JP3055973B2 - Antibiotic aldecarmycin and its production method, and its derivative and its production method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a new antibiotic having antibacterial activity, aldecarmycin and an acetal derivative, and to a method for producing aldecarmycin and an acetal derivative thereof. Further, the present invention relates to Aldeca
The present invention relates to a novel microorganism, Streptomyces strain MJ147-72F6, which has properties capable of producing lumycin. Further, the present invention relates to an antibacterial agent containing aldecarmycin and an acetal derivative thereof as an active ingredient.

[0002]

BACKGROUND OF THE INVENTION In chemotherapy of bacterial infections, the emergence of multidrug-resistant bacteria is a serious problem. It is always desirable to find or create new compounds that have a different chemical structure backbone than previously known or used known antibacterial compounds and that exhibit excellent antibacterial activity and properties.
Research for that purpose is being conducted.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel antibiotic having an antibacterial activity capable of meeting the above demand.

[0004]

Means for Solving the Problems The present inventors conducted research to find a useful new antibiotic, and as a result, succeeded in isolating a strain belonging to the genus Streptomyces as a novel microorganism, They have also found that this strain produces antibiotics with a new structural skeleton. And succeeded in isolating this substance, MJ147-7
It was initially named 2F6 substance. And the name of this substance
Aldecalmycin and now
Times, renamed. The aldehyde duocarmycin has been found to exhibit antimicrobial activity on bacteria and their drug-resistant bacteria Gram-positive (methicillin-resistant bacteria and the like). Furthermore A continued research
Acetal derivative of Le decal mycin {hereinafter of the formula (I)
Of R Make it derivatives when} By By analyzing Al
The chemical structure of decarmycin was determined. And Aldeka
It was confirmed that rumycin was a novel compound and contained a hexapyranose ring, and it was found that it could be represented by the formula (I) described below.

[0005] Thus, the present invention is, A by obtained by the present inventors by culturing a microorganism belonging to the genus Streptomyces
A substance named rudecarmycin and its production method,
And an acetal derivative thereof and a production method thereof. Further, the present invention provides
An antimicrobial agent comprising syn and an acetal derivative thereof as an active ingredient.

That is, according to the first invention, the following formula (I) [Wherein R is an aldehyde group -CHO], or an antibiotic aldecarmycin or a hydrate thereof.

According to the present invention, aldecarmish of formula (I)
Physicochemical properties of the emissions is as follows.

[0008] (1) Appearance White powder (2) Molecular formula _{C 33 H 54 O 9 (3} ) Elemental analysis Found Calculated (as _{C 33 H 54 O 9 · 1/2} H 2 O) C 65.48 65. 64% H 9.29 9.18% O 25.47 25.18% (4) high resolution mass spectrometry (HRFABMS: negative ion mode) Found: 593.3687 (M-H) ^- calcd (C ₃₃ H _{(As 53} O ₉ ): 593.3689 (5) Melting point 125-128 ° C. (6) Specific rotation [α] _D ²⁶ -78.7 ° (c 1.0, methanol) (7) Ultraviolet absorption spectrum As shown in FIG.

(8) Infrared absorption spectrum (KBr tablet method) As shown in FIG. 2 of the accompanying drawings.

(9) Proton nuclear magnetic resonance spectrum The proton NMR spectrum measured at room temperature in heavy methanol at 400 MHz is as shown in FIG. 3 of the accompanying drawings.

(10) Carbon-13 nuclear magnetic resonance spectrum The carbon-13 NMR spectrum measured at room temperature in deuterated methanol at 100 MHz is as shown in FIG. 4 of the accompanying drawings.

(11) Solubility It is soluble in methanol, ethanol, dioxane, and dimethyl sulfoxide, but insoluble in water and hexane.

(12) TLC Silica gel 60F ₂₅₄ (manufactured by Merck) has an Rf value of 0.39 when developed with chloroform-methanol (20: 3) as a developing solvent.

