluble in DMSO and methanol. Poorly soluble in hexane, ethyl acetate and chloroform.

  In the present specification, the salt of the compound represented by the formula (I) of the present invention means a complex salt formed by coordination of a metal or the like to the compound represented by the formula (I). In addition, the hydrate or solvate of the compound represented by the formula (I) of the present invention and the hydrate or solvate of the salt of the compound represented by the formula (I) of the present invention are also included in the present invention. . Preferably, the salt of the compound represented by the formula (I) of the present invention, and the hydrate or solvate of the compound represented by the formula (I) of the present invention and the compound represented by the formula (I) of the present invention Hydrates or solvates of salts of the following are pharmacologically acceptable. In addition, the compound represented by the formula (I) of the present invention or a salt thereof may be a crystal or an amorphous.

  The compounds of the present invention include, in addition to the above-mentioned compounds, pharmacologically acceptable ester derivatives of these compounds. Here, the “pharmaceutically acceptable ester derivative” is a compound containing a group which is metabolized in vivo to give the compound of the present invention, and the pharmaceutically acceptable ester of the ester is acceptable for administration in the body. It is. In the present specification, an ester includes, in addition to an ester-linked compound, an amide-linked compound. The esters may be degraded by in vivo esterases to give active compounds. For example, as the ester, substituted or unsubstituted lower alkyl ester, lower alkenyl ester, lower alkylamino lower alkyl ester, acylamino lower alkyl ester, acyloxy lower alkyl ester, aryl ester, aryl lower alkyl ester, amide, lower Alkyl amide, hydroxide amide can be mentioned. As ester, preferred is propionic acid ester or acyl ester.

  Since the compounds of the present invention may have asymmetric carbons, optical isomers may exist. The compounds of the present invention may be either dextrorotatory (+) or levorotatory (-) compounds, or may be a mixture of these isomers such as racemates. The compounds of the present invention also include any tautomers or geometric isomers (eg, E-form, Z-form and the like) unless otherwise specified.

  Alternatively, the present invention relates to a pharmaceutical composition comprising the compound represented by the above formula (I) or an ester derivative thereof or a salt or hydrate thereof as an active ingredient.

  Furthermore, the present invention relates to a pharmaceutical composition comprising, as an active ingredient, the compound represented by the above formula (I) or an ester derivative thereof or a salt or hydrate thereof. In another aspect, the present invention is to damage cells in which epithelial-mesenchymal transition has been induced, which comprises the compound represented by the above formula (I) or an ester derivative thereof or a salt or hydrate thereof as an active ingredient. Related to the drug. As used herein, “agent for damaging cells in which epithelial-mesenchymal transition has been induced” specifically damages and kills cells in which conversion of epithelial cells into mesenchymal cells has been induced, Or, it means an agent for inducing apoptosis.

  The pharmaceutical composition of the present invention preferably specifically damages cells induced to convert epithelial cells to mesenchymal cells. In the present specification, to specifically damage the cells induced to be transformed from epithelial cells to mesenchymal cells does not cause any damage to the epithelial cells, but it has been transformed from epithelial cells to mesenchymal cells It means to injure cells. Here, hardly causing harm does not mean that there is no toxicity at all, but means that there is no toxicity to the extent that it does not cause any problem when used as a medicine, for example, it exhibits no toxicity at 30 μM. No, or 100 μM is meant to indicate a survival rate of 85% or more.

  Further, in the present invention, the kind of the pharmaceutical composition, the cytotoxic agent, the therapeutic agent, the preventive agent, the metastasis inhibitor, and the infiltration suppressor (hereinafter referred to as “pharmaceutical composition etc.”) is not particularly limited. The dosage forms include tablets, capsules, granules, powders, syrups, suspensions, suppositories, ointments, creams, gels, patches, inhalants, injections and the like. These formulations can be prepared according to a conventional method. In addition, the liquid preparation may be in the form of being dissolved or suspended in water or other suitable solvent when used. The tablets and granules may also be coated by known methods. In the case of injection, the compound of the present invention is prepared by dissolving it in water, but it may be dissolved in physiological saline or glucose solution if necessary, and even if a buffer or preservative is added. Good. It is provided in any dosage form for oral or parenteral administration. For example, pharmaceutical compositions for oral administration in the form of granules, fine granules, powders, hard capsules, soft capsules, syrups, emulsions, suspensions or solutions, for intravenous administration, for intramuscular administration, Alternatively, it can be prepared as a pharmaceutical composition for parenteral administration in the form of injections such as subcutaneous administration and the like, drips, percutaneous absorptions, transmucosal absorptions, nasal drops, inhalations and suppositories. The injections, drips and the like may be prepared as a powdery dosage form such as a lyophilised form, and may be dissolved in a suitable aqueous medium such as physiological saline at the time of use.

