JPH07215989A - New antibiotic substance mi481-42f-a related substance and its production - Google Patents

Abstract

PURPOSE:To obtain a new substance excellent in antimicrobial activities. CONSTITUTION:These antimicrobial substances M1481-42F4-B1 (molecular weight: 1030 and molecular formula: C43H42N12O7S6), M1481-42F4-B2 (molecular weight: 1016 and molecular formula: C42H40N12O7S6) and M1481-42F4-B3 (molecular weight: 1072 and molecular formula: C46H48N12O7S6) are obtained by decomposing a new antibiotic substance M1481-42F4-A. Since, these antimicrobial substances have antimicrobial activities in strongly inhibiting the growth of Gram-positive and Gram-negative bacteria, they are useful as medical antimicrobial agents.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel antibiotic MI481-42F4
The present invention relates to novel antibacterial substances, MI481-42F4-B1, -B2 and -B3, obtained by decomposing -A, and then esterifying if desired, and salts thereof, and a process for producing these substances.

[0002]

BACKGROUND ART A large number of antibiotics produced by microorganisms have been discovered so far, and their decomposition products or derivatives are widely used for the treatment of bacterial infections.

[0003]

In the treatment of bacterial infections, the emergence of resistant bacteria is unavoidable, and new antibacterial antibiotics effective against resistant bacteria are constantly being sought.

[0004]

[Means for Solving the Problems] As a result of the above research, the present inventors have found that the MI481-42F4 strain belonging to the genus Amycolatopsis (F
ERM P-12739), a novel antibiotic MI481-42F4-
A was obtained (see Japanese Patent Application No. 4-142045, filed on May 8, 1992). Then, the antibiotic MI481-42F4-A is hydrolyzed at room temperature in dilute hydrochloric acid or hydrochloric acid-methanol solution to give the antibacterial substance MI481-42F4-A1 (molecular weight 1182, molecular formula C ₅₀ H ₅₀ O ₉ N ₁₄ S ₆ ). And MI481-42F4-A2 (molecular weight
1217) and MI481-42F4-A3 (molecular weight 1199) were obtained (see Japanese Patent Application No. 5-78537, filed on Mar. 15, 1993).
This time, the present inventors decomposed the antibiotic MI481-42F4-A by the method described below, and if desired, then esterified it to obtain novel antibacterial substances MI481-42F4-B1, -B2 and -B3 substances. Was successful.

Therefore, according to the first aspect of the present invention, an antibiotic
MI481-42 New antibacterial substance obtained by decomposing F4-A MI481-
42F4-B1 or a salt thereof is provided.

The first antibacterial substance MI481-42F4-B1 according to the present invention obtained by degrading the antibiotic MI481-42F4-A, for example, methanolysis under acidic conditions has the following physicochemical properties.

(1) Color and shape: colorless crystals (2) Classification of acidic, neutral and basic: basic substance (3) Mass spectrum: FAB-MS m / z 1031 (M + H) ⁺ (4) Molecular formula: C ₄₃ H ₄₂ N ₁₂ O ₇ S ₆ (Molecular weight 1030) The antibacterial substance MI481-42F4-B1 is the antibacterial substance MI481-42F4 described below.
-Corresponds to the methyl ester of B2.

Furthermore, according to the second invention, the antibiotic MI
There is provided a novel antibacterial substance MI481-42F4-B2 or a salt thereof obtained by degrading 481-42F4-A, for example, by acid hydrolysis.

The antibacterial substance MI481-42F4-B2 is a substance having one free carboxyl group.
MI481-42F4-B1 can also be produced by alkaline hydrolysis and then acid treatment, and can also be produced simultaneously with the antibiotic MI481-42F4-A in the culture solution of the MI481-42F4 strain.

Second antibacterial substance MI481-42F4-B according to the present invention
2 has the following physicochemical properties.

(1) Color and shape: colorless powder (2) Classification of acidic, neutral and basic: amphoteric substance (3) Mass spectrum: FAB-MS m / z 1017 (M + H) ⁺ (4) molecular formula : C ₄₂ H ₄₀ N ₁₂ O ₇ S ₆ (molecular weight 1016)

Furthermore, according to the third aspect of the present invention, the antibiotic MI
There is provided a novel antibacterial substance MI481-42F4-B3 obtained by butyl esterification of 481-42F4-B2, or a salt thereof.