The aldecarmay of the formula (I) of the present invention
The hydrate of syn is an aldehyde group -C as R in the formula (I).
HO is changed to the form of the group -CH (OH) ₂ . This is seen from NMR analysis in aqueous organic solvents.

The following formula (I'-a) The physicochemical properties of the acetal derivative of aldecarmycin represented by are shown.

[0016] (1) Appearance White powder (2) Molecular formula _{C 35 H 58 O 10 (3} ) High resolution mass spectrometry (HRFABMS: negative ion mode) Found: 637.3935 (M-H) ^- calcd (C ₃₅ as _{H 57 O 10): 637.3952 (} 4) mp 113 to 117 ° C. (5) specific rotation ^{_{[α] D 26 -73.6 ° (}} c 1. 0, methanol) (6) ultraviolet absorption spectrum terminus Only absorption.

(7) Infrared absorption spectrum (KBr tablet method) As shown in FIG. 5 of the accompanying drawings.

(8) Proton nuclear magnetic resonance spectrum The proton NMR spectrum measured at room temperature in heavy methanol at 400 MHz is as shown in FIG. 6 of the accompanying drawings.

(9) Carbon-13 nuclear magnetic resonance spectrum A carbon-13 proton NMR spectrum measured at room temperature in heavy methanol at 100 MHz is as shown in FIG. 7 of the accompanying drawings.

(10) Solubility It is soluble in methanol, ethanol, dioxane, and dimethyl sulfoxide, but insoluble in water and hexane.

[0021] (11) chloroform as the developing solvent a thin layer chromatography TLC on silica gel 60F ₂₅₄ (Merck) - methanol (20: 3) Rf value when developed in a 0.46.

Aldeca , a novel antibiotic according to the invention
The fact that lumycin has the chemical structure represented by the above formula (I) is based on the fact that the alkyne represented by the above formula (I'-a)
Proton NMR of acetal derivative of mycin , carbon 1
It was determined as described above by examining the analysis such as 3NMR in detail.

According to the present invention, the aldehyde carmishi of the formula (I)
Aldehyde duocarmycin-acetamide <br/> Lumpur derivatives of emissions and Formula (I'-a) has a later biological properties.

That is, aldecarmycin (I) and its acetal derivative (I'-a) exhibit antibacterial activity against Gram-positive bacteria including drug-resistant bacteria (such as methicillin-resistant bacteria). Aldecarmycin and its acetal derivatives also have low acute toxicity to mammals.

The antimicrobial spectrum of aldehyde duocarmycin for Test Example 1 various microorganisms (I) and its acetal derivatives (I'-a) is based on the Japanese Society of Chemotherapy standard method, by a serial dilution method on Murat Hinton agar It was measured. Table 1 shows the results.

[0026] Table 1 Minimum Inhibitory Concentration (MIC.) (Μg / ml ) aldehyde duocarmycin (I) aldehyde duocarmycin acetal derivative of the trial bacteria (I'-a) Staphylococcus aureus FDA209P 6.25 6. 25 Staphylococcus aureus Smith 12.5 6.25 Staphylococcus aureus (multidrug resistance) MS9610 6.25 6.25 Staphylococcus aureus (methicillin resistance) No. 5 12.5 6.25 Staphylo Coccus aureus (methicillin resistance) No. 17 12.5 6.25 Micrococcus luteus FDA16 12.5 12.5 Micrococcus luteus IFO3333 12.5 12.5 Micrococcus luteus PCI1001 50 12.5 Bacillus anthracis 6.25 6.25 Bacillus subtilis NRRL B-558 6.2 6.25 Bacillus subtilis PCI219 6.25 6.25 Bacillus cereus ATCC10702 6.25 6.25 Corynebacterium bovis 1810 12.5 6.25 Escherichia coli NIHJ>100> 100 Escherichia coli K-12>100> 100 Shigella discenterier JS11910>100> 100 Salmonella typhi T-63>100> 100 Proteus brigaris OX19>100> 100 Serratia marcessens>100> 100 Cyudomonas aeruginosa A3> 50 100 Shiudomonas eruginosa 100 GN Arudekaru Klebsiella Niu Monnier PCI602> 100> 100 Mycobacterium smelling Gumatisu ATCC607 100 12.5 Candida albicans 3147 100 25 test example 2 mice using the formula (I) In testing the leucine and wherein the acute toxicity of the acetal derivative of the aldehyde duocarmycin the (I'-a), dissolved aldehyde duocarmycin or its nitrous acetal derivative 10% dimethyl sulfoxide and a small amount of Tween 80 containing saline The solution was injected into the tail vein and the mice were observed for 14 days. As a result, LD ₅₀ of aldehyde duocarmycin 10
At 0 mg / kg or the acetal derivative of aldecarmycin, no toxicity was observed at 100 mg / kg.