  The compounds of the present invention can specifically damage epithelial-mesenchymal transformed cells.

FIG. 1 is a graph showing the growth suppressive / toxic effects (MTS assay) on MDCK cells of nanaomycin H in a confluent state and a thin state. The absorbance is 490 nm on the vertical axis, DMSO (control) or nanaomycin H-treated group on the horizontal axis, BSF is added on the left, and TGF is added on the right, resulting in an epithelial-mesenchymal transition induced group. FIG. 2 is a photograph showing the results of observing the influence of the drug on the migrating cells. (A: DMSO (control), B: nanaomycin H), scale bar: 100 μm FIG. 3 is a graph showing the cell-killing effect of nanaomycins on MDCK cells induced to undergo epithelial-mesenchymal transition by TGF-β. As a control, BSA was added to the group to which TGF-β was not added. The upper row shows the results of addition of the nanaomycins at 5 μg / ml, and the lower row shows the results of the addition of the nanaomycins at 50 μg / ml (however, only nanaomycin H at 100 μg / ml). The vertical axis represents the cell viability when the number of cells before addition of the nanaomycins is 1. Open graphs show the results of wells seeded with 6000 cells of MDCK cells, and filled graphs show results of wells seeded with 2000 cells of MDCK cells. In the horizontal axis, BSA represents a control group without TGF-β addition, and TGF represents a TGF-β addition group. “H”, “I”, “J” and “K” each represent the type of nanaomycin added.

  The compound represented by the formula (I) of the present invention or its ester derivative or a salt or hydrate thereof is obtained by culturing a microorganism belonging to an actinomycete having an ability to produce naomycin H in a culture medium, and Can be produced by collecting (separating / extracting / purifying) nanaomycin from the culture, or by further chemically converting or modifying the obtained nanaomycin.

In the method for producing the naomycins (Nanaomycin H, Nanaomycin I, Nanaomycin J, and Nanaomycin K) of the present invention, “a microorganism belonging to an actinomycete having the ability to produce naomycins” is a bacterium belonging to an actinomycete. It is not particularly limited as long as it is a microorganism having an ability to produce naomycins. The strains that can be used in the method for producing a naomycin of the present invention include all the bacteria having the ability to produce naomycin H belonging to actinomycetes, as well as the above strains, as well as the above-mentioned strains. Whether or not the microorganism is "a microorganism belonging to an actinomycete having the ability to produce naomycin H" can be determined, for example, by the following method. Liquid medium (pH 7.0) consisting of 2.4% starch, 0.1% glucose, 0.3% peptone, 0.3% bonito extract, 0.5% yeast extract and 0.4% calcium bicarbonate (pH 7.0) Inoculate 1 mL of the test microorganism cultured in the liquid medium in a 500 mL Erlenmeyer flask containing the above, shake culture at 27 ° C. for 3 days, and then obtain the seed culture solution obtained by 5.0% starch, 0.5% glycerol, Inoculate 1 mL of a 500 mL Erlenmeyer containing 100 mL of a liquid medium (pH 7.0) consisting of 1.0% of defatted wheat germ, 1.0% of dry yeast, and 0.5% of calcium hydrogen carbonate for 8 days at 27 ° C. If Naomycin H is present in the culture obtained by shaking culture, it can be determined that the microorganism belongs to an actinomycete having an ability to produce naomycin H. Preferably, a microorganism belonging to an actinomycete having the ability to produce naomycin H is Streptomyces sp. Strain K15-0591 isolated from the soil of Nanao City, Ishikawa Prefecture. This microorganism has been deposited as of Sep. 26, 2017, at the Patent Microorganisms Depositary Center of the National Institute of Technology and Evaluation, Patent Microorganisms Depositary (2-8, Kazusa, Kisarazu City, Chiba) under the receipt number NITE ABP-02304. There is.

