WO2002020816A1

WO2002020816A1 - Novel compound f-15949

Info

Publication number: WO2002020816A1
Application number: PCT/JP2001/007589
Authority: WO
Inventors: Toshio Takatsu; Tatsuya Yano; Shiho Kozuma; Yasunori Ono
Original assignee: Sankyo Company, Limited; Ohnuki, Takashi
Priority date: 2000-09-06
Filing date: 2001-09-03
Publication date: 2002-03-14
Also published as: AU2001282599A1

Abstract

Novel compounds represented by the following general formula (I) or salts thereof: (I) wherein R1 represents C1-6 alkyl or hydrogen; novel compounds represented by the following general formula (II) or salts thereof: (II) a process for producing these compounds by using a microorganism belonging to the genus Haplographium, etc. Because of having an excellent antifungal effect, these compounds are useful as preventives or remedies for various fungal infections.

Description

Specification

New compound F— 1 5 9 4 9

"Technical field"

The present invention relates to a compound having antifungal activity or a salt thereof, a method for producing the compound, a medicament containing the compound as an active ingredient, an antifungal agent containing the compound as an active ingredient, and a microorganism producing the compound. Etc.

"Background technology"

Examples of antifungal compounds currently used in clinical practice include amphotericin B, flucytosine, and azole compounds. Many of these compounds have become problematic in terms of cytotoxicity and emergence of resistant bacteria.

'' Examples of compounds having antifungal activity produced by microorganisms include pneumocandins (Schnetz, D.M., eta 1., Eta 1., J.) produced by the genus Zarelion (Za 1 erion) and the like. A ntibiot., 4 5,

Echinocandin (Nyfe 1 er, R. and K eller — Schierlein, W., H) produced by the genus Aspergillus (A spergi 11 us), etc. elv. C hi m.

Acta, 57, 2549 (1974)), Aureobasidium genus S-producing aureobasidin (aureobasidin) species (Ikai, K., eta 1. , J. Antibiot., 44, 925 (1991)), an unidentified fungus MF6200, produced by Kefrefundin (Manda 1a, S., M., eta 1., J. Bio then C he m. 2 7 2, 3

27 14 (19997)) etc., but these drugs have not been applied to clinical applications.

"Disclosure of the invention"

The inventors conducted intensive searches to find novel compounds having antifungal activity As a result, a novel compound was found in a fungal culture, and it was confirmed that the compound had antifungal activity. Thus, the present invention was completed.

That is, the present invention provides a compound having an antifungal activity or a salt thereof, a method for producing the compound, a medicament containing the compound as an active ingredient, an antifungal agent containing the compound as an active ingredient, and production of the compound. The purpose of the present invention is to provide microorganisms and the like that perform the treatment. The present invention

(1)

The following general formula (I)

(I)

(Wherein, R represents an alkyl group having 1 to 6 carbon atoms or a hydrogen atom.) Or a salt thereof,

(2)

The compound or a salt thereof according to (1), wherein R i is a methyl group or a hydrogen atom,

(3) 'A compound having the following physicochemical properties or a salt thereof:

1). Property of the substance: Colorless powdery substance ·

2) Solubility: Soluble in methanol and acetone, insoluble in water and normal hexane

3) Molecular formula: C _{3 3} H _6. 0 ₉

4) Molecular weight: 600 (measured by FAB mass spectral method)

5) 髙 Resolution The exact mass, [M + N a] · ⁺ , measured by the LC-TOF mass spectrum method is as follows.

Measured value: 6 2 3. 4 1 22 ·

Calculated value: 6 2 3. 4 1 3 5

6) Ultraviolet absorption spectrum: The ultraviolet absorption spectrum measured in methanol is Shows the following 'maximum absorption': 238 nm (ε 59-70)

7) Optical rotation: The optical rotation measured in methanol shows the following values:

_{^{[Α] D 22: - 2}} 0. 0 ° (c 2. 0)

8) Infrared absorption spectrum: Potassium bromide (KBr) The infrared absorption spectrum measured by the tablet method shows the maximum absorption shown below:

3 4 7, 3 1 3 7, 2 9 2 8, 2 8 5 6, 1 6 7 9, 1 4 9, 1 3 8 1, 1 3 1 6, 1 2 1 3, 1 1 4 6, 1 0 3 7, 8 4 5 , 8 0 3, 7 2 6 cm- 1 9) 1 H- nuclear magnetic resonance scan Bae click Honoré: signal in heavy methanol methano Ichiru 3.3 as a 1 ppm The measured nuclear magnetic resonance spectrum is as follows:

0.89 (3 H, m), 0.92 (3 H, m), 1.06 (3 H, d, J = 6.6 Hz), 1.12 (2 H, m ), 1.25—1.46 (23H, m), 1.31 (3H, m), 1.32 (2H, m), 1.80, (3H, d, J = 1. 1 Hz), 2.77 (1 H, m), 3.49 (1 H, m), 3.88 (1 H, dd, J = 1. 9, 1 2.lHz), 3.76 (1H, dd.J = 4.5,12.1Hz), 4.05 (1H, d, J = 2.5Hz), 4.09 (1H, dd, J = 2.5, 9.3Hz), 4.47 (1H, d, 1 = 7.1Hz), 4.97 (1H, ddd, J = 1.9, 4. 5, 9.3 Hz), 6.64 (1H, dd, J = 1.1, 9.3 Hz) ppm '10) ^{1 3} C Nuclear Magnetic Resonance Spectrum: The ¹³ C—Nuclear Magnetic Resonance spectrum, measured in heavy methanol at a methanol signal of 49.0 ppm, is as follows: '

12.0 (q), 14.6 (q), 15.0 (q), 20.1 (q), 20.6

(q), 23.9 (t), 26.9 (t), 27.2 (t), 28. 3 (t), 30. 6 (t), 30. 9 (t X 4), 31.0 (t), 32.1 (d), 33.0 (d), 33.2 (t), 38.5 (t), 38.6 (t ), 38.0 (t) _: 45.6 (t), 47.6 (d), 61.7 (t), 71.4 (d), 71.6

(d), 75.5 (d), 75.6 (d), 135.5 (s), 153.3 (d), 172.1 "),, 16.7 (s), 20.9 (s) ppm 10) High-performance liquid chromatography:

Column: Capsule pack C ₈ UG 1 2 0

(4.6 mm in diameter X 150 mm in length: manufactured by Shiseido Co., Ltd.)