Furthermore, according to the second aspect of the present invention, aldehyde Karma Ishi of the formulas belonging to the genus Streptomyces (I)
Culturing the emissions of producing bacteria, antibiotics Arudeka of formula (I) and recovering the aldehyde duocarmycin from the culture
A method for producing lumycin is provided.

Aldecarmica used in the method of the present invention
If emission-producing bacterium those having the ability to produce antibiotics with physicochemical and biological properties described above,
It can be used regardless of its species and can be selected from a wide range of microorganisms. Among such microorganisms, a specific preferred example of an aldecarmycin- producing bacterium was disclosed by the present inventors in 1989.
This is an actinomycete isolated from soil in Setagaya-ku, Tokyo at the Institute for Microbial Chemistry in the month, and has a strain number of MJ147-72F6.

Hereinafter, various bacteriological properties of the MJ147-72F6 strain will be described.

1. Morphology MJ147-72F6 strain shows 6
Elongate aerial hyphae with ~ 8 turns of helix. Rotating branches and spores are not found. A chain of 50 or more spores is observed at the tip of the aerial hyphae, and the size of the spores is
It was 0.7 × 0.7 to 1.2 microns. The spore surface is smooth.

2. Growing state in various media Regarding the description of the color The standard shown in [] is the color harmony manual (Container Corporation) of Container Corporation of America.
coloration of the nation of America
armonymanual) was used.

(1) Sucrose / nitrate agar medium (2
(Culture at 7 ° C.) On colorless growth, aerial hyphae of yellowish ash [1 1/2 ec, Putty] are slightly formed, and no soluble pigment is observed.

(2) Glucose-asparagine agar medium (cultured at 27 ° C.) The growth is poor, light yellow and aerial hypha do not form, and no soluble pigment is observed.

(3) Glycerin-asparagine agar medium (ISP-medium 5, cultured at 27 ° C.) The growth is bright brown ash [2ge, Covert Tan ~ 2
ig, Slate Tan], aerial mycelium does not form, and the soluble pigment slightly browns.

(4) Starch / inorganic salt agar medium (ISP
-Medium 4, cultivated at 27 ° C) The growth is good, and the light olives [1 1 / 2ie, Lt Ol
ive]-light yellow tea [2ie, Lt Mustard]
Tan ~ 2le, Mustard] and yellowish ash [1
ba, Yellow Tint].
No soluble dye is observed.

(5) Tyrosine agar medium (ISP-medium 7, cultured at 27 ° C.) The growth is light yellow tea [2ie, Lt Mustard Ta].
n] to olive ash [1 li, Lt Olive Dra
b), no aerial mycelia formed and no soluble pigment was observed.

(6) Nutrient agar medium (cultured at 27 ° C.) The growth is poor, and yellow tea [3ni, Close Brow]
n], aerial mycelia did not form and no soluble pigment was observed.

