JP2002326952A - Fungus infection treating agent containing new compound f-15078 - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a fungus infection treating agent containing a new compound as an active ingredient. SOLUTION: A medicine containing a compound expressed by the formula (I) or its salt as an active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a medicament containing a novel compound having an antifungal activity as an active ingredient, and a therapeutic and preventive agent for a fungal infection containing the compound as an active ingredient.

[0002]

2. Description of the Related Art Antifungal compounds currently in clinical use include amphotericin B, flucytosine, azole compounds and the like. Many of these compounds are becoming problematic in terms of cytotoxicity and emergence of resistant bacteria.

[0003] Antifungal compounds produced by microorganisms include:
Pneumocandins (Schmatz, DM, et al., J.A.) produced by the genus Zalerion and the like.
ntibiotics 45, 1886 (1992)), Aspergillus (Asper
gillus) produced by genus Echinocandin (echinocandi)
n) (Nyfeler, R. and Keller-Schierlein, W., Hel
v. Chim. Acta 57, 2459 (1974)) and Aureobasidin produced by the genus Aureobasidium
(aureobasidin) (Ikai, K., et al., J. Antibiotics
44, 925 (1991)), but these have not been applied to clinical use.

[0004]

DISCLOSURE OF THE INVENTION The inventors of the present invention have proposed a method of producing a spoma collected from soil collected at Chichijima, Ogasawara-mura, Tokyo.
A novel compound was found in cultures of nine strains, and it was confirmed that the compound had antifungal activity. Thus, the present invention was completed.

That is, the present invention relates to (1) the following formula (I)

[0006]

Embedded image A pharmaceutical containing a compound represented by the following formula or a salt thereof as an active ingredient; (2) a pharmaceutical containing a compound having the following physicochemical properties or a salt thereof as an active ingredient: 1) Properties of the substance: basic fat-soluble powder 2 ) Molecular formula: C ₅₅ H ₉₈ N ₈ O ₁₄ 3) Molecular weight: 1094 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₅ H ₉₉ N ₈ O)
₁₄ ): Actual value: 1095.7365 Calculated value: 1095.7281 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3434, 3335, 2962, 2937, 2875, 2806, 1750, 16
84, 1641, 1509, 1469, 1412,1371, 1314, 1294, 1271,
1204, 1156, 1128, 1074, 1020 7) Optical rotation: [α] _D ²⁵ -120 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.82 (3H), 0.87 (3H),
0.88 (1H), 0.92 (3H), 0.93 (3H), 0.94 (3H), 0.96 (3H),
0.97 (3H), 0.98 (3H), 1.01 (3H), 1.02 (3H), 1.03 (3H),
1.06 (3H), 1.21 (1H), 1.41 (3H), 1.41 (1H), 1.48 (1H),
1.48 (1H), 1.49 (1H), 1.52 (3H), 1.55 (1H), 1.65 (1H),
1.66 (1H), 1.70 (2H), 1.73 (1H), 1.81 (1H), 1.87 (1H),
2.28 (1H), 2.31 (1H), 2.37 (1H), 2.48 (3H), 2.89 (3H),
2.94 (3H), 2.96 (1H), 3.29 (3H), 3.56 (1H), 4.06 (1H),
4.14 (1H), 4.77 (1H), 4.78 (1H), 4.84 (1H), 4.91 (1H),
4.96 (1H), 5.21 (1H), 5.25 (1H), 5.53 (1H), 6.39 (1H),
7.83 (1H), 7.94 (1H), 8.28 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured in heavy chloroform using tetramethylsilane as an internal standard.
Is as follows. 10.9 (q), 11.9 (q), 15.0 (q), 15.1 (q), 16.0 (q), 16.6
(q), 17.4 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.1 (q), 2
1.0 (q), 21.4 (q), 22.1 (q), 23.1 (q), 23.51 (q), 23.54
(q), 24.2 (t), 24.6 (d), 24.8 (d), 25.4 (d), 25.5 (t),
27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 36.1 (q), 36.5
(t), 37.7 (t), 38.3 (d), 38.4 (d), 39.7 (t), 40.9 (q),
46.2 (d), 51.8 (d), 53.1 (d), 54.7 (d), 55.1 (d), 63.9
(d), 64.7 (d), 68.1 (d), 70.1 (d), 73.4 (d), 74.3 (d),
77.1 (d), 169.03 (s), 169.04 (s), 169.6 (s), 169.8 (s),
169.9 (s), 170.3 (s), 172.0 (s), 173.4 (s), 173.8 (s),
174.0 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter x 250 mm in length: manufactured by Showa Denko KK) Mobile phase: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 10. 20 minutes 11) Solubility: Soluble in dimethyl sulfoxide, methanol and chloroform. 12) Amino acid analysis: Threonine, alanine and isoleucine were recognized as hydrolysates. (3) Formula (I)
I)

Embedded image A drug containing a compound represented by the following as an active ingredient,
(4) Pharmaceutical containing a compound having the following physicochemical properties as an active ingredient: 1) Property of substance: neutral fat-soluble colorless powder 2) Molecular formula: C ₅₇ H ₁₀₀ N ₈ O ₁₅ 3) Molecular weight: 1136 (FAB) 4) High-resolution FAB-MS [M + H] ⁺ (C ₅₇ H ₁₀₁ N ₈₎
O ₁₅ ): Actual value: 1137.7410 Calculated value: 1137.7387 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3433, 3333, 2963, 2937, 2875, 1751, 1686, 16
42, 1516, 1469, 1409, 1388,1372, 1311, 1292, 1272,
1201, 1156, 1128, 1074, 1017 7) Optical rotation: [α] _D ²⁵ -131 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.83 (3H), 0.87 (1H),
0.87 (3H), 0.90 (3H), 0.92 (3H), 0.93 (3H), 0.95 (3H),
0.95 (3H), 0.98 (3H), 0.98 (3H), 1.01 (3H), 1.01 (3H),
1.03 (1H), 1.05 (3H), 1.28 (3H), 1.37 (1H), 1.40 (1H),
1.46 (1H), 1.47 (1H), 1.49 (1H), 1.51 (3H), 1.64 (1H),
1.65 (1H), 1.66 (1H), 1.86 (1H), 1.72 (1H), 1.78 (1H),
2.12 (3H), 2.13 (1H), 2.26 (1H), 2.31 (1H), 2.37 (1H),
2.88 (3H), 2.93 (3H), 2.97 (3H), 3.28 (3H), 3.56 (1H),
4.03 (1H), 4.15 (1H), 4.73 (1H), 4.78 (1H), 4.82 (1H),
4.83 (1H), 4.91 (1H), 4.97 (1H), 5.15 (1H), 5.28 (1H),
5.50 (1H), 6.37 (1H), 6.87 (1H), 7.86 (1H), 8.29 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) In deuterated chloroform, tetramethylsilane was used as an internal standard. The measured nuclear magnetic resonance spectrum (500 MHz)
It is as follows. 10.5 (q), 10.9 (q), 14.9 (q), 15.1 (q), 15.6 (q), 16.6
(q), 16.7 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.0 (q), 2
0.8 (q), 21.4 (q), 22.0 (q), 22.1 (q), 23.1 (q), 23.6
(q), 23.6 (q), 24.1 (t), 24.6 (t), 24.7 (d), 24.8 (d),
25.4 (d), 27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 31.6
(d), 31.8 (q), 36.1 (t), 37.6 (t), 38.4 (d), 39.6 (t), 4
0.9 (q), 46.1 (d), 51.8 (d), 53.1 (d), 54.7 (d), 54.7
(d), 61.2 (d), 63.9 (d), 64.6 (d), 68.1 (d), 73.1 (d), 7
4.3 (d), 77.0 (d), 168.9 (s), 168.9 (s), 169.1 (s), 16
9.9 (s), 169.9 (s), 170.3 (s), 170.6 (s), 171.7 (s), 17
2.0 (s), 173.3 (s), 173.8 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter × 250 mm in length: manufactured by Showa Denko KK) Moving bed: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 9. 05 min 11) Solubility: soluble in dimethyl sulfoxide, methanol, chloroform 12) Amino acid analysis: threonine, alanine, isoleucine were recognized as hydrolysates, (5) Formula (I) below