Solvent: 0.04% Trifluoroacetic acid in water 72% Acetonitrile Flow rate: 0.6 m1 / min

Detection: UV absorption wavelength 240 nm

Retention time: 14.3 minutes

(Four )

Compounds having the following physicochemical properties or salts thereof:

1) Properties of the substance: colorless powdery substance

3) Molecular formula: C ₃₄ H _{S 2} O _s

4) Molecular weight: 6 1 4 (measured by FAB mass spectral method)

5) High-resolution FAB—exact mass, measured by mass spectrometry, [M + N a] ⁺ is as follows. '

Measured value: 6 3 7. 4 2 9 2

Calculated value: 6 3 7. 4 2 9 1

6) Ultraviolet absorption spectrum: The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption: 2336 nm (ε 79-70)

_{^{Non] D 26: - 5 8 °}} (c 1. 1) '

8) Infrared absorption spectrum: 1 iq uid f i The infrared absorption spectrum measured by the 1 m method shows the following maximum absorption:

3 4 2 8, 2 9 5 3, 2 9 2 5, 2 8 5 4, 2 7 2 9, 1 7 4 4, 1 6 6 3, 1 4 5 6, 1 3 7 6, 1 2 5 1, 1 2 1 6, 1 1 3 0, 1 0 7 3, 1 0 3 6, 9 5 1 cm ¹ '

9) — Nuclear magnetic resonance spectrum: in heavy dimethyl sulfoxide, The rufoxide signal was measured as 2.50 ppm, and the nuclear magnetic resonance spectrum is as follows:

0.89 (3H, m), 0.92 (3H, m), 1.03 (1H, m), 1.06 (3H, d, J = 6.6Hz ), 1.17 (2 H, m), 1.23 (2 H, m), 1.21-1.30 (22 H, m), 1.31 (1 H, m ), 1.32 (3H, m), 1.35 (1H, m), 1.80 (3H, d, J = 1.1Hz):

I.33 (2H, m), 2.64 (1H, m), 3.40 (1H, m), 3.63 (3H, s), 3.76 (1 H, dd, J = 1.9, 12.1 Hz), 3.88 (1H, dd, J = 4.5, 12.1Hz), 4.05 (1H, d, J = 23 Hz), 4.09 (1 H, dd, J = 2.5, 9.3 Hz), 4.19 (1 H d, J = 5.0 Hz) , 4.42 (1H, d, J = 7.0Hz), 4.57 (1H, t, J = 5.4Hz), 4.97 (1H, ddd, J = 1. 9, 4. 5, 93 Hz),, 5.02 (1H, d, J = 2.8Hz), 5.04 (1H, d, J = 7.4 Hz), 6.6 1 (1H, dd, J = 1.1, 9.3Hz) ppm

1 0) ^{1 3} C-NMR spectrum: in deuterated dimethyl sulfoxide, signals for heavy-dimethylcarbamoyl Rusuruhokishi de 3 9. measured as a 5 ppm, ^{1 3} C-nuclear magnetic co-sounding spectrum is less As shown in:

II.3 (q), 14.0 (q), 14.1 (q), 1 9-.3 (q), 19.9 (q), 22.1 (t), 25 2 (t), 25.6 (t), 26.5 (t),

28.8 (t), 29.1 (tX4), 29.2 (t), 30.1 (d), 310 (d), 31.3 (t), 3 6.2 (t), 37.1 (t), 37.2 (t) 43.6 (t), 45.8 (d), 51.6 (q), 59.3 (t), 69.4 (d), 69.5 (d), 70.4 (d), 73.7 (d), 133.4 (s) 150.9 ( d), 1 70.2 (s), 1,73.3 (s), 19.7.8 (s) ppm

1 1) High-speed liquid mouth chromatography:

Column: YMC—Pac OD S—AM S—3

(Diameter 4.6 mm X length 150 mm: manufactured by YMCS Corporation) Solvent: 90% acetate water containing 0.04% trifluoroacetic acid Flow rate: 1.0 O ml / min

Detection: UV absorption 240 nm

Retention time: 8.26 minutes,

( Five ) '

The following formula (I I)

(I I)

A compound represented by or a salt thereof,

(6)

Compounds having the following physicochemical properties or salts thereof: '

1) Properties of the substance: colorless powdery substance

3) Molecular formula: C _{3 3} H _6. 〇 ₉

4) Molecular weight: 600 (measured by FAB mass spectrometry)

5) The high-precision mass, [M + Na] ⁺ , measured by the high-resolution FAB mass spectrum method is as follows.

Measured value: 6 2 3. 4 1 3 4

Calculated value: 6 2 3. 4 1 3 5

6) Ultraviolet absorption spectrum: The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption: 2336 nm (ε8588)

_{^{[Α] D 2 6: -}} 3 3. 0. (C 1, 0)

8) Infrared absorption spectrum: Infrared absorption spectrum measured by potassium bromide (KBr) tablet method shows the maximum absorption shown below:

3 3 5 3, 2 9 5 5, 2 9 1 9, 2 8 5 0, 1 7 8 5, 1 7 3 7, 1 6 6 6, 1 46 5, 14 56, 1 3 7 7, 1 3 1 7, 1 28 3, 1 2 5 1, 1 20 1, 1 1 3 1, 1 0 9 3, 1 0 5 2 cm- ¹ '

9) — Nuclear magnetic resonance spectrum: In heavy pyridin, the signal of heavy pyridin

The nuclear magnetic resonance spectrum, measured at 74 ppm, is as follows: '.

0.88 (3H, m), 0.90 (3H, m), 1.03 (3H, d, J = 6.

3 H z), 1.14 (1 H, m), 1.22 (1 H, m), 1.25 (2 H, m), 1.27 (2 H, m), 1. 2 1 i. 40 (1 2 H, m), 1.34

(1H, m), 1.41 (2H, m), 1.44 (1H, m), 1.46 (2H, m), 1.51 (3H, d, J = 6 9 H z), 1.6 2 (l H, m), 1.6

8 (2H, m), 1.69 (2H, m), 1.74 (2H, m), 1.97 (3H, brs), 2.70 (1H, m), 3.90 (1 H, m), 4.56 (1 H: d, J = 6.9 Hz), 4.82 (1 H, m), 4.83 (1 H, m) , 4. 9

4 (1H, dd, J = 5.1, 10.9 Hz), 5.12 (1H, dd, J = l, 9, 10.9 Hz), 5.46 (1 H, brs), 6.77 (1H, d, J = 9, 4Hz) ppm

10) ^13C- nuclear magnetic resonance spectrum: ^13C- nuclear magnetic resonance spectrum measured in heavy methanol at 49.O ppm in heavy methanol is shown below:

12.5 (q), 14.8 (q), 15.2 (q), 20.3 (q), 20.6 (q), 23.4 (t), 27.0 (t) ), 27.3 (t), 28.0 t), 3

0.1 (t), 30.4 (t), 30.5 (tX2), 30.6 (t), 30.

8 (t), 31.4 (d), 32.2 (d), 32.6 (t), 37.7 (t):

39.0 (t), 39.1 (t), 44.9 (t), 47.9 (d), 69.3 (t) ', 70.3 (d), 71.5 (d), 72.4 (d), 74.5 (d),

13.5 (s), 151.5 (d), 172.8 (s), 17.8 (s),

1 9 8.6 (s) p p m

1 1) High performance liquid chromatography:. Column: YMC—Pack OD S—AM S 3

(4.6 mm in diameter X 150 mm in length: manufactured by PMC Inc.) Solvent: 90% acetonitrile water containing 0.04% trifluoroacetic acid Flow rate: 1.0 m1 / min

Detection: UV absorption 240 nm

Retention time: 6.92 minutes,

(7)

A bacterium producing the compound according to any one of (1) to (6), which belongs to the genus Hapographium, is cultured, and (1) to (6) The method for producing a compound according to any one of (1) to (6), wherein the compound is collected in any one of:

(8)

A bacterium producing the compound according to any one of (1) to (6), which belongs to the genus Nopographium (Hap 1 graphium), is a haplograph 'Helicose Fanolem' Lao 'and' de 'Hog (H aplographi um he 1 iocephalum V asant Rao & de Hoog) cs ANK 2 689 (FE RM BP-724) strain (7), the production method according to (7), (9) '

(1) a medicament comprising the compound according to any one of (1) to (6) or a salt thereof as an active ingredient;

( Ten )

(1) An antifungal composition comprising the compound according to any one of (1) to (6) or a salt thereof as an active ingredient.