The antibacterial substance MI481-42F4-B3 has the following physicochemical properties.

(1) Color and shape: colorless powder (2) Classification of acidic, neutral and basic: basic (3) Mass spectrum: FAB-MS m / z 1073 (M + H) ⁺ (4) Molecular formula : C ₄₆ H ₄₈ N ₁₂ O ₇ S ₆ (Molecular weight 1072) The antibacterial substance MI481-42F4-B3 corresponds to the butyl ester of the antibacterial substance MI481-42F4-B2.

The second antibacterial substance MI481-42F4-B2 of the present invention has one free carboxyl group. Therefore, various groups such as a lower alkyl group such as a methyl group, an ethyl group and a propyl group as well as an ester forming group can be ester-bonded thereto by a known method.

Further, the antibacterial substance MI481-42F4-B2 can be obtained by acid hydrolysis of the above-mentioned antibiotic MI481-42F4-A.
It can also be obtained by acid treatment of MI481-42F4-B1 after alkaline hydrolysis.

After the reaction such as acid hydrolysis, alkali hydrolysis, esterification, etc., an extract using an organic solvent such as alcohol or acetonitrile, adsorption chromatography, gel filtration chromatography, thin layer chromatography, countercurrent partition chromatography, high speed The antibacterial substances MI481-42F4-B1, -B2 and -B3 can be collected purely by appropriately combining or repeating known methods such as liquid chromatography.

Antibacterial substances MI481-42F4-A, -B1, -B2 and -B
The biological properties of 3 are as follows.

Minimum inhibitory concentration of each of the above substances against various bacteria on nutrient agar medium (by agar plate dilution method)
Was measured and the results are shown in Table 1 below.

[0020]

[Table 1]

As is clear from Table 1, the antibacterial substance MI481-42
F4-A, -B1 and -B2 showed strong antibacterial activity against multidrug-resistant bacteria including methicillin-resistant Staphylococcus aureus (MRSA). Also, -B1 and -B2 substances are mycobacterium
It showed strong antibacterial activity against Smegmatis ATCC 607.

Further, the toxicity of the MI481-42F4-B group antibacterial substance provided by the present invention is weak, and LD ₅₀ is
It was over 150 mg / kg.

From the above, the antibacterial substances MI481-42F4-B1 and -B2 of the present invention can be used as therapeutic agents for bacterial infections in humans and animals. Antibacterial substance MI481-42
The physicochemical properties and biological properties of F4-B1, -B2 and -B3 are as described above in detail, and compounds similar to these properties have not been known so far and the antibacterial substance MI48
1-42F4-B1, -B2 and -B3 are new substances.

Novel antibacterial substance MI481-42F4-B1, -B2 of the present invention
And the antibiotic MI used as raw material for the production of -B3
481-42F4-A is a substance having the following characteristics.

(1) Color and shape: colorless powder (2) Melting point: 223-228 ° C (decomposition) (3) Elemental analysis: C 49.46%, H 4.79%, N 16.76%, O
12.23%, S 15.72% (4) Ultraviolet absorption spectrum: Lambda maximum (nm) (in methanol) 203 309 221 350 (shoulder) 245 (shoulder) (in methanol-hydrochloric acid) 205 308 222 353 (shoulder) 250 (shoulder) (In methanol-sodium hydroxide) 205 310 221 353 (shoulder) 250 (shoulder)

(5) Infrared absorption spectrum (KBr method): 330
0, 3120, 2980, 1660, 1550, 1500, 1250, 990, 940, 7
60 cm ^-1 (6) Specific rotation: [α] _D ²⁸ = + 133 ° (c 0.745, dimethyl sulfoxide) (7) Acidic, neutral, basic classification: basic substance (8) molecular weight: 1182.44 ( 9) Molecular formula: C ₅₀ H ₅₁ O ₈ N ₁₅ S ₆

The antibiotic MI481-42F4-A is obtained by culturing a MI481-42F4-A producing bacterium belonging to the genus Amycolatopsis and separating and collecting the antibiotic MI481-42F4-A from the culture. It is obtained by the method for producing the antibiotic MI481-42F4-A, which is characterized by

[0028] MI481- belonging to the genus Amycolatopsis
As an example of 42F4-A-producing bacteria, the present inventors show
Amycolatopsis sp. ( Amycolatopsis sp.) Isolated from a soil sample collected in Tokyo in June 1987
There is MI481-42F4. The mycological properties of this strain are as follows.

