JPH10330363A - Compound CRPF-2721 - Google Patents

Abstract

(57) [Problem] To provide a novel physiologically active substance having excellent antibacterial activity. SOLUTION: Formula (I) Compound CRPF-2721s represented by

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel piericidin compound CRPF-2721 having an antibacterial activity and a method for producing the same.

[0002]

[Prior art]

 Formula (II)

[0003]

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[0004] Piericidin A [Agricultural and biological chemistry (Agricu
ltural and Biological Chemistry, 27, 576 (1963)] and the formula (III)

[0005]

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[0006] Glucopyerisidine A [Journal of Antibotic
iotics), Vol. 40, p. 149 (1987)]. Piericidin A is reported as a substance having an insecticidal action, and glucopielicidin A is reported as a substance that suppresses antibody production in mouse spleen cultured cells.

[0007]

An object of the present invention is to provide a novel physiologically active substance having antibacterial activity and a method for producing the same.

[0008]

According to the invention, the formula (I)

[0009]

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(Wherein, R ¹ represents a hydroxyl group, β-glucose residue or 6-O-acetyl-β-glucose residue, R ² represents hydrogen or a methyl group, and R ³ represents a methyl group or an ethyl group. Provided that when R ¹ is a hydroxyl group or a β-glucose residue, at least one of R ² and R ³ represents a group other than a methyl group.) Or a pharmacologically acceptable compound represented by the formula: A salt is provided. Further, a compound CRPF-characterized in that it belongs to the genus Streptomyces and is cultivated in a medium having a capacity to produce the compound CRPF-2721, and the compound CRPF-2721 is collected from the culture.
A method of manufacturing 2721 is provided.

[0011]

BEST MODE FOR CARRYING OUT THE INVENTION The present invention is described in detail below.
I will tell. Specific examples of the compound represented by the formula (I) include:
R¹Is a hydroxyl group, R²Is hydrogen, R ³Is represented by a methyl group
Compound (hereinafter referred to as CRPF-2721-3), R¹Is 6-O-A
In the case of a cetyl-β-glucose residue, R², R³Together
A compound represented by a methyl group (hereinafter referred to as CRPF-2721-5
U), R¹Is a β-glucose residue, R²Is methyl
Group, R³Is a compound represented by an ethyl group (hereinafter referred to as CRPF-272
1-6) is an example. Also, these compounds
Are collectively referred to as CRPF-2721. Here, β-glucose
And 6-O-acetyl-β-glucose residues are
It shows a group obtained by removing hydrogen from the hydroxyl group at the 1-position of Lucose.

The pharmacologically acceptable salts of the compounds of the CRPF-2721 class include pharmaceutically acceptable acid addition salts, metal salts, ammonium salts, organic amine addition salts, amino acid addition salts and the like. . Examples of the acid addition salt include inorganic acid salts such as hydrochloride, sulfate and phosphate, and organic acid salts such as acetate, maleate, fumarate, tartrate, citrate and lactate, and metal salts. Examples of the salt include alkali metal salts such as lithium salt, sodium salt and potassium salt, alkaline earth metal salts such as magnesium salt and calcium salt, aluminum salt and zinc salt, and ammonium salt includes ammonium,
Examples thereof include salts such as tetramethylammonium, examples of organic amine addition salts include addition salts such as morpholine and piperidine, and examples of amino acid addition salts include addition salts such as glycine, phenylalanine, aspartic acid and lysine.