(1 1)

Haplographium 'Heliosef Rum' Lao 'and de Hog (H aplographiumhelioceph al um V asant Rao & de Hoog) S ANK 2 689 9 (FE RM BP— 7 2 4 3) 1 2) (1) A method for preventing or treating mycosis, comprising using the compound or the salt thereof according to any one of (6) to (6).

( 13 )

Use of the compound or the salt thereof according to any one of (1) to (6) for producing an antifungal agent,

About. ·

In the present invention, the compound described in any one of the above (1) to (6) is generically referred to as a substance F-15949.

In the general formula (I) of the above (1), is a linear or branched alkyl or carbon atom having 1 to 6 carbon atoms. Examples of the straight-chain or branched-chain alkyl having 1 to 6 carbon atoms include methyl, ethyl, propyl, butyl, pentyl, hexyl, isopropyl, isobutyl, t-butyl, isopentyl, and isobutyl. A hexyl group and the like can be exemplified. '

Further, in the present invention, a compound represented by the above formula (1) represented by the general formula (I) and R i in the general formula represents a hydrogen atom or the compound described in the above (3) is represented by F-15949. —1 is a compound represented by the general formula (I) of the above (1), wherein R i is a methyl group or the compound of the above (4). 9 4 9 4 referred to as _c as described above, straight鎮又compounds showing a branched chain alkyl group having the general formula Ri has 1 to 6 carbon atoms represented and in said general formula (I) is, according to conventional methods It can be synthesized by reacting F-15949-1 or F-15949-4 with various alkylating agents and the like.

Further, in the present invention, the compound described in the above (5) or (6) is referred to as F-15949-12.

F—1 5 9 4 9—1, F—1 5 9 4 9—2 and F—1 5 949—4 are represented by the following general formula (III)

(The compound represented by the above general formula (III) and in which is a substituent i in the following Table 1 is F—15949-9-1, and the compound represented by the above general formula

The compound represented by the formula (III), wherein R i in the general formula is the substituent ii of the following Table 1, is F-15949-2, represented by the general formula (III) and The compound in which R in the formula is the substituent iii in the following table is F—15949—4. Table 1. Substituents in general formula (III)

R l

The F-15949 substance can be converted to a salt according to a conventional method. Salts of F—15949 substances can be used for pharmaceutical or non-pharmaceutical applications. Salts of the F15949 substance used in medicine are not particularly limited as long as they are pharmacologically acceptable.Examples include sodium salts, potassium salts, ammonium salts and the like. it can. The salt of the F-15949 substance used for non-pharmaceutical uses (for example, as a synthetic intermediate) is not particularly limited. Alkali metal salts such as tritium salts and calcium salts, alkaline earth metal salts such as calcium salts, barium salts and magnesium salts, aluminum salts, iron salts, zinc salts, copper salts, nickel salts, Metal salts such as cobalt salts; inorganic salts such as ammonium salts, tert-octylamine salts, dibenzylamine salts, morpholine salts, dalcosamine salts, phenylglycine alkyl ester salts, ethylenediamine salts, and N-methyldal. Kamin salt, Guanidine salt, Getylamine salt, Triethylamine salt, Dicyclohexylamine salt, N, N 'Dibenzinoleethylenediamine salt, Salt, pro force in salt, diethanolamine salt, N- benzyl Hue phenethyl Rua Min salts, piperidines Rajin Salts, organic amine salts such as tetramethylammonium salt, tris (hydroxymethyl) amino methane salt; and amino acids such as glycine, lysine, arginine, ordin, and asparagine. Salts and the like can be mentioned.

In addition, the F-159499 substance can be converted into an ester according to a conventional method. Examples of the ester of the F-159499 substance include an ester formed of an alkyl group having 1 to 6 carbon atoms. Such esters include, for example, methyl ester, ethyl ester, propyl ester, isopropyl ether, butyl ester, isobutyl ester, s_butyl ester, t-butynoleestenol, pentynoleestenol, isopentinole Examples include esdenole and neopentinole ester.

Further, the F-159949 substance has various isomers. In the present invention, these isomers and mixtures of these isomers are all represented by a single formula. In this onset Ming, isomers thereof and _c also includes Te base that is also advised mixtures of these isomers, F- 1 5 94 9 substances and their salts, it may become solvates. For example, water may be absorbed when left in the air or recrystallized to form adsorbed water or form a hydrate. Such a solvate is also included in the present invention.

Further, the present invention also includes all of so-called prodrugs, which are metabolized in the living body and converted into F-15949 substances.

The F-15949 substance is present in a fungal culture that produces the F-159499 substance. Examples of the F—15949 substance-producing bacterium include fungi belonging to the genus Haplographium. Preferably, the haplopro- gram “Helicosepharum Lao” and “De” are used. Hog (Hap 1 ographlumheliocephalum Vasant Rao & de Hoog), more preferably haplography film 'Heliosefalum' Rao 'and'de'hog S ANK 268 899 (F ERM BP-7243) strain ( Hereinafter, it is simply referred to as “S ANK 26 89 9 shares.”) S 'ANK 26 8 9 9 share, 1 9 July 1998, has been separated from the soil that was collected in the pine forest of Shimane Prefecture _c

The bacteriological properties of the S ANK 26 89 9 strain are described below. The composition of the medium used for culturing the bacteria is as shown below.

[Potatodextros agar medium (hereinafter referred to as “PDA medium”)] Nissipotedextrox agar 39 g

Agar 5 g

Distilled water 100 000 ml

[Improved Weitz manandSilva-Hutner medium (hereinafter referred to as "WSH medium")]

Taiki Tominore 10 g

MgS0 ₄ -7H ₂ 0 1

KH ₂ P0 ₄ 1 g

Na.N0 ₃ '1 g

Agar 20 g

Distilled water 0 0 0 m

[Corn meal agar medium (hereinafter referred to as "CMA medium")]

Nissi Corn Minore Agar 1 7 g

Agar 5 g

Distilled water 100 000 ml

[Miura medium]

Gunolet Course

0.2 g of yeast extract

KH ₂ P0 ₄ 1 g

MgS0 ₄ -7H ₂ 0 0. ₂ g

K C 1 0.2 g

Na.N0, 2 g .

, 14 agar 1 3 g

100 ml of distilled water Growth on a plate medium prepared using the medium having the above composition is as follows. The display of the color tone is "Metune Handbook of Color" (A. Kornerupand J. H. Wansher, Methuen H andbook of Color, Eyre Methuen, London, 25 2 pp (1 984) ) Color chip pick-up.

When the S ANK 2689 9 strain is cultured on WSH medium at 23 ° C for 7 days, the culture diameter reaches 21 to 22 mm. The flora is flat, consisting of a thin mycelium, colorless, white at all edges, without exudate and odor. It produces sparse conidia in the center. The back is colorless.

When S ANK 2689 9 strain is cultured on CMA medium at 23 ° C for 7 days, the culture diameter reaches 27 to 29 mm. The flora is flat, consists of a thick hyphal layer, colorless, white at all edges, without exudate and phlegm. The back is colorless.

When S ANK 2689 9 strain is cultured on Miura medium at 23 ° C for 7 days, the culture diameter reaches 24 to 25 mm. The flora is flat, consists of a thin mycelium, is colorless, appears white at all edges, and has no exudate and smell. The back is colorless.