Embedded image (6) A therapeutic agent for a fungal infection comprising a compound having the following physicochemical properties or a salt thereof as an active ingredient: 1) a substance Properties: basic fat-soluble powder 2) Molecular formula: C ₅₅ H ₉₈ N ₈ O ₁₄ 3) Molecular weight: 1094 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₅ H ₉₉ N ₈ O)
₁₄ ): Actual value: 1095.7365 Calculated value: 1095.7281 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3434, 3335, 2962, 2937, 2875, 2806, 1750, 16
84, 1641, 1509, 1469, 1412,1371, 1314, 1294, 1271,
1204, 1156, 1128, 1074, 1020 7) Optical rotation: [α] _D ²⁵ -120 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.82 (3H), 0.87 (3H),
0.88 (1H), 0.92 (3H), 0.93 (3H), 0.94 (3H), 0.96 (3H),
0.97 (3H), 0.98 (3H), 1.01 (3H), 1.02 (3H), 1.03 (3H),
1.06 (3H), 1.21 (1H), 1.41 (3H), 1.41 (1H), 1.48 (1H),
1.48 (1H), 1.49 (1H), 1.52 (3H), 1.55 (1H), 1.65 (1H),
1.66 (1H), 1.70 (2H), 1.73 (1H), 1.81 (1H), 1.87 (1H),
2.28 (1H), 2.31 (1H), 2.37 (1H), 2.48 (3H), 2.89 (3H),
2.94 (3H), 2.96 (1H), 3.29 (3H), 3.56 (1H), 4.06 (1H),
4.14 (1H), 4.77 (1H), 4.78 (1H), 4.84 (1H), 4.91 (1H),
4.96 (1H), 5.21 (1H), 5.25 (1H), 5.53 (1H), 6.39 (1H),
7.83 (1H), 7.94 (1H), 8.28 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured in heavy chloroform using tetramethylsilane as an internal standard.
Is as follows. 10.9 (q), 11.9 (q), 15.0 (q), 15.1 (q), 16.0 (q), 16.6
(q), 17.4 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.1 (q), 2
1.0 (q), 21.4 (q), 22.1 (q), 23.1 (q), 23.51 (q), 23.54
(q), 24.2 (t), 24.6 (d), 24.8 (d), 25.4 (d), 25.5 (t),
27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 36.1 (q), 36.5
(t), 37.7 (t), 38.3 (d), 38.4 (d), 39.7 (t), 40.9 (q),
46.2 (d), 51.8 (d), 53.1 (d), 54.7 (d), 55.1 (d), 63.9
(d), 64.7 (d), 68.1 (d), 70.1 (d), 73.4 (d), 74.3 (d),
77.1 (d), 169.03 (s), 169.04 (s), 169.6 (s), 169.8 (s),
169.9 (s), 170.3 (s), 172.0 (s), 173.4 (s), 173.8 (s),
174.0 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter x 250 mm in length: manufactured by Showa Denko KK) Mobile phase: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 10. 20 minutes 11) Solubility: Soluble in dimethyl sulfoxide, methanol and chloroform. 12) Amino acid analysis: Threonine, alanine and isoleucine were recognized as hydrolysates. (7) Formula (I)
I)

Embedded image And (8) a fungal infection containing a compound having the following physicochemical properties as an active ingredient: 1) Nature of substance: neutral Fat-soluble colorless powder 2) Molecular formula: C ₅₇ H ₁₀₀ N ₈ O ₁₅ 3) Molecular weight: 1136 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₇ H ₁₀₁ N ₈₎
O ₁₅ ): Actual value: 1137.7410 Calculated value: 1137.7387 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3433, 3333, 2963, 2937, 2875, 1751, 1686, 16
42, 1516, 1469, 1409, 1388,1372, 1311, 1292, 1272,
1201, 1156, 1128, 1074, 1017 7) Optical rotation: [α] _D ²⁵ -131 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.83 (3H), 0.87 (1H),
0.87 (3H), 0.90 (3H), 0.92 (3H), 0.93 (3H), 0.95 (3H),
0.95 (3H), 0.98 (3H), 0.98 (3H), 1.01 (3H), 1.01 (3H),
1.03 (1H), 1.05 (3H), 1.28 (3H), 1.37 (1H), 1.40 (1H),
1.46 (1H), 1.47 (1H), 1.49 (1H), 1.51 (3H), 1.64 (1H),
1.65 (1H), 1.66 (1H), 1.86 (1H), 1.72 (1H), 1.78 (1H),
2.12 (3H), 2.13 (1H), 2.26 (1H), 2.31 (1H), 2.37 (1H),
2.88 (3H), 2.93 (3H), 2.97 (3H), 3.28 (3H), 3.56 (1H),
4.03 (1H), 4.15 (1H), 4.73 (1H), 4.78 (1H), 4.82 (1H),
4.83 (1H), 4.91 (1H), 4.97 (1H), 5.15 (1H), 5.28 (1H),
5.50 (1H), 6.37 (1H), 6.87 (1H), 7.86 (1H), 8.29 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) In deuterated chloroform, tetramethylsilane was used as an internal standard. The measured nuclear magnetic resonance spectrum (500 MHz)
It is as follows. 10.5 (q), 10.9 (q), 14.9 (q), 15.1 (q), 15.6 (q), 16.6
(q), 16.7 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.0 (q), 2
0.8 (q), 21.4 (q), 22.0 (q), 22.1 (q), 23.1 (q), 23.6
(q), 23.6 (q), 24.1 (t), 24.6 (t), 24.7 (d), 24.8 (d),
25.4 (d), 27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 31.6
(d), 31.8 (q), 36.1 (t), 37.6 (t), 38.4 (d), 39.6 (t), 4
0.9 (q), 46.1 (d), 51.8 (d), 53.1 (d), 54.7 (d), 54.7
(d), 61.2 (d), 63.9 (d), 64.6 (d), 68.1 (d), 73.1 (d), 7
4.3 (d), 77.0 (d), 168.9 (s), 168.9 (s), 169.1 (s), 16
9.9 (s), 169.9 (s), 170.3 (s), 170.6 (s), 171.7 (s), 17
2.0 (s), 173.3 (s), 173.8 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter × 250 mm in length: manufactured by Showa Denko KK) Moving bed: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 9. 05 min 11) Solubility: soluble in dimethyl sulfoxide, methanol, chloroform 12) Amino acid analysis: related to the presence of threonine, alanine, and isoleucine as hydrolysates.

The compounds of the present invention are described in the above (1) to (8). Of these, (1), (3),
The compounds described in (5) and (7) have the following general formula (II)
I)

[0008]

Embedded image (Wherein, R represents a hydrogen atom or COCH ₃ ), and in the present invention, the above general formula (II)
The compounds represented by I) are collectively referred to as F-15078.

The compounds described in the above (1) and (5) are compounds represented by the above general formula (III) and wherein R is a hydrogen atom. In the present invention, the above (1) and (5)
The compound described above or a compound having the physicochemical properties described in the above (2) and (6) is referred to as F-15078A.

The compounds described in the above (3) and (7) are compounds represented by the above general formula (III) and wherein R = COCH ₃ . In the present invention, the compound described in the above (3) and (7) or the compound having the physicochemical property described in the above (4) and (8) is prepared using F-15078B
That.

[0011] F-15078A can be converted into a salt according to a conventional method. The salt of F-15078A is for medical use,
It can be applied to both veterinary applications and other applications.

When the salt of F-15078A is applied to medical use and / or veterinary use, the salt of the compound is not particularly limited as long as it is pharmacologically acceptable. Examples thereof include inorganic salts such as salts, and organic acid salts such as pamoate.

[0013] There is no particular limitation when the salt of F-15078A is used for purposes other than medical and veterinary uses, for example, as a synthetic intermediate. Such salts include, for example, inorganic salts such as acetate, bromide, chloride, hydrochloride, bromate, iodide, sulfate, phosphate, diphosphate; citrate, Organic acid salts such as maleate, pamoate and tartrate can be exemplified.

The F-15078 of the present invention has various isomers. In the present invention, all of these isomers are represented by a single formula. Accordingly, the present invention includes all these isomers and mixtures of these isomers.

Furthermore, the present invention includes all of the cases where F-15078 forms solvates (eg, hydrates).

For example, when the F-15078 of the present invention is left in the atmosphere or recrystallized, it may absorb moisture, adhere to adsorbed water, or form a hydrate. The present invention includes such a solvate.

The present invention also includes all compounds that are metabolized in vivo and converted into F-15078, so-called prodrugs.

