JPH08198880A - New antibiotic - Google Patents

Abstract

PURPOSE: To obtain a new antibiotic which is easily degraded in the environment, has a high activity and excellent selective toxicity, and is hence useful as a bactericidal and herbicidal agent for agricultural and gardening purposes, by culturing actinomyces which belongs to the genus Streptomyces and produces antibiotic RS-22.

CONSTITUTION: This antibiotic shown by the formula [R is an unsubstituted or one or more lower alkyl group-substituted guanidino group] is obtained by culturing actinomyces which belongs to the genus Streptomyces and produces antibiotic RS-22 [e.g. Streptomyces sp. RS-22 (FERMP-14441)] followed by isolation/purification of a cultured product. Antibiotic RS-22 is a new macrolide antibiotic which is easily degraded in the environment, hence does not cause environmental pollution, has a high activity and excellent selective toxicity, and is hence useful, for example, as an active ingredient of bactericidal and herbicidal agents for agricultural and gardening purposes.

COPYRIGHT: (C)1996,JPO

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel antibiotic, a method for producing the antibiotic, and its use. More specifically, the present invention relates to a novel macrolide antibiotic, RS-22
A substance, an actinomycete belonging to the genus Streptomyces and producing the antibiotic, a method for producing the antibiotic using the actinomycete,
In addition, the present invention relates to an agricultural and horticultural fungicide / herbicide containing the antibiotic as an active ingredient.

[0002]

2. Description of the Related Art In recent years, environmental pollution caused by pesticides used for agricultural production has become a serious social problem. Therefore, there is an increasing demand for pesticides that are easily degraded in the environment and have excellent selective toxicity. Certain antibiotics made by microorganisms are known to exhibit high utility as pesticides having such desirable properties. Representative examples include blasticidin S, which is used as an epoch-making antibiotic for pesticides to control blast, and polyoxin that has been developed to control sheath blight. However, from the viewpoint of the range of action and the problem of resistant bacteria, it cannot be said that a sufficient type of agricultural and horticultural fungicide / herbicide has been developed, and it has excellent activity as an agricultural / horticultural fungicide / herbicide. The search for new antibiotics with is needed.

[0003]

SUMMARY OF THE INVENTION An object of the present invention is to provide a novel antibiotic which is useful as an agricultural and horticultural fungicide / herbicide. Another object of the present invention is to provide a fungicide for agricultural and horticultural use containing such an antibiotic as an active ingredient. Still another object of the present invention is to provide a novel actinomycete which produces a novel antibiotic useful as an agricultural and horticultural fungicide / herbicide. One of the objects of the present invention is a method for producing the above antibiotics using such actinomycetes.

[0004]

The present inventors have made intensive efforts to solve the above-mentioned problems, and as a result, a novel antibiotic produced by a novel actinomycete belonging to the genus Streptomyces has been disinfected for agricultural and horticultural use. It was found to be extremely useful as an agent. The present invention has been completed based on the above findings.

That is, the present invention provides:

Embedded image (Wherein R represents an unsubstituted or guanidino group substituted by one or more lower alkyl groups). In a preferred embodiment of the present invention, the guanidino group is unsubstituted guanidino group or HN = C (NHCH ₃₎ -NH- or _{CH 3 N = C (NHCH 3} ), -N
An antibiotic RS-22, which is a guanidino group represented by H-, is provided.

According to another aspect of the present invention, there is provided a bacterium producing an RS-22 antibiotic belonging to the genus Streptomyces. As a preferred embodiment, a novel actinomycete RS-22 strain belonging to the genus Streptomyces is provided. According to still another aspect of the present invention, the antibiotic RS-22 producing bacterium belonging to the genus Streptomyces is cultured, and the antibiotic RS-
A method for producing an antibiotic RS-22, characterized in that 22 is collected. As a preferred embodiment, there is provided a production method using the above-mentioned novel actinomycete RS-22 belonging to the genus Streptomyces. Further, the present invention provides a fungicide for agricultural and horticultural use containing the above antibiotic RS-22 as an active ingredient.

The antibiotic RS-22 of the present invention represented by the above formula
In the above, as the guanidino group represented by R 2, an unsubstituted guanidino group as well as a guanidino group substituted by one or more linear or branched lower alkyl groups can be used. As the lower alkyl group, for example, 1-6 carbon atoms
A degree of linear or branched lower alkyl groups can be used. Among these, for example, a methyl group, an ethyl group,
N-propyl, isopropyl, n-butyl, sec-butyl, tert-butyl and the like are preferred lower alkyl groups, and methyl is the most preferred lower alkyl group.

