JPH07184666A - New antibiotics, nf00659a1, a2 b1 and b2, their production and use thereof - Google Patents

Abstract

PURPOSE:To obtain new antibiotics, NF00659A1, A2, B1 and B2 having antitumor activity, insecticidal activity and nematocidal activity. CONSTITUTION:These antibiotics, NF00659A1 A2 B1 and B2 are obtained by culturing Aspergillus sp. NF00659 strain (FERM p-13,834). NF00659A1 is a compound having the molecular formula C36H52O7 and 596 molecular weight, NF00659A2 is a compound having the molecular formula C38H54O7 and 622 molecular weight, F00659B1 is a compound having the molecular formula C36H52O6 and 580 molecular weight and NF00659B2 is a compound having the molecular formula C38H54O6 and 606 molecular weight.

Description

Detailed Description of the Invention

[0001]

The present invention relates to a novel antibiotic NF006.
59A ₁ , A ₂ , B ₁ and B ₂ , a method for producing the same, and an antitumor agent, insecticide or nematicide containing the same as an active ingredient.

[0002]

2. Description of the Related Art Conventionally, adriamycin, actinomycin and the like have been used as antitumor agents. As the insecticide, organic phosphorus agents, carbamate agents and the like are used. Monensin, lasalocid, salinomycin and the like are used as nematicides.

[0003]

The antitumor agents, insecticides, or nematicides that have hitherto been used are insufficient in potency, toxicity, etc., and have problems such as resistance. There is a need for anti-tumor agents, insecticides and nematicides.

[0004]

As a result of various searches for microbial metabolites, the inventors have found that one strain belonging to the genus Aspergillus is a novel antibiotic NF00659A ₁ ,
It has been found that it produces A ₂ , B ₁ and B ₂ and that the antibiotic has excellent antitumor, insecticidal and nematicidal activity. The present invention has been completed based on the above findings. That is, the present invention is a novel antibiotic NF0065.
The present invention relates to an antitumor agent, an insecticide and a nematicide containing 9A ₁ , A ₂ , B ₁ and B ₂ as active ingredients.

The above novel antibiotics NF00659A ₁ , A
₂ , B ₁ and B ₂ are NF0 belonging to the genus Aspergillus
A new antibiotic NF00659 was cultured by culturing a 0659-producing bacterium.
A _1, A _2, B ₁ and B ₂ to afford storage, new antibiotics NF00659A _1, A ₂ from the culture, B ₁
And B ₂ are obtained. A typical example of a bacterium producing the novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B ₂ has the following mycological properties.

Mycological properties The potato-glucose agar medium (25 ° C.) grows extremely well, and the diameter of the colony reaches 51 to 52 mm after 7 days.
On the surface of the village, aerial mycelium develops, becomes fluffy, and rises up concentrically. Depending on the number of days of culturing, the color is white to yellow, and the reverse side is cream to pale yellowish brown.

The malt extract agar medium (25 ° C.) grows well, and the colony diameter reaches 49 to 50 mm after 7 days. Aerobic hyphae develop on the surface of the village and become fluffy and swell.
Depending on the number of days of culturing, the color is white to yellow, and the reverse side is cream to pale yellowish brown.

The Tuapeck agar medium (25 ° C.) does not grow well, and after 7 days, the diameter of the colony reaches 22 to 23 mm. The surface of the settlement becomes fluffy. Depending on the number of days of culturing, the color is white to yellow, and the reverse side is cream to pale yellowish brown.

Czapek Dock yeast extract agar medium (25 ° C.) grew well, and the diameter of the colony was 56 after 7 days.
Reach ~ 57 mm. On the surface of the village, aerial mycelium develops, becomes fluffy, and rises up concentrically. Depending on the number of days of culturing, the color is white to yellow and the back side is cream colored. Causes radial wrinkles.