The physicochemical properties of the compounds CRPF-2721 are as follows. The physicochemical properties were measured using the following instruments. Specific Rotation: JASCO DIP370 Model Digital Polarimeter Mass Spectrum: JEOL HX110A Mass Spectrometer Infrared Absorption Spectrum: JEOL JIR-RFX3001 Infrared Absorption Spectrometer UV Absorption Spectrum: Shimadzu UV-2200 UV Absorption Spectrometer Nuclear magnetic resonance spectrum: JEOL α400 type nuclear magnetic resonance apparatus

Physicochemical properties of CRPF-2721-3 : pale yellow candy Thin layer chromatography (Rf value): 0.75 Developing solvent: chloroform-methanol = 9: 1 Thin layer: Kieselgel 60 F254 (manufactured by Merck) 5628) Thin layer chromatography (Rf value): 0.61 Developing solvent: hexane-isopropanol = 3: 1 Thin layer: Kieselgel 60 F254 (Art.5628 manufactured by Merck) Thin layer chromatography (Rf value): 0.14 Developing solvent: acetonitrile - water = 7: 3 thin layer: RP-18 F254s (Merck Art.13724) molecular formula: C ₂₄ H ₃₅ NO ₄ molecular weight: 401 FAB-MS: m / z 402 [M + H] + high-resolution FAB- MS: measured value 402.2643 theoretical value 402.2644 (C ₂₄ H ₃₆ NO ₄ ) specific rotation: [α] _D ²³ = + 4.6 ° (c = 0.1, CH ₃ OH) UV spectrum (CH ₃ OH): λ _max nm ( ε) 230 (24,100),
240 (20,100), 268 (3,900) IR spectrum (KBr): ν _max cm ^-1 3430, 2935, 1585,
1475, 1410, 1190, 1125, 1045, 970 ¹ H-NMR: δ (CDCl ₃ ) 6.15 (1H, br s), 6.08 (1H, d, J = 15.6Hz), 5.70 (1H,
dq, J = 15.4, 6.3Hz), 5.60 (1H, dt, J = 15.6, 6.8Hz),
5.46 (1H, ddd, J = 15.4, 7.8, 1.7Hz), 5.42 (1H, m),
5.21 (1H, d, J = 9.3Hz), 3.95 (3H, s), 3.86 (3H,
s), 3.76 (1H, t, J = 7.8Hz), 3.37 (2H, d, J = 6.8Hz),
2.79 (2H, d, J = 7.1Hz), 2.56 (1H, m), 2.09 (3H, s),
1.77 (3H, d, J = 1.2Hz), 1.75 (3H, d, J = 0.7Hz), 1.7
2 (3H, dd, J = 1.7, 6.3Hz), 0.93 (3H, d, J = 6.8Hz) ¹³ C-NMR: δ (CDCl ₃ ) 154.0 (s), 153.5 (s), 150.9 (s), 135.8 (d), 135.7
(s), 134.8 (s), 132.5 (d), 132.0 (d), 128.7 (d), 12
8.0 (s), 126.7 (d), 122.3 (d), 112.0 (s), 77.4
(d), 60.6 (q), 53.0 (q), 43.1 (t), 39.4 (d), 34.4
(t), 17.8 (q), 17.0 (q), 16.7 (q), 13.1 (q), 10.5
(q)