When the S ANK26899 strain is cultured on PDA medium at 23 ° C for 7 S, the culture diameter reaches 24 to 28 mm. The flora is flat, consisting of a thin mycelium, colorless, wool-like in the center, light yellow (2A3) at all edges, without exudate and smell. The back was colorless.

The micromorphology of the S ANK 2 689 9 strain was as follows.

The bristles are elongate, smooth and smooth, 4440-650 / im in length, 4-6 μm in width at the base, tapering to the top and dark. The conidium is 45 to 105 μm in length and 4 to 6 μm in width and stands upright from the hypha, straight or slightly bent, solitary, smooth, with septum, near the base It becomes dark on thick walls and colorless on thin walls towards the top. The conidium-forming cells are multibudding, forming primary metres at the top of the conidiophores, Form conidia or secondary metres. The primary metrology is 5 to 9 m in length and 3.5 to 5 / m in width. Thin wall ♦ Smooth and colorless. Secondary metres are colorless and form conidia. Conidia are 6 to 9 / m in length and 2 to 3 m in width, are aerobically linked with or without branching, are single-celled, smooth and thin-walled. Conidia are cylindrical to oblong, rarely slightly bent, have flat emergent roots at both ends, and are colorless.

Based on the above bacteriological characteristics, the strain was compared with the strains described in recent literature.The strain was found to be Zucconi & Pagano Coneerningthegenericli mits ιnHap 1 ographiu m., Mycotaxon 66, 11 (1993), and Rao & de Hoogeworcritical Hyphomycetes from India.Stud.Myco 1., 28, 56 (19986) Has been matched with the haplography game 'Heliosef Alm. Lao and de Hog. Therefore, this strain was identified as haplografium 'Heliosefalum-Lao' and de hog, and transferred to haplografum 'Loseofalum. Lao-and-de-hog S ANK2698 It was named 9 strains. This strain was established on July 26, 2000 at 1-1-1 Higashi, Tsukuba City, Ibaraki Prefecture, Japan 1 The 6th National Institute of Advanced Industrial Science and Technology (AIST) International Depositary with the Faculty of Engineering, Biotechnology and Industrial Technology Research Institute) and given the deposit number FE RM BP—.

As is well known, fungi are susceptible to mutation in the natural world or by artificial manipulation (for example, ultraviolet irradiation, irradiation, chemical treatment, etc.), and the SANK 26899 strain of the present invention is used. Is also presumed to be prone to such mutations. In the present invention, the SANK26889 strain includes all its mutants.

These mutants also include those obtained by genetic methods, for example, recombination, transduction, transformation, and the like. That is, SANK 28699 strains that produce F-15949 substances, their mutant strains, and all strains that are not clearly distinguished from them are S ANK 28699 strains. Is included.

As described above, the F_15949 substance can be produced by culturing a bacterium producing the substance. F— 1 5 9 4 9 It can be performed using a medium such as that used for production of the product. Such media contain carbon and nitrogen sources and inorganic salts that can be assimilated by microorganisms.

Carbon sources include, for example, glucose, fructose, maltose, sucrose, mannitol, glycerin, dextrin, oats, rye, corn starch, potatoes, corn flour, soybean oil, cottonseed oil, Molasses, citric acid, tartaric acid and the like can be mentioned. These carbon sources can be contained in the medium alone or in combination of two or more.

The amount of the carbon source contained in the medium depends on other components in the medium, but is usually 1 to 10% by weight of the medium.

Nitrogen sources include, for example, soybean flour, bran, peanut flour, cottonseed powder, casein hydrolyzate, pharmamine, corn steep liquor, peptone, meat extract, yeast, yeast extract, malt extract, and sodium nitrate Examples thereof include lithium, ammonium nitrate, and ammonium sulfate. These nitrogen sources can be contained in the medium alone or in combination of two or more.

The amount of nitrogen source to be contained in the culture medium depends on other components in the culture medium, but is usually 0.2 to 6% by weight of the culture medium.

The carbon source and the nitrogen source need not be of high purity, but may be of low purity. Low purity carbon and nitrogen sources may contain trace amounts of growth factors, vitamins, inorganic nutrients, and the like. '

^{(1) The} inorganic salt is not particularly limited as long as an ion can be obtained.Examples thereof include a sodium salt, a potassium salt, a magnesium salt, an ammonium salt, a calcium salt, a phosphate, and a sulfate. , Hydrochloride, carbonate and the like.

Also, vitamin B1 which can be assimilated by the F ^ 15949 substance-producing bacteria, vitamins such as biotin, cell growth promoting substances such as thiamine, metal salts such as manganese and molybdenum, etc. May also be included in the medium as needed.

Further, when performing liquid culture, an antifoaming agent such as silicon oil, polyalkylene glycol ether, vegetable oil, animal oil, or a surfactant may be contained in the medium. It is preferable that the pH of the medium used for the culture of the F-15949 substance producing bacterium is pH 4.0 to 6.5. ''

The upper limit of the culture temperature of the F-159 4 9 substance-producing bacterium is 23 to 37 ° C, the lower limit is 15 to 18 ° C, and the preferred range is 18 to 30 ° C. The temperature is more preferably 18 ° C to 23 ° C.

The culture method is not particularly limited as long as it is a method usually used for culturing microorganisms, and examples thereof include a culture method using a solid medium, a stirring culture method, a shaking culture method, and an aeration culture method. The stirring culture method, the shaking culture method, and the aeration culture method are preferable, and the shaking culture method is more preferable. The aeration and stirring culture method is most suitable for industrial production.

The following is an example of the culture of the F-15949 substance producing bacterium.

Seed culture is performed by shaking at 20 to 23 ° C for several days in an Erlenmeyer flask containing the liquid medium as described above. Seed culture is divided into two or more stages, if necessary, and scaled up in stages. The obtained seed culture is used for main culture. The main culture is basically performed under the same conditions as the seed culture. That is, the seed culture is inoculated into the main culture medium, and cultured with shaking at a constant temperature for several days.

If the main culture is to be performed on a larger scale, use a jar armmenter or tank equipped with a stirrer and aeration device. In that case, the medium is prepared in a jar fermenter or tank. Sterilize the medium at 121 ° C for 20 minutes, cool, and inoculate the seed culture. Culture is performed at 20 ° C by aeration and stirring.

The amount of F-15949 substance in the culture solution can be measured by high performance liquid chromatography (HPL) analysis. The production of the nine substances usually reaches a maximum after 100 to 240 hours from the start of the culture.

After the cultivation, the F-15949 substance can be extracted from the obtained culture, the cells contained therein and / or the culture supernatant thereof. Centrifugation or filtration using diatomaceous earth as a filter aid can be used to separate cells and other solids from the culture supernatant. Can be used.

The physical and chemical properties of the compound can be used for the extraction of F-155949. The F-15949 substance present in the culture filtrate or culture supernatant is an organic solvent that is immiscible with water under acidic or neutral conditions, such as ethyl acetate, chloroform, chloride, methylene chloride. , Butanol, and a mixed solvent of two or more thereof. The F-15949 substance present in the cells is extracted from the cells using 50 to 90% aqueous acetone or methanol, and the organic solvent is distilled off. Extraction can be carried out in the same manner as when it is present in a liquid or culture supernatant. The F-15949 substance present in the whole culture is 20 to 80%, preferably 40 to 60%, more preferably 50 to 50% acetone or methanol in the whole culture. % Can be added and extracted. After completion of the extraction, diatomaceous earth is filtered as a filter aid, and the resulting soluble matter can be extracted in the same manner as in the culture filtrate or culture supernatant. Purification to isolate the F-15949 substance from the above extract is performed by adsorption chromatography, ion exchange chromatography, partition chromatography, reverse phase chromatography, HPLC, etc. This can be done by any means.