The present bacterium rapidly grows on Czapek Dock yeast extract agar medium and forms a large number of conidia on the entire surface of the colony. Good conidial head formation, length 200-500
μm, diameter 60-100 μm, loose cylindrical shape, colorless to yellow. Conidia peduncle length 1-2 mm, diameter 7-12 μm,
It is colorless to yellowish white in color and has a slightly rough surface. The apex is 10-16 μm in diameter and oval to hemispherical. Metre is 3.4-4.
4 × 1.7 to 2.6 μm. Fear ride is 6.0-7.2 x 1.4
~ 1.7 μm. Conidia diameter 1.7-2.5 μm, spherical, colorless to yellow. The surface is rough. Hule cells are formed in the latter stage of culture. Optimal growth temperature is around 25 ℃, 37 ℃
Does not grow in. The growth pH range is as wide as 2.0 to 8.0, and the optimum pH is around 6.0.

Based on the above-mentioned mycological properties, this bacterium is "Ainsworth
and Bisby's Dictionary of the Fungi "(by DL Ha
wksworth, BC Ainsworth, 7th ed., CMI Kew,
1983), the strain was identified as a genus of Aspergillus (Aspergillus) belonging to the genus Fungi, Subgenus Mycelia, and Aspergillus (Aspergillus sp.) NF.
It was named 00659. The strain has been deposited at the Institute of Biotechnology, Institute of Industrial Science and Technology as FERM P-13834.

The novel antibiotic of the present invention can be advantageously produced using the Aspergillus sp. Strain NF00659 strain. In general, filamentous fungi are naturally or artificially susceptible to mutation in their properties, and the novel antibiotic of the present invention can be produced by using a mutant strain of NF00659 strain, and is not limited to the above. It can also be produced by using various known bacteria belonging to the genus.

The novel antibiotic NF00659 according to the invention
In order to produce A ₁ , A ₂ , B ₁ and B ₂ , Aspergillus sp. Strain NF00659 strain is first aerobically cultured in a medium containing nutrients that can be utilized by filamentous fungi. As the nutrient source, known ones conventionally used for culturing filamentous fungi can be used.For example, as the carbon source, glucose, fructose, glycerin, sucrose, dextrin, galactose, organic acid, etc., alone or in combination. Can be used. Inorganic and organic nitrogen sources include ammonium chloride, ammonium sulfate, urea, ammonium nitrate, sodium nitrate, peptone, meat extract, yeast extract, dry yeast, corn steep liquor, soybean powder, cottonseed oil residue, casamino acid, bactoiton, soluble, vegetable , Protein, oatmeal and the like can be used alone or in combination. In addition, inorganic salts such as sodium chloride, calcium carbonate, magnesium sulfate, copper sulfate, iron sulfate, zinc sulfate, manganese chloride and phosphate can be added as necessary, and organic substances such as amino acids, vitamins, nucleic acids and Inorganic substances can be added appropriately. It is also effective to prevent foaming by adding silicon, vegetable oil or synthetic antifoaming agent to the medium as needed. The most suitable culture method is solid static culture, liquid static culture, liquid shaking culture, particularly deep agitation culture. The culturing temperature is preferably 20 to 45 ° C., and the pH is preferably weakly acidic to slightly alkaline. Usually 1 to 1 in liquid culture
After culturing for 2 days, the new antibiotic NF00659A ₁ ,
A ₂ , B ₁ and B ₂ substances are produced and accumulated in the culture medium. When the production amount in the culture solution reaches the maximum, the culture is stopped, and the desired product is isolated and purified from the bacterial cells.

For the isolation / purification of the substance from the culture medium, a separation / purification method generally used for isolating a microbial metabolite from the culture medium is used. For example, the culture is separated into a filtrate and cells by a filtration method. The cells are extracted with acetone or an organic solvent mixed with water. The obtained organic solvent extraction solution was distilled under reduced pressure to remove the organic solvent, then transferred to a general organic solvent immiscible with water and concentrated under reduced pressure to obtain a novel antibiotic NF00659A ₁ ,
An oily substance containing A ₂ , B ₁ and B ₂ is obtained. The crude substance is a known method usually used for purification of a fat-soluble substance,
For example, it is purified by chromatography using silica gel, alumina, florisil, Sephadex, or the like, or by recrystallization methods alone or in combination. Silica gel is used as a suitable example for purification, and n-hexane-ethyl acetate or n-hexane- is used as an eluent.
A column chromatography method using acetone can be used. By concentrating and drying the active fractions obtained by these methods, the novel antibiotics NF00659A ₁ , A ₂ ,
A colorless powder of B ₁ and B ₂ can be obtained.