Mycological properties of MI481-42F4 strain 1. Well-branched basal hyphae are often zigzag,
In addition, division is recognized. The aerial hyphae are straight or irregularly curved, with fragmentation, forming cylindrical spores. The spore size is about 0.6-0.7 x 0.7-1.2 microns and the surface is smooth. No helix formation, limbus or sporangia are observed.

2. Regarding the description of the growth state color in various media, the standard shown in [] is the Container Harmony Manual (Container Corporation of America).
Color harmony manual).

(1) Sucrose / nitrate agar medium (30
(° C culture) On the development of light yellow [1 ea, Canary Yellow], white aerial hyphae were slightly settled and no soluble pigment was observed. (2) Glucose / asparagine agar medium (30 ° C culture) On the development of yellow [1 1/2 ia, Sunlight Yellow], aerial hyphae of light yellow [1 ca, Pale Yellow] grow on the surface, and the amount of soluble pigment is small. Yellowish. (3) Glycerin-asparagine agar medium (ISP-medium 5, 30 ° C culture) To develop light yellow [1 1/2 ga, Butter Yellow], use aerial hyphae of light yellow [1 1/2 ca, Cream] Once settled, the soluble dyes have a slight yellow tint. (4) Starch / inorganic salt agar medium (ISP-medium 4, 30 ℃ culture) Light yellow [1 1/2 ic, Lt Antique Gold ~ 2gc, Bamboo]
White aerial hyphae grew lightly on the growth of the, and no soluble pigment was observed.

(5) Tyrosine agar medium (ISP-medium 7, 30
℃ culture) Brown [2 ca, Lt Ivory] on the growth of yellow [1 1/2 lc, Gold]
] The aerial hyphae of [] are settled, and the soluble pigment becomes yellowish. (6) Nutrient agar medium (30 ° C culture) On the development of light yellow [2 gc, Bamboo], white aerial hyphae grew slightly and no soluble pigment was observed. (7) Yeast-malt agar medium (ISP-medium 2, culture at 30 ° C.) White aerial hyphae grew on the development of light yellow tea [2 ne, Mastard Gold], and no soluble pigment was observed. (8) Oatmeal-agar medium (ISP-medium 3, 30 ° C culture) White aerial hyphae grew slightly on the growth of light yellow [1 1/2 ic, Lt Antique Gold], and soluble dye Slightly yellowish. (9) Glycerin / nitrate agar medium (30 ° C. culture) White aerial hyphae grow slightly on the development of light yellow [1 ga, Lt Lemon Yellow], and no soluble pigment is observed. (10) Starch agar medium (30 ° C culture) White aerial hyphae grow slightly on the development of yellow [1 la, Lemon Yellow]. No soluble dye is found.

(11) Lime malate agar medium (cultured at 30 ° C.) On the development of light yellow [1 ga, Lt Lemon Yellow], white aerial mycelium slightly grew and no soluble pigment was observed. (12) Cellulose (synthetic solution with filter paper pieces added, cultivated at 30 ° C) Growth develops pale yellow and white aerial mycelium, and no soluble pigment is observed. (13) Gelatin stab culture In a 15% simple gelatin medium (cultured at 20 ° C), growth was pale yellow and white aerial hyphae were faintly settled, and no soluble pigment was observed. Glucose / peptone / gelatin medium (27 ℃
In the case of culture), the growth is pale yellow, aerial hyphae do not grow, and soluble pigments are not observed. (14) Skim milk (37 ℃ and 30 ℃ culture) No growth was observed at 37 ℃ culture. In the case of cultivation at 30 ° C, the growth is pale yellow, aerial hyphae do not grow, and soluble pigments are not observed.