(7) Yeast / malt agar medium (ISP-
Medium 2, culture at 27 ° C.) The growth is light yellow tea [2 ng, Dull Gold to 2 p.
i, Mustard Brown], aerial mycelium does not form,
The soluble pigment is slightly brownish.

(8) Oatmeal / agar medium (ISP-
(Medium 3, culture at 27 ° C.) The growth is colorless, slightly forming aerial hyphae of yellowish ash [2ba, Pearl], and no soluble pigment is observed.

(9) Glycerin / nitrate agar medium (27
The growth is colorless, aerial mycelia do not form, and no soluble pigment is observed.

(10) Starch agar medium (cultured at 27 ° C.) Light yellow tea [2ie, Lt Mustard Tan]
White aerial mycelium develops slightly on the growth of ashy yellow tea [2 pl, Mustard Brown]. No soluble dye is observed.

(11) Lime malate agar medium (cultured at 27 ° C.) Growth is colorless, aerial mycelia do not form, and no soluble pigment is observed.

(12) Cellulose (Synthetic solution containing filter paper pieces,
(Culture at 27 ° C.) Growth is colorless, aerial mycelia do not form, and no soluble pigment is observed.

(13) Gelatin Puncture Culture In a 15% simple gelatin medium (cultured at 20 ° C.), the growth is pale yellow, aerial mycelia do not form, and no soluble pigment is observed.
In the case of the glucose / peptone / gelatin medium, no growth was observed in the culture at 27 ° C, and very slight colorless growth was observed in the culture at 20 ° C, but no aerial hyphae and no soluble pigment were observed.

(14) Skim milk (cultured at 37 ° C.) The growth is colorless to pale yellow, no aerial mycelia form, and no soluble pigment is observed.

3. Physiological properties (1) Growth temperature range Glucose / asparagine agar medium (glucose 1.0
%, Asparagine 0.05%, dipotassium phosphate 0.0
5%, string agar 3.0%, pH 7.0) and starch
Using an inorganic salt agar medium (ISP-medium 4), 6 ° C, 10
° C, 20 ° C, 24 ° C, 27 ° C, 30 ° C, 37 ° C, 50 ° C
As a result of the test at each temperature, growth was observed at any temperature except 50 ° C., and the optimum growth temperature was considered to be around 24 ° C.

(2) Liquefaction of gelatin (15% simple gelatin medium, cultivation at 20 ° C .; glucose / peptone gelatin medium, cultivation at 20 ° C. and 27 ° C.) In a 15% simple gelatin medium, liquefaction starts about 7 days after culturing. However, its effect is moderate. The growth was extremely poor in the glucose / peptone / gelatin medium, and no liquefaction was observed even after 20 days from the culture.

(3) Hydrolysis of starch (starch / inorganic salt agar medium and starch agar medium, both at 27 ° C.)
Cultivation) In any medium, water degradability is observed from about 10 days after culturing, and the action is moderate.

(4) Coagulation and peptone conversion of skim milk (defatted milk, cultivation at 37 ° C.) No coagulation or peptone formation was observed after culturing for 35 days.

(5) Production of melanin-like pigment (trypton yeast broth, ISP-medium 1; peptone yeast iron agar medium, ISP-medium 6; tyrosine agar medium, ISP-medium 7; all cultured at 27 ° C. ) Both media are negative.

(6) Utilization of carbon source (Pridham-Godlieve agar medium, ISP-medium 9); starch / inorganic salt agar medium (ISP-medium 4) without starch;
(Culture at 7 ° C) D-glucose, L-arabinose, D-glucose
-It grows using xylose, rhamnose and starch and does not use sucrose, inositol and raffinose. D-fructose, D-mannitol, lactose are likely to be utilized.

(7) Dissolution of malate lime (maleic acid lime agar medium, cultivation at 27 ° C.) Negative.

(8) Nitrate reduction reaction (peptone water containing 0.1% potassium nitrate, ISP-medium 8, cultured at 27 ° C.) Negative.