As used herein, the term "mutant" refers to Streptomyces sp. The K15-0591 strain is a strain having a mycological property or gene different from that of the K15-0591 strain, and such mutant strains include Streptomyces sp. Besides the strain derived from the K15-0591 strain, Streptomyces sp. The original strain derived K15-0591 is also included. In the present specification, a mutant strain does not ask for the presence or absence of an actual derivative trace, and, for example, Streptomyces sp. Strains having a gene having high homology (eg, 80% or more, 85% or more, 90% or more, 95% or more, etc.) with the K15-0591 strain gene (eg, 16S rRNA gene) are also included as mutant strains. In addition, such mutant strains may be artificially produced or collected from nature as long as they maintain the ability to produce nanaomycins.

  A medium for culturing a microorganism belonging to actinomycetes having the ability to produce nanaomycins can contain, as a nutrient source, one that can be used as a nutrient source for actinomycetes. For example, commercially available peptone, meat extract, corn steep liquor, cottonseed powder, peanut powder, soybean powder, yeast extract, NZ-amine, casein hydrate, nitrogen source such as sodium nitrate, ammonium nitrate, ammonium sulfate, glycerin Carbohydrates such as starch, glucose, galactose and mannose, or carbon sources such as fat, and inorganic salts such as sodium chloride, phosphate, calcium carbonate and magnesium sulfate can be used alone or in combination. In addition, trace amounts of metal salts may be added to the medium, and as a defoamer, animal / plant / mineral oil may be added. Any of these materials may be used as long as they play a role in the production of mangromycins using production bacteria, and all known culture materials of actinomycetes can be used.

  In addition, the culture of the microorganism belonging to actinomycetes having the ability to produce nanaomycins is a temperature range in which the producing bacteria can grow and produce nanaomycins (for example, 10 ° C to 40 ° C, preferably 25 to 30 ° C) And shaking culture for several days to two weeks. The culture conditions can be appropriately selected and carried out according to the nature of the nanaomycin-producing strain to be used, with reference to the description of the present specification.

  The collection of nanaomycins can be performed by extraction from the culture solution using a water immiscible organic solvent such as ethyl acetate. In addition to this extraction method, known methods used for collection of fat-soluble substances, such as adsorption chromatography, partition chromatography, gel filtration chromatography, scraping from thin layer chromatography, centrifugal countercurrent distribution chromatography, high-speed liquid Purification can be carried out to purity by combining or repeating chromatography etc. as appropriate.

The compound represented by the above formula (I) of the present invention or an ester derivative thereof or a salt or hydrate thereof can be synthesized by appropriately converting or modifying a nanomycin. Alternatively, the compound represented by the above-mentioned formula (I) of the present invention can be prepared by using Streptomyces sp. It can be obtained by isolation using the K15-0591 strain as a production strain and in the same manner as the production and purification of the above-mentioned nanaomycins.

  The pharmaceutical composition of the present invention can be formulated in a conventional manner using a conventional pharmaceutically acceptable carrier. There is no limitation on the dosage form for formulating a pharmaceutical composition containing the compound or the like represented by the above-mentioned formula (I) of the present invention as an active ingredient, and solid preparations such as tablets, pills, capsules, powders and granules The composition can be used orally as a liquid preparation such as a solution, suspension or emulsion, or parenterally as an intravenous, intramuscular, subcutaneous injection, suppository, patch or the like. When preparing a solid preparation for oral use, after adding an excipient and, if necessary, a binder, a disintegrant, a lubricant and the like to the main drug, solvents, granules, powders, capsules and the like are prepared by a conventional method. I assume. When preparing an injection, a pH adjuster, a buffer, a stabilizer, a solubilizer, etc. may be added to the main drug as necessary, and a subcutaneous or intravenous injection can be prepared by a conventional method.