[0018]

BEST MODE FOR CARRYING OUT THE INVENTION [Production bacteria] F-15078 can be obtained from a culture of a fungus producing the compound. F
As a producing microorganism of -15078, for example, Houma (Ph
oma) genus and the like, and, preferably, Poma sp.
13899 strains (hereinafter simply referred to as “SANK 13899 strains”). SANK13899 strain was isolated from soil collected at Chichijima, Ogasawara-mura, Tokyo.

In order to observe the mycological properties of the bacterium, cultivation was performed on each of the following media.

The composition of each medium used is described below. PDA medium (potato dextrose agar medium) Nissui potato dextrose agar medium (manufactured by Nissui Pharmaceutical Co., Ltd.) 39 g distilled water 1000 ml CMA medium (corn meal agar (Corn Meal Agar) medium) corn meal agar Pharmaceutical Co., Ltd.) 17 g Distilled water 1000 ml WSH medium Quaker oatmeal 10 g Magnesium sulfate heptahydrate 1 g Potassium dihydrogen phosphate 1 g Sodium nitrate 1 g Agar 20 g Distilled water 1000 ml Miura medium glucose 1 g Potassium dihydrogen phosphate 1 g Magnesium sulfate heptahydrate 0.2 g of potassium chloride 0.2 g of yeast extract 0.2 g of sodium nitrate 2 g of agar 20 g of distilled water 1000 ml of CYA medium (Czapek Yeast Extract Agar medium) dipotassium hydrogen phosphate 1.0 g of Zapec concentrate * 10 ml of yeast extract 5 g Sucrose 30g Agar 15g Distilled water 1000 ml MEA medium (Malt Extract Agar medium) Malt extract 20 g Peptone 1 g Glucose 20 g Agar 20 g Distilled water 1000 ml G25N medium (25% Glycerol Nitrate Agar medium) Dipotassium hydrogen phosphate 0.75 g Zapec concentrated Liquid * 7.5 ml Yeast extract 3.7 g Glycerol 250 g Agar 12 g Distilled water 750 ml * Zapec concentrate refers to a solution having the following composition.

Sodium nitrate 30 g Potassium chloride 5 g Magnesium sulfate heptahydrate 5 g Iron (II) sulfate heptahydrate 0.1 g Zinc sulfate heptahydrate 0.1 g Copper sulfate pentahydrate 0.05 g Distilled water 100 ml `` Metune Handbook of Color '' (Kornerup, A. and Wanscher, JH (1978) Meth
uen handbook of color (3rd. edition) .EryeMethuen,
London.)

The bacteriological properties of the SANK 13899 strain in culture are as follows.

The growth on the PDA medium is 13 to 17 mm in diameter at 23 ° C. for 2 weeks. The colonies are fluffy, wobbly in the center, and slightly marginal at the margins of the colonies. The colony surface is gray (3B1) to white. The back surface is brown (6E5 to 4).

The growth on CMA medium is 15 to 17 mm in diameter at 23 ° C. for 2 weeks. The colonies are powdery and the margins of the colonies are toothy. Colony surface is grayish green (27E5) to dull green '
(27E4). The back is dark green (27F
4).

The growth on the Miura medium is 22 to 33 mm in diameter at 23 ° C. for 2 weeks. The colonies are compacted and the margins of the colonies are whole. The colony surface is white. The back is yellowish white (3A2)
To white.

The growth on WSH medium is 33 to 34 mm in diameter at 23 ° C. for 2 weeks. The colonies are compacted and the margins of the colonies are whole. The colony surface is pale yellow (3A3) to yellowish white (3A2). The back is the same color as the colony front.

The growth on the CYA medium is 34 to 35 mm in diameter at 23 ° C. for 2 weeks. The colonies are fluffy, wobbly in the center and secrete a weak soluble pigment. The margins of the colonies are constricted and are all-edge. The colony surface is grayish orange (6B3). The back side is brownish orange (6C8) to brown (6D8).

The growth on the MEA medium is 15 to 23 mm in diameter at 23 ° C. for 2 weeks. The colony is fluffy and wobbly in the center. The margins of the colonies are constricted and are full-edge or slightly tooth-like. The colony surface is white to gray (3B1). The back is dark green (2
8F4 to 3F).

It does not grow on G25N medium.

The hyphae are 0.5 to 3 μm in diameter, often bundled, thin to thick walled, smooth to rough, colorless to brown.

The SANK13899 strain can be obtained by continuously culturing on an ODA medium or Miura medium for one month or more.
Sectors were formed, and conidial shells buried in the agar medium were formed. The bacteriological properties are as follows.

The conidial shells are 100 to 200 μm in diameter, all buried or partially buried in an agar medium, and are spherical or subspherical brown or black. No openings were observed. Conidia forming cells are 7.5-9 μm × 1.3-1.8 μm
, Formed on spherical conidia inner lining cells, single cells, cylindrical or nib-shaped, and colorless. Phyaro-type conidia are 3.5 to 4.7 μm × 1.3 to 1.8 μm
It is cylindrical, single cell and colorless. Mycelia are 0.5 to 3 μm in diameter, often bundled, thin or thick walled, smooth to rough, colorless to brown. Kobayashi, M., J. Antibact. Antifung. Agents, 2
2, 757 (1994)).
99 strains were identified as Houma SP.

The SANK13899 strain was
1-1-3 Higashi, Tsukuba, Ibaraki, Japan on August 20, 2008
Of the Ministry of International Trade and Industry
It has been internationally deposited with the National Institute of Advanced Industrial Science and Technology (AIST), 1-1-1-1 Higashi, Tsukuba, Ibaraki, Japan, and has been assigned the accession number FERM BP-6851.

As is well known, fungi can be produced in nature or by man-made operations (eg, UV irradiation, irradiation,
Mutation easily occurs due to chemical treatment, etc., and the same applies to the SANK13899 strain of the present invention. In the present invention, the SANK13899 strain includes all mutants thereof.
These mutants also include those obtained by genetic methods, for example, recombination, transduction, transformation and the like. That is, SANK producing F-15078
13899 strains, their mutants and strains not clearly distinguished from them are all included in the SANK13899 strain. [Culture method and purification method] Culture of the F-15078-producing bacterium as described above is generally performed in a medium used for producing a fermentation product. Such a medium contains a carbon source, a nitrogen source, an inorganic salt, a trace amount of a growth factor, a trace metal, and the like, which can be assimilated by the microorganism.

Examples of the carbon source include glucose, fructose, maltose, sucrose, mannitol, glycerin, dextrin, oats, rye, corn starch, potato, corn flour, soybean oil, cottonseed oil, molasses, citric acid, tartaric acid and the like. Can be mentioned. These carbon sources can be used alone or in combination of two or more.

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

Examples of the nitrogen source include soybean flour, bran, fallen flour, cottonseed powder, casein hydrolyzate, pharmamine, corn steep liquor, peptone, meat extract, yeast, yeast extract, malt extract, and sodium nitrate , Ammonium nitrate, ammonium sulfate and the like. These nitrogen sources can be used alone or in combination of two or more nitrogen sources.

The amount of the carbon source used in the medium depends on other components in the medium, but is usually 0.2 to 6% by weight.

The carbon source and the nitrogen source need not be of high purity, but may be of lower purity containing trace amounts of growth factors, vitamins, inorganic nutrients and the like.

Also, sodium, potassium, magnesium, ammonium, calcium, phosphoric acid, sulfuric acid, hydrochloric acid,
Salts capable of obtaining ions such as carbonate may be added to the medium.

Furthermore, vitamin B1, which can be assimilated by bacteria, vitamins such as biotin, cell growth promoting substances such as thiamine, and salts of metals such as manganese and molybdenum may be added to the medium.

In the case of a liquid medium, an antifoaming agent such as silicone oil, polyalkylene glycol ether, vegetable oil, animal oil, or a surfactant may be added to the medium to prevent foaming of the medium. .

The pH of the liquid medium used for culturing the F-15078-producing bacterium depends on the strain, the pH stability of the F-15078, etc. When the producing bacterium is SANK13899, the pH of the liquid medium is adjusted to pH 5.0 to 5.0. Adjust to pH 7.0.

The cultivation temperature of the F-15078-producing bacterium depends on the strain, the thermal stability of the F-15078, etc., and when the producing bacterium is SANK13899, the temperature is 15 to 37 ° C, preferably 22 to 35 ° C. It is. The culture temperature of the SANK13899 strain for producing F-15078 is as follows:
Preferably it is 22 to 26 ° C, more preferably 23 ° C.