Adsorption chromatography involves contacting the above-mentioned extract containing the F-15949 substance with an adsorbent and adsorbing and removing the foreign substances, or adsorbing the F-15949 substances and contaminating them. After removing the substance, the elution is performed by eluting the F-15949 substance. Examples of the adsorbent include activated carbon, Amberlite XAD-2, XAD-4 (hereinafter, manufactured by Rohm and Haas), Diaion HP-10, HP-20, and CH P—20 P; HP—50; Sepabeads SP—207 (manufactured by Mitsubishi Chemical Corporation). Examples of the solvent used to elute the adsorbed F-15949 substance include methanol water, acetone water, and butanol water.

Ion-exchange chromatography uses the fact that F-15949 acts as an acidic substance. That is, the above-mentioned extract containing the F-15949 substance is brought into contact with an anion exchange carrier to adsorb the F-15949 substance to the carrier, and after removing impurities, the solvent-based F— 1 5 9 4 by changing ionic strength, pH, etc. Elute 9 substances. Examples of anion exchange carriers include DEAE-cellulose (Brown), DEAE-Sephadex, DEAE-Sepharose, QAE-Sephadex (above, manufactured by Pharmacia's Fine 'Chemicals), DEAE-G Opal C (manufactured by Tohsoichi Co., Ltd.), Duo Light A-2 (manufactured by Diamond 'Shamrock' Chemical), Amber Light IRA-68 (manufactured by Rohm & Norse) And Dowex 1X4, 21K, and SBR-P (manufactured by Dow Chemical Company).

Carriers used for distribution chromatography include, for example, silica gel, TS Κgelt Yopearl HW-40F (manufactured by Tosoichi Co., Ltd.), and Sephadex LH-20 (manufactured by Amersham Pharmacia). And the like.

Examples of the carrier used for the reverse phase chromatography include Cosmo Seal 140C18 (manufactured by Nacalai Tester Co., Ltd.).

Columns used for HPLC include, for example, Shodex Asahi Pack C8P50-4E (manufactured by Showa Denko KK), YMC Pack OD S-AM (manufactured by IMC Corporation), capsules Reverse-phase columns such as Pack C ₈ UG 1220 (manufactured by Shiseido Co., Ltd.) and Gasil ODS (manufactured by Sensyu Kagaku Co., Ltd.). These purification means can be used alone or in appropriate combination to isolate the F-15949 substance. If necessary, the same purification procedure may be repeated. To carry out each purification means, a method suitable for purification, such as column chromatography, batch chromatography, thin-layer chromatography, etc., can be selected.

The F-15949 substance or its salt has an antifungal effect and is therefore useful as a prophylactic or therapeutic agent for fungal infections.

When F-155949 or a salt thereof is used as a prophylactic or therapeutic agent for fungal infection, it can be administered in various forms. The dosage form depends on the formulation, age, sex, disease and the like. For example, tablets, pills, powders, granules, syrups, solutions, suspensions, emulsions, granules, capsules and the like are orally administered. Injections are administered intravenously, intramuscularly, intradermally, subcutaneously or intraperitoneally. Suppositories are administered rectally.

Formulations containing various substances as active ingredients are formed according to standard methods. It can be produced using known auxiliaries usually used in the field of pharmaceutical preparations such as agents, binders, disintegrants, lubricants, dissolving agents, flavoring agents, coating agents and the like. For molding into tablets, widely known carriers in the art can be used, such as lactose, sucrose, sodium chloride, glucose, urea, starch, carbonated calcium, kaolin, crystalline cellulose, and the like. Binding of excipients such as silicic acid, water, ethanol, propanol, simple syrup, dextrose solution, starch solution, gelatin solution, carboxymethyl cenorellose, shellac, methinoresenorelose, potassium phosphate, polyvinylinolepyrrolidone, etc. Agent, dried starch, sodium alginate, powdered agar, powdered laminarane, powdered sodium hydrogencarbonate, calcium carbonate, polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, monoglyceride stearate, starch, Disintegrators such as lactose, sucrose, stealine, cacao batata , Decay inhibitor such as hydrogenated oil, quaternary ammonium base, absorption enhancer such as sodium lauryl sulfate, humectant such as glycerin and starch, starch, lactose, kaolin, bentonite, colloid Adsorbents such as silica, lubricating agents such as purified talc, stearate, powdered boric acid, polyethylene glycol and the like can be mentioned. Tablets can be made into tablets coated with a usual coating, if necessary, for example, sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets or double tablets, or multiple tablets.

For molding into pill form, a wide variety of carriers known in the art can be used as a carrier, for example, excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, and talc. And binders such as gum arabic powder, tragacanth powder, gelatin and ethanol, and disintegrants such as laminaran and agar.

When prepared as an injection, the liquid and suspensions are preferably sterilized and isotonic with blood. When forming these liquids, emulsions and suspensions, they should be used as diluents. Those widely known in the field can be widely used, and examples thereof include water, ethyl alcohol, propylene glycol, epoxidized isostearyl alcohol, poloxysylated isostearyl alcohol, and polyoxetylene sorbitan fatty acid esters. it can. They may contain sufficient amounts of salt, sugar, or glycerin to maintain isotonicity. Solubilizer, buffer, soothing sugar, Coloring agents, preservatives, flavoring agents, flavoring agents, sweetening agents, and other agents may be included. When an injection is administered intravenously, it is administered alone, mixed with a normal liquid sampling agent such as glucose or amino acid, or as an emulsion with polyoxyethylene sorbitan fatty acid esters and the like.

For molding into suppositories, a wide variety of carriers known in the art can be used as carriers, such as polyethylene glycol, cocoa butter, higher alcohols, higher alcohol esters, gelatin, and semi-synthetic. Glyceride and the like can be mentioned. The amount of the F-1,5,949 substance contained in the above-mentioned pharmaceutical preparation is not particularly limited, but the upper limit is 30 to 70% by weight, the lower limit is 1% by weight, and the preferable range is 1 to 3%. 0% by weight. .

The dosage of a pharmaceutical preparation containing F-15949-substance depends on the symptoms, age, body weight, administration method, dosage form, etc., but is usually administered to adults daily. the amount of 9 material, the upper limit is 2 0 to 2 0 0 0 mg, the lower limit is 0.0 0 1 to 0. lm _g, the preferred range is 0.0 1 to 2 0 0 mg, more preferred range Is between 0.1 and 20 mg.

The number of doses of pharmaceutical preparations containing F-155949 is once every few days, once a day, or several times a day.