Physicochemical properties of CRPF-2721-5 Properties: colorless candy thin layer chromatography (Rf value): 0.47 developing solvent: chloroform-methanol = 9: 1 thin layer: Kieselgel 60 F254 (Art.5628 manufactured by Merck & Co., Inc.) ) Thin layer chromatography (Rf value): 0.17 Developing solvent: hexane-isopropanol = 3: 1 Thin layer: Kieselgel 60 F254 (Merck & Co. Art.5628) Thin layer chromatography (Rf value): 0.29 Developing solvent: acetonitrile- Water = 7: 3 Thin layer: RP-18 F254s (Merck Co. Art.13724) Molecular formula: C ₃₃ H ₄₉ NO ₁₀ Molecular weight: 619 FAB-MS: m / z 620 [M + H] ⁺ high resolution FAB-MS : Actual value 620.3420 Theoretical value 620.3434 (C ₃₃ H ₅₀ NO ₁₀ ) [α] _D ²⁴ = + 6.7 ° (c = 0.2, CH ₃ OH) UV spectrum (CH ₃ OH): λ _max nm (ε) 230 (29 , 40
0), 237 (28,800), 266 (2,800) IR spectrum (KBr): ν _max cm ^-1 3420, 2925, 1735,
1585, 1475, 1415, 1250, 1125, 1080, 1045, 975 ¹ H-NMR: δ (CDCl ₃ ) 6.19 (1H, br s), 6.05 (1H, d, J = 15.1 Hz), 5.61 (1H,
dt, J = 15.1, 6.8Hz), 5.40 (1H, q, J = 6.3Hz), 5.39
(1H, dt, J = 1.0, 5.9Hz), 5.23 (1H, d, J = 9.5Hz), 4.3
4 (1H, dd, J = 4.9, 12.0Hz), 4.20 (1H, dd, J = 1.7, 1
2.0Hz), 4.09 (1H, d, J = 7.8Hz), 3.95 (3H, s), 3.86
(3H, s), 3.49 (1H, t, J = 9.0Hz), 3.43 (1H, d, J = 9.8
Hz), 3.39-3.35 (2H, m), 3.37 (2H, d, J = 5.9Hz), 3.2
5 (1H, dd, J = 7.8, 9.0Hz), 2.78 (3H, m), 2.089 (3H,
s), 2.087 (3H, s), 1.78 (3H, d, J = 1.0Hz), 1.74 (3
H, s), 1.63 (3H, dd, J = 0.7, 6.3Hz), 1.61 (3H, d, J
= 1.0Hz), 0.75 (3H, d, J = 6.8Hz) ¹³ C-NMR: δ (CDCl ₃ ) 171.6 (s), 154.5 (s), 153.6 (s), 150.9 (s), 135.2
(d), 135.1 (s), 134.9 (d), 134.8 (s), 134.7 (s), 12
7.9 (s), 127.0 (d), 123.7 (d), 122.3 (d), 112.1
(s), 103.5 (d), 94.8 (d), 76.1 (d), 74.3 (d), 73.9
(d), 70.2 (d), 63.6 (t), 60.7 (q), 53.1 (q), 43.1
(t), 35.2 (d), 34.5 (t), 20.9 (q), 16.9 (q), 16.6
(q), 13.4 (q), 13.2 (q), 10.9 (q), 10.5 (q)

Physicochemical properties of CRPF-2721-6 Properties: colorless candy thin layer chromatography (Rf value): 0.35 Developing solvent: chloroform-methanol = 9: 1 Thin layer: Kieselgel 60 F254 (Art.5628 manufactured by Merck) ) Thin layer chromatography (Rf value): 0.11 Developing solvent: hexane-isopropanol = 3: 1 Thin layer: Kieselgel 60 F254 (Merck Co. Art.5628) Thin layer chromatography (Rf value): 0.35 Developing solvent: acetonitrile- Water = 7: 3 Thin layer: RP-18 F254s (Merck Co. Art.13724) Molecular formula: C ₃₂ H ₄₉ NO ₉ Molecular weight: 591 FAB-MS: m / z 592 [M + H] ⁺ high resolution FAB-MS : Measured value 592.3506 Theoretical value 592.3486 (C ₃₂ H ₅₀ NO ₉ ) [α] _D ²⁴ = -7.2 ° (c = 0.2, CH ₃ OH) UV spectrum (CH ₃ OH): λ _max nm (ε) 231 (35 , 50
0), 238 (34,900), 266 (3,400) IR spectrum (KBr): ν _max cm ^-1 3420, 2960, 1605,
1470, 1410, 1125, 1080, 1045, 1015, 970 ¹ H-NMR: δ (CDCl ₃ ) 6.18 (1H, br s), 6.06 (1H, d, J = 15.6Hz), 5.62 (1H,
dt, J = 15.6, 7.1Hz), 5.40 (1H, td, J = 6.8, 1.2Hz),
5.34 (1H, td, J = 7.1, 1.0Hz), 5.23 (1H, d, J = 9.5H
z), 4.16 (1H, d, J = 7.8Hz), 3.95 (3H, s), 3.86 (3H,
s), 3.81 (1H, m), 3.67 (1H, m), 3.50 (1H, t, J = 7.
8Hz), 3.47 (1H, t, J = 7.8Hz), 3.42 (1H, d, J = 9.8Hz),
3.37 (2H, d, J = 6.8Hz), 3.29 (1H, m), 3.24 (1H, t,
J = 7.8Hz), 2.78 (3H, m), 2.09 (3H, s), 2.04 (2H,
m), 1.79 (3H, d, J = 1.2Hz), 1.74 (3H, s), 1.62 (3H,
s), 0.97 (3H, t, J = 7.6Hz), 0.77 (3H, d, J = 6.8Hz) ¹³ C-NMR: δ (CDCl ₃ ) 154.0 (s), 153.5 (s), 150.9 (s) , 135.2 (d), 134.8
(s), 134.72 (d), 134.67 (s), 134.0 (s), 131.5 (d),
127.9 (s), 127.1 (d), 122.3 (d), 112.0 (s), 103.9
(d), 94.8 (d), 76.5 (d), 75.3 (d), 74.4 (d), 71.0
(d), 62.9 (t), 60.7 (q), 53.1 (q), 43.1 (t), 35.1
(d), 34.5 (t), 20.9 (t), 16.9 (q), 16.6 (q), 14.0
(q), 13.4 (q), 11.0 (q), 10.5 (q) Next, the biological activity of compounds CRPF-2721 will be described in the following test examples.