3. Physiological properties (1) Growth temperature range Glucose / asparagine agar medium (glucose 1.0
%, L-asparagine 0.05%, dipotassium phosphate 0.05
%, String agar 3.0%, pH 7.0), 6 ℃, 20 ℃, 24
Tested at each temperature of ℃, 27 ℃, 30 ℃, 37 ℃, 50 ℃,
It grows at any temperature except 6 ° C, 37 ° C, and 50 ° C, and the optimum growth temperature seems to be around 27 ° C to 30 ° C.

(2) Liquefaction of gelatin (15% simple gelatin medium, 20 ° C. culture; glucose / peptone / gelatin medium, 27 ° C. culture) In 15% simple gelatin medium, liquefaction started about 3 days after culturing and about 3 Completed a week later. Glucose peptone
In gelatin medium, a slight liquefaction was observed from the 7th day after culturing, but the progress was extremely slow, and it was not completed in 1 month of culturing.

(3) Hydrolysis of starch (starch / inorganic salt agar medium and starch agar medium, both at 30 ° C.) Water-degradability was observed from about 6 days after culturing, and its action was moderate. It is degree. (4) Coagulation / peptonization of skim milk (skim milk, 37 ° C and 30 ° C culture) No growth was observed at 37 ° C culture. In the case of culturing at 30 ° C, peptone formation begins without coagulation around 10 days after culturing,
It was completed after about one month of culture. (5) Formation of melanin-like pigment (tryptone, yeast,
Broth, ISP-medium 1; peptone-yeast-iron agar medium, ISP-medium 6; tyrosine agar medium, ISP-medium 7; all cultures at 30 ° C) Both media are negative.

(6) Utilization of carbon source (Pridham Godleybe agar medium, ISP-medium 9; 30 ° C. culture) D-glucose, L-arabinose, D-xylose,
It grows using inositol, D-mannitol and lactose, but does not use sucrose. D-fructose, rhamnose and raffinose are likely to be utilized.

(7) Dissolution of lime malate (lime malate agar medium, culturing at 30 ° C.) Solubility was recognized around 10 days after culturing, and its action was moderate. (8) Nitrate reduction reaction (0.1% potassium nitrate-containing peptone water, ISP-medium 8, culture at 30 ° C) Positive. (9) Decomposition of cellulose (synthesis solution with filter paper pieces added, culturing at 30 ° C) No decomposition is observed after 6 months of observation.

In summary of the above properties, the MI481-42F4 strain often exhibits zigzag-like basal hyphae due to its morphology and fragmentation is observed. The aerial mycelium is straight or irregularly curved, with fragmentation, forming cylindrical spores, and its surface is smooth. No helix formation, limbal branch or sporangium is observed. In various media, white to light yellow aerial hyphae grow on the development of light yellow to yellow, and the soluble pigment is slightly yellowish or not observed. The formation of melanin-like pigment is negative, the water-degradability of starch is moderate, the reduction reaction of nitrate is positive, and the proteolytic activity is moderate.

By the way, in the bacterial cell component of the MI481-42F4 strain, 2,6-diaminopimelic acid contained in the cell wall is mesotype, and the reducing sugar in all the bacterial cells is A type containing arabinose and galactose. Also, it contains no mycolic acid, the phospholipid is PII type (containing phosphatidylethanolamine and no phosphatidylcholine and unknown glucosamine-containing phospholipid), and the major menaquinone is MK-9 (H).
-9 (H) was also recognized.

From the above results, the MI481-42F4 strain was identified as Amycolatopsis (Reference, International Jo
urnal of Systematic Bacteriology, 36, 29-37,
1986) considered to belong to the genus. When searching for known bacterial species of the genus Amycolatopsis , Amycolatopsis orientalis (Reference 1, Ibid .; Reference 2, International Journal of Systematic Ba
cteriology, 37, 292-295, 1987) and Amycolatopsis medi terran.
ei , references 1 and 2, ibid.) were mentioned as closely related species. Therefore, the MI481-42F4 strain and the above-mentioned two strains preserved by this institute are currently being compared and examined. Currently MI481-42F4
Amycolatopsis sp.
) MI481-42F4.