The method for culturing the F-15078-producing bacterium is not particularly limited, and examples thereof include a culturing method using a solid medium, a stirring culturing method, a shaking culturing method, an aeration culturing method, and an aeration-stirring culturing method. Preferably, stirring culture method,
A shaking culture method, aeration culture method, and aeration stirring culture method, and more preferably a shaking culture method. For culturing on an industrial scale, the aeration stirring culture method is more preferable.

The cultivation of the compound-producing bacterium for producing F-15078 usually starts with a seed culture in which a slant in which the bacterium has grown is inoculated into a small amount of medium, and if necessary, the seed culture is repeated on a larger scale. After the cultivation, the final stage is performed by conducting a main culture on a larger scale.

When small-scale cultivation of the F-15078-producing bacterium is performed, seed culture is performed using an Erlenmeyer flask or the like, and if necessary, a larger-scale seed culture is performed again. Perform culture.

When cultivating the F-15078-producing bacteria on a large scale, it is preferable to use a jar fermenter or a tank equipped with a stirrer and an aerator. With such a device, the medium can be prepared and sterilized in a jar fermenter or tank. This method is suitable for large-scale production of F-15078.

The production of F-15078 by the strain SANK 13899 usually reaches its maximum value after 144 to 192 hours of culture.

F-15078 present in the culture can be obtained by subjecting the culture and the culture to a filtration operation using diatomaceous earth or the like as an auxiliary agent, and subjecting the culture to a centrifugation operation. Extraction and purification can be performed from the obtained culture filtrate and / or cells using the physicochemical properties of F-15078.

F-1 present in culture filtrate or culture supernatant
5078 can be extracted with an organic solvent that is immiscible with water under neutral pH conditions, for example, ethyl acetate, chloroform, ethylene chloride, methylene chloride, butanol alone or a mixed solvent of two or more thereof. In addition, the culture filtrate or the culture supernatant is treated with an adsorbent, for example, activated carbon, Amberlite XAD-2, Amberlite XAD-4 (all manufactured by Rohm and Haas), Diaion HP
-10, DIAION HP-20, DIAION CHP
-20P, Diaion HP-50, Sepabeads SP-
207 (manufactured by Mitsubishi Chemical Corporation) or the like to adsorb and remove impurities, or after adsorbing F-15078 and separating from impurities, methanol water, acetone water, butanol water Can be used to elute F-15078. .

F-15078 present in the cells contains 50
Extraction with 90% aqueous acetone or aqueous methanol, filtration using diatomaceous earth or the like as an auxiliary agent, removal of the organic solvent from the obtained filtrate, and extraction in the same manner as in the case of the culture filtrate or culture supernatant. It can be obtained by performing a purification operation.

After completion of the culture, F-15078 present in the culture can be extracted by adding an appropriate amount thereof, preferably 50% (vol / vol) final concentration of acetone or methanol. After completion of the extraction, a filtration operation using diatomaceous earth as a filter aid is performed, and the obtained filtrate can be subjected to the same extraction and purification operation as in the case of the culture filtrate or the culture supernatant.

The crude fraction of F-15078 thus obtained was subjected to distribution column chromatography using TSK Geltoyopearl HW-40F (manufactured by Tosoh Corporation), Sephadex LH-20 (manufactured by Amersham Pharmacia), or the like. It can be further purified by chromatography, reverse layer column chromatography using Cosmoseal 140C18 (manufactured by Nacalai Tesque, Inc.) or the like. If necessary, the thus-purified F-15078-containing fraction was subjected to Sh
high-performance liquid chromatography using a reverse-layer column such as Odex asahipak C8P50-4E (manufactured by Showa Denko KK), YMC Pack ODS-AM (manufactured by YMC), Capsule Pack UG120 (manufactured by Shiseido), etc. (High performance li
liquid chromatography: Hereinafter, "H
PLC ". ) Can be used for further purification.

These extraction means and isolation means can be used alone or in an appropriate combination, and if necessary, repeated for the same means, and used for the isolation of F-15078.

Culture step of the above-mentioned F-15078-producing bacterium,
And F-15078A in the purification step of A or B
The behavior of 15078A or B can be tracked by the following method 1) or 2). 1) Method by HPLC analysis: The method is not particularly limited as long as it is a method using HPLC. For example, the following HPLC conditions can be applied.

Column: Shodex Asahipa
k C8P 504E (4.6 mm diameter x 250 m
m: manufactured by Showa Denko KK Moving bed: acetonitrile: 10 mM aqueous ammonium bicarbonate = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time of F-15078A: 10.20 min F-15078B Retention time: 9.05 minutes 2) Method by measuring antifungal activity: The method is not particularly limited as long as it is a method for measuring antifungal activity.
The broth dilution method of Yamagichi et al. (Yamaguchi, H., etal., J.
Med. Mycol. 36, 61 (1995)) can be applied. The F-15078 or a salt thereof of the present invention has an antifungal activity and is useful as a preventive or therapeutic agent for fungal infections.

When F-15078 or a salt thereof is used as a prophylactic or therapeutic agent for fungal infection, the form of administration is not particularly limited, and may vary depending on the preparation, age, sex, type of disease, degree of disease and the like. It can be appropriately selected depending on the situation.

For example, tablets, pills, powders, granules, syrups, solutions, suspensions, emulsions, granules, capsules and the like can be administered orally. Injectables are used alone or as a mixture with a replacement fluid such as glucose and amino acids, or as an emulsion mixed with polyoxyethylene sorbitan fatty acid esters, etc., for intravenous administration, intramuscular administration, intradermal administration, subcutaneous administration and / or It can be administered intraperitoneally. Suppositories may be administered rectally.

These various preparations are produced using excipients, binders, disintegrants, lubricants, solubilizers, flavoring agents, coating agents and the like, in addition to the main drug, in the art. be able to.

For forming tablets, carriers known in the art such as excipients such as lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silicic acid, water, Ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone and other binders, dried starch, sodium alginate, agar powder, laminaran powder, sodium hydrogen carbonate, Disintegrants such as calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, lactose, sucrose, stearin,
Disintegration inhibitors such as cocoa butter and hydrogenated oil; absorption promoters such as quaternary ammonium bases and sodium lauryl sulfate; humectants such as glycerin and starch; adsorbents such as starch, lactose, kaolin, bentonite and colloidal silicic acid Lubricants such as purified talc, stearates, powdered boric acid, and polyethylene glycol can be used. For tablets,
If necessary, tablets coated with a usual coating, such as sugar-coated tablets, gelatin-coated tablets, enteric-coated tablets, film-coated tablets, double tablets, and multiple tablets, can be prepared.

For forming pills, carriers known in the art such as excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, talc, gum arabic powder, tragacanth powder, gelatin And binders such as ethanol, disintegrants such as laminaran and agar.

In molding suppositories, carriers known in the art, such as polyethylene glycol, cocoa butter, higher alcohols, esters of higher alcohols, gelatin, and semi-synthetic glycerides can be used as carriers.

When the injection is a solution, emulsion or suspension, it is preferably sterile and isotonic with blood. When the solution, emulsion and suspension are formed, they are known in the art as diluents. For example, water, ethyl alcohol, propylene glycol, epoxidized isostearyl alcohol, polyoxylated isostearyl alcohol, polyoxyethylene sorbitan fatty acid esters and the like can be used. The injection may contain a sufficient amount of salt, glucose, glycerin and the like to make the injection isotonic with blood, and may contain a usual solubilizing agent, a buffer and a soothing sugar.

These preparations may contain coloring agents, preservatives, fragrances, flavors, sweeteners, other drugs, and the like, if necessary.

F-150 contained in these various preparations
The amount of 78 depends on the dosage form, administration method and the like, but is usually 1 to 70% by weight, preferably 1 to 30% by weight.

The dose of F-15078 depends on the type of disease,
Depending on the degree of the disease, age, weight, administration method, dosage form, etc., for adults, the upper limit of the daily dose is 20 to 2000 mg, the lower limit is 0.001 to 0.1 mg, A preferred range is 0.01 to 200 mg, and a more preferred range is 0.1 to 20 mg.

The number of administrations of the drug containing F-15078 depends on the dosage form, the type of disease, the degree of disease, the weight, etc., but is not particularly limited.
Or several times a day.