Test Example: Antibacterial activity against various bacteria Regarding the minimum growth inhibitory concentration (MIC) against various bacteria, bactotripton (manufactured by Difco) 3 g / L, meat extract 3 g / L, yeast extract 1 g / L, glucose 1 g / L
It was measured by an agar dilution method using a medium (pH 7.0) consisting of L and 16 g / L of agar. Table 1 shows the results.

[0018]

[Table 1]

Next, a method for producing CRPF-2721s will be described. CRPF-2721 belong to the genus Streptomyces, and CRP
A microorganism having the ability to produce F-2721s is cultured in a medium, and CRPF-2721s are produced and accumulated in the culture.
Manufactured by collecting 2721. CRPF-2721
As the microorganism having the ability to produce fungi, any strain that belongs to the genus Streptomyces and has the ability to produce CRPF-2721 can be used. In addition, these strains are subjected to artificial mutation methods such as ultraviolet irradiation, X-ray irradiation,
Mutant strains mutated by treatment with a mutagenic agent or naturally mutated mutant strains can be used in the present invention as long as they have CRPF-2721-producing ability. A particularly suitable example is Streptomyces sp.
( Streptomyces sp. ) CRPF-2721 strain. Streptomyces sp. CRPF-2721
Describe the characteristics of strains in morphology, culture, and physiological properties.

1. Morphological properties 1) Hyphae Aerial mycelium formation; aerial mycelium fragmentation and motility; abasal mycelium fragmentation and motility; no The presence or absence of sporangia formation and the epithelial position; the number of spores linked on the asporeless spore; the shape when 10 or more spores are linked; the characteristic surface structure of straight spores; smooth shape and size ; Rod, about 0.8-1.0μm x 1.8-2.0
μm Motility and presence of flagella; None 3) Others Chlamydospores; Unfocused hyphae; No pseudosporangia; No hyphal division pattern; Simple branching

2. Culture property CRPF-2721 strain shows vigorous growth in commonly used synthetic and natural media, and the basal mycelium shows white to brown color. A brownish soluble pigment may be produced by the medium. The characteristics of growth and color when cultured at 28 ° C. for 28 days on various media are shown below. The color display is Col
or Harmony Manual (Container Corporation of Americ
According to the color classification according to a).