The MI481-42F4 strain was applied for deposit to the Institute of Microbial Science and Technology of the Agency of Industrial Science and Technology, and on February 3, 1992, it was entrusted as Microorganism Research Institute No. 12739.

The above-mentioned antibiotic MI481-42F4-A is produced by inoculating the above-mentioned strain into a medium suitable for producing the antibiotic MI481-42F4-A and culturing (see Reference Example 1 described later). The medium may be a nutrient-source-containing medium used for culturing ordinary actinomycetes. As its nutritional source, for example, commercially available peptone, meat extract, corn steep liquor, cottonseed flour, peanut flour, soybean flour, yeast extract, MZ-amine, casein hydrolyzate, sodium nitrate, ammonium nitrate,
Nitrogen sources such as ammonium sulfate and commercially available glycerin, sucrose, starch, glucose, galactose, mannose, molasses and other carbohydrates, or carbon sources such as fat, and inorganics such as salt, phosphate, calcium carbonate and magnesium sulfate. Salt can be used. In addition, if necessary, a trace amount of a metal salt, or an antifoaming agent such as animal / vegetable / mineral oil may be added. Any of these may be used as long as it can be used by the producing strain and is useful for the production of the antibiotic MI481-42F4-A, and all known culture materials for actinomycetes can be used.

Liquid culture is preferred for large-scale production of the antibiotic MI481-42F4-A, and the culture temperature can be applied within a range where the antibiotic MI481-42F4-A can be produced. Cultivation can be performed by applying the above-mentioned conditions and selecting appropriately according to the properties of the MI481-42F4-A-producing bacterium. The antibiotic MI481-42F4-A is mainly present in the cells and can be extracted with an organic solvent such as alcohol, acetonitrile, or acetone that is miscible with water. The cell extract can be extracted with a water-immiscible organic solvent having a relatively high polarity such as butanol. In addition to the above-mentioned extraction methods, adsorption chromatography, gel filtration chromatography, thin layer chromatography, countercurrent partition chromatography, high performance liquid chromatography, etc. may be combined as appropriate or repeated to obtain a pure sample. You can

[0045]

EXAMPLES The present invention will now be described with reference to reference examples and examples, but the present invention is not limited thereto.

Reference Example 1 Production of Antibiotic MI481-42F4-A Glycerol 2.0%, Dextrin (Wako Pure Chemical Industries) 2.0
%, Polypeptone (Wako Pure Chemical Industries) 1.0%, yeast extract (Wako Pure Chemical Industries) 0.3%, ammonium sulfate 0.2%, calcium carbonate 0.2%, antifoaming agent (KM70, Shin-Etsu Chemical) 0.01% (pH 7.4) Place 110 ml in a 500 ml Erlenmeyer flask with waffles, sterilize at 120 ° C for 20 minutes, and then add MI481-42F4 belonging to the genus Amycolatopsis belonging to the genus Amycolatopsis.
One platinum loop of a strain (FERM P-12739) was inoculated, and the culture solution obtained by shaking culture at 27 ° C. for 3 days was used as an inoculum source. 2 ml of this inoculum was transplanted to the same medium as above and cultured at 27 ° C. for 4 days to accumulate the antibiotic MI481-42F4-A.

The culture solution (pH 8.2, 10.1 liter) was centrifuged to collect the cells, and this was extracted with 5 liters of methanol.
Concentrated methanol was distilled off from the extract, 5 liters of water was added, and the mixture was extracted with an equal amount of butanol. After concentration, methanol was added and the precipitate was collected to obtain 4.3 g of a dried product. This was dissolved in dimethylformamide and suspended in dimethylformamide and packed into Sephadex LH-20 (Pharmacia) (outer diameter 50 mm × 100 m
It was put on a column of m) and gel-filtered with dimethylformamide.
Toyopearl HW-40 (tosaw) (outer diameter 40 mm x 420 mm) packed with active fractions (fractions showing antibacterial activity against Bacillus thermophilus ) concentrated under reduced pressure, dissolved in dimethylformamide and suspended in dimethylformamide The mixture was passed through a column No. 3, and the gel was filtered through dimethylformamide, and the active fraction was concentrated and dried under reduced pressure to obtain 780 mg of a dried product.