[0069]

Next, the present invention will be described more specifically with reference to examples, test examples and production examples, but the present invention is not limited to these examples. Embodiment 1 FIG. Production of F-15078A (1) [Culture of SANK13899 strain (1)] SANK138
One loopful of the 99 strains was inoculated into three 500 ml Erlenmeyer flasks containing 100 ml of a seed culture medium having the composition shown in Table 1 and sterilized at 121 ° C. for 30 minutes.
The culture was performed for 5 days under the condition of pm (rotation radius: 7 cm). The obtained culture solution was used for 2 L containing 400 ml of a medium having the same composition.
5% inoculated in 9 Erlenmeyer flasks, 23 ℃, 210rpm
m (rotational radius 7 cm) for 2 days. The obtained seed culture was sterilized at 121 ° C. for 30 minutes.
30 containing 15 L of main culture medium (1) having the described composition
4 L jar incubators are inoculated with 5%, 23 ° C., aeration rate 1 vvm, 100 to 420 rpm, dissolved oxygen amount 5.
The cells were cultured for 7 days under the condition of 0 ppm.

[Table 1] Medium composition of seed culture medium ―――――――――――――――――――――――――――――――――― Glycerin 30g Glucose 30g Soluble starch 20g Soy flour 10g Gelatin 2.5g Yeast extract (Difco) 2.5g NH4NO3 2.5g Defoamer * 0.1ml Tap water 1000ml (pH is not adjusted.) ――――――――――――――――――――――――――

[Table 2] Medium composition of main culture medium (1) ―――――――――――――――――――――――――――――――― Dextrin (Difco) 10 g glycerin 20g glucose 30g malt extract (Difco) 10 g yeast extract (Difco) 2 g tryptone _{(Difco) 1g NH 4 NO 3} 1g NaNO 3 1g KH 2 PO 4 1g MgSO 4 · 7H 2 O 1g antifoam * 0 ．1ml tap water 1000ml (pH is not adjusted.) ―――――――――――――――――――――――――――――――――― Nissan Deform CB-442 (manufactured by NOF CORPORATION) [Isolation of F-15078A (1)] The obtained culture 60
60 L of acetone was added to L, and the mixture was stirred for 40 minutes, and 3 kg of a filter aid (Celite 545: manufactured by Celite Corporation) was added to the extract, followed by filter press filtration. Further, 50 L of ethyl acetate was added to the filtrate to perform extraction, and 49 L of the obtained organic layer was washed with 50 L of saturated saline and 50 L of purified water, and then 5 kg of anhydrous sodium sulfate was added thereto and dehydrated for 1 hour. After removing sodium sulfate by filter press filtration, the filtrate was concentrated under reduced pressure to dryness to obtain 95.9 g of an oily substance. The oily substance was dissolved in methanol: 0.04% aqueous trifluoroacetic acid = 8: 2100
The resulting solution was supplied to a 22 L Toyopearl HW-40F (manufactured by Tosoh Corporation) column which had been equilibrated with the same solvent in advance. The column was developed with the same solvent, and the eluate was
The resulting mixture was fractionated by ml, and F-15078A was recovered in fraction numbers 13 to 25 (6.5 L in total). The obtained fraction was concentrated under reduced pressure to 2 L, and then adjusted to pH 7 with 6.25 N sodium hydroxide. 3.1 L of ethyl acetate was added to the concentrate to extract the active substance. Extract 3
After washing with saturated sodium chloride solution and dehydration with anhydrous sodium sulfate, the solution was concentrated under reduced pressure to dryness to obtain 66.5 g of an oily substance. This oily substance was treated with methanol: 0.05
% Trifluoroacetic acid aqueous solution = 8: 2 dissolved in 160 ml,
Acetonitrile: 0.05% aqueous trifluoroacetic acid = 4:
6L Cosmo Seal 140C18 pre-equilibrated in 6
(Nakarai Tesque Co., Ltd.). The column was washed with 10 L of the same solvent and acetonitrile: 0.
Developed with 05% aqueous trifluoroacetic acid = 6: 4. The eluate was fractionated in 2 L portions, and F No. 2 (2 L)
-15078A was recovered. 800 fractions obtained
After concentration under reduced pressure to a volume of 7 ml, the pH was adjusted to pH 7 with 6.25 N sodium hydroxide, 1 L of ethyl acetate was added to extract the active substance, washed, dehydrated, and concentrated to dryness under reduced pressure to obtain 234.3 mg of a crude product. Obtained. This crude product was dissolved in 4 ml of methanol and subjected to HPLC under the following HPLC conditions (1) to obtain a fraction containing F-15078A. HPLC conditions (1): Column: YMC Pack ODS-AM 30 mm in diameter x 300 mm in length (manufactured by YMC) Solvent: acetonitrile: 1% aqueous triethylamine phosphate (pH 6.0) = 3: 1 Detection wavelength: UV 210 nm flow rate: 10.4 ml / min Temperature: room temperature Retention time: 77 to 98 minutes The obtained F-15078A-containing fraction is desalted using ethyl acetate as described above, and concentrated to dryness under reduced pressure. As a result, 34.1 mg of F-15078A was obtained. Embodiment 2. FIG. Production of F-15078A (2) [Culture (2) of SANK13899] SANK138
Example 1 99 strains were sterilized at 121 ° C. for 30 minutes.
6 loops of Erlenmeyer flasks containing 500 ml of a seed culture medium having the composition shown in
The cells were cultured for 6 days under the conditions of 10 rpm (rotation radius: 7 cm). The obtained seed culture solution was inoculated with 5% inoculation into two 60 L tank incubators containing 30 L of medium having the same composition, and was incubated for 2 days at 23 ° C. under aeration of 1 vvm, 100 rpm, and dissolved oxygen of 5.0 ppm. Cultured. The obtained culture solution is heated at 121 ° C.
5% was inoculated into one 600 L tank incubator containing 300 L of the main culture medium (2) having the composition shown in Table 3 and sterilized for 30 minutes at 23 ° C., aeration volume 1 vvm, 83 to 2
The cells were cultured for 7 days under the conditions of 40 rpm and 5.0 ppm of dissolved oxygen.

[Table 3] Medium composition of main culture medium (2) ―――――――――――――――――――――――――――――――――― glucose 80g malt extract (Difco) 20 g yeast extract (Difco) 2 g tryptone _{(Difco) 10g NH 4 NO 3} 1g NaNO 3 1g KH 2 PO 4 1g MgSO 4 · 7H 2 O 1g antifoam * 0.1 ml tap water 1000 ml ( (pH is not adjusted.) ―――――――――――――――――――――――――――――――――― * Nissan Deform CB- 442 (manufactured by NOF Corporation) [Isolation of F-15078A (2)] The obtained culture 37
15 kg of a filter aid (Celite 545: manufactured by Celite Corporation) was added to 0 L, and the mixture was subjected to filter press filtration. Methanol: water = 1: 14
After adding 00 L and stirring, the obtained extract was adjusted to pH 2 by adding 6 N hydrochloric acid. This extract was filtered with a filter press.

Of 465 L of the obtained filtrate, 270 L of Cosmo Seal 140 C18- having a volume of 30 L which had been equilibrated with methanol: 0.05% aqueous trifluoroacetic acid = 1: 1 in advance.
The sample was supplied to an OPN (Nacalai Tesque, Inc.) column.
The column was washed with 270 L of the same solvent, and acetonitrile: 0.05% aqueous trifluoroacetic acid = 4: 6 100 L
, And developed with acetonitrile: 0.05% aqueous trifluoroacetic acid = 6: 4. The first 15 L of the eluate was fractionated as fraction No. 1, and then the eluate was fractionated into 5 fractions (fraction numbers 2 to 6) in 10 L increments.
15078A was recovered.

The remaining 195 L of the above-mentioned filtrate was washed with a 30 L volume of Cosmoseal 140 C18-OPN which had been pre-equilibrated with methanol: 0.05% aqueous trifluoroacetic acid = 1: 1.
(Manufactured by Nacalai Tesque, Inc.), the column was washed with 200 L of the same solvent, and acetonitrile: 0.0
After washing with 5% aqueous trifluoroacetic acid = 4: 6 100 L, acetonitrile: 0.05% aqueous trifluoroacetic acid =
It developed at 6: 4. The first 10 L of the eluate was fractionated as fraction No. 1, then 5 L of the eluate was fractionated as fraction No. 2, then the eluate was fractionated by 15 L into two fractions (fractions 3 and 4). Then, 10 L of the eluate was collected as fraction No. 5, and F-15078A was recovered in fractions 3 to 5 (40 L in total).