NF00659A ₁ , A ₂ , B ₁ and B ₂ which are novel antibiotics having the following physicochemical properties, or salts thereof. a) NF00659A ₁ 1) Appearance; colorless powder 2) melting point; 224-226 ° C. 3) molecular weight; HRFAB-MS m / z 597.37
43 (M + H) ⁺ 4) molecular formula; C ₃₆ H ₅₂ O ₇ 5) solubility; easy solubility: dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.5 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 265 ± 5 nm (ε28,000) 8) Infrared absorption spectrum; The spectrum measured with a potassium bromide tablet is shown in FIG. 9) Hydrogen nuclear magnetic resonance spectrum: The spectrum measured in deuterated DMSO is shown in FIG. 10) Carbon nuclear magnetic resonance spectrum; the spectrum measured in deuterated DMSO is shown in FIG. 11) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, Ninhydrin 12) Distinction between basic, acidic and neutral; Weakly acidic b) NF00659A ₂ 1) Appearance; Colorless powder 2) Melting point: 184-187 ° C 3) Molecular weight; HRFAB-MS m / z 623.39
^{36 (M + H) + 4} ) molecular _{formula; C 38 H 54 O 7 5} ) Solubility; easily dissolved: Dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform-soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.5 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 300 ± 5 nm (ε43,000) 8) Infrared absorption spectrum; The spectrum measured with potassium bromide tablets is shown in FIG. 9) Hydrogen nuclear magnetic resonance spectrum: The spectrum measured in deuterated DMSO is shown in FIG. 10) Carbon nuclear magnetic resonance spectrum; the spectrum measured in deuterated DMSO is shown in FIG. 11) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, ninhydrin 12) Distinction between basic, acidic and neutral; Weakly acidic c) NF00659B ₁ 1) Appearance; Colorless powder 2) Melting point; 139-145 ° C 3) Molecular weight; HRFAB-MS m / z 581.38
34 (M + H) + 4 ) Molecular _{formula; C 36 H 52 O 6 5} ) Solubility; easily dissolved: Dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform-soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.4 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 265 ± 5 nm (ε28,000) 8) Infrared absorption spectrum; The spectrum measured with a potassium bromide tablet is shown in FIG. 7. 9) Hydrogen nuclear magnetic resonance spectrum; FIG. 8 shows the spectrum measured in deuterated DMSO. 10) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, Ninhydrin 11) Distinction between basic, acidic and neutral; Weakly acidic d) NF00659B ₂ 1) Appearance; Colorless powder 2) Melting point; 176-178 ° C 3) Molecular weight; HRFAB-MS m / z 607.39
^{87 (M + H) + 4} ) molecular _{formula; C 38 H 54 O 6 5} ) Solubility; easily dissolved: Dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform-soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.4 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 300 ± 5 nm (ε43,000) 8) Infrared absorption spectrum; The spectrum measured with potassium bromide tablets is shown in FIG. 9. 9) Hydrogen nuclear magnetic resonance spectrum; the spectrum measured in deuterated DMSO is shown in FIG. 10) Carbon nuclear magnetic resonance spectrum; FIG. 11 shows the spectrum measured in deuterated DMSO. 11) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, ninhydrin 12) Distinction between basic, acidic and neutral; Weakly acidic Next, a test example will be shown.