This was dissolved in 15 ml of chloroform, 25 g of silica gel (Merck Art. 7734) was suspended in this solution and charged into a packed column, which was washed with 200 ml of chloroform and then chloroform / methanol = 10. The active fraction was collected by elution with a mixed solution of / 1, and concentrated under reduced pressure to dryness to obtain 232 mg of pure antibiotic MI481-42F4-A colorless powder.

Example 1 Production of antibacterial substance MI481-42F4-B1 Antibiotic substance MI481-42F4-A obtained in Reference Example 1 5% to 1.0 g
100 ml of anhydrous hydrochloric acid methanol (Tokyo Kasei) was added, and refluxed at 80 ° C. for 2 hours for methanolysis. Thereafter, the reaction solution was concentrated under reduced pressure and dried to remove hydrochloric acid. The powder obtained 1.123
g was dissolved in 5 ml of methanol, 6 g of silica gel (Merck, Art 7734) was added thereto, and the mixture was dried under reduced pressure. Separately, put 25 g of silica gel in chloroform and pack the column (inner diameter 32 mm x height 100 mm) with the above dried silica gel, wash with chloroform-methanol mixture (50: 1, volume ratio), and then chloroform-methanol. Mixed liquid (10:
(1, volume ratio). The eluate was concentrated under reduced pressure, dried and crystallized from a chloroform solution. Pure antimicrobial MI
481-42F4-B1 531 mg was obtained.

Example 2 Production of antibacterial substance MI481-42F4-B2 200 mg of the antibacterial substance MI481-42F4-B1 obtained in Example 1
0.1N sodium hydroxide solution dissolved in 30% methanol 30
ml was added, and the mixture was allowed to stand at room temperature (25 ° C) for 2 hours for alkaline hydrolysis. After that, 150 ml of water and 1N hydrochloric acid were added to the reaction solution.
3.5 ml was added. Add 150 ml of chloroform to this solution,
The mixture was shaken in a separating funnel and extracted into a chloroform layer. Similarly, 100 ml of chloroform was added again for extraction. The extract was dried over anhydrous sodium sulfate, concentrated and dried to 242mg.
Of pale yellow powder was obtained.

The obtained powder was dissolved in 0.5 ml of methanol, 1.0 g of silica gel was added, and the mixture was dried under reduced pressure. Separately prepare silica gel on a column (10 g, inner diameter 28 mm x height 60 mm), wash with chloroform-methanol mixture (10: 1, v / v), and butanol-methanol-water mixture (4: 1:
1, v / v / v). Concentrate the eluate under reduced pressure and dry to 3
Dissolve ml of methanol and use Sephadex LH-20 column.
200 ml (internal diameter 24 mm x height 520 mm) was applied and gel filtration was performed with methanol. Active fractions were collected, concentrated under reduced pressure and dried to obtain 162 mg of antibacterial substance MI481-42F4-B2.

Example 3 Production of antibacterial substance MI481-42F4-B2 400 mg of the antibacterial substance MI481-42F4-A obtained in Reference Example 1 was placed in a techno-vacuum hydrolysis tube and suspended by adding 20 ml of 1N hydrochloric acid. I sealed it. Place this decomposition tube in an oil bath at 110 ° C.
It was left to stand for 24 hours for acid hydrolysis. The reaction solution was concentrated under reduced pressure and dried to obtain 45 mg of a dry solid.

The above dry solid was dissolved in 10 ml of acetonitrile water and purified by high performance liquid chromatography (HPLC). The conditions of the reversed-phase silica gel HPLC used are as follows.

Column: Capsule Pak C
₁₈ , Shiseido), inner diameter 4.6 mm × height 250 mm, eluent: 40% acetonitrile water → 48% acetonitrile water (17 minutes linear gradient), flow rate 1.4 ml / min, detection: UV ₂₅₄ or higher chromatography results The antibacterial substance MI481-42F4-B2 (8 mg) was eluted at a retention time of 11.3 minutes.