  The present invention further comprises administering an effective amount of the compound represented by the above formula (I) of the present invention or its ester derivative or a salt or hydrate thereof to a patient in need thereof. Can be used in a method for treating or preventing a disease or disorder associated with or contributing to the onset or exacerbation. For example, when the compound or the like represented by the formula (I) of the present invention is used for therapeutic or prophylactic purposes, a pharmaceutical composition containing the compound or the like represented by the formula (I) or the like of the present invention as an active ingredient is orally It can be administered in a dosage form or a parenteral dosage form such as an injection, an infusion and the like. When the compound etc. represented by the said Formula (I) of this invention are administered to a mammal etc., you may orally administer as a tablet, a powder, a granule, a syrup etc. It may be administered orally. Although the dose varies depending on symptoms, age, sex, body weight, administration form and the like, for example, when orally administered to adults, the daily dose is usually 0.1 to 1000 mg.

  EXAMPLES Although an Example is given to the following and this invention is concretely demonstrated to it, this invention is not limited to this. All references cited throughout the present application are incorporated herein by reference in their entirety.

Example 1 Bacteriological Properties of Streptomyces sp. Strain K15-0591 By the present inventors, Streptomyces sp. K15 was newly prepared from the soil of Nanao City, Ishikawa Prefecture. The strain -0591 was isolated. The bacteriological properties of the Streptomyces sp. Strain K15-0591 were as follows.

(I) Morphological Properties The vegetative mycelium develops well on various agar media, and no shedding is observed. The aerial mycelium is abundantly deposited on starch mineral salt agar and exhibits a yellow shade. It forms aerial mycelium of about 0.1 μm. No sporangia and zoospores are found.

(II) Properties on Various Media Methods of EB Shirling and EB Gottlieb (International Journal of Systematic Bacteriology, Vol. 16, p. 313) The culture characteristics of the present strain examined by 1966) are shown in Table 1. The color tone can be determined using the color harmony manual 4th edition (Container Corporation of America Chicago, 1958) as a standard color, and the color chart name is described together with the code in parentheses. ing. The following are the results of observation in each culture medium at 27 ° C. for one week unless otherwise stated.

(III) Physiological properties (1) Melanin pigment formation: Positive (2) Tyrosinase reaction: Positive (3) H ₂ S production: Negative (4) Starch hydrolysis: Positive (5) Gelatin liquefaction (simple gelatin) Medium) (21-23 ° C.): Positive (6) Peptonization of skimmed milk (37 ° C.): Positive (7) Coagulation of skim milk (37 ° C.): Positive (8) Degradation of cellulose: weak positive (9) Growth Temperature range: 15 to 45 ° C
(10) Availability of carbon source (Predham and Gotribe agar)
Use: D-glucose, D-xylose, D-mannitol, L-arabinose, sucrose, rhamnose, raffinose Do not use: D-fructose, i-inositol

(IV) Chemical composition of cells Diaminopimelic acid in the cell wall is LL type.

(V) 16S rRNA gene analysis The sequence of about 1400 bases of the 16S rRNA gene was determined, and it was registered by the DNA database, using the data of the strain belonging to the genus Streptomyces and other actinomycetes by the neighbor connection method using data From the result of phylogenetic analysis, it is appropriate to classify this strain into the genus Streptomyces, which is most closely related to Streptomyces sp.

(VI) Conclusion The summary of the bacteriological properties of this bacterium is as follows. Diaminopimelic acid in the cell wall is LL type. Enriched aerial mycelium forms a helix. The colonies have a yellowish color and produce melanin pigment. From these results and the analysis results of the 16S rRNA gene, it was determined that this strain is a strain belonging to the genus Streptomyces. This strain is Streptomyces sp. K15-0591, and as of September 26, 2017, National Institute of Technology and Evaluation, National Institute of Technology and Evaluation, Patent Microorganisms Depositary Center, Kisarazu City, Chiba Prefecture 2-5-5. 8) (Accession No. NITE ABP-02304).