(9) Decomposition of Cellulose (Synthetic Solution with Filter Paper Strips, Cultivation at 27 ° C.) No decomposition was observed in the observation for 40 days of culture.

To summarize the above properties, MJ147-7
The 2F6 strain has, in its form, aerial hyphae forming a helix,
Rotating branches and spores are not observed. More than 50 spores are linked to the tip of the aerial hyphae, and the surface is smooth. In various media, growth is colorless or pale yellowish brown, aerial mycelia are less adherent, and no soluble pigment is observed. The optimum growth temperature is around 24 ° C., and growth is observed even at relatively low temperatures. Production of melanin-like pigment-negative, water-degradable starch is moderate, coagulation and peptone of skimmed milk was not observed, the liquefaction of a 15% simple gelatin is moderate. This producing bacterium is isolated at 30 ° C. on a special medium obtained by adding glycerin, yeast extract and inorganic salt to an aloe (Alliaceae, Aloe spp.), And grows best on a medium having that composition. The next best growth is the starch / inorganic salt agar medium (ISP-medium 4), and when the growth is good, the aerial mycelium of yellowish ash grows abundantly. In addition, although not remarkable in the description of the growth state described above, Cover Tan to Slate Tan on glycerin-asparagine agar medium (ISP-medium 5) or the like.
This strain is characterized by a dark color.

The 2,6-diaminopimelic acid contained in the cell wall was of the LL-type.

From these properties, MJ147-72F6
The strain is Streptomyces
It is considered to belong to the genus.

The MJ147-72F6 strain was applied for deposit with the Research Institute of Microbial Industry and Technology of the National Institute of Advanced Industrial Science and Technology, and was deposited on April 9, 1991 as Microkoken Kenkyu No. 12174.

In the second method of the present invention, Aldeca
The production of lumycin is performed as follows.

[0060] In other words, production of aldehyde Cal clarithromycin is A
Rudecarmycin producing bacteria were inoculated into a nutrient medium and
The culture is performed by aerobically shaking at a temperature of ° C. to obtain a culture containing aldecarmycin . As a nutrient medium used for such a purpose, a medium that can be used as a nutrient source for actinomycetes is used. As a nutrient source, for example, commercially available nitrogen sources such as peptone, meat extract, corn steep liquor, peanut flour, soy flour, yeast extract, ammonium sulfate, etc., and glycerin, starch, glucose, galactose, dextrin, etc. Carbon sources such as carbohydrates or fats can be used. Further, inorganic salts such as salt, phosphate and calcium carbonate can be added for use. In addition, a trace amount of a metal salt can be added as needed. These things, A
Le decal mycin producing strain using, as long as it helps in the production of aldehyde duocarmycin <br/>, aldehyde duocarmycin raw <br/> Sankin that can be used all culture materials known actinomycetes may , aldehyde karma Issy belonging to the genus Streptomyces
Microorganism capable of producing emissions are used. In particular,
Our isolated Streptomyces MJ147-
72F6 strain is able to produce the aldehyde duocarmycin has been clarified by the present inventors, but for other strains, can be separated from nature by conventional methods of isolation of the antibiotic-producing bacteria. In addition, Aldecalmy, including Streptomyces strain MJ147-72F6,
Sin- producing bacteria were subjected to irradiation and other mutation treatments,
There is still room for increasing the productivity of aldecarmycin . In addition, Arudekarumai by genetic engineering techniques
Thin production is also possible.

[0061] Aldecarmycin can be produced by aerobically culturing aldecarmycin- producing bacteria belonging to the genus Streptomyces in a suitable medium, and collecting the target substance from the culture. The culture temperature is Aldecal
There is no particular limitation as long as this antibiotic can be produced without substantially inhibiting the growth of the mycin- producing bacterium, and it can be selected according to the producing bacterium to be used. The temperature in the range of can be mentioned.