The R-substituents of the preferred compounds encompassed by the antibiotic RS-22 of the present invention are illustrated by way of example.
RS-22 is not limited to these. HN = C (NH ₂ ) -NH
-; HN = C (NHCH ₃ ) -NH-; CH ₃ N = C (NHCH ₃ ) -NH-; HN = C [N (CH ₃ )
₂ ] -NH-; CH ₃ N = C [N (CH ₃ ) ₂ ] -NH-; HN = C (NH ₂ ) -N (CH ₃ )-; HN =
C (NHCH ₃ ) -N (CH ₃ )-; CH ₃ N = C (NHCH ₃ ) -N (CH ₃ )-; HN = C (N (CH
_{3) 2] -N (CH 3} ) -; CH 3 N = C [N (CH 3) 2] -N (CH 3) -; HN = C (NHCH 2 CH
₃ ) -NH-; CH ₃ N = C (NHCH ₂ CH ₃ ) -NH-; HN = C [N (CH ₂ CH ₃ ) ₂ ] -NH
-; CH ₃ N = C [N (CH ₂ CH ₃ ) ₂ ] -NH-; CH ₃ CH ₂ N = C (NHCH ₃ ) -NH-; C
H ₃ CH ₂ N = C [N (CH ₂ CH ₃ ) ₂ ] -NH-;

Of these, the guanidino group is an unsubstituted guanidino group, or HN HC (NHCH ₃ ) —NH— or CH ₃ N = C
Antibiotic RS which is a guanidino group represented by (NHCH ₃ ) -NH-
-22 is a particularly preferred substance. Hereinafter, a substance in which the guanidino group is an unsubstituted guanidino group is referred to as an antibiotic RS-22A,
The substance in which the guanidino group is represented by HN = C (NHCH ₃ ) -NH- is an antibiotic RS-22B, and the substance in which the guanidino group is CH ₃ N = C (NHCH ₃ ) -N
The substance indicated by H- may be called antibiotic RS-22C.

The above antibiotic RS-22 was introduced in China in 1990.
It was isolated and collected from a culture of the actinomycetes RS-22 strain isolated from soil collected in Wuhan. Based on the following biological properties, the actinomycetes RS-22 strain of the present invention were found to be actinomycetes belonging to the genus Streptomyces. In addition, Streptomyces sp.RS-22
Deposited with the National Institute of Advanced Industrial Science and Technology on July 21, 1994 under the accession number FERMP-14441.

Actinomycetes producing the antibiotic RS-22 of the present invention
The RS-22 strain has the following biological properties: Growth temperature: 12.0-42.5 ℃ Morphological features a) Spore stalk: forming spiral b) Spore surface: Warty (wrinkle-wart) c) Aerial hypha color: gray to black d) Base hypha color : Brown soluble pigment: None Chemical taxonomic properties a) Cell wall component: Contains L, L-diaminopimelic acid

Culturing properties: as shown in Table 1 below.

[Table 1] Medium Aerial mycelium Growth Back color Soluble pigment ────────────────────────────────── ISP-2 3cb (sand color) ++ 3lc (brown) − 35fe (gray) +++ 3ge (beige) − 45ih (lead gray) ++ 2ie (brown) − 55ih (lead gray) ++ 2ie (brown) − 6 None + 2gc (light yellow) − 7 3cb (sand) +++ 3le (brown) − ────────────────────────── ────────

Physiological and biochemical properties a) Formation of melanoid pigment: almost none b) Utilization of carbon source D-glucose + L-arabinose + D-xylose ++ i-inositol + D-mannitol + D-fructose + L -Rhamnose + sucrose + raffinose ++ Note) ++ often use + use ± rarely use-do not use

To produce the antibiotic RS-22 of the present invention,
In addition to the above Streptomyces sp. RS-22 strain, an RS-22 producing bacterium belonging to the genus Streptomyces can be used. Among such bacteria, the RS-22 strain is a preferred strain. The antibiotic RS-22-producing bacteria belonging to the genus Streptomyces can be cultured and the antibiotic RS-22 of the present invention can be separated and collected from the culture by a method well known to those skilled in the art. Examples are shown below as examples, but the method for producing the antibiotic RS-22 of the present invention is not limited to these examples. Further, for example, by producing RS-22A, RS-22B, RS-22C or the like according to the examples of the present specification, and performing a chemical conversion known by those skilled in the art to convert the chemical structure, Antibiotic RS-22.

Since the antibiotic RS-22 of the present invention has a bactericidal action and a herbicidal action, the antibiotic RS-22 of the present invention is useful as a fungicide and / or herbicide for agricultural and horticultural use.
When the antibiotic RS-22 of the present invention is used for sterilization for agricultural and horticultural use and / or for herbicidal use, the antibiotic itself may be used, but it is preferably produced using an agricultural chemical adjuvant commonly used in the art. An agricultural and horticultural fungicide and / or herbicide in the form of a prepared composition can be used. The form of the fungicide for agricultural and horticultural use is not particularly limited. For example, an emulsion, a wettable powder, a powder, a flowable, a fine granule, a granule, a tablet, an oil, a spray, an aerosol and the like are preferable. is there. One or more of the antibiotics RS-22 can be incorporated into agricultural and horticultural fungicides and / or herbicides.

The pesticide adjuvant used for producing the above-mentioned agricultural and horticultural fungicide and / or herbicide includes, for example, improvement, stabilization and dispersibility of the agricultural and horticultural fungicide and / or herbicide. It can be used for the purpose of improvement and the like, and for example, a carrier (diluent), a spreading agent, an emulsifier, a wetting agent, a dispersant, a disintegrant and the like can be used.