(Example 2) Acquisition of nanaomycin H 2.4% starch, 0.1% glucose, peptone (manufactured by Kyoto Pharmaceutical Industry Co., Ltd.) 0.3%, bonito extract (manufactured by Kyouto Pharmaceutical Co., Ltd.) 0.3% , 100 mL of a 500 mL Erlenmeyer flask containing 100 mL of a liquid medium (pH 7.0) containing 0.5% of yeast extract (manufactured by Oriental Yeast Co., Ltd.) and 0.4% of calcium hydrogen carbonate was cultured in the liquid medium Streptomyces sp. 1 ml each of K15-0591 (Accession No. NITE ABP-02304) was inoculated and shake cultured at 27 ° C. for 3 days. The obtained seed culture solution is 5.0% of starch, 0.5% of glycerol, 1.0% of defatted wheat germ (manufactured by Nisshin Pharma Co., Ltd.), 1% of dry yeast (manufactured by JT Foods Co., Ltd.), hydrogen carbonate 1 mL each was inoculated in 100 500 mL Erlenmeyer flasks containing 100 mL of a liquid medium (pH 7.0) consisting of 0.5% of calcium, and shake culture was carried out at 27 ° C. for 6 days.

  100 mL of ethanol was added to 100 500 mL Erlenmeyer flasks in which the culture was completed, and vigorously stirred for 1 hour. Next, ethanol in the extract was distilled off under reduced pressure, 10 L of ethyl acetate was added to the obtained aqueous solution, and after stirring well, the ethyl acetate layer was recovered. Concentration to dryness was performed using an evaporator to obtain 4.7 g of crude purified product 1. This is dissolved in a small amount of methanol, and step elution is carried out using a silica gel (manufactured by MERCK) open column chromatography with a chloroform-methanol solvent system (100: 0, 100: 1, 50: 1, 10: 1, 1: 1,0: 100) to obtain 1774 mg of 1: 1 fractions containing nanaomycin H and 0: 100 fractions (crude product 2).

  The crude product 2 is dissolved in a small amount of methanol, and stepwise elution is performed in a methanol-water system using ODS (manufactured by Fuji Silysia Chemical Ltd.) open column chromatography (methanol concentration 0%, 20%, 30%, 40% , 50%, 60%, 70%, 80%, 90%, 100%) to obtain 392 mg of a 20% fraction (crude product 3) containing nanaomycin H. The crude product 3 is dissolved in methanol and injected into an octadecylsilyl column (Inertsil ODS-4, φ14 × 250 mm, flow rate 6.5 mL / min, detection 254 nm) by high performance liquid chromatography, and 30% containing 0.1% formic acid Elution was with 2% methanol water. The peak with a retention time of about 22 minutes was separated, and concentration under reduced pressure gave 4.1 mg of nanaomycin H as a pale yellow powder.

  The physicochemical properties of the obtained nanaomycin H were measured and found to be as follows.

Nanaomycin H
(1) Properties: White powder or pale yellow powder (2) Molecular weight: 804
(3) Molecular formula: C ₃₃ H ₄₄ N ₂ O ₁₉ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 805.2307, actual value (m / z) 805.2337
(5) Specific rotation: [α] _D ^25.7 = -7.675 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 231 (40 200), 270 (16643, sh), 353 (13346)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3412, 1637 cm ^-1 (8) Proton nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy methanol are shown in Table 1. (In the table, s is a singlet, d is a doublet, m is a multiplet, and H is the number of protons.)
(9) Carbon nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy methanol are shown in Table 1.
(10) Solubility in solvents: Easily soluble in water, ethanol and methanol. Poorly soluble in acetone and hexane.

  As described above, a compound corresponding to nanaomycin H from which various physicochemical properties have been obtained has not been reported so far, so nanaomycin H is considered to be a novel substance.

(Example 3) Effect on MDCK (Canine Epithelial Cell Line) Inducing Epithelial-Mesenchymal Transition Induced by TGF-β The canine renal tubular epithelium-derived cell line MDCK (hereinafter referred to as “MDCK cell”) is used as a cell culture dish Eagle's minimal essential medium (hereinafter referred to as "MEM medium") (10% fetal bovine serum albumin, 1% MEM non-essential amino acid solution, 1% sodium pyruvate, 1% penicillin-streptomycin solution, 1% glutamine) Subculture was performed so as to be maintained in a semi-confluent state at 37 ° C. in a 5% CO ₂ incubator. The cells were treated with a trypsin-EDTA solution, seeded in the above-mentioned medium containing TGF-β at a final concentration of 10 ng / mL and 0.001% BSA, and cultured for 3 days. As a blank, cells were similarly prepared for 3 days in a medium supplemented with only BSA, without TGF-β added. These cells were adjusted to 96 wells with MDCK cells at 20,000 cells / well (confluent) and 2,000 cells / well (thin).