"Best mode for carrying out the invention" '

Hereinafter, the present invention will be described in more detail with reference to Examples, Reference Examples, Test Examples, and Formulation Examples, but the present invention is not limited thereto. Example 1. Culture of F—1 5 9 4 9 substance producing bacteria

Put 100 ml of each of the seed culture media having the composition shown in Table 2 below into two 500 ml Erlenmeyer flasks, sterilize them at 21 ° C for 20 minutes, and add S ANK 286 9 Nine strains were inoculated with one loop of platinum and cultured for 5 days at 23 ° C and 210 rpm (rotational radius 7 cm). The obtained culture solution is inoculated into four 2 L Erlenmeyer flasks containing a medium of the same composition, 40 Om1, at 5% (volume by volume), at 23 ° C and 210 rpm (radius of rotation). 7 cm) for 2 days. (The resulting culture is hereinafter referred to as “seed culture.”) Table 2. Medium composition of seed culture medium Glycerin 30 g

Gunore course 30 g

Soluble starch 20 g

Soy flour 10 g

2.5 g gelatin

Yeast extract (Difco 2.5 g

NH ₄ N0 ₃ 2. 5 g

Antifoam .0.1 ml

Tap water 1 0 0 0 ml

* ¹ Nissan. Disform CB-442 (manufactured by NOF Corporation) The pH was adjusted to 6.5 before sterilization. Two 30 L jar incubators containing 15 L of the main culture medium with the composition shown in Table 3 below were inoculated with 5% (by volume) of the above-described seed culture solution, at 20 ° C, and aeration volume lv vm The cells were cultured for 5 days under the conditions of rotation speed of 100 to 180 rpm and dissolved oxygen of 5.0 ppm. Table 3. Medium composition of main culture medium Gunorecose 60 g

Manolet extract (Difco) 10 g

2 g of yeast extract (D ifco) Tripton (D ifco 1

NH ₄ N0 ₃ 1 g

NaNO,

KH ₂ P0 ₄ 1

MgS0 ₄ -7H ₂ 0 丄 g

Antifoam agent ¹ 0.1 ml

Tap water 100 ml Disoform CB-442 (manufactured by NOF Corporation) The pH was not modified and sterilized at 121 ° C for 20 minutes. Example 2. Isolation of F—1 5 9 4 9—1

The behavior of F-15949-l in this example was monitored by using HP stripes under the following conditions.

Column: Capsule pack C _s UG 1 2 0

(4.6 mm diameter x 150 mm length: manufactured by Shiseido Co., Ltd.)

Solvent: 0.02% 72% cetonitrile water containing 4% trifluoroacetic acid Flow rate: 0.6 m1 / min

Detection: UV absorption wavelength 240 nm

Retention time: 14.3 minutes

To 28.5 L of the culture obtained in Example 1, 1.5 kg of celite (diatomaceous earth cerite 5445) was added as a filter aid, and the mixture was filtered with a filter press. 6 kg of the obtained cells were extracted with 30 L of 80% acetone water. After adding 20 L of water to 30 L of the extract, the mixture was extracted with 30 L of ethyl acetate. The obtained organic layer (38 L) was washed with water (20 L), concentrated under reduced pressure, and the precipitated insoluble material was filtered using a filter paper and a funnel. The obtained filtrate was concentrated under reduced pressure to remove ethyl acetate, thereby obtaining 28.4 g of an oily substance. ' The whole amount of the oily substance was dissolved in 200 ml of acetone, and 15 ml of this was subjected to the following purification.

An HPLC column (capsule pack C ₈ UG 12 0: diameter: 200 μl) was previously prepared by equilibrating the acetone solution (300 μl) with 72% acetonitrile water containing 0.04% trifluoroacetic acid. mm X length 250 mm: manufactured by Shiseido Co., Ltd.) and developed at a flow rate of 13. O ml / min with the solvent system used for equilibration. The ultraviolet absorption of the active fraction was detected at a wavelength of 240 nm, and a peak eluted at a retention time of 25 to 26 minutes was collected. This HPLC preparative operation was repeated 50 times, the obtained eluted fractions were combined, concentrated under reduced pressure, and 200 ml of ethyl acetate was added to 50 ml of the obtained suspension to partition. I went. The obtained organic layer was concentrated under reduced pressure to 1 ml, suspended in 2 O ml of water, and lyophilized to obtain 26.6 mg of a crude purified powder.

The coarsely purified powder was dissolved in 500 μl of acetone, and a thin gel of thin gel was used (TLC platessi 1 i ₍ cagel 60 F 25 4: 10 cm × 10 cm, Thickness: 2 mm: Merck Co., Ltd.) and developed with 10 ml of a mixed solvent of methylene chloride: methanol: formic acid = 9: 1: 0.01. A silica gel containing spots having an R f value of 0.3 was cut out and eluted with 40 m 1 of a mixed solvent of methylene chloride: methanol: formic acid = 8: 2: 0.01. After concentration under reduced pressure to m 1, the suspension was suspended in 2 O ml of water and freeze-dried to obtain 10 mg of F-159, 49-1 as a colorless powder. Isolation of 5 9 4 9—2

The behavior of F-155949-2 in this example was monitored by HPLC under the following conditions.

Column: YMC—Pac OD S—AM S—3

(4.6 mm diameter x 150 mm length: manufactured by JM Corporation) Solvent: 90 containing 0.04% trifluoroacetic acid. /. Acetonitrile water Flow rate: 1.0 m1 / min Detection: UV absorption wavelength 240 nm

Retention time: 6.9 2 minutes

To 28.5 L of the culture solution obtained in Example 1, 1.5 kg of celite (diatomaceous earth cerite 5445) was added as a filter aid, followed by filtration with a filter press. 6 kg of the obtained cells were extracted with 30% L of 80% aqueous acetone. After adding 20 L of water to 30 L of the extract, this was extracted with 30 L of ethyl acetate. The obtained organic layer (38 L) was washed with water (20 L), concentrated under reduced pressure, and the insoluble matter was filtered using a filter paper and a funnel. The obtained filtrate was subjected to concentration under reduced pressure to distill off ethyl acetate to obtain an oily substance 28.

4 g were obtained. The total amount of this oily substance was dissolved in 200 ml of acetone, and 1

5 ml was provided for the following purification. An HPLC column (Pegasil ODS: 20 mm diameter) was prepared by preliminarily equilibrating the aceton solution (500 μl) with 90% acetonitrile water containing 0.04% trifluoroacetic acid. Length 250 mm: manufactured by Senshu Ichigaku Co., Ltd.) and developed at a flow rate of 10 Oml / min, depending on the solvent system used for equilibration. An ultraviolet absorption of 240 nm in the active fraction was detected, and a peak eluted at a retention time of 21 minutes was collected. Such an HPLC preparative operation was repeated 30 times, and the obtained eluted fractions were combined, and 20 ml of the suspension obtained by concentration under reduced pressure was lyophilized to give F-1559 9-2 Was obtained as a colorless powder (2 O mg). '' Example 4. Isolation of F—1 5 9 4 9—4,

The behavior of F-1 595 49-4 in the present embodiment was monitored by monitoring HPLC under the following conditions.

Column: YMC—Pac OD S-AM S-3

(4.6 mm in diameter X 150 mm in length: manufactured by JMC Corporation) Solvent 90% acetonitrile water containing 0.04% of trifrenoloacetic acid Flow rate 1. Oml / min

Detection UV absorption wavelength λ 24 0 n m

Retention time: 8.26 minutes To 28.5 L of the culture solution obtained in Example 1, 1.5 kg of celite (diatomaceous earth celite 545) was added as a filter aid, followed by filtration with a filter press. 6 kg of the obtained cells were extracted with 30 L of 80% aqueous acetone. After adding 20 L of water to 30 L of the extract, this was extracted with 30 L of dimethyl acetate. The obtained organic layer (38 L) was washed with water (20 L), concentrated under reduced pressure, and the insoluble matter was filtered using a filter paper and a funnel. The obtained filtrate was subjected to concentration under reduced pressure to distill off ethyl acetate, thereby obtaining 28.4 g of an oily substance. The whole amount of the oily substance was dissolved in 200 ml of acetone, and 15 ml of this was supplied to the following purification.