Example 4 Production of antibacterial substance MI481-42F4-B3 Antibacterial substance MI481-42F4-B2 obtained in Example 2 1 to 14.9 mg
10 ml of anhydrous 10% hydrochloric acid-butanol was added, and the mixture was refluxed at 86 ° C. for 2.5 hours for butyl esterification. The reaction solution was concentrated under reduced pressure and dried to dryness, the dried product was dissolved in 0.5 ml of methanol, and 1 g of silica gel was added thereto and dried under reduced pressure to obtain a dried solid product.

The dried solid substance was placed on a silica gel column (5 g, inner diameter 28 mm × height 40 mm) prepared separately, washed with a chloroform-methanol mixture (100: 1, v / v), and then the same mixture (50: 1, 50: 1). v / v) was eluted. The eluate was concentrated under reduced pressure and dried to obtain 10.1 mg of the antibacterial substance MI481-42F4-B3.

Example 5 Preparation of antimicrobial substance MI481-42F4-B3 Antimicrobial substance MI481-42F4-A obtained in Reference Example 1
% Anhydrous hydrochloric acid-butanol (2 ml) was added, and the mixture was refluxed at 80 ° C. for 1 hour to perform butanolysis. Concentrate the reaction mixture under reduced pressure and dry to dryness.
4.5 mg of dried product was obtained. This is applied to HPLC (YMC packed column, inner diameter 20 mm × 250 mm, Yamamura Chemical), washed with 70% acetonitrile water, and then eluted with acetonitrile,
2.7 mg of the antibacterial substance MI481-42F4-B3 was obtained.

[0058]

As described above in detail, the present invention provides a novel antibacterial substance and a method for producing the same.

[Brief description of drawings]

FIG. 1 represents the ¹ H-NMR spectrum of the antibacterial substance MI481-42F4-B1 measured in deuterated chloroform at 400 MHz.

FIG. 2 represents the ¹³ C-NMR spectrum of the antibacterial substance MI481-42F4-B1 measured at 100 MHz in deuterated chloroform.

FIG. 3 represents the ¹ H-NMR spectrum of the antimicrobial MI481-42F4-B2 measured at 400 MHz in deuterated dimethylsulfoxide.

FIG. 4 represents the ¹³ C-NMR spectrum of the antimicrobial MI481-42F4-B2 measured in deuterated dimethyl sulfoxide at 100 MHz.

FIG. 5 represents the ¹ H-NMR spectrum of the antimicrobial MI481-42F4-B3 measured in deuterated dimethylsulfoxide at 500 MHz.

FIG. 6 represents the ¹³ C-NMR spectrum of the antimicrobial MI481-42F4-B3 measured in deuterated dimethylsulfoxide at 125 MHz.

Claims (3)

[Claims] 1. The following characteristics :-( 1) Color and shape: colorless crystals (2) Classification of acidic, neutral and basic: basic substance (3) Mass spectrum: FAB-MS m / z 1031 (M + H) ⁺ (4) Novel antibacterial substance MI481-42F4-B1 characterized by having the molecular formula: C ₄₃ H ₄₂ N ₁₂ O ₇ S _6.
, Or its salt. 2. The following characteristics :-( 1) Color and shape: colorless powder (2) Classification of acidic, neutral and basic: amphoteric substance (3) Mass spectrum: FAB-MS m / z 1017 (M + H) ⁺ (4) A novel antibacterial substance MI481-42F4-B2 characterized by having a molecular formula of C ₄₂ H ₄₀ N ₁₂ O ₇ S _6.
, Or its salt. 3. The following characteristics :-( 1) Color and shape: colorless powder (2) Classification of acidic, neutral and basic: basic substance (3) mass spectrum: FAB-MS m / z 1073 (M + H) ⁺ (4) New antibacterial substance MI481-42F4-B3 characterized by having molecular formula: C ₄₆ H ₄₈ N ₁₂ O ₇ S ₆
, Or its salt.