Add nanaomycin H (final concentration 0.5% DMSO) or nanaomycin methyl ester (final concentration 0.5% DMSO) dissolved in DMSO to a final concentration of 50 μg / ml to each well and add 37 The cells were cultured for 24 hours in a 5% CO ₂ incubator at 25 ° C., and toxicity / proliferation tests were performed using a morphology observation and live cell quantification kit (MTS assay). The results are shown in FIG. Although nanaomycin H showed almost no toxicity at confluence, it became thin and became about 40% cytotoxic (DMSO ratio). On the other hand, in the cells treated with TGFβ to induce EMT, most of the cells were killed.

(Example 4) Toxicity to mobile cells Using a functional substrate (Biomaterials, 2012, Voi. 33, pp. 2409-2428; Shimizu et al. Analytical Sciences, under printing) capable of cell patterning in response to light irradiation. After forming colonies of MDCK cells in geometrically defined circular area (φ = 150 μm), cell migration was induced by secondary irradiation. In this case, prepare a medium to which final concentrations of 50 μg / ml nanaomycin H and 0.5% DMSO, 50 μg / ml nanaomycin H methyl ester and 0.5% DMSO, or only DMSO were added. The migration behavior of cells in culture medium was observed under a phase contrast microscope equipped with a culture device.

  In the medium containing only DMSO, it is observed that the colony is expanded, but when the nanaomycin H or the nanaomycin H methyl ester is added, it is observed that the cells around the colony are selectively killed. The Since MDCK cells migrate in a population with the appearance of epithelial-mesenchymal transformed cells at the leading end, it was found that the result was specifically effective on epithelial-mesenchymal transformed cells (FIG. 2).

  From the above results, it was shown that nanaomycin H and nanaomycin H methyl ester have a growth inhibitory effect on cells in which epithelial-mesenchymal transition was induced.

(Example 5) Acquisition of nanaomycin I, J and K 2.4% of starch, 0.1% of glucose, 0.3% of peptone (manufactured by Kyoto Pharmaceutical Co., Ltd.), 0.3% of bonito extract (manufactured by Kyoto Pharmaceutical Co., Ltd.) In a 500 mL Erlenmeyer flask containing 60 mL of a 100 mL liquid medium (pH 7.0) consisting of 0.3%, 0.5% yeast extract (manufactured by Oriental Yeast Co., Ltd.) and 0.4% calcium bicarbonate Streptomyces sp. Cultured in culture medium. 1 ml each of K15-0591 (Accession No. NITE ABP-02304) was inoculated and shake cultured at 27 ° C. for 3 days. The obtained seed culture solution is 5.0% of starch, 0.5% of glycerol, 1.0% of defatted wheat germ (manufactured by Nisshin Pharma Co., Ltd.), 1% of dry yeast (manufactured by JT Foods Co., Ltd.), hydrogen carbonate 1 mL each was inoculated in 60 mL of a 500 mL Erlenmeyer flask containing 100 mL of a liquid medium (pH 7.0) consisting of 0.5% of calcium, and shake cultured at 27 ° C. for 6 days.

  Sixty 500 mL Erlenmeyer flasks that had been cultured were separated into cells and supernatant by centrifugation, and the resulting supernatant was passed through an HP-20 column and washed with water. The adsorbate was eluted with 3 L of methanol and concentrated to dryness using an evaporator to obtain 3.2 g of crude product 1. This is dissolved in a small amount of methanol and adsorbed onto silica gel, and step elution (50: 1, 25: 1, 10: 1, 6) in a chloroform-methanol solvent system using silica gel (manufactured by MERCK) open column chromatography. 4: 4: 6, 0: 1), and the 4: 6 fraction (crude product 1) containing nanaomycin I and J and the 0: 100 fraction (crude product 2) containing nanaomycin K, respectively 1.02 g, 0.65 g were obtained.