1) Sucrose / nitrate agar medium Growth condition; vigorous color of base mycelium; white (a) Aerial mycelial growth and color; poor, white (a) soluble pigment; no 2) glucose asparagine Agar medium Growing state; Vigorous color of basal hyphae; Natural (2dc) Aerophyte hyphal growth state and its color tone; Soluble dye that hardly grows; Slightly produced (brown) 3) Glycerin-asparagine agar medium Growth state; vigorous Color of basal hyphae: Olive gray (1 ig) Epiphytic state of aerial mycelia and its color tone: Soluble pigment that rarely grows; slightly produced (brown)

4) Starch / inorganic salt agar medium Growth state: vigorous color of basal hyphae; olive gray (1 1/2 ig) Vegetative state of aerial hyphae and its color tone; soluble pigment that rarely grows; production (brown) 5) Tyrosine agar medium Growing condition; vigorous color of base mycelium; light olive drab (1 1/2 li)-dark olive (11/2 nl) Produced in brown (brown) 6) Nutrient agar medium Growth condition; Vigorous color of basal hyphae; Master-Dotan (2lg) Germ state of aerial hyphae and its color tone; Soluble pigment that hardly occurs; Production (brown) 7) Yeast・ Malt agar medium Growth condition; vigorous color of basal hyphae; dark olive (1 1/2 pn) Growth condition of aerial mycelium and its color tone: Soluble pigment that hardly grows; production (brown) 8) Oatmeal agar medium growth condition ; Color of basic hyphae ; Citron gray (1ge) epiphytic state of aerial mycelium and its color; hardly epiphytic soluble dye; slightly production (brown)

3. Physiological Properties The physiological properties of the CRPF-2721 strain are shown below. The results are shown after culturing for 28 days at the growth temperature range and at 28 ^° C for 2-3 weeks for the rest. 1) Growth temperature range; 5-44 ° C 2) Liquefaction of gelatin; Yes 3) Hydrolysis of starch; Yes 4) Coagulation and peptonization of skim milk powder; No 5) Melanin-like pigment formation (1) Peptone yeast Iron agar; No (2) Tyrosine agar; Yes

6) Utilization of carbon source As the basal medium, Pleedham-Gotrieve agar medium was used. Hereinafter, + means to use, and − means not to use. L-arabinose;-D-xylose;-D-glucose; + sucrose;-raffinose;-D-fructose; + rhamnose;-inositol;-D-mannitol;

4. Chemotaxonomic properties 1) Optical isomers of diaminopimelic acid in cells; LL type 2) Main quinone of cell lipids; MK-9 (H ₆ ) and above, aerial hyphae morphologically The formation of spore chains in the
Chemotaxonically, the cell wall is type I (LL-diaminopimelic acid), and the major quinone is menaquinone-9.
Since it is a 3-saturated type (MK-9 (H ₆ )), this strain is classified into the genus Streptomyces among actinomycetes.

Therefore, this strain was designated as Streptomyces sp. CRPF-2721 ( Streptomyces sp. CRPF-2721) and named to the Institute of Biotechnology, Institute of Industrial Science and Technology (1-3, Higashi 1-3, Tsukuba City, Ibaraki Prefecture). Deposited on April 16, 1997 as FERM BP-5918. Compound CRPF-2721 of the present invention
In culturing the bacterium-producing bacterium, a usual culturing method used for culturing actinomycetes is applied. As the medium, any natural medium or synthetic medium can be used as long as it contains a carbon source, a nitrogen source, an inorganic substance, and the like that can be assimilated by bacteria.

Examples of carbon sources include carbohydrates such as glucose, starch, dextrin, mannose and maltose, organic acids such as citric acid, malic acid, acetic acid and fumaric acid, alcohols such as methanol and ethanol, methane, ethane, propane, n. -Hydrocarbons such as paraffin, amino acids such as glutamic acid, and glycerol are used.

As the nitrogen source, ammonium salts such as ammonium chloride, ammonium sulfate, ammonium nitrate and ammonium phosphate, aspartic acid, glutamine,
Amino acids such as cystine and alanine, urea, peptone,
Meat extract, yeast extract, dry yeast, corn steep
Liquor, soluble vegetable protein, soy bean meal, tryptone, cottonseed meal, soybean casein, casamino acid, and Pharmamedia are used.