Two Cosmoseal 140 C18-OP
A total of 70 L of the fraction containing F-15078A obtained by N (Nakarai Tesque) column fractionation was adjusted to pH 7 with 6N sodium hydroxide, and ethyl acetate 5
The active substance was extracted by adding 0 L. The extract was washed with 50 L of saturated saline, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure to dryness, and 32.3 g of an oily substance was obtained. This oily substance was dissolved in 200 ml of methanol, and 50 ml of the obtained solution was subjected to HPLC shown below.
Was supplied to HPLC according to the condition (2). The remaining 150 ml of the solution was also divided into five 30 ml portions and provided to HPLC under the same conditions. As a result, F-1
A fraction 26L containing 5078A was obtained. 1 for this fraction
0 L of water was added and an additional 10 L of ethyl acetate was added to extract the active. The extract was washed, dehydrated and concentrated to dryness under reduced pressure to obtain 7.78 g of a crude product. 326 mg of this crude product was dissolved in 2 ml of methanol.
This solution was divided into 10 portions of 200 μl each and provided to HPLC according to the following HPLC condition (3). The obtained fraction was concentrated and freeze-dried to obtain F-15.
275 mg of 078A were obtained. HPLC conditions (2) Column: YMC Pack ODS-AM Diameter 100 mm x Length 500 mm (manufactured by YMC) Solvent: Acetonitrile: 1% aqueous triethylamine phosphate (pH 6.0) = 3: 1 Detection wavelength: UV Flow rate: 210 nm Flow rate: 240 ml / min Temperature: room temperature Retention time: 64 minutes HPLC conditions (3) Column: Shodex Asahipak C8P 90 2F 20 mm in diameter × 250 mm in length (manufactured by Showa Denko KK) Solvent: acetonitrile: 10 mM carbonic acid Aqueous ammonium hydrogen solution = 6: 4 Detection wavelength: UV 210 nm Flow rate: 14 ml / min Temperature: room temperature Retention time: 19.2 minutes Production of F-15078B [Culture of SANK13899 strain (3)] SANK138
99 strains were sterilized at 121 ° C. for 30 minutes and contained 500 ml of a seed culture medium having the composition shown in Table 1 of Example 1.
One platinum loop was inoculated into 6 L-volume Erlenmeyer flasks at 23 ° C and 21
The cells were cultured for 6 days under the condition of 0 rpm (rotation radius: 7 cm). The obtained seed culture solution was inoculated with 5% inoculation into two 60 L tank incubators containing 30 L of medium having the same composition, and was incubated for 2 days at 23 ° C. under aeration of 1 vvm, 100 rpm, and dissolved oxygen of 5.0 ppm. Cultured. The obtained culture solution was added to 121
5% was inoculated into one 600 L tank incubator containing 300 L of the main culture medium (2) having the composition shown in Table 3 of Example 2 and sterilized at 30 ° C. for 30 minutes.
The cells were cultured for 7 days under the conditions of 83 to 240 rpm and 5.0 ppm of dissolved oxygen. [Isolation of F-15078B] To 370 L of the obtained culture, 15 kg of a filter aid (Celite 545: manufactured by Celite Corporation) was added, followed by filtration with a filter press. The obtained cells are mixed with methanol: water = 1: 1 400 L
Was added and stirred, and 6N hydrochloric acid was added to the obtained extract to adjust the pH to 2. This extract was filtered with a filter press.

Of 465 L of the obtained filtrate, 270 L of Cosmo Seal 140 C18- having a volume of 30 L which had been pre-equilibrated with methanol: 0.05% aqueous trifluoroacetic acid = 1: 1 was used.
The sample was supplied to an OPN (Nacalai Tesque, Inc.) column.
The column was washed with 270 L of the same solvent, and acetonitrile: 0.05% aqueous trifluoroacetic acid = 4: 6 100 L
, And developed with acetonitrile: 0.05% aqueous trifluoroacetic acid = 6: 4. The first 15 L of the eluate was fractionated as fraction No. 1, and then the eluate was fractionated into 5 fractions (fraction numbers 2 to 6) in 10 L increments.
15078B was recovered.

The remaining 195 L of the above filtrate was used to equilibrate 30 L of Cosmoseal 140 C18-OPN which had been pre-equilibrated with methanol: 0.05% aqueous trifluoroacetic acid = 1: 1.
(Manufactured by Nacalai Tesque, Inc.), the column was washed with 200 L of the same solvent, and acetonitrile: 0.0
After washing with 5% aqueous trifluoroacetic acid = 4: 6 100 L, acetonitrile: 0.05% aqueous trifluoroacetic acid =
It developed at 6: 4. The first 10 L of the eluate was fractionated as fraction No. 1, then 5 L of the eluate was fractionated as fraction No. 2, then the eluate was fractionated by 15 L into two fractions (fractions 3 and 4). Then, 10 L of the eluate was fractionated as fraction No. 5, and F-15078B was recovered in fractions 3 to 5 (40 L in total).

Two Cosmoseal 140 C18-OP
A total of 70 L of the fraction containing F-15078B obtained by N (Nakarai Tesque) column fractionation was adjusted to pH 7 with 6N sodium hydroxide, and ethyl acetate 5
The active substance was extracted by adding 0 L. The extract was washed with 50 L of saturated saline, dehydrated with anhydrous sodium sulfate, concentrated under reduced pressure to dryness, and 32.3 g of an oily substance was obtained. This oily substance was dissolved in 200 ml of methanol, and 50 ml of the resulting solution was used for the HP described in Example 2.
Provided to HPLC according to LC conditions (2). The remaining 150 ml of the solution was also divided into five 30 ml portions and provided to HPLC under the same conditions. As a result, F-
A fraction 26L containing 15078B was obtained. To this fraction was added 10 L of water, and further 10 L of ethyl acetate was added to extract the active substance. The extract was washed, dehydrated and concentrated to dryness under reduced pressure to obtain 7.78 g of a crude product. 2.10 g of the crude product was dissolved in 5 ml of methanol, and 5 g of Cosmosil 140 C18-OPN (manufactured by Nacalai Tesque, Inc.) was added. After distilling off the solvent, the remaining Cosmosil 140 C18-OPN (manufactured by Nacalai Tesque, Inc.) was previously equilibrated with acetonitrile: 0.05% aqueous trifluoroacetic acid = 4: 6 to a 170 ml volume Cosmosil 140. It was overlaid on a C18-OPN (manufactured by Nakarai Tesque Co., Ltd.) column. Use the same solvent 30
After washing with 0 ml, acetonitrile: 0.05% aqueous trifluoroacetic acid = 6: 4 300 ml and acetonitrile: 0.05% aqueous trifluoroacetic acid = 9: 120
When F-150 was collected in fractions of 64 to 72 (total 90 ml), the fractions were collected in fractions of 10 ml each.
78B was recovered. After concentrating the obtained fraction under reduced pressure,
Lyophilization gave 109 mg of a yellow-white powder. 100 mg of this powder was dissolved in 1 ml of methanol, and this solution was divided into 200 μl aliquots five times, and the following HPLC was used.
Was supplied to HPLC according to condition (4). The obtained fraction was concentrated under reduced pressure and freeze-dried to obtain F-1507.
69.5 mg of 8B white powder was obtained. HPLC conditions (4) Column: Shodex Asahipak C8P 90 2F 20 mm in diameter × 250 mm in length (manufactured by Showa Denko KK) Solvent: acetonitrile: 10 mM ammonium bicarbonate water = 6: 4 Detection wavelength: UV 210 nm Flow rate: 14 ml / Temperature: Room temperature Holding time: 17.8 minutes Test example 1. Anti-fungal activity of F-15078A and B The anti-fungal activity of F-15078A and B, ie, the minimum inhibitory concentration measurement for test bacteria, was 0.165M 3-
Using an RPMI1640 medium containing [N-morpholino] propanesulfonic acid (Sigma) buffer, 96
Broth dilution method using a well microtiter plate (Yama
guchi, H., et al., J. Med. Mycol. 36, 61 (1995)). Table 4 shows the measurement results.

[0076]

[Table 4] Antifungal activity of F-15078A and B ------------------------------------------------------------------------- Bacteria test Minimum growth inhibitory concentration (μg / ml) ―――――――――――――――――――― F-15078A F-15078B ―――――――――――― ――――――――――――――――――――――― Candida albicans ATCC90028 2.5> 50 Aspergillus fumigatus IAM2034 1.3> 50 Cryptococcus neoformans IAM4772 0.31 1.56 ―――――――――― As shown in Table 4, F-15078A and B exhibited antifungal activity. Formulation Example 1 Oral Capsules F-15078A 30mg Lactose 170mg Corn Starch 150mg Magnesium Stearate 2mg ――――――――――――――――――――――――― After passing through a 30 mesh sieve, this powder is placed in a gelatin capsule to form a capsule.