  The crude product 1 is dissolved in a small amount of methanol and eluted with methanol-water system using ODS (manufactured by Fuji Silysia Chemical Co., Ltd.) open column chromatography (methanol concentration 0% to 100% for 60 minutes) There were obtained 416 mg of a fraction containing nanaomycin I (crude product 3) and 163 mg of a fraction containing nanaomycin J (crude product 4). The crude product 3 is dissolved in methanol, eluted with methanol-water system using ODS (manufactured by Fuji Silysia Chemical Co., Ltd.) open column chromatography (methanol concentration 0% to 100% for 60 minutes, gradient), 28 mg of a fraction containing crude I (crude product 5) was obtained. The crude product 5 is injected into an octadecylsilyl column (Inartsil ODS-4, φ10 × 250 mm, flow rate 4.0 mL / min, detection PDA) by high performance liquid chromatography, and eluted with 20% methanol water containing 0.1% formic acid did. The peak with a retention time of around 15 minutes was separated, and concentration under reduced pressure gave 9.4 mg of nanaomycin I as a pale yellow powder. The crude product 4 is dissolved in methanol, eluted with methanol-water system using ODS (manufactured by Fuji Silysia Chemical Co., Ltd.) open column chromatography (gradient from 0% to 100% of methanol for 60 minutes), 95 mg of a fraction containing crude J (crude product 6) was obtained. The crude product 6 is injected into an octadecylsilyl column (YMC-Triart C-18, φ 20 x 250 mm, flow rate 10.0 mL / min, detection PDA) by high performance liquid chromatography, and 15% methanol water containing 0.1% formic acid The reaction mixture was eluted with to give a fraction containing nanaomycin J (crude product 7). The crude product 7 is injected into an octadecylsilyl column (YMC-Triart PFP, φ10 × 250 mm, flow rate 4.0 mL / min, detection PDA) by high performance liquid chromatography, and with 20% methanol water containing 0.1% formic acid It eluted. The peak with a retention time of about 10 minutes was separated, and concentration under reduced pressure gave 5.5 mg of nanaomycin J as a pale yellow powder. The crude product 2 is dissolved in a small amount of methanol and eluted with methanol-water system using ODS (manufactured by Fuji Silysia Chemical Ltd.) open column chromatography (methanol concentration 0% to 100% for 60 minutes) 74 mg of a fraction containing nanaomycin K (crude product 8) was obtained. The crude product 8 is injected into an octadecyl silyl column (YMC-Triart PFP, φ10 × 250 mm, flow rate 4.0 mL / min, detection PDA) by high performance liquid chromatography, and with 15% methanol water containing 0.1% formic acid It eluted. The peak with a retention time of about 12 minutes was separated, and concentration under reduced pressure gave 16.8 mg of nanaomycin K as an orange powder.

  The physicochemical properties of the obtained nanaomycins H, I, J and K were measured and found to be as follows.

Nana omycin I
(1) Properties: light yellow powder or light yellow amorphous solid (2) molecular weight: 481
(3) Molecular formula: C ₂₁ H ₂₃ NO ₁₀ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 482.1115, actual value (m / z) 482.1129
(5) Specific rotation: [α] _D ²⁶ = -142.7 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 204 (15392), 231 (15969), 267 (sh), 353 (5050)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3451, 1646, 1527 cm ^-1 (8) Proton nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy dimethyl sulfoxide are shown in Table 3 . (In the table, s is a singlet, d is a doublet, m is a multiplet, and H is the number of protons.)
(9) Carbon nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy dimethyl sulfoxide are shown in Table 3.
(10) Solubility: Easily soluble in DMSO and methanol. Poorly soluble in hexane, ethyl acetate and chloroform.

As used herein, nanaomycin J is a compound having the following physical properties:
(1) Properties: light yellow powder or light yellow amorphous solid (2) molecular weight: 643
(3) Molecular formula: C ₂₇ H ₃₄ N ₂ O ₁₄ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 643.1803, found value (m / z) 643.1833
(5) Specific rotation: [α] _D ²⁶ = -125.5 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 204 (240 75), 232 (18746), 267 (sh), 353 (5778)
(7) Infrared absorption maximum ν _max (KBr tablet): 3463, 1643, 1527 cm ⁻¹ (8) Proton nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy methanol are shown in Table 4. (In the table, s is a singlet, d is a doublet, m is a multiplet, and H is the number of protons.)
(9) Carbon nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy methanol are shown in Table 4.
(10) Solubility: Easily soluble in DMSO and methanol. Poorly soluble in hexane, ethyl acetate and chloroform.