Examples of the liquid carrier include water, aromatic hydrocarbons such as toluene and xylene, alcohols such as methanol, butanol and glycol, ketones such as acetone, amides such as dimethylformamide, sulfoxides such as dimethylsulfoxide, and the like. Examples include methylnaphthalene, cyclohexane, animal and vegetable oils, fatty acids, fatty acids and the like. Further, as the solid carrier, clay, kaolin, talc, diatomaceous earth, silica, calcium carbonate, montmorillonite, bentonite, feldspar, quartz, alumina, sawdust, nitrocellulose, starch, gum arabic and the like can be used.

As the emulsifier and dispersant, ordinary surfactants can be used. For example, higher alcohol sodium sulfate, stearyltrimethylammonium chloride, polyoxyethylene alkylphenyl ether,
Anionic surfactants such as lauryl betaine, cationic surfactants, nonionic surfactants, amphoteric surfactants and the like can be used. A spreading agent such as polyoxyethylene nonyl phenyl ether and polyoxyethylene lauryl ether; a wet spreading agent such as polyoxyethylene nonyl phenyl ether and dialkyl sulfosuccinate; a fixing agent such as carboxymethyl cellulose and polyvinyl alcohol; lignin sulfonic acid Disintegrants such as sodium and sodium lauryl sulfate can be used.

The content of the active ingredient (antibiotic RS-22 or a salt thereof) in the agricultural and horticultural fungicide and / or herbicide of the present invention can be determined as appropriate according to various conditions such as the form of preparation and the method of application. For example, in the case of emulsion, 0 to 50 parts by weight of antibiotic RS-22 as an active ingredient, 10 to 80 parts by weight of a solvent, and 3 to 20 parts by weight of a surfactant can be blended. . An emulsion of a stock solution can be prepared by mixing a predetermined amount of the active ingredient antibiotic RS-22, a solvent, and a surfactant, and the stock solution is diluted with water to a predetermined concentration before use. Can be.

In the case of a wettable powder, the active ingredient is contained in an amount of 5 to 80 parts by weight,
The solid carrier may be blended in a proportion of 10 to 90 parts by weight and the surfactant in a proportion of 1 to 20 parts by weight. A stock solution is prepared by mixing a predetermined amount of the active ingredient, the solid carrier, and the surfactant, and the stock solution can be diluted with water to a predetermined concentration before use. In the case of a powder, the active ingredient can be mixed and used as it is in a proportion of 1-5 parts by weight and a solid carrier in an amount of 95-99 parts by weight. Granules can be produced by mixing 5 to 15 parts by weight of an active ingredient, 82 to 94 parts by weight of a solid carrier, and 1 to 3 parts by weight of a surfactant and granulating. Such a granule can be applied as it is. However, the mixing ratio of the components in each of the above-mentioned preparation forms is not limited to the above, and can be appropriately selected by those skilled in the art.

The fungicide and / or herbicide for agricultural and horticultural use of the present invention includes, in addition to the antibiotic RS-22 as an active ingredient, fungicides, insecticides, herbicides, fertilizers, soil conditioners, acaricides, and the like. It may contain any active ingredient. The agricultural and horticultural fungicide of the present invention can be applied by any of foliage application and water surface application. The application rate for foliage application is usually 20 to
A 1,000 ppm solution can be about 100-200 liters per 10 ares. In the case of surface application,
Usually, granules containing 5 to 15% of active ingredient
It can be 1 to 10 Kg. As a method of applying the agricultural and horticultural herbicide of the present invention, for example, foliage as a place of use,
Sites such as roots, seeds, seedlings, soil, etc. may be used before or after germination of weeds, for example, in an amount of 0.1 to 100 ppm, preferably 1 to 20 ppm, and as an application method, irrigation.

[0022]

【Example】

Production example glucose 2%, soluble starch 1%, meat extract 0.1%, yeast 0.4%, salt 0.2%, dipotassium phosphate 0.005%,
And 70 ml of medium containing 2.5% soy flour (pH 7.3) 50
Twenty-four 0 ml flasks were inoculated with the RS-22 strain, and cultured with shaking at 28 ° C. for 96 hours at 300 rpm. The culture was centrifuged (6000 rpm) to separate the cells from the supernatant, and the obtained cells were extracted with 80% acetone (1 liter). After concentration under reduced pressure, the obtained aqueous solution was extracted with 120 ml of butanol, and the butanol layer was concentrated under reduced pressure to obtain 2.2 g of a crude extract.

This crude extract was dissolved in 30 ml of chloroform to obtain 1.0 g of a chloroform-insoluble fraction. This oil was subjected to centrifugal liquid-liquid chromatography (flow rate 3 ml /
Min, rotation speed 100 rpm), butanol: ethanol: water =
The antibiotic of the present invention can be obtained by eluting 10: 2.5: 10 with the upper layer as the fixed layer and the lower layer as the mobile layer by the reverse partition method.
725 mg of an active fraction containing RS-22A (hereinafter sometimes simply referred to as “RS-22A” in the examples; the same applies hereinafter), RS-22B, and RS-22C as main components were obtained. 98 mg of this powder
DS column (2 cm diameter, 25 cm length; Pegacil ODS,
(Manufactured by Senshu Kagaku Co., Ltd.). The fractionation was performed using 80% methanol as the developing solvent and at a flow rate of 9.9 ml / min.