[0077]

The novel compound F-150 provided by the present invention
78A and its salts and F-15078B have antifungal activity and are useful for the prevention or treatment of various fungal infections.

[Sequence listing free text] SEQ ID NO: 1 <223> Explanation of unknown species: Houma Esp (Phom
a sp. ) Oligopeptide which is a part of a novel compound F-15078 produced by SANK13899 strain <220> Sequence characteristics <221> Sequence type: abnormal amino acid <222> Characteristic position: 2 <223> Description of characteristics: Xaa represents N-methylleucine. <220> Sequence characteristics <221> Type of sequence: abnormal amino acid <222> Position indicating characteristics: 4 <223> Description of characteristics: Xaa represents N-methylvaline. <220> Sequence characteristics <221> Type of sequence: abnormal amino acid <222> Position indicating characteristics: 6 <223> Description of characteristics: Xaa represents N-methylleucine.

[0078]

[Sequence List] SEQUENCE LISTING <110> Sankyo Co., Ltd. <120> Pharmaceutical composition for treating mycosis comprising F-15078 s, novel compounds. <130> 2001234SW <140> <141> <150> JP 2001-056197 < 151> 2001-03-01 <160> 1 <170> PatentIn Ver. 2.1 <210> 1 <211> 6 <212> PRT <213> Unknown Organism <220> <223> Description of Unknown Organism: An oligopeptide which is a part of novel componuds F-15078s produced by Phoma sp.SANK 13899. <220> <221> PEPTIDE <222> 2 <223> Xaa represents N-Methylleucine. <220> <221> PEPTIDE <222> 4 <223> Xaa represents N-Methylvaline. <220> <221> PEPTIDE <222> 6 <223> Xaa represents N-Methylleucine. <400> 1 Ile Xaa Thr Xaa Ala Xaa 1 5

 ──────────────────────────────────────────────────続 き Continued on the front page (72) Inventor Tatsuya Yano 1-58, Hiromachi, Shinagawa-ku, Tokyo Sankyo Co., Ltd. (72) Inventor Kazunori Tanaka 33 Miyukigaoka, Tsukuba, Ibaraki Prefecture Sankyo Co., Ltd. F-term (reference) 4C084 AA02 AA07 BA01 BA17 BA27 CA05 DA43 NA14 ZB352

Claims (8)