An HPLC column (Pegasil ODS: 20 mm in diameter X 250 mm in length: manufactured by Senshu Kagaku Co., Ltd.) previously equilibrated with 500 μl of the acetone solution with 90% acetonitrile water containing 0.04% trifluoroacetic acid. And developed at a flow rate of 10 Oml / min, depending on the solvent system used for equilibration. The UV absorption of the active fraction was detected at 240 nm, and the peak eluted at a retention time of 26 minutes was collected. Such an HP LC fractionation operation was repeated 30 times. The obtained eluted fractions were combined, and 40 ml of the suspension obtained by concentration under reduced pressure was freeze-dried. 3.2 mg of 4 was obtained as a colorless powder. Test Example 1. Antifungal activity of F-1 595 9

16 5 mM 3 -— [N-morpholino] propanesulfonic acid (manufactured by Sigma) RPMI 1640 medium containing buffer and 96-well microtiter plate for microbial dilution (Yamaguchi, H., According to J. Med. Myco 1., 36, 61 (1995), the antibacterial activity of F—15949-11 and F—15949-4 against test bacteria Was measured as the minimum inhibitory concentration. Table 4 shows the results. Table 4. Antifungal activity of F-15949 substances Test bacteria Minimum growth inhibitory concentration (μgZm 1) F-15949-1 F-15949-4

Candida albicans ATCC24433 16

Candida albicans ATCC90028> 64 Candida glabrata ATCC90030 16 64 Aspergillus fumigatus ATCC26430> 16> 64 Cryptococcus neof ormans T0 M185b> 16 64

As shown in Table 4, F-15949-1 and F-15949-4 exhibited growth inhibitory activity against various fungi.

Also, the antifungal activity of F-15949-2 can be confirmed and confirmed by the method of this example.

Formulation example 1. Oral capsule

F-15949-1, F-15949-2 or F-15949-4 330 mg

Lactose 170 mg

Corn starch 150 mg

Magnesium stearate 2 mg

3 52 2 mg Mix the above ingredients, collect the powder through a 30 mesh sieve,

Put into capsules to make capsules.

"Possibility of industrial use" The novel compound F-15949 or the salt thereof provided by the present invention has an excellent antifungal activity and is useful as a preventive or therapeutic agent for various fungal infections.

Claims

Claims The following general formula (I

(I)

(In the formula, R represents an alkyl group having 1 to 6 carbon atoms or a hydrogen atom.) Or a salt thereof.

2. The compound according to claim 1, wherein R is a methyl group or a hydrogen atom, or a salt thereof.

3. Compounds having the following physicochemical properties or salts thereof:

1) Properties of the substance: colorless powdery substance

3) Molecular formula: C _{3 3} H _6. 0 ₉

4) Molecular weight: 600 (measured by FAB mass spectrum method)

5) The exact mass, [M + N a.] +, Measured by the high-resolution LC-TOF mass spectrum method is as follows. '

Measured value: 6 2 3. 4 1 22

Calculated value: 6 2 3. 4 1 3 5

6) Ultraviolet absorption spectrum: The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption: 238 nm (£ 5970)

[ _a ] _D ²² : -20.0 ° (c 2.0)

340 7, 3 1 3 7, 2 9 28, 28 5 6, 1 6 7 9, 1 44 9, 1 38 1, 1 3 1 6, 1 2 1 3 , 1 1 4 6, 1 0 3 7, 8 4 5, 8 0 3, 7 2 6 cm- 1 9) 1 H- nuclear magnetic resonance scan Bae click Honoré: heavy methanol one Honoré The H-nuclear magnetic resonance spectrum, measured with the signal of methanol at 3.31 ppm, is as follows:

0.89 (3H, m), 0.92 (3H, m), 1.06 (3H, d, J = 6.6Hz), 1.12 (2H, m ), 1.25—1.46 (23H, m), 1.31 (3H, m), 1.32 (2H, m), 1.80 (3H, d) , J = l. 1 H z), 2.77 (1 H, m), 3.49 (1 H, m), 3.88 (1 H, dd, J = 1.9, 1 2 1 H z), 3.76 (1 H, dd. J = 4.5, 1 2., 1 H z), 4.05 (1 H, d, J = 2.5 H z), 4.09 (1H, dd, J = 2, 5, 9.3Hz), 4.47 (1H, d, J = 7.1Hz), 4.97 (1H, d, dd, J = 1.9, 4.5, 9.3 Hz), 6.64 (1H, dd, J = 1.1, 9.3 Hz) ppm

¹ 0) 1 ³ C mononuclear magnetic resonance scan Bae-vector: heavy methanol in our measured signal methanol 4 9. and 0 ppm, ^{1 3} C-NMR spectra are in shown below is there :

12.0 (q), 14.6 (q), 15.0 (q), 20.1 (q), 20.6 (q), 23.9 (t), 26 9 (t), 27. 2 (t), 28. 3 (t), 30. 6 (t), 30. 9 (t X 4), 31. 0 (t), 32 1 (d), 33.0 (d), 33.2 (t), 38.5 (t), 38.6 (t), 38.0 (t), 45.6 (t), 47.6 (d), 61.7 (t), 71.4 (d), 71.6 (d), 75.5 (d) ', 75.6 ( d), 1 35.5 (s), 153.3 (d), 172.1 (s), 16.7. (s), 20.9 (s) ppm 10 ) High performance liquid chromatography:

Column: Capsule pack C ₈ UG 1 20

(Diameter 4.6 mm x length 150 mm: manufactured by Shiseido Co., Ltd.) Solvent: 0.02% trifluoroacetic acid containing 72% acetonitrile Water flow rate: 0.6 m 1 Z min Detection: UV absorption wavelength 240 nm

Retention time: 14.3 minutes. '

4. Compounds having the following physicochemical properties or salts thereof:

1) Properties of the substance: colorless powdery substance

3) Molecular formula: C ₃₄ H ₆₂ 0 ₉

4) Molecular weight: 6 14 (measured by the FAB mass spectrum method),

5) High-resolution FAB—exact mass, measured by mass spectrometry, [M + Na] ⁺ is as follows.