As a result, the purified RS-
22A about 14.5 mg, RS-22B about 27 mg, RS-22C about 28.7
mg was obtained. The physicochemical properties of these substances are as follows. 1. Melting point (decomposition point): RS-22A: 132-137 ° C RS-22B: 136-139 ° C RS-22C: 129-132 ° C 2. Properties of substances: All substances are white powders. 3. Specific rotation: RS-22A: [α] ²⁸ _D + 36.7 ° (c = 0.45, MeOH) RS-22B: [α] ²⁸ _D + 36.7 ° (c = 0.52, MeOH) RS-22C: [α] ] ²⁸ _D + 36.4 ° (c = 0.51, MeOH)

4. Molecular weight, molecular formula: FAB mass spectrum of RS-22B and high-resolution FAB mass spectrum analysis, and
It was determined as follows by FAB mass spectrum and NMR spectrum analysis of RS-22A and RS-22C. RS-22A (M + H) ⁺ : m / z 1054.6 (FAB): C ₅₄ H ₉₁ N ₃ O ₁₇ RS-22B (M + H) ⁺ : m / z 1068.6603 (high-resolution FAB):
C ₅₅ H ₉₃ N ₃ O ₁₇ RS-22C (M + H) ⁺ : m / z 1082.7 (FAB): C ₅₆ H ₉₅ N ₃ O ₁₇ 5. Solubility: All substances are readily soluble in butanol and methanol . Insoluble in ethyl acetate, chloroform and water.

[0026] 6 Color reaction:. Either substances, anisaldehyde sulfuric, 10% sulfuric acid, positive for color reaction by I _2. 7. UV absorption spectrum λ _max (MeOH) (ε): RS-22A: 233 nm (22300, sh), 238 nm (23900), 265
(14900) RS-22B: 234 nm (26500, sh), 240 nm (28200), 266
(18400) RS-22C: 233 nm (22400, sh), 238 nm (30900), 265
(21200) 8. Infrared absorption spectrum:

RS-22A: 3380, 2990, 2960, 1700, 1670,
1640, 1450, 1380, 1255, 1140, 1068, 1000, 970 RS-22B: 3380, 2990, 2960, 1700, 1670, 1640, 1450,
1380, 1255, 1140, 1068, 1000, 970 RS-22C: 3380, 2990, 2960, 1700, 1670, 1630, 1450,
1380, 1255, 1140, 1067, 1000, 970 9. Nuclear magnetic resonance spectrum (a) ¹³ C NMR (100 MHz, CD ₃ OD)

[Table 2] 位置 Position RS-22A RS-22B RS-22C ───────── ────────────────── 1 169.0s 168.9s 169.1s 2 120.8d 120.7d 120.8d 3 146.7d 146.7d 146.8d 4 130.4d 130.4d 130.4d 5 147.4d 147.4 d 147.5d 6 46.5d 46.5d 46.6d 6-Me 16.9q 16.9q 16.9q 7 75.5d 75.5d 75.6d 8 39.4t 39.3t 39.4t 9 74.8d 74.7d 74.7d 10 44.3d 44.4d 44.4d 10-Me 10.4q 10.4q 10.4q 11 72.4d 72.2d 72.3d 12 33.6t 33.6t 33.6t 13 30.6t 30.6t 30.67t 14 41.7d 41.6d 41.6d 14-Me 14.9q 14.8q 14.91q 15 72.3d 72.3d 72.4d 16 41.8t 41.7t 41.8t 17 99.8s 99.7s 99.8s 18 77.3d 77.1d 77.3d 19 69.7d 69.7d 69.8d 20 41.2t 41.2t 41.3t 21 65.5d 65.5d 65.5d 22 44.6t 44.6t 44.6t 23 70.6d 70.6d 70.7d 24 43.8t 43.9t 44.0t 25 65.5d 65.5d 65.5d 26 44.1t 44.1t 44.1t 27 66.3d 66.1d 66.1d 28 40.8t 40.8t 40.8t 29 74.3d 74.3d 74.3d 30 139.9 s 139.9s 140.0s 30-Me 13.4q 13.3q 13.3q 31 125.1d 125.1d 125.1d 32 128.6d 128.6d 128.7d 33 136.2d 1 36.2d 136.2d 34 41.0d 41.0d 41.0d 34-Me 17.5q 17.5q 17.6q 35 80.4d 80.3d 80.4d 36 35.0d 34.9d 35.0d 36-Me 14.2q 14.0q 14.1q 37 34.4t 34.5t 34.5t 38 27.9t 27.9t 27.9t 39 33.7t 33.6t 33.7t 40 132.7d 132.5d 132.6d 41 130.2d 130.3d 130.3d 42 30.6t 30.6t 30.7t 43 29.8t 29.8t 29.8t 44 41.9t 41.9t 42.1t 1 '171.7s 171.6s 171.7s 2' 49.7t * 49.6t * 49.6t * 3 '174.1s 174.2s 174.1s N-Me-28.3s 28.5s N-Me'--28.5s N-CN ₂ 158.7s 158.2s 157.3s ─────────────────────────── * Measured in CD ₃ OH (b) ¹ H NMR (400 MHz, CD ₃ OD)

