JP2780828B2 - Polyether antibiotic MI215-NF3 substance, method for producing the same, and agent for controlling chicken coccidiosis - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION (Field of Industrial Application) The present invention relates to a novel antibiotic MI215-NF3 substance and a salt thereof, and also relates to a method for producing the MI215-NF3 substance or a salt thereof. Further, the present invention relates to an agent for controlling coccidiosis in chickens, which comprises an MI215-NF3 substance or a salt thereof as an active ingredient. The present invention also relates to a novel microorganism, Actinomadura sp. MI215-NF3, which has the property of producing a MI215-NF3 substance.

(Prior art) It is known that various polyether antibiotics have bactericidal activity. In addition, monensin (see Japanese Patent Publication No. 45-113) or salinomycin (see "The Journal of Antibiotics", Vol. 27, pp. 814 to 821) belonging to the polyether antibiotics, etc. are known to cause coccidiosis in chickens. Used as a control agent.

(Problems to be solved by the invention) Coccidiosis is a serious problem in poultry farming,
There is always a need to find or create new compounds that exhibit better anticoccidial activity and properties than previously known or used known compounds, and research is being done to do so.

(Means for Solving the Problems) The present inventors conducted research to find useful new antibiotics, and as a result, actinomadulla (Actinomadura)
ra) A new strain belonging to the genus was successfully isolated, and it was found that this strain produces a new polyether antibiotic, which was named MI215-NF3 substance. It has also been found that this MI215-NF3 substance or a salt thereof (carboxylate) exhibits a control activity against chicken coccidiosis. Further research was continued, and the chemical structure of the MI215-NF3 substance was determined to confirm that it was a novel compound, and it was found that the compound could be represented by the formula (I) described below.

Wherein R is a hydrogen atom.

Thus, the present invention relates to Actinomadur
a) A substance obtained by culturing a microorganism belonging to the genus and named by the present inventors as MI215-NF3 substance, a salt thereof, a method for producing the same, and further, a use as an agent for controlling coccidiosis in chickens To provide.

That is, according to the first invention, there is provided a MI215-NF3 substance having the above formula (I) and a salt thereof.

The MI215-NF3 substance of formula (I) is in the form of a free carboxylic acid [where R of formula (I) is a hydrogen atom], or alternatively, R of formula (I) is generally a metal or ammonium group. As it is possible, the present invention also covers salts of the MI215-NF3 substance. This salt can also be in the form of an alkali metal salt (where R in formula (I) is an alkali metal atom), for example a sodium, potassium or lithium salt, or in the form of an alkaline earth metal salt [formula When R in (I) is an alkaline earth metal atom], it can be, for example, a calcium salt or a magnesium salt, and can be another metal salt, for example, an aluminum salt or an iron salt, and furthermore, an ammonium salt. There can be.

The physicochemical properties of the MI215-NF3 substance according to the present invention are as follows.

The physicochemical properties of the MI215-NF3 substance sodium salt are shown below.

(1) Appearance Colorless plate-like crystal (2) Calculated value of elemental analysis experimental value (as C ₃₇ H ₆₃ O ₁₁ Na) C 62.47% 62.87% H 8.96% 8.98% O 24.60% 24.90% Na 3.53% 3.25% (3) Mass spectrometry (SIMS) m / z 707 (MH ⁺ ) (4) Melting point 217 to 218 ° C. (5) Specific rotation ▲ [α] ²⁵ _D ▼ = + 30.6 ゜ (c 1.0, chloroform) (6) Infrared absorption Spectra (KBr tablet method) As shown in FIG. 1 of the accompanying drawings.

(7) Proton nuclear magnetic resonance spectrum A proton NMR spectrum measured at room temperature in heavy chloroform at 400 MHz is as shown in FIG. 2 of the accompanying drawings.

(8) Solubility Soluble in benzene, chloroform, acetone, ethyl acetate, ethanol, and methanol, but insoluble in water.

The biological properties of the MI215-NF3 substance according to the present invention are as follows.

The biological properties of the MI215-NF3 substance sodium salt are shown below.

(1) Antibacterial spectrum The antibacterial spectrum of the MI215-NF3 substance sodium salt against various microorganisms was measured by a multiple dilution method on an agar medium by a method known per se. Table 1 shows the results.

(2) Toxicity To test the acute toxicity of the sodium salt of the MI215-NF3 substance using mice, the MI215-NF3 substance was suspended in a physiological saline solution containing 10% dimethyl sulfoxide, and the suspension was intraperitoneally injected. Mice were observed for 14 days. As a result, MI215-N
The LD ₅₀ value of the F3 substance was estimated to be 56 mg / kg.

(3) Anticoccidial activity The activity of MI215-NF3 for controlling coccidiosis in chickens was tested as shown below.

1. Preliminary rearing of chicks (males) for one week,
Two week old chicks were used for testing.

2. At the start of the test, the chicks were weighed individually and selected so as to have little variation, and the chicks were divided into 10 birds / group.

3. 5 × 10 ⁴ oocysts per bird were orally administered using a gastric tube.

4. After administration of the oocysts, a special diet (without adding an anticoccidial agent) was allowed to freely ingest a diet containing a fixed amount of MI215-NF3 substance (sodium salt). In addition, drinking water was also allowed freely.

5. From the oocyst administration to the 7th day, the number of oocysts in feces and the condition of feces were observed every day.