Measured value: 6 3 7. 4 2 9 2

Calculated value: 6 3 7. 4 2 9 1

6) Ultraviolet absorption spectrum: The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption: 236 nm (ε7970),

[α] _D ²⁶ : ^{-58 c} '(c 1.1),

34 28, 2 9 5 3, 2 9 25, 28 54, 2 7 2 9, 1 744, 1 6 6 3, 1

4 5 6, 1 3 7 6, 1 2 5 1, 1 2 1 6, 1 1 30, 1 0 7 3, 1 0 3 6, 9 5

1 cm— ¹

9) ¹ H—nuclear magnetic resonance spectrum: the signal of heavy dimethyl sulfoxide was measured at 2.5 O ppm in heavy dimethyl sulfoxide. —The nuclear magnetic resonance spectrum is as shown below. :

0.89 (3H, m), 0.92 (3H, m), 1.03 (1H, m), 1.06 (3 0, d, J = 6.6Hz ), 1.17 (2 H, m), .l. 23 (2 H, m), 1.2 1—1.30 (22 H, m), 1.31 (l H, m) , 1.32 (3H, m), 1.35 (1H, m), 1.80 (3H, d, J = 1. 1Hz) 1.33 (2H, m) , 2.64 (1 H, m), 3.40 (1 H, m), 3.6 3 (3 H, s), 3.76 (1 H, dd, J = 1.9, 12.1 Hz), 3.88 (1 H, dd, J = 4.5, 1 2 1 H z), 4.05 (1 H, d, J = 2, 3 H z), 4.09 (1 H, dd, J = 2.5, 9.3 H z), 4. 1 9 (1H _; d, J = 5.0Hz), 4.42 (1H, d, J = 7.OHz), 4.57 (1H, t, J = 5.4 Hz), 4.97 (1H, ddd, J = 1.9, 4.5, 93Hz), 5.02 (1H, d, J = 2.8Hz), 5 0 4 (1 H, d, J = 7.4 H z), 6.6 1 (1 H, dd, J = l. 1, 9.3 H z) ppm

¹ 0) 1 ³ C- nuclear magnetic resonance spectra: in deuterated dimethyl sulfoxide was measured signals for the heavy dimethylcarbamoyl Rusuruhokishido as ³ 9. 5 ppm, ¹ 3 C- nuclear magnetic co-sounding spectrum is shown below As: C:

11.3 (q), 14.0 (q), 14.1 (q), 19.3 (q), 19.9 (q), 2.2.1 (t), 25 2 (t), 25.6 (t), 26.5 (t), 28.8 (t), 29.1 (t X 4), 29, 2 (t), 30 1 (d), 3 10 (d), 3 1. 3 (t), 36.2 (t), 37.1 (t), 37.2 (t) 4 3.6 (t ), 45.8 (d), 51.6 (q), 59.3 (t), 69.4 (d), 69.5 (d), 70.4 (d), 73.7 (d), 13.3.4 '(s) 150.9 (d), 170.2 (s), 173.3 (s), 197.8 ( s) ppm

1 1) High performance liquid chromatography:

Column: YMC—Pac ODS—AMS—3

(Diameter 4.6 mm X length 150 mm: manufactured by PMC Inc.) Solvent: 90% acetonitrile water containing 0.04% trifluoroacetic acid Flow rate: 1.0 ml / min

Detection: UV absorption 240 nm

Retention time: 8.26 minutes.

5. The following formula (II)

Or a salt thereof.

6. Compounds having the following physicochemical properties or salts thereof:

1) Properties of the substance: colorless powdery substance ·

3) Fraction formula: C a 3 H ₆ O O ₉

4) Molecular weight: 600 (measured by FAB mass spectral method),

5) The high-precision mass, [M + Na] ⁺ , measured by the high-resolution FAB mass spectrum method is as follows. .

Measured value: 6 2 3 4 1 3 4.

Calculated value: 6 2 3. 4 1 3 5

6) Ultraviolet absorption spectrum: The ultraviolet absorption spectrum measured in methanol shows the following maximum absorption: 2336 nm ( _{£ 8.588} )

7) Optical rotation: The optical rotation measured in methanol shows the following values: [H] _D ²⁶ : —33 · 0 ° (c 1.0)

8) Infrared absorption spectrum: Infrared absorption spectrum measured by potassium bromide (KBr) tablet method shows the following maximum absorption: '

3 3 5 3, 2 9 5 5, 2 9 19, 2 8 5 0, 1 7 8 5, 1 7 3 7, 1 6 6 6, 1 4 6 5, 1 4 5 6, 1 3 7 7, 1 3 1 7, 1 2 8 3, 1 2 5 1, 1 2 0 1, 1 1 3 1, 1 0 9 3, 1 0 5 2 cm ¹

9) ¹ H—nuclear magnetic resonance spectrum: In heavy pyridine, the signal of heavy pyridine is

The ¹ H—NMR spectrum, measured at 74 ppm, is as follows:

0.88 (3H, m), 0.90 (3H, m), 1.03, (3 (, d, J = 6.3Hz), 1.1.4 (1H, m), 1.22 (1H, m), 1.25 (2H, m), 1.27 (2H, m), 1.21-1.40 (12H, m), 1.3.4 (1 H, m), 1.4 1 (2 H, m), 1.44 (1 H, m), 1.46 (2 H, m), 1.5 1 (3 H, d, J = 6.9 Hz,) 1.62 (1H, m),, 1.68 (2H, m), 1.69 (2H, m), 1.74 (2H, m) , 1.97 (3 H, brs), 2.70 (1 H, m), 3.90 (1 H, m), 4.56 (1 H, d, J = 6.9 Hz ), 4.82 (1H, m), 4.83 (1H, m), 4.94 (1H, dd, J = 5.1, 10.9Hz), 5. 1 2 (1 H, dd, J = l. 9, 10.9 Hz), 5.46 (1 H, br s.), 6.77 (1 H, d, J = 9.4 H z) ppm

1 0) ¹³ C-Nuclear magnetic resonance spectrum ': in deuterated methanol circuit the signal of deuterated methanol were measured as 49. O ppm, ¹³ C-nuclear magnetic resonance spectrum is as shown below :

12.5 (q), 14.8 (q), 15.2 (q), 20.3 (q), 20.6 (q), 23.4 (t), 27.0 ( t), 27.3 (t), 28.0 t), 3

0.1 (t), 30.4 (t), 30.5 (tX2), 30.6 (t), 30.

8 (t), 31.4 (d), 32, 2 (d), 32.6 (t), 37.7 (t),

39.0 (t), 39.1 (t), 44.9 (t), 47.9 (d), 69.3 (t), 70.3 (d), 71.5 ( d), 72.4 (d), 74.5 (d),

1 35.1 (s), 15 1.5 (d), 17 2.8 (s), 17.8 (s),

1 9 8.6 (s) p p m

1 1) High performance liquid chromatography:

Column: YMC—Pac OD S-AM S 3,

(Diameter 4.6 mm x length 150 mm: manufactured by JMC Corporation) Solvent: 90% acetonitrile water containing 0.04% trifluoroacetic acid Flow rate: 1. Oml / min

Detection: UV absorption 240 nm

Retention time: 6.92 minutes.

7. Culturing a bacterium producing the compound according to any one of claims 1 to 6 belonging to the genus Haplographium, and culturing the bacterium; 7. The production of a compound according to any one of claims 1 to 6, characterized in that the compound is collected from the product according to any one of claims 1 to 6. Method.

8. The bacterium according to any one of claims 1 to 6, which belongs to the genus Haplographium, is a bacterium producing the compound according to any one of claims 1 to 6, wherein the haplodaradium 'Heliocephalum' Lao 'and' De 'Hog is used. (H aplographiumhelioceph alum V asant Rao & denoog) S ANK2 689 9 (FE RM BP—7243) strain, the production method according to claim 7,

9. A medicament comprising the compound according to any one of claims 1 to 6 or a salt thereof as an active ingredient. .

1 Q. An antifungal composition comprising the compound according to any one of claims 1 to 6 or a salt thereof as an active ingredient.

1 1. Haplographic 'Heliosefalum' Lao 'and de Hog (Haplograaph i umheel lcocephaalumvas antRao & deHoog) S ANK26899 (FERMBP-7243) strain.

1 2. A method for preventing or treating mycosis, comprising using the compound according to any one of claims 1 to 6 or a salt thereof.

1 3. Use of the compound or a salt thereof according to any one of claims 1 to 6 for producing an antifungal agent.