[Table 3] 位置 Position RS-22A RS-22B RS-22C ─ ────────────────────────────────── 2 5.82 (1H, d, J = 15Hz) 5.82 (1H, d , J = 15Hz) 5.82 (1H, d, J = 15Hz) 3 7.17 (1H, dd, 7.18 (1H, dd, 7.18 (1H, dd, J = 10Hz, J = 15Hz) J = 10Hz, J = 15Hz) J = 10Hz, J = 15Hz) 4 6.26 (1H, dd, 6.26 (1H, dd, 6.26 (1H, dd, J = 10Hz, J = 15Hz) J = 10Hz, J = 15Hz) J = 10Hz, J = 15Hz ) 5 6.2 (1H, dd, 6.1 (1H, dd, 6.1 (1H, dd, J = 8.8Hz, J = 15Hz) J = 8.8Hz, J = 15Hz) J = 8.8Hz, J = 15Hz) 6 2.42 ( 1H, m) 2.42 (1H, m) 2.42 (1H, m) 6-Me 1.1 (1H, d, J = 6.8Hz) 1.09 (1H, d, J = 6.8Hz) 1.09 (1H, d, J = 6.8 Hz) 7 3.75 (1H, m) 3.75 (1H, m) 3.75 (1H, m) 8 1.42, 1.72 1.46, 1.72 1.46, 1.72 (each 1H, m) (each 1H, m) (each 1H, m) 9 3.78 (1H, m) 3.8 (1H, m) 3.8 (1H, m) 10 1.44 (1H, m) 1.44 (1H, m) 1.44 (1H, m) 10-Me 0.87 (1H, d, J = 6.8Hz ) 0.86 (1H, d, J = 6.8Hz) 0.86 (1H, d, J = 6.8Hz) 11 3.88 (1H, m) 3.88 (1H, m) 3.88 (1H, m) 12 1.32, 1.59 1.32, 1.59 1.32 , 1.59 (ea ch 1H, m) (each 1H, m) (each 1H, m) 13 1.28 (2H, m) 1.28 (2H, m) 1.28 (2H, m) 14 1.58 (1H, m) 1.58 (1H, m) 1.58 (1H, m) 14-Me 0.91 (1H, d, J = 6.8Hz) 0.91 (1H, d, J = 6.8Hz) 0.91 (1H, d, J = 6.8Hz) 15 3.86 (1H, m) 3.86 ( 1H, m) 3.86 (1H, m) 16 1.83 (2H, m) 1.83 (2H, m) 1.83 (2H, m) 18 3.34 (1H, d, J = 9.3Hz) 3.35 (1H, d, J = 9.3 Hz) 3.35 (1H, d, J = 9.3Hz) 19 3.86 (1H, m) 3.86 (1H, m) 3.86 (1H, m) 20 1.3, 1.9 1.3, 1.9 1.3, 1.9 (each 1H, m) (each 1H, m) (each 1H, m) 21 4.08 (1H, m) 4.08 (1H, m) 4.08 (1H, m) 22 1.5, 1.67 1.5, 1.67 1.5, 1.67 (each 1H, m) (each 1H, m) ) (each 1H, m) 23 5.23 (1H, m) 5.21 (1H, m) 5.21 (1H, m) 24 1.65-1.7 (2H, m) 1.65-1.7 (2H, m) 1.65-1.7 (2H, m ) 25 3.86 (1H, m) 3.85 (1H, m) 3.85 (1H, m) 26 1.58 (2H, m) 1.58 (2H, m) 1.58 (2H, m) 27 3.86 (1H, m) 3.85 (1H, m) 3.85 (1H, m) 28 1.58 (2H, m) 1.58 (2H, m) 1.58 (2H, m) 29 4.16 (1H, m) 4.16 (1H, m) 4.16 (1H, m) 30-Me 1.64 (3H, m) 1.64 (3H, m) 1.65 (3H, m) 31 5.97 (1H, d, 5.97 (1H, d, 5.97 (1H, d, J = 10.7Hz) J = 10.7Hz) J = 10.7Hz ) 32 6.2 (1H, dd, 6.2 (1H, dd, 6.21 (1H, dd, J = 10.7Hz, J = 15.1Hz) J = 10.7Hz, J = 15. 1Hz) J = 10.7Hz, J = 15.1Hz) 33 5.4 (1H, dd, 5.4 (1H, dd, 5.4 (1H, dd, J = 8.8Hz, J = 15.1Hz) J = 8.8Hz, J = 15.1Hz) ) J = 8.8Hz, J = 15.1Hz) 34 2.52 (1H, m) 2.51 (1H, m) 2.52 (1H, m) 34-Me 0.99 (1H, d, J = 6.9Hz) 0.99 (1H, d, J = 6.9Hz) 0.99 (1H, d, J = 6.9Hz) 35 4.76 (1H, dd, 4.75 (1H, dd, 4.76 (1H, dd, J = 3.8Hz, J = 8.8Hz) J = 3.8Hz, J = 8.8Hz) J = 3.9Hz, J = 8.3Hz) 36 1.77 (1H, m) 1.77 (1H, m) 1.77 (1H, m) 36-Me 0.92 (3H, d, J = 6.8Hz) 0.92 ( 3H, d, J = 6.8Hz) 0.92 (3H, d, J = 6.8Hz) 37 1.1 (2H, m) 1.1 (2H, m) 1.1 (2H, m) 38 1.34 (2H, m) 1.38 (2H, m) 1.38 (2H, m) 39 1.85-2.0 (2H, m) 1.85-2.0 (2H, m) 1.85-2.0 (2H, m) 40 5.44 (1H, m) 5.44 (1H, m) 5.44 (1H, m) 41 5.42 (1H, m) 5.42 (1H, m) 5.42 (1H, m) 42 2.06 (2H, m) 2.06 (2H, m) 2.06 (2H, m) 43 1.62 (2H, m) 1.62 (2H , m) 1.62 (2H, m) 44 3.15 (2H, t, J = 6.9Hz) 3.14 (2H, t, J = 6.9Hz) 3.16 (2H, t, J = 6.4Hz) 2 '3.25 (2H, m ) 3.25 (2H, m) 3.25 (2H, m) N-Me-2.83 (3H, s) 2.83 (3H, s) N-Me '--2.83 (3H, s) ───────── ──────────────────────────