[0016]

Action: Cell growth inhibitory activity Novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B
_The growth inhibitory activity against human colon cancer cells (SW480) and human ovarian cancer cells (A2780) of 2 was examined. The human colon carcinoma cells 5.0 × 10 ⁴ cells / ml 2 or human ovarian cancer cells in a 96-well test plate at a rate of 2 × 10 ⁴ cells / ml,
Inoculate 00 ml and incubate at 37 ° C, 5% CO ₂ incubator for 24 hours.
After culturing for a period of time, the novel antibiotics NF00659A ₁ , A
₂ , B ₁ and B ₂ were added to the culture medium at various concentrations. 72 hours after addition, the viable cell count was measured by the MTT method,
Novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B
The growth inhibition rate of human colon cancer cells or human ovarian cancer cells at various concentrations was determined. The results are shown in Tables 1 and 2.
It was shown to.

[0017]

[Table 1]

[0018]

[Table 2]

As shown in these tables, the novel antibiotic N
F00659A ₁ , A ₂ , B ₁ and B ₂ are human colon cancer cells (SW480) and human ovarian cancer cells (A278
It has a strong antiproliferative activity against 0).

Insecticidal activity Novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B
_The insecticidal activity of ₂ mosquito larvae was examined. Novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B ₂ were diluted with dimethylsulfoxide to a predetermined concentration to prepare a reagent. 199.8 ml of well water was put in a plastic container with a diameter of 9 cm, and 20 3-4 instar larvae of Chica squid (Ageo) were released,
0.2 ml of the reagent solution was dropped and applied with a pipette. After 48 hours, the number of live and dead insects was investigated and the mortality rate was calculated. The results are shown in Table 3.
It was shown to.

[0021]

[Table 3]

As shown in this table, the novel antibiotic NF
[00659] A1, A2, B1 and B2 clearly show insecticidal activity against Culex pipiens.

Next, specific pest names to which the insecticide of the present invention can be applied are listed. Hemiptera (Hemiptera), such as leafhopper (Nephotettix cincti-ceps), brown planthopper (Nilaparvata lugens), Lepidoptera (Lepidoptera), such as diamondback moth (Plutella xylostella), and Kobunomeiga (C
naphaloc-rocis medinalis), Diptera [Diptera] and the like. However, it is not limited to these.

Nematicidal activity The nematicidal activity of the novel antibiotic NF00659A ₁ against the phagocytic soil self-sustaining nematode (Caenorhabditis elegance var. Bristol) was examined. New antibiotic NF00659A1
Was diluted to a predetermined concentration with dimethyl sulfoxide to give a reagent. 1% agar was dispensed into a 96-well test plate.
After solidification, 10 μl of the reagent solution and 20 μl of nematode suspension were dispensed, and after 3 days, the results of observation with a microscope are shown in Table 4.

[0025]

[Table 4]

As shown in this table, the novel antibiotic NF
00659A ₁ clearly shows the nematicidal activity against nematodes.

Next, specific nematode names to which the nematicide of the present invention can be applied are listed. Haricenchu [Tylenchida] For example, soybean cyst nematode (Heteroderaglycines),
Rice cyst nematode (Heterodera elachista) and the like. However, it is not limited to these.

Acute toxicity The acute toxicity of the novel antibiotic NF00659A _{1 to} mice survived 7 days at 8 and 37.5 mg / Kg, respectively, by intraperitoneal (ip) and oral (po) administration.

[0029]

As described above, the novel antibiotic NF0065 is used.
Since 9A ₁ , A ₂ , B ₁ and B ₂ have anticancer action, insecticidal action and nematicidal action, they can be used as medicines such as anticancer agents, insecticides and nematicides, agricultural chemicals or veterinary drugs.

Various conventionally known methods can be applied to the formulation and administration method when used as a drug. That is, as the administration method, injection, oral administration, rectal administration and the like are possible. The dosage form may be injections, powders, granules, tablets, suppositories and the like.

A novel antibiotic NF00659 was used in the formulation.
As long as it does not adversely affect A ₁ , A ₂ , B ₁ and B ₂ .
Various auxiliaries used for medicine, that is, carriers and other auxiliaries such as stabilizers, preservatives, soothing agents, and emulsifiers can be used as necessary. In the formulation, the contents of the novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B ₂ can be varied over a wide range depending on the formulation form and the like.
In general, new antibiotics NF00659A ₁ , A ₂ , B ₁
And B ₂ 0.01 to 100% (by weight), preferably 0.10 to 70
% (Weight), and the rest consists of carriers and other auxiliaries usually used for medicinal purposes.