[Claims] (1) The following formula (I): Or a salt thereof as an active ingredient. 2. A medicine containing a compound having the following physicochemical properties or a salt thereof as an active ingredient: 1) Properties of substance: basic fat-soluble powder 2) Molecular formula: C ₅₅ H ₉₈ N ₈ O ₁₄ 3) Molecular weight : 1094 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₅ H ₉₉ N ₈ O)
₁₄ ): Actual value: 1095.7365 Calculated value: 1095.7281 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3434, 3335, 2962, 2937, 2875, 2806, 1750, 16
84, 1641, 1509, 1469, 1412,1371, 1314, 1294, 1271,
1204, 1156, 1128, 1074, 1020 7) Optical rotation: [α] _D ²⁵ -120 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.82 (3H), 0.87 (3H),
0.88 (1H), 0.92 (3H), 0.93 (3H), 0.94 (3H), 0.96 (3H),
0.97 (3H), 0.98 (3H), 1.01 (3H), 1.02 (3H), 1.03 (3H),
1.06 (3H), 1.21 (1H), 1.41 (3H), 1.41 (1H), 1.48 (1H),
1.48 (1H), 1.49 (1H), 1.52 (3H), 1.55 (1H), 1.65 (1H),
1.66 (1H), 1.70 (2H), 1.73 (1H), 1.81 (1H), 1.87 (1H),
2.28 (1H), 2.31 (1H), 2.37 (1H), 2.48 (3H), 2.89 (3H),
2.94 (3H), 2.96 (1H), 3.29 (3H), 3.56 (1H), 4.06 (1H),
4.14 (1H), 4.77 (1H), 4.78 (1H), 4.84 (1H), 4.91 (1H),
4.96 (1H), 5.21 (1H), 5.25 (1H), 5.53 (1H), 6.39 (1H),
7.83 (1H), 7.94 (1H), 8.28 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured in heavy chloroform using tetramethylsilane as an internal standard.
Is as follows. 10.9 (q), 11.9 (q), 15.0 (q), 15.1 (q), 16.0 (q), 16.6
(q), 17.4 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.1 (q), 2
1.0 (q), 21.4 (q), 22.1 (q), 23.1 (q), 23.51 (q), 23.54
(q), 24.2 (t), 24.6 (d), 24.8 (d), 25.4 (d), 25.5 (t),
27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 36.1 (q), 36.5
(t), 37.7 (t), 38.3 (d), 38.4 (d), 39.7 (t), 40.9 (q),
46.2 (d), 51.8 (d), 53.1 (d), 54.7 (d), 55.1 (d), 63.9
(d), 64.7 (d), 68.1 (d), 70.1 (d), 73.4 (d), 74.3 (d),
77.1 (d), 169.03 (s), 169.04 (s), 169.6 (s), 169.8 (s),
169.9 (s), 170.3 (s), 172.0 (s), 173.4 (s), 173.8 (s),
174.0 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter x 250 mm in length: manufactured by Showa Denko KK) Mobile phase: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 10. 20 minutes 11) Solubility: Soluble in dimethyl sulfoxide, methanol and chloroform. 12) Amino acid analysis: Threonine, alanine and isoleucine were found as hydrolysates. 3. A compound represented by the following formula (II): A pharmaceutical comprising the compound represented by the formula (1) as an active ingredient. 4. A medicament containing a compound having the following physicochemical properties as an active ingredient: 1) Properties of substance: neutral fat-soluble colorless powder 2) Molecular formula: C ₅₇ H ₁₀₀ N ₈ O ₁₅ 3) Molecular weight: 1136 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₇ H ₁₀₁ N ₈
O ₁₅ ): Actual value: 1137.7410 Calculated value: 1137.7387 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3433, 3333, 2963, 2937, 2875, 1751, 1686, 16
42, 1516, 1469, 1409, 1388,1372, 1311, 1292, 1272,
1201, 1156, 1128, 1074, 1017 7) Optical rotation: [α] _D ²⁵ -131 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.83 (3H), 0.87 (1H),
0.87 (3H), 0.90 (3H), 0.92 (3H), 0.93 (3H), 0.95 (3H),
0.95 (3H), 0.98 (3H), 0.98 (3H), 1.01 (3H), 1.01 (3H),
1.03 (1H), 1.05 (3H), 1.28 (3H), 1.37 (1H), 1.40 (1H),
1.46 (1H), 1.47 (1H), 1.49 (1H), 1.51 (3H), 1.64 (1H),
1.65 (1H), 1.66 (1H), 1.86 (1H), 1.72 (1H), 1.78 (1H),
2.12 (3H), 2.13 (1H), 2.26 (1H), 2.31 (1H), 2.37 (1H),
2.88 (3H), 2.93 (3H), 2.97 (3H), 3.28 (3H), 3.56 (1H),
4.03 (1H), 4.15 (1H), 4.73 (1H), 4.78 (1H), 4.82 (1H),
4.83 (1H), 4.91 (1H), 4.97 (1H), 5.15 (1H), 5.28 (1H),
5.50 (1H), 6.37 (1H), 6.87 (1H), 7.86 (1H), 8.29 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) tetrachlorosilane as an internal standard in deuterated chloroform The measured nuclear magnetic resonance spectrum (500 MHz) is as shown below. 10.5 (q), 10.9 (q), 14.9 (q), 15.1 (q), 15.6 (q), 16.6
(q), 16.7 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.0 (q), 2
0.8 (q), 21.4 (q), 22.0 (q), 22.1 (q), 23.1 (q), 23.6
(q), 23.6 (q), 24.1 (t), 24.6 (t), 24.7 (d), 24.8 (d),
25.4 (d), 27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 31.6
(d), 31.8 (q), 36.1 (t), 37.6 (t), 38.4 (d), 39.6 (t), 4
0.9 (q), 46.1 (d), 51.8 (d), 53.1 (d), 54.7 (d), 54.7
(d), 61.2 (d), 63.9 (d), 64.6 (d), 68.1 (d), 73.1 (d), 7
4.3 (d), 77.0 (d), 168.9 (s), 168.9 (s), 169.1 (s), 16
9.9 (s), 169.9 (s), 170.3 (s), 170.6 (s), 171.7 (s), 17
2.0 (s), 173.3 (s), 173.8 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter × 250 mm in length: manufactured by Showa Denko KK) Moving bed: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 9. 05 min 11) Solubility: Soluble in dimethyl sulfoxide, methanol, chloroform 12) Amino acid analysis: Threonine, alanine and isoleucine were recognized as hydrolysates. 5. A compound represented by the following formula (I): A therapeutic agent for a fungal infection, comprising a compound represented by the formula (I) or a salt thereof as an active ingredient. 6. A therapeutic agent for fungal infections containing a compound having the following physicochemical properties or a salt thereof as an active ingredient: 1) Substance properties: basic fat-soluble powder 2) Molecular formula: C ₅₅ H ₉₈ N ₈ O ₁₄ 3) Molecular weight: 1094 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₅ H ₉₉ N ₈ O)
₁₄ ): Actual value: 1095.7365 Calculated value: 1095.7281 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3434, 3335, 2962, 2937, 2875, 2806, 1750, 16
84, 1641, 1509, 1469, 1412,1371, 1314, 1294, 1271,
1204, 1156, 1128, 1074, 1020 7) Optical rotation: [α] _D ²⁵ -120 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured with methylsilane as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.82 (3H), 0.87 (3H),
0.88 (1H), 0.92 (3H), 0.93 (3H), 0.94 (3H), 0.96 (3H),
0.97 (3H), 0.98 (3H), 1.01 (3H), 1.02 (3H), 1.03 (3H),
1.06 (3H), 1.21 (1H), 1.41 (3H), 1.41 (1H), 1.48 (1H),
1.48 (1H), 1.49 (1H), 1.52 (3H), 1.55 (1H), 1.65 (1H),
1.66 (1H), 1.70 (2H), 1.73 (1H), 1.81 (1H), 1.87 (1H),
2.28 (1H), 2.31 (1H), 2.37 (1H), 2.48 (3H), 2.89 (3H),
2.94 (3H), 2.96 (1H), 3.29 (3H), 3.56 (1H), 4.06 (1H),
4.14 (1H), 4.77 (1H), 4.78 (1H), 4.84 (1H), 4.91 (1H),
4.96 (1H), 5.21 (1H), 5.25 (1H), 5.53 (1H), 6.39 (1H),
7.83 (1H), 7.94 (1H), 8.28 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) Nuclear magnetic resonance spectrum (500 MHz) measured in heavy chloroform using tetramethylsilane as an internal standard.
Is as follows. 10.9 (q), 11.9 (q), 15.0 (q), 15.1 (q), 16.0 (q), 16.6
(q), 17.4 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.1 (q), 2
1.0 (q), 21.4 (q), 22.1 (q), 23.1 (q), 23.51 (q), 23.54
(q), 24.2 (t), 24.6 (d), 24.8 (d), 25.4 (d), 25.5 (t),
27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 36.1 (q), 36.5
(t), 37.7 (t), 38.3 (d), 38.4 (d), 39.7 (t), 40.9 (q),
46.2 (d), 51.8 (d), 53.1 (d), 54.7 (d), 55.1 (d), 63.9
(d), 64.7 (d), 68.1 (d), 70.1 (d), 73.4 (d), 74.3 (d),
77.1 (d), 169.03 (s), 169.04 (s), 169.6 (s), 169.8 (s),
169.9 (s), 170.3 (s), 172.0 (s), 173.4 (s), 173.8 (s),
174.0 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter x 250 mm in length: manufactured by Showa Denko KK) Mobile phase: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 10. 20 minutes 11) Solubility: Soluble in dimethyl sulfoxide, methanol and chloroform. 12) Amino acid analysis: Threonine, alanine and isoleucine were found as hydrolysates. 7. A compound represented by the following formula (II): A therapeutic agent for fungal infection comprising a compound represented by the formula (1) as an active ingredient. 8. A therapeutic agent for fungal infections containing a compound having the following physicochemical properties as an active ingredient: 1) Properties of substance: neutral fat-soluble colorless powder 2) Molecular formula: C ₅₇ H ₁₀₀ N ₈ O ₁₅ 3 ) Molecular weight: 1136 (FAB-MS method) 4) High resolution FAB-MS [M + H] ⁺ (C ₅₇ H ₁₀₁ N ₈
O ₁₅ ): Actual value: 1137.7410 Calculated value: 1137.7387 5) Ultraviolet absorption spectrum: terminal absorption 6) Infrared absorption spectrum (in potassium bromide tablet, cm
^-1 ): 3433, 3333, 2963, 2937, 2875, 1751, 1686, 1642, 15
16, 1469, 1409, 1388,1372, 1311, 1292, 1272, 1201,
1156, 1128, 1074, 1017 7) Optical rotation: [α] _D ²⁵ -131 ° (c 1.0, methanol) 8) ¹ H-nuclear magnetic resonance spectrum: (δ, ppm) Tetramethylsilane in heavy chloroform Nuclear Magnetic Resonance Spectrum (500 MHz) measured by using as internal standard
Is as follows. 0.78 (3H), 0.79 (3H), 0.80 (3H), 0.83 (3H), 0.87 (1H),
0.87 (3H), 0.90 (3H), 0.92 (3H), 0.93 (3H), 0.95 (3H),
0.95 (3H), 0.98 (3H), 0.98 (3H), 1.01 (3H), 1.01 (3H),
1.03 (1H), 1.05 (3H), 1.28 (3H), 1.37 (1H), 1.40 (1H),
1.46 (1H), 1.47 (1H), 1.49 (1H), 1.51 (3H), 1.64 (1H),
1.65 (1H), 1.66 (1H), 1.86 (1H), 1.72 (1H), 1.78 (1H),
2.12 (3H), 2.13 (1H), 2.26 (1H), 2.31 (1H), 2.37 (1H),
2.88 (3H), 2.93 (3H), 2.97 (3H), 3.28 (3H), 3.56 (1H),
4.03 (1H), 4.15 (1H), 4.73 (1H), 4.78 (1H), 4.82 (1H),
4.83 (1H), 4.91 (1H), 4.97 (1H), 5.15 (1H), 5.28 (1H),
5.50 (1H), 6.37 (1H), 6.87 (1H), 7.86 (1H), 8.29 (1H) 9) ¹³ C-nuclear magnetic resonance spectrum: (δ, ppm) In deuterated chloroform, tetramethylsilane was used as an internal standard. The measured nuclear magnetic resonance spectrum (500 MHz)
It is as follows. 10.5 (q), 10.9 (q), 14.9 (q), 15.1 (q), 15.6 (q), 16.6
(q), 16.7 (q), 18.3 (q), 18.6 (q), 18.7 (q), 19.0 (q), 2
0.8 (q), 21.4 (q), 22.0 (q), 22.1 (q), 23.1 (q), 23.6
(q), 23.6 (q), 24.1 (t), 24.6 (t), 24.7 (d), 24.8 (d),
25.4 (d), 27.7 (d), 29.5 (q), 29.8 (d), 30.2 (q), 31.6
(d), 31.8 (q), 36.1 (t), 37.6 (t), 38.4 (d), 39.6 (t), 4
0.9 (q), 46.1 (d), 51.8 (d), 53.1 (d), 54.7 (d), 54.7
(d), 61.2 (d), 63.9 (d), 64.6 (d), 68.1 (d), 73.1 (d), 7
4.3 (d), 77.0 (d), 168.9 (s), 168.9 (s), 169.1 (s), 16
9.9 (s), 169.9 (s), 170.3 (s), 170.6 (s), 171.7 (s), 17
2.0 (s), 173.3 (s), 173.8 (s) 10) High performance liquid chromatography: Column: Shodex Asahipak C8P
504E (4.6 mm in diameter × 250 mm in length: manufactured by Showa Denko KK) Moving bed: acetonitrile: 10 mM ammonium bicarbonate water = 13: 7 Flow rate: 0.7 ml / min Detection wavelength: λ 210 nm Retention time: 9. 05 min 11) Solubility: Soluble in dimethyl sulfoxide, methanol, chloroform 12) Amino acid analysis: Threonine, alanine and isoleucine were recognized as hydrolysates.