As the inorganic substance, potassium monohydrogen phosphate,
Potassium dihydrogen phosphate, sodium dihydrogen phosphate, magnesium phosphate, magnesium sulfate, ferrous sulfate, manganese sulfate, copper sulfate, cobalt sulfate, zinc sulfate, calcium pantothenate, ammonium molybdate, potassium aluminum sulfate, barium carbonate. , Calcium carbonate, cobalt chloride, nickel sulfate, salt and the like are used.

In addition, if necessary, a substance such as vitamins such as thiamine that promotes the growth of bacterial cells or the production of CRPF-2721s is added to the medium. When the microorganism used requires a specific substance, the substance is added. Culture is performed at 15 to 35 ° C. by shaking culture method, aeration stirring culture method, etc.
At a pH of around neutral, at a temperature of. Usually, after 3 to 15 days of culture, the maximum accumulation of CRPF-2721s is reached and the culture is completed.

When the CRPF-2721s accumulated in the culture solution are isolated and collected from the culture solution, an ordinary method for collecting microbial metabolites from the culture solution is applied. That is, acetone, extraction of cell components with an organic solvent such as methanol, filtration, cell removal by centrifugation, adsorption resin, silica gel, silanized silica gel, reverse phase silica gel,
CRPF-2721s by adsorption / desorption treatment of active substance by column chromatography or thin layer chromatography using aluminum, cellulose, diatomaceous earth, magnesium silicate, gel filtration agent, ion exchange resin, etc., distribution by an appropriate solvent system, etc. Is isolated.

The detection of CRPF-2721s during the above purification step
Thin-layer chromatography using silica gel containing a fluorescent agent (Kieselgel 60F254, manufactured by Merck) followed by an iodine reaction or an ultraviolet irradiation method at 253.7 nm. Examples of the present invention will be shown below.

[0034]

[Example] Streptomyces sp.
Using CRPF-2721 (FERM BP-5918), 10 g / L of glucose, 10 g / L of soluble starch, 5 g / L of bactotryptone, 5 g / L of yeast extract, 3 g / L of meat extract, 3 g / L of phosphate Potassium hydrogen 1 g / L, magnesium sulfate heptahydrate 0.5 g / L, magnesium phosphate-8
A medium having a composition of hydrate 0.5 g / L (pH 7.0) was used. One loopful of the inoculum was inoculated into 10 ml of a first-type medium placed in a thick test tube, and cultured at 28 ° C. for 5 days with shaking (first-type culture).

5 ml of the first culture broth was inoculated into 50 ml of the second culture medium contained in a 300 ml Erlenmeyer flask. The composition of the second type medium is the same as that of the first type medium. The second seed culture was performed at 28 ° C. for 4 days. The obtained second species culture solution 50
ml was inoculated into 500 ml of the third type medium in a 2 L Erlenmeyer flask. The composition of the third medium is the same as the composition of the first medium. The third seed culture was carried out at 28 ° C. for 4 days.