Novel antibiotics NF00659A ₁ , A ₂ ,
The dose of B ₁ and B ₂ varies depending on the symptoms and the like, but is about 0.01 to 800 mg per adult per day. When continuous casting is required, it is desirable to reduce the daily usage.

Novel antibiotics NF00659A ₁ , A ₂ ,
When B ₁ and B ₂ are used as pesticides, no special conditions are required for formulation. That is, according to the method generally performed in the agricultural chemicals manufacturing field,
It can be used in any formulation form such as powder, granules, emulsions and microcapsules. In addition, a carrier (diluent) and other auxiliary agents such as an emulsifier and a fixing agent can be used as an auxiliary agent during formulation.

Novel antibiotics NF00659A ₁ , A ₂ ,
The amount of B ₁ and B ₂ used as a pesticide varies depending on the dosage form, the method of application, etc., but usually 10 to 300 g, preferably 15 to 200 g is used as the amount of active ingredient per 10 ares.

Novel antibiotics NF00659A ₁ , A ₂ ,
When B ₁ and B ₂ are used as veterinary drugs, no special conditions are required for formulation. That is,
By the method generally used in the veterinary drug manufacturing field, it can be used in the form of any preparation such as injections, emulsions and microcapsules. In addition, a carrier (diluent) and other auxiliary agents such as a developing agent, an emulsifying agent, and a fixing agent can be used as an auxiliary agent during formulation.

In the formulation, the novel antibiotic NF0065
The content of 9A ₁ , A ₂ , B ₁ and B ₂ can be varied over a wide range depending on the formulation form and the like. Generally, the novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B ₂ are added in an amount of 0.01
% To 100% (weight), preferably 0.10 to 70% (weight), and the rest consists of carriers and other auxiliaries usually used for veterinary medicine.

New antibiotics NF00659A ₁ , A ₂ ,
The dose of B ₁ and B ₂ varies depending on the symptoms and the like, but is about 1 mg to 1 g per animal per day. When continuous casting is required, it is desirable to reduce the daily usage.

[0038]

EXAMPLES Examples of the present invention will be shown below, but these are merely examples and do not limit the present invention in any way, and various modifications are possible.

(1) Fermentation In a 500 ml Erlenmeyer flask for rotary type shaker, glucose 1%, sucrose 2%, adipron (manufactured by Ajinomoto Co.) 1.5%, peptone 0.3%, yeast extract 0.2% , Potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.025%, pronal ST 0.01%,
Iron sulfate heptahydrate 0.00011%, copper sulfate pentahydrate 0.00064%, zinc sulfate heptahydrate 0.00015
% And manganese chloride tetrahydrate 0.00079% medium (pH 6.5) 100 ml was dispensed, and autoclave sterilization was performed at 120 ° C. for 20 minutes. New antibiotic NF0
One platinum loop of strain 0659 (FERM P-13834) was inoculated and shaken at 27 ° C., 220 rpm for 2 days for seed culture.

The main culture was carried out in a 500 ml Erlenmeyer flask for rotary shaker with 1% glucose, 2% sucrose,
Adipron (manufactured by Ajinomoto Co.) 2%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.025%, pronal ST 0.01%, iron sulfate heptahydrate 0.00011%,
Dispense 100 ml of a medium (pH 6.5) containing copper sulfate pentahydrate 0.00064%, zinc sulfate heptahydrate 0.00015%, and manganese chloride tetrahydrate 0.00079% to give 120 2 ml of the above seed culture solution was transferred to a flask sterilized by autoclaving at 20 ° C for 20 minutes, then at 25 ° C and 220 revolutions /
Shaking culture was carried out for 7 days under the condition of minutes. The culture broth was suction-filtered to separate into bacterial cells and filtrate.