Test Example 1 (Test for antibacterial activity) Sabouraud agar medium as a fungal medium and sen as a bacterial medium
Using a sitivity disk agar-N (Nissui Pharmaceutical) medium, the minimum inhibitory concentration (MIC, unit in the table: μ
g / ml) are shown in Table 4.

[Table 4] ────────────────────────────────── Strain RS-22A RS-22B RS-22C ─ ───────────────────────────────── Cryp. Neoformans KC-201 3.13 3.13 3.13 C. albicans KC-07 3.13 3.13 6.25 C. tropicalis KC-104 1.56 3.13 3.13 C. parapsilosis KC-110 6.25 6.25 6.25 C. glabrata KC-308 6.25 6.25 6.25 Asp. Fumigatus KA-01 12.5 12.5 25 Asp. Flavus KA-06 12.5 12.5 12.5 T. mentagropytes KD -04 25 25 25 T. rubrum KD-114 12.5 12.5 12.5 M. Canis KD-305 12.5 12.5 12.5 M. gypseum KD-318 12.5 12.5 12.5 S. aureus FDA 209P JC-1 6.25 6.25 12.5 S. aureus Smith 12.5 12.5 25 S. pyogenes Cook 25 12.5 25 E. faecalis 1373> 50> 50> 50 E. coli NIHJ JC-2> 50> 50> 50 K. pneumoniae No.42> 50> 50> 50 P. vulgaris No.33> 50 > 50> 50 P. aeruginosa PAO-1> 50> 50> 50 ──────────────────────────────── ─

As a result, it was found that RS-22A, RS-22B and RS-22C all exhibited strong antibacterial activity against Gram-positive bacteria, filamentous fungi and yeast. The bacterial names in Table 4 are as follows. (Fungi) Cryp. Neoformans: Cryptococcus neoformans C. albicans: Candida albi
cans) C. tropicalis: Candida tr.
opicalis) C. parapsilosis: Candid
a parapsilosis) C. glabrata: Candida glabra
ta) ASP. fumigatus: Aspergillus fumigatus
rgillus fumigatus) ASP. flavus: Aspergillus flavus
s flavus) T. mentagrophytes: Trichophyton mentagrophytes T. rubrum: Trichophyton rubrum
rubrum) M. canis: Microsporum canis
nis) M. gypseum: Microsporu
m gypseum) (Bacteria) S. aureus: Staphylococcus aureus
lococcus aureus) S. pyogenes: Staphylococcus pyogenes
hylococcus pyogenes) E. faecalis: Enterococcus faecalis
rococcus faecalis) E. coli: Escherichia coli K. pneumoniae: Klebsiella pneumoniae
ella pneumoniae) P. vulgaris: Proteus vulgaris (Proteus vulg)
aris) P. aeruginosa: Pseudonas aeruginosa (Pseu
domonas aeruginosa)

Test Example 2 (Control Activity against Plant Pathogenic Bacteria) A paper disk assay was performed on eight kinds of plant pathogenic bacteria in accordance with a conventional method. R. solani as inoculation source
As for V. dahliae, a spore formed by culturing with shaking in a PD medium was used for V. dahliae. For other strains, spores cultured in PDA medium were used. The diameter of the blocking circle was measured by measuring its diameter (mm). As a result, RS-22B and RS-22C were obtained from Alternaria mali, Botrytis sineria (Botryt).
is cinerea), Rhizoctonia solani (Rhizoctonia sol)
ani) (stock), Verticillium dahlia (Verti)
cillium dahliae) (Table 5). The shape of the blocking circle is clear cut for RS-90-22B,
C was a double stop circle.