[0062] As the seed for the production of aldehyde duocarmycin, on an agar medium, using a growth product obtained from slant culture of MJ147-72F6 strain.

[0063] Although the highest in the growth of MJ147-72F6 share is usually 3 to 4 days is reached, continue to generally sufficient antimicrobial activity is imparted to the culture areas. Alde in this culture
The titer of aging of Karumaishin can be measured by a cylindrical plate method to test bacteria Bacillus stearothermophilus.

In the second method of the present invention, aldecarmycin is collected from the culture obtained as described above. The method for collecting aldecarmycin is a method used for collecting metabolites produced by microorganisms. As appropriate. For example, means of extraction with a solvent that does not mix with water,
Aldecarmycin can be collected using a method utilizing the difference in adsorption affinity for various adsorbents, gel filtration, chromatography using countercurrent distribution, or the like, alone or in combination.

The isolated cells can be extracted from the cells by a solvent extraction method using an appropriate organic solvent or an elution method by crushing the cells, isolated and purified in the same manner as described above, and collected. Thus, the above-mentioned antibiotic aldecarmyci
Emissions can be obtained. Al represented by the above formula (I'-a)
Acetal derivative decal clarithromycin is of formula (I)
It is obtained by dissolving decarmycin in dioxane as shown in Example 2 below and reacting it with ethylene glycol as a dihydric alcohol under acidic conditions. From the reaction solution, extraction with an organic solvent immiscible with water, adsorption, chromatography using countercurrent distribution, gel filtration, and the like are combined to obtain a compound of the formula (I'-a).
An acetal derivative of aldecarmycin represented by the following formula was obtained.

On the other hand, in general, the aldehydes of the formula (I)
The aldehyde group (-CHO) of mycin is replaced by the following formula (II)
A monohydric alcohol represented by the following formula, for example, methanol, ethanol, n-propanol, n-butanol, benzyl alcohol, or a dihydric alcohol represented by the following formula (III), such as ethylene glycol, propylene glycol, butylene glycol or catechol, solvents, such as dioxane, under acidic conditions in dimethylformamide, for example, is reacted at room temperature or with heating in the presence of Kanfua sulfonic acid or hydrogen chloride, the aldehyde duocarmycin represented by below formula (I ") The present inventors have found that an acetal derivative can be produced.

Therefore, according to the third aspect of the present invention, the following formula (I ') [Wherein R 'is (Although the two R ^a is optionally in each aliphatic or aromatic group of carbon numbers, for example, a lower alkyl group or an aralkyl group, or, alternatively the two R ^a is an arbitrary linked in particular benzyl group one group of aliphatic or aromatic carbon atoms, an acetal derivative of aldehyde duocarmycin is provided represented by a group] represented by example form a lower alkylene group).

[0068] Furthermore, according to the fourth invention, the aldehyde group of the aldehyde duocarmycin of the formula (I) (-C
HO) is a monohydric alcohol represented by the following formula: ^Rb- OH (II) wherein ^Rb is an aliphatic or aromatic group having an arbitrary carbon number, for example, an alkyl group, or HO- ^Rc -OH (III) wherein R ^c is an aliphatic or aromatic group having an arbitrary number of carbon atoms, for example, a lower alkylene group. [Where R ″ is the following equation Wherein R ^b is as defined above, or
Or: (Where R ^c is as defined above)
A method for producing an acetal derivative of aldecarmycin represented by the formula:

Further, in the fifth invention, the above-mentioned formula (I)
Streptomyces MJ147-72F6 strain is provided having the ability of producing the aldehyde duocarmycin.

[0070] In the present invention the sixth, Al of the formula (I)
Antibacterial agent comprising as an active ingredient <br/> acetal derivative of aldehyde duocarmycin decal mycin or formula (I ') is provided.

In this antibacterial agent, the active ingredient compound can be mixed with a conventional pharmaceutically acceptable solid or liquid carrier such as ethanol, water, starch and the like.