(2) Purification 6 liters of acetone-methanol (1: 1) was added to the obtained cells to extract them. The extract was concentrated under reduced pressure,
1.2 l of the obtained concentrated liquid was added to ethyl acetate 2.25.
Extracted twice with liters. The ethyl acetate solution was concentrated under reduced pressure to obtain 11.2 g of a brown oily substance. This was subjected to silica gel chromatography (silica gel 60 g, developing solvent n-hexane-acetone). The active fractions were collected and concentrated under reduced pressure to obtain 956 mg of a solid substance.

The obtained solid substance was further subjected to silica gel chromatography (silica gel 72 g, developing solvent n-hexane-diethyl ether-methanol). Two active fractions were obtained. The active fraction that eluted first was the active section A, and the active fraction that eluted later was the active section B, which were collected and concentrated under reduced pressure. 135m of solid substance from active area A
g, and 291 mg of a solid substance was obtained from each of active areas B.

The solid substance in the active zone A was further subjected to medium pressure liquid chromatography (diaion CHP-20, developing solvent acetonitrile-5 mM ammonium acetate buffer pH 4.0.
), 21 mg of NF00659A ₁ and NF00
9 mg of 659A ₂ was obtained.

The solid substance in the active section B was further subjected to medium pressure liquid chromatography (diaion CHP-20, developing solvent acetonitrile-5 mM ammonium acetate buffer pH 4.0.
), 22 mg of NF00659B ₁ and NF00
27 mg of 659B ₂ was obtained.

[Brief description of drawings]

FIG. 1 shows an infrared absorption spectrum measured with a potassium bromide tablet of NF00659A ₁ .

FIG. 2: NF00659A ₁ in heavy DMSO (400M
The hydrogen nuclear magnetic resonance spectrum measured by (Hz) is shown.

FIG. 3: NF00659A ₁ in heavy DMSO (100M
The carbon nuclear magnetic resonance spectrum measured by (Hz) is shown.

FIG. 4 shows an infrared absorption spectrum measured with a potassium bromide tablet of NF00659A ₂ .

FIG. 5: NF00659A ₂ in heavy DMSO (400M
The hydrogen nuclear magnetic resonance spectrum measured by (Hz) is shown.

FIG. 6: NF00659A ₂ in heavy DMSO (100M
The carbon nuclear magnetic resonance spectrum measured by (Hz) is shown.

FIG. 7 shows an infrared absorption spectrum measured with a potassium bromide tablet of NF00659B ₁ .

FIG. 8: NF00659B ₁ in heavy DMSO (500M
The hydrogen nuclear magnetic resonance spectrum measured by (Hz) is shown.

FIG. 9 shows an infrared absorption spectrum measured with a potassium bromide tablet of NF00659B ₂ .

FIG. 10: NF00659B ₂ in heavy DMSO (500
The hydrogen nuclear magnetic resonance spectrum measured by (MHz) is shown.

FIG. 11: NF00659B ₂ in heavy DMSO (125
The carbon nuclear magnetic resonance spectrum measured by (MHz) is shown.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification number Internal reference number FI Technical indication C07G 11/00 A // (C12P 1/02 C12R 1:66) (72) Inventor Taizo Nakagawa Saitama 4-671-3 Mitsuhashi, Omiya City, Japan

Claims (6)