1.5 L of the obtained third culture was inoculated into 18 L of the main fermentation medium in a 30 L culture tank. As the main fermentation medium, soluble starch 20 g / L, glucose 10 g / L, bactotryptone 5 g / L, meat extract 3 g / L, yeast extract 3 g / L, potassium dihydrogen phosphate 1 g / L, magnesium phosphate.8 Hydrate 0.5g / L
(PH 7.0) was used. This main fermentation culture is 28 ° C., aeration rate 1 vvm, stirring speed 300 rpm
For 3 days with aeration and stirring. A filter aid (Radiolite # 60) is added to 54 L of the obtained main fermentation broth.
0, manufactured by Showa Kagaku Co., Ltd., and then centrifuged (rotation speed: 1,
(800 rpm). The separated culture supernatant was passed through a 2 L Diaion HP-20 column (manufactured by Mitsubishi Chemical Co., Ltd.) packed with deionized water. The column was washed with 6 L of an 80% aqueous methanol solution and eluted with 6 L of methanol.
The eluate was concentrated under reduced pressure and extracted with 1 L of 2-butanol (extract A). 50 L of 2-butanol was added to the cells separated by the above centrifugation, followed by stirring and filtration (extract B). The extract A and extract B were concentrated to dryness under reduced pressure, dissolved in 1 L of a 60% aqueous methanol solution, and filled with a 500 mL Diaion HP-20ss column (manufactured by Mitsubishi Chemical Corporation) using a 60% aqueous methanol solution. Passed through. The column was washed with 1.5 L of an 80% aqueous methanol solution and eluted with 2 L of methanol. The eluate was dried under reduced pressure to obtain 2 g of a brown oily substance. This substance was dissolved in 25 ml of 80% methanol solution, layered on a 1.4 L reversed-phase silica gel column (YMC, ODS-AQS50) packed with a solvent of the same composition, and eluted with a solvent of the same composition. . When 45 ml of the eluate was collected into a test tube, CRPF-2721s were eluted in fraction numbers 40 to 60. As a result of repeating this operation for all amounts,
110 mg of a crude product (1) of CRPF-2721 was obtained. 10 mg of the obtained crude purified product (1) was taken and 0.5 ml of 80
% Methanol solution. This was applied to a reverse phase silica gel column (YMC ODS AQ SH-3235-A).
Q, manufactured by YMC). Preparative HPLC was performed at room temperature, a flow rate of 10 ml / min, a mobile phase of 80% aqueous methanol solution, and the absorbance was detected at 220 nm. As a result, CRPF-2721 was found to be HPL.
C was eluted 24 to 25 minutes after the start of C. After collecting the eluted fractions and drying under reduced pressure, oily CRPF-2721
A crude product (2) was obtained. This operation was repeated for the entire amount of the crude purified product (1) to obtain 41 mg of the crude purified product (2) of CRPF-2721s. The obtained crude product (2) was dissolved in 1 ml of an aqueous 70% acetonitrile solution. Reverse phase silica gel column (YMC ODS AQ
SH-323 5-AQ, manufactured by YMC)
Fractionated by PLC. Preparative HPLC at room temperature, flow rate 10 ml
Per minute, mobile phase 70% acetonitrile aqueous solution, as a result of detection by measuring the absorbance at 220 nm, CRPF-2721-3, -5, and -6 were 32 to 34 minutes and 20 to 22 minutes, respectively. , And 17 to 18 minutes. After collecting the eluate fractions and drying under reduced pressure,
Oily CRPF-2721s were obtained. As a result of repeating the above operation for all 41 mg of the crude purified product (2), the compounds CRPF-2721-3, -5, and -6 were each 10.9.
mg, 6.1 mg and 10.2 mg were obtained.

[0037]

According to the present invention, compounds CRPF-2721 having excellent antibacterial activity can be provided.

────────────────────────────────────────────────── ─── Continuation of front page (51) Int.Cl. ⁶ Identification symbol FI (C12N 1/20 C12R 1: 465) (C12P 17/12 C12R 1: 465) (72) Inventor Satoshi Nakanishi Nakamachi, Machida, Tokyo 3-9-11

Claims (5)

[Claims] 1. Formula (I): (Wherein R ¹ is a hydroxyl group, β-glucose residue or 6-O-
An acetyl-β-glucose residue, R ² represents hydrogen or a methyl group, and R ³ represents a methyl group or an ethyl group. However, when R ¹ is a hydroxyl group or a β-glucose residue, R ^1
At least one of ² and R ³ represents a group other than a methyl group. ) Compound CRPF-2721 or a pharmacologically acceptable salt thereof. 2. The compound according to claim 1, wherein R ¹ is a hydroxyl group, R ² is hydrogen and R ³ is a methyl group. 3. The compound according to claim 1, wherein R ¹ is a 6-O-acetyl-β-glucose residue, and R ² and R ³ are both methyl groups. 4. The compound according to claim 1, wherein R ¹ is a β-glucose residue, R ² is a methyl group, and R ³ is an ethyl group. 5. A compound CR belonging to the genus Streptomyces
The method for producing the compound CRPF-2721 according to claims 1 to 4, wherein a microorganism having the ability to produce PF-2721 is cultured in a medium, and the compound CRPF-2721 is collected from the culture.