Hereinafter, the present invention will be described in more detail by way of examples.

[0073] Streptomyces were cultured prepared agar slant medium of Example 1 Antibiotic aldehyde duocarmycin MJ147
-72F6 strain (Microtechnical Laboratories No. 12174) was prepared using galactose 2%, dextrin 2%, soypeptone 1%, corn steep liquor 0.5%, ammonium sulfate 0.2%, calcium carbonate 0.2%, A liquid medium containing 1 drop of silicon (adjusted to pH 7.4) was placed in an Erlenmeyer flask (50
0 ml) and dispensed in an amount of 110 ml by a conventional method.
The mixture was inoculated at 20 ° C. for 20 minutes and then cultured with shaking at 27 ° C. for 3 days. Thereby, a seed culture was obtained.

The seed culture was dispensed into an Erlenmeyer flask and sterilized with 2.0% glucose and 3.0% meat extract.
%, Ammonium sulfate 0.6%, magnesium sulfate 0.
3%, dipotassium phosphate 0.6%, calcium carbonate 0.1%
6 liters of liquid medium (pH uncorrected) containing 2% silicon and 1 drop
Was inoculated with a 2% amount and cultured with shaking at 27 ° C. for 4 days.

The culture thus obtained was centrifuged to separate the cells. One liter of methanol was added to the cells, stirred and filtered. After concentrating the methanol under reduced pressure, the filtrate is combined with the centrifuged supernatant to obtain a pH.
After adjusting to 2.0, aldecarmide with 6.5 l of butyl acetate
Syn was extracted. The extract is concentrated under reduced pressure,
The obtained residue (6.4 g) was subjected to silica gel column chromatography (50 g), and eluted with chloroform-methanol (10: 1). The eluted active fractions were collected and concentrated under reduced pressure, and the resulting crude powder was eluted by centrifugal liquid chromatography using a solvent system of chloroform-methanol-water (5: 6: 4).

The active fraction containing aldecarmycin was concentrated under reduced pressure, purified by Sephadex LH-20 column chromatography (500 ml), and developed with methanol. The crude product was developed on a silica gel plate with chloroform-methanol-water (4: 1: 0.1), the active fraction was scraped off the silica gel plate, and extracted with methanol. Next, the residual active substance from which methanol was removed under reduced pressure was subjected to Sephadex LH-20 column chromatography (300 ml) and developed with methanol. The active fraction was concentrated under reduced pressure to give 154 mg of Aldeca.
Lumycin was obtained as a white powder.

To further increase the purity, a part (about 100 mg) of the aldecarmycin obtained here was purified by high-performance liquid chromatography on reversed-phase silica gel.
The solution was developed with a% acetonitrile aqueous solution. Ardekarmashi
Collected and concentrated to a single fraction of emissions, again hexane by centrifugation liquid-liquid chromatography - purified using a solvent system of methanol, a white powder 48.4mg of aldehyde duocarmycin high purity of the formula (I) Obtained.

Example 2 Production of Acetal Derivative of Antibiotic Aldecarmycin The production of an acetal derivative of aldecarmycin was carried out by a known method. That is, Embodiment 1
The aldehyde duocarmycin 58.0mg obtained by 1
The resultant was dissolved in 5.8 ml of a dioxane solution containing camphor sulfonic acid at a concentration of mg / ml, and 145 μl of ethylene glycol as a dihydric alcohol was added thereto, followed by reaction at room temperature for 2 days.

The reaction mixture was neutralized with 20 ml of a saturated aqueous solution of sodium hydrogen carbonate and extracted with 40 ml of ethyl acetate. This extract is dehydrated with anhydrous sodium sulfate,
The residue was dried under reduced pressure to obtain 55.3 mg of a coarse powder. This was subjected to silica gel column chromatography (5 g),
Ethyl acetate - methanol: developed with (15 1), Al de
The fractions containing the acetal derivative of carmycin were collected and concentrated to dryness (24.6 mg).