[Claims] 1. A novel antibiotic N having the following physicochemical properties.
F00659A ₁ , A ₂ , B ₁ and B ₂ , or salts thereof. a) NF00659A ₁ 1) Appearance; colorless powder 2) melting point; 224-226 ° C. 3) molecular weight; HRFAB-MS m / z 597.37
43 (M + H) ⁺ 4) molecular formula; C ₃₆ H ₅₂ O ₇ 5) solubility; easy solubility: dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.5 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 265 ± 5 nm (ε28,000) 8) Infrared absorption spectrum; The spectrum measured with a potassium bromide tablet is shown in FIG. 9) Hydrogen nuclear magnetic resonance spectrum: The spectrum measured in deuterated DMSO is shown in FIG. 10) Carbon nuclear magnetic resonance spectrum; the spectrum measured in deuterated DMSO is shown in FIG. 11) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, Ninhydrin 12) Distinction between basic, acidic and neutral; Weakly acidic b) NF00659A ₂ 1) Appearance; Colorless powder 2) Melting point: 184-187 ° C 3) Molecular weight; HRFAB-MS m / z 623.39
^{36 (M + H) + 4} ) molecular _{formula; C 38 H 54 O 7 5} ) Solubility; easily dissolved: Dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform-soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.5 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 300 ± 5 nm (ε43,000) 8) Infrared absorption spectrum; The spectrum measured with potassium bromide tablets is shown in FIG. 9) Hydrogen nuclear magnetic resonance spectrum: The spectrum measured in deuterated DMSO is shown in FIG. 10) Carbon nuclear magnetic resonance spectrum; the spectrum measured in deuterated DMSO is shown in FIG. 11) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, ninhydrin 12) Distinction between basic, acidic and neutral; Weakly acidic c) NF00659B ₁ 1) Appearance; Colorless powder 2) Melting point; 139-145 ° C 3) Molecular weight; HRFAB-MS m / z 581.38
^{34 (M + H) + 4} ) Molecular _{formula; C 36 H 52 O 6 5} ) Solubility; easily dissolved: Dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform-soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.4 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 265 ± 5 nm (ε28,000) 8) Infrared absorption spectrum; The spectrum measured with a potassium bromide tablet is shown in FIG. 7. 9) Hydrogen nuclear magnetic resonance spectrum; FIG. 8 shows the spectrum measured in deuterated DMSO. 10) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydon Smith, Ninhydrin 11) Distinction between basic, acidic and neutral; Weakly acidic d) NF00659B ₂ 1) Appearance; Colorless powder 2) Melting point; 176-178 ° C 3) Molecular weight; HRFAB-MS m / z 607.39
^{87 (M + H) + 4} ) molecular _{formula; C 38 H 54 O 6 5} ) Solubility; easily dissolved: Dimethyl sulfoxide, dimethylformamide, tetrahydrofuran, chloroform-soluble: methanol, acetone, ethyl acetate, ethyl ether insoluble: water, n-hexane 6) Rf value by silica gel thin layer chromatography;
0.4 with a developing solvent of n-hexane-ethyl acetate (1: 1)
Indicates 0. 7) Ultraviolet absorption spectrum; methanol λmax 300 ± 5 nm (ε43,000) 8) Infrared absorption spectrum; The spectrum measured with potassium bromide tablets is shown in FIG. 9. 9) Hydrogen nuclear magnetic resonance spectrum; the spectrum measured in deuterated DMSO is shown in FIG. 10) Carbon nuclear magnetic resonance spectrum; FIG. 11 shows the spectrum measured in deuterated DMSO. 11) Color reaction; Positive: Sulfuric acid-vanillin reagent Negative: Rydney Smith, ninhydrin 12) Distinction between basic, acidic and neutral; weakly acidic 2. A microorganism belonging to the genus Aspergillus and capable of producing the novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B ₂ of claim 1 is cultured in a medium, and the novel antibiotic NF00659A is added to the culture. _1. A process for producing a novel antibiotic NF00659A ₁ , A ₂ , B ₁ and B ₂ which comprises producing ₁ , accumulating ₁ , A ₂ , B ₁ and B ₂ and collecting them. 3. The novel antibiotic NF00659A according to claim 1.
_An antitumor agent containing ₁ or A ₂ or B ₁ or B ₂ as an active ingredient. 4. The novel antibiotic NF00659A according to claim 1.
An insecticide or nematicide containing ₁ or A ₂ or B ₁ or B ₂ as an active ingredient. 5. A bacterium which has the ability to produce the novel antibiotics NF00659A ₁ , A ₂ , B ₁ and B ₂ of claim 1 belonging to the genus Aspergillus. 6. The producing bacterium of claim 5, which is Aspergillus SP-NF0659 strain.