[0031]

[Table 5] 円 Inhibitory circle of pathogenic bacteria (diameter: mm) RS-22A RS-22B RS-22C ─────────────────────────────────── Alternaria mali + 17 16 Botrytis cinerea 13 + Ceratocystis fimbriata + F. oxysporum f.sp.cucumerinum Glomerella cingulata + Pyricularia oryzae Rhizoctonia solani 15 10 Verticillium dahliae 20 18 ────────────────────────── ───────── * Tested at 100 ppm. A blank column indicates "no blocking circle".

A mold pot test was conducted using RS-22A, RS-22B and RS-22C. Each test substance was dissolved in 10% methanol and further diluted with distilled water to adjust to 100 ppm. Details of the test method are as follows.

Target disease: cucumber gray mold, sclerotium disease Test crop: cucumber Varieties: Sagami Hanshiro Growth degree: 1 leaf stage (2 weeks after seeding) Treatment method: sprayed 1 day before inoculation Control drug: benrate hydration Agent (250ppm) Inoculation source: A disk obtained by punching out a soda cultivated in a PDA medium at 20 ° C for 3 days with a cork borer. Inoculation method: One day after the treatment with the agent, the fungal surface is attached to the leaves. Inoculate two discs per leaf. Pathogenic condition: Keep in a dark room at 20 ° C in a dark room for 3 days. Investigation method: Measure the diameter of the lesion and calculate the control value based on it. Evaluation: A, B and C are determined as follows according to the control value. A: (90 to 100%) B: (60 to 89.9%) C: (0 to 59.
9%)

Target disease: Tomato epidemic Test crop: Tomato Varieties: Momotaro Growing degree: 1-2 leaf stage (17 days after seeding) Treatment method: Spray 1 day before inoculation Control agent: Orthoside wettable powder (1333 ppm) inoculation Source: zoospore suspension (1 × 10 ⁵ cells / ml) of diseased tomato leaves inoculated inoculation: Inoculation method: 1 day after drug treatment, inoculation of spores with an air brush Infection condition: Keep in darkness at 20 ° C. in a humid chamber for 3 days I do. Investigation method: The disease area ratio is investigated in five stages, and the control value is calculated. Evaluation: A, B and C are determined as follows according to the control value. A: (80 to 100%) B: (60 to 79.9%) C: (0 to 59.
9%)

Target disease: rice blast Test crop: rice Varieties: Asahi Aichi Growth: four-leaf stage (18 days after sowing) Treatment method: sprayed on the day of inoculation Control agent: plaes (20 ppm) Inoculation source: oatmeal After culturing for 2 weeks in the medium, 3 days B
Spore suspension formed after irradiation with LB lamp (2 × 10 ⁶ cells / m
l) Inoculation method: On the day of drug treatment, inoculate spores with an airbrush. Infection condition: Keep in a dark room at 20 ° C for 1 day and then in an artificial weather room (25-30 ° C) for 4 days. Investigation method: The number of lesions is measured, and the control value is calculated. Evaluation: A, B and C are determined as follows according to the control value. A: (90 to 100%) B: (60 to 89.9%) C: (0 to 59.
9%)

Target disease: rice sheath blight Test crop: rice Variety: Aichi Asahi Growing degree: 1 to 4 leaf stage (2 weeks after seeding) Treatment method: sprayed 1 day before inoculation Control agent: Validacin solution (60 ppm) inoculation Source: Bacteria grown on PDA medium at 28 ° C for 3 days Inoculation method: One day after treatment with the drug, the bacteria are stuck to the rice plant. Disease onset: Keep in the dark in a wet room (25-30 ° C) for 4 days. Investigation method: The disease area ratio is investigated in five stages, and the control value is calculated. Evaluation: A, B and C are determined as follows according to the control value. A: (80 to 100%) B: (60 to 79.9%) C: (0 to 59.
9%)

Target disease: wheat powdery mildew Test crop: wheat Variety: Chihoku Growth: 2 leaf stage (5 days after sowing) Treatment method: sprayed 1 day after inoculation Control drug: inoculation of bileton wettable powder (125 ppm) Source: Passaged diseased wheat leaves Inoculation method: One day before drug treatment, spores of diseased wheat leaves are directly applied to test wheat leaves. Disease onset: Keep in a climate chamber (25-30 ° C) for 6 days. Investigation method: The number of lesions is measured, and the control value is calculated. Evaluation: A, B and C are determined as follows according to the control value. A: (90 to 100%) B: (60 to 89.9%) C: (0 to 59.
9%)

As a result, RS-22A, RS-22B and RS-22C
100p against cucumber gray mold, sclerotium, tomato blight, rice blast, rice sheath blight, wheat powdery mildew
It was confirmed that the compound exhibited a controlling activity at a concentration of pm or more.