For further purification, Sephadex LH
The -20 column chromatography (200ml) and eluted with methanol collected single fraction acetal derivative of aldehyde duocarmycin to give a white powder 21.2 mg. Finally, centrifugal liquid chromatography is performed using a hexane-methanol system, and the acetal derivative of aldecarmycin represented by the above formula (I'-a) is used.
9 mg was obtained as a white powder.

[Brief description of the drawings]

FIG. 1 is an ultraviolet absorption spectrum of ardecarmycin in a methanol solution.

Figure 2 is an infrared absorption spectrum measured by KBr tablet method of aldehyde duocarmycin.

FIG. 3 is a proton nuclear magnetic resonance spectrum at 400 MHz measured in a deuterated methanol solution of aldecarmycin at room temperature.

FIG. 4 is a carbon-13 nuclear magnetic resonance spectrum at 100 MHz measured in a deuterated methanol solution of aldecarmycin at room temperature.

Figure 5 is an infrared absorption spectrum measured by KBr tablet method acetal derivatives of aldehyde duocarmycin.

FIG. 6 shows 400 MHz measured at room temperature in a heavy methanol solution of an acetal derivative of aldecarmycin .
3 is a proton nuclear magnetic resonance spectrum of the sample.

FIG. 7 shows 100 MHz measured at room temperature in a deuterated methanol solution of an acetal derivative of aldecarmycin .
13 is a carbon-13 nuclear magnetic resonance spectrum of the sample.

──────────────────────────────────────────────────の Continuation of the front page (51) Int.Cl. ⁷ Identification symbol FI C12R 1: 465) (C12P 19/44 C12R 1: 465) (72) Inventor Ryuichi Sawa 4-6-7-1 Ayanishi, Ayase-shi, Kanagawa Prefecture No. (72) Inventor Masaru Hamada 1-26 Naitocho, Shinjuku-ku, Tokyo (58) Field surveyed (Int.Cl. ⁷ , DB name) C07H 1/00-23/00 A61P 31/04 C12N 1/00 -1/38 C12P 19/00-19/64 BIOSIS (DIALOG) CA (STN) REGISTRY (STN) WPI (DIALOG)

Claims (6)

(57) [Claims] 1. The following formula (I) [Wherein R is an aldehyde group -CHO] an antibiotic aldecarmycin or a hydrate thereof. Wherein culturing the producing strain of aldehyde duocarmycin of formula (I) according to claim 1 belonging to the genus Streptomyces, antibiotics aldehyde duocarmycin for and collecting the aldehyde duocarmycin from the culture Manufacturing method. 3. The following formula (I ′) [Wherein R 'is (However, two R ^a are each an aliphatic or aromatic group having an arbitrary number of carbons, or two R ^a are linked to form an aliphatic or aromatic group having an arbitrary number of carbons. ) Is an acetal derivative of aldecarmycin . 4. Aldeca of the formula (I) according to claim 1.
A monohydric alcohol represented by the following formula R ^b —OH (II) wherein R ^b is an aliphatic or aromatic group having an arbitrary number of carbon atoms, or the following formula HO— Reacting a dihydric alcohol of R ^c —OH (III) wherein R ^c is an aliphatic or aromatic group having any number of carbon atoms.
The following formula (I ″) [Where R ″ is the following equation (Where R ^b has the above-mentioned meaning) or a group represented by the following formula: (Where R ^c is as defined above)
A method for producing an acetal derivative of aldecarmycin represented by the formula : 5. Ardecal of formula (I) according to claim 1.
Streptomyces MJ with mycin- producing properties
147-72F6 strain. 6. A compound represented by the formula (I) according to claim 1.
Antimicrobial agent containing as an active ingredient an acetal derivative of aldehyde duocarmycin of formula (I ') according to aldehyde duocarmycin or claim 2.