As used herein, nanaomycin K is a compound having the following physical properties:
(1) Properties: orange powder or orange amorphous solid (2) molecular weight: 547
(3) Molecular formula: C ₂₅ H ₂₉ N ₃ O ₉ S
(4) High resolution mass spectrometry [M + H] ⁺ theoretical value (m / z) 548.1697, found value (m / z) 548.1709
(5) Specific rotation: [α] _D ²⁶ = −200.1 (c = 0.1, methanol)
(6) Ultraviolet absorption maximum (in methanol) λ _max (ε): 204 (18488), 235 (26967), 356 (5907)
(7) Infrared absorption maximum ν _max (KBr tablet): maximum absorption at 3432, 1677, 1527 cm ^-1 (8) Proton nuclear magnetic resonance spectrum: chemical shifts (ppm) in heavy dimethyl sulfoxide are shown in Table 5 . (In the table, s is a singlet, d is a doublet, m is a multiplet, and H is the number of protons.)
(9) Carbon nuclear magnetic resonance spectrum: Chemical shifts (ppm) in heavy dimethyl sulfoxide are shown in Table 5.
(10) Solubility: Easily soluble in DMSO and methanol. Poorly soluble in hexane, ethyl acetate and chloroform.

  As described above, since compounds corresponding to nanaomycins H, I, J and K for which various physicochemical properties were obtained have not been reported so far, these nanaomycins are considered to be novel substances.

(Example 6) Killing activity of nanaomycin analog to MDCK (canine epithelial cell line) induced to epithelial-mesenchymal transition by TGF-β MDCK cells were cultured in a cell culture dish with MEM medium (10% fetal bovine serum albumin, Subculture was carried out to maintain semiconfluence in a 1% MEM nonessential amino acid solution, 1% sodium pyruvate, 1% penicillin-streptomycin solution, 1% glutamine) at 37 ° C., 5% CO ₂ incubator . The cells were treated with a trypsin-EDTA solution, seeded in the above-mentioned medium containing TGF-β at a final concentration of 10 ng / mL and 0.001% BSA, and cultured for 3 days. As a blank, cells were similarly prepared for 3 days in a medium supplemented with only BSA, without TGF-β added. These cells were adjusted to 96 wells with MDCK cells at 6,000 cells / well and 2,000 cells / well.

Nanaomycin H (final concentration 0.5% DMSO) dissolved in DMSO to a final concentration of 100 μg / ml and 5 μg / ml in each well or dissolved in DMSO to a final concentration of 50 μg / ml and 5 μg / ml Cultured at 37 ° C in a 5% CO ₂ incubator for 24 hours with the addition of modified nanaomycin I, J and K (final concentration 0.5% DMSO), and with the morphology observation and live cell quantification kit (MTS assay) The proliferation test was done. The results are shown in FIG. In the test adjusted at 6,000 cells / well, nanaomycin H showed higher killing activity at 100 μg / ml and 5 μg / ml against cells in which epithelial-mesenchymal transition was induced by TGF-β. Similarly, nanaomycin I, J and K also showed the same effect as nanaomycin H.

Claims (6)

The compound represented by following formula (I).
[Wherein, R ¹ represents a C1-6 alkyl group, R ² represents a C1-6 alkoxycarbonyl group, a carboxy group, or a C1-6 alkyl group substituted with a hydroxyl group, and R ³ represents The group represented by either is shown. ]
The compound according to claim 1, wherein a microorganism belonging to an actinomycete having the ability to produce a compound wherein R ¹ is a methyl group and R ² is a carboxy group is cultured in a culture medium, and the compound is accumulated in the culture. The method of producing a compound according to claim 1, wherein R ¹ is a methyl group and R ² is a carboxy group in the compound according to claim 1, wherein the compound is collected from the culture. The compound according to claim 1, wherein the microorganism belonging to an actinomycete having the ability to produce a compound wherein R ¹ is a methyl group and R ² is a carboxy group is Streptomyces sp. The method according to claim 2, which is 0591 (reception number NITE ABP-02304).   Streptomyces sp. K15-0591 (reception number NITE ABP-02304).   A pharmaceutical composition comprising the compound according to claim 1 as an active ingredient.   An agent for damaging cells in which epithelial-mesenchymal transition has been induced, comprising the compound according to claim 1 as an active ingredient.