Test Example 3 (herbicidal activity) The results of measuring the herbicidal activity of a mixture of RS-22A, RS-22B and RS-22C are shown in Table 6. As is clear from these results, the mixture of RS-22A, RS-22B, and RS-22C exhibited activity in the paddy field weeding evaluation test (in vivo). The details of the test method are as follows. (Paddy field herbicidal efficacy test) Scale 30 cm ² cups, greenhouse soil Clay loam Test plant Tainubie (A in Table 6): Before emergence Tamabayatsuri (B in Table 6): Before emergence Broadleaf (C in Table 6) ): Before emergence Onion (D in Table 6): Before emergence Firefly (E in Table 6): Before emergence Pine tree (F in Table 6): Before emergence Rice (Koshihikari: G in Table 6): 2.3 L Comparative drug KPP-314 Treatment method Dropping treatment The day after transplantation Water management Depth of water 3 cm Depth of water No restriction 2 Survey method Observational survey (1-5) 14 days later

[0040]

[Table 6] Paddy field weeding spectrum ─────────────────────────────────── Test ai Sample g / a ABCDEFG ─────────────────────────────────── RS-22 20 2.5 3 3.5 5 1 1 1.5 * 10 1.5 2 2.5 3 1 1 1.3 * 5 1 1 1 1.2 1 1 1 ─────────────────────────────────── KPP-314 1 5 5 5 5 4.8 5 1.3B 0.5 5 5 5 5 4.3 5 1.1B ─────────────────────────────本 Number per CONT.1 POT 30 50 20 5 12 + 1 ─────────────────────────────── ──── *: Leaf tip withering B: Leaf sheath browning 1─ No effect ~ 5─ Complete withering, days after treatment

Test Example 4 (Toxicity test) When the toxicity of RS-22 (a mixture of RS-22A, RS-22B and RS-22C) was tested, the LD ₅₀ value was 25 mg / kg. As is clear from these results, RS-22 has low toxicity and is useful as a fungicide and herbicide for agricultural and horticultural use. The test method is as follows. Method Animal used: Mouse Jcl / ICR strain, 4 weeks old, male Drug preparation method: Drug was acetamide (final concentration 5%
) And 5% glucose solution to a final concentration of 10, 20,
30, 40, and 50 mg / kg. The route of administration is 0.1 ml / 10 g b.
wip.

The novel antibiotic RS provided by the present invention
-22 is useful as an agricultural and horticultural fungicide / herbicide. Further, the novel actinomycete Streptomyces RS-22 strain provided by the present invention is useful for producing the above antibiotic.

 ────────────────────────────────────────────────── ─── Continuing on the front page (72) Inventor Shin Nobukata, 2-1 Hirosawa, Wako-shi, Saitama Prefecture, RIKEN (72) Inventor Kimie Kimie 2-1 Hirosawa, Wako-shi, Saitama Prefecture, RIKEN (72 Inventor Yumiko Kobayashi 2-1 Hirosawa, Wako City, Saitama Prefecture Inside the RIKEN (72) Inventor Kiyoshi Isono 324-1 Orito, Shimizu City, Shizuoka Prefecture (72) Inventor Tomei Tomei 2354 Shadoji, Shanghai, Shanghai, China Inside the Agricultural Chemical Research Institute of the City (72) Inventor Ni Changchun No. 2354, Shado Road, Shanghai, China Inside the Agrochemical Research Institute, Shanghai (72) Inventor Zhang Yunba, No. 2354, Shado Road, Shanghai, the People's Republic of China Inside the Shanghai Agrochemical Research Center (72) Inventor Shen No.2354, Shado-ro, Shanghai, China Inside the Agricultural Chemicals Research Institute in Shanghai (72) Inventor Shen Tora-first, No.2354, Shado-ro, Shanghai, Shanghai, China In agricultural chemicals Institute

Claims (9)

[Claims] 1. The following formula: (Wherein R represents unsubstituted or a guanidino group substituted by one or more lower alkyl groups). 2. The antibiotic RS-22A, wherein the guanidino group is an unsubstituted guanidino group, wherein the guanidino group is HN = C (NHCH ₃ )
The antibiotic RS-22B represented by -NH-, and the guanidino group
_{CH 3 N = C (NHCH 3} ) according to claim 1, wherein the -NH- in selected from the group consisting of antibiotic RS-22C represented antibiotic RS-22. 3. An actinomycete belonging to the genus Streptomyces and producing the antibiotic RS-22 according to claim 1 or 2. 4. The RS-22-producing bacterium according to claim 3, which is a Streptomyces sp. RS-22 strain. 5. Streptomyces sp. RS-22
stock. 6. The method according to claim 1, wherein the RS-22-producing bacterium according to claim 1 or 2 belonging to the genus Streptomyces is cultured, and the antibiotic RS-22 is collected from the culture. Or the method for producing the antibiotic RS-22 according to 2. 7. The method according to claim 6, wherein the Streptomyces RS-22 strain is cultured. 8. The antibiotic RS- according to claim 1 or 2.
An agricultural and horticultural fungicide containing 22 or a salt thereof as an active ingredient. 9. The antibiotic RS- according to claim 1 or 2.
A herbicide containing 22 or a salt thereof as an active ingredient;