6. Dissected 7 days after infection. The dissection procedure is shown below.

1) Body weight was measured for each chick.

2) Chicks were dislocated and killed.

3) The abdomen was opened to remove the cecum, and the lesions outside and inside the cecum were visually inspected.

4) The cecum examined was disintegrated for each individual with a homogenizer to prepare a cecal suspension, and the number of oocysts in the suspension was calculated.

5) ACI (Anticoccidial Ind.) Based on the evaluation method proposed by Merck, USA
ex) values were rated.

ACI = (relative weight gain + chick survival rate) − (lesion value + oocyst value) In evaluating the above ACI value, the weight gain rate of each group of chicks during the drug efficacy test period, and The relative weight gain used in the calculation of ACI is calculated by the following equation.

 Gain rate = [(average weight of chick at end of test-average weight of chick at start of test) / average weight of chick at start of test] x 100.

 Relative weight gain = (weight gain rate of dose group or non-infected drug group / weight gain rate of non-infected non-medicated group) x 100.

The lesion value is a value obtained by indexing the result of a macroscopic examination of a lesion in the excised cecum, and the oocyst value is a value obtained by indexing the actually measured number of oocysts.
The details of the method for evaluating the ACI value are described, for example, in the literature, "chicken coccidiosis", pp. 95-101, supervised by Kiyoshi Tsunoda (Chikusan Shuppan, published May 1983).

 Table 2 shows the obtained results.

As is clear from the above results, the MI215-NF3 substance or a salt thereof can be used as a control agent for coccidiosis in chickens.

Further, according to a second aspect of the present invention, a bacterium producing the MI215-NF3 substance of the above formula (I) belonging to the genus Actinomadura is cultured, and the MI215-NF3 substance or a salt thereof is collected from the culture. And a method for producing the antibiotic MI215-NF3 or a salt thereof.

The mycological properties of the MI215-NF3 substance producing bacterium used in the method of the present invention are as follows.

The MI215-NF3 substance-producing bacterium used in the present invention can be used irrespective of species as long as it has an ability to produce an antibiotic having the above-mentioned physicochemical properties and biological properties, and is selected from a wide range of microorganisms. be able to. Among such microorganisms, a specific preferable example of the MI215-NF3 substance-producing bacterium is an actinomycete isolated from the soil of Bunkyo-ku, Tokyo-country by the present inventors, and a strain with the strain number of MI215-NF3. It is.

The bacteriological properties of the MI215-NF3 strain are described below.

1. Morphology Under a microscope, elongate aerial hyphae from branched mycelia. No division of the mycelium in the base was observed. It forms spore chains on aerial hyphae and takes on a key or spiral shape. The chain of mature spores is usually 7 to 15, and spores are oval (about 0.7 × 1.0 to 1.2
Microns) and its surface is wart-like. It should be noted that no ring branch and spore sac were observed.

2. Growth conditions in various media Regarding color description Standards shown in [] are the Color Harmony Manual (Container Corporation of Ameri) of Container Corporation of America.
ca harmony manual) was used.

(1) Sucrose / nitrate agar medium (cultured at 30 ° C.) White aerial mycelia grow slightly on the growth of colorless to light [3 gc, Lt Tan]. No soluble dye is observed.

(2) Glucose and asparagine agar medium (cultured at 30 ° C.) Slightly develop white aerial hyphae on the growth of light yellow tea [3ic, Lt Amber to 3le, Cinnamon] to bright tea [4 ng, Lt Brown] . The soluble pigment has a slightly yellow tint.

(3) Glycerin-asparagine agar medium (ISP-medium
(5, 30 ° C. culture) On growth of light yellow to light yellow tea [2le, Mustard to 3le, Cinnamon], aerial mycelia of white to gray white [2dc, Natural] are slightly formed. The soluble pigment is slightly yellowish from around 21 days after the culture.

(4) Starch / inorganic salt agar medium (ISP-medium at 4,30 ° C) Light yellow to light yellow tea [3ng, Yello Maple] to yellow tea [3pi, G
olden Brown] and pink white [5ba, Shell Pin
k] to aerial hyphae of gray [2dc, Natural]. The soluble pigment has a slight yellow tint.

(5) Tyrosine agar medium (ISP-medium at 7,30 ° C) Light yellow to yellow tea [3ng, Yellw Maple] to grayish yellow tea [3ni, C
love Brown), yellow ash (1dc, Putty-lec, Lt)
To form aerial mycelium of Citron Gray]. The soluble pigment has a slight yellow tint.

(6) Nutrient agar medium (cultured at 30 ° C) Light yellow tea [3ic, Lt Amber] to grayish yellow tea [3ni, Clove Br
own], on which white aerial hyphae partially form. The soluble pigment has a faint yellow tint.

(7) Yeast / malt agar medium (ISP-medium, culture at 30 ° C) Light yellow tea [2le, Mustard]-grayish yellow tea [2nl, Covert Br
own] and develop white-brown aerial mycelia. The soluble pigment has a faint brownish tinge.

(8) Oatmeal agar medium (ISP-medium, cultivation at 30 ° C) Light brown to light brown [4le, Maple] to yellow tea [3ng, Yel
low Maple], white to pink white [5ba, Shell Pin
k ~ 7ba, Pink Tint]. The soluble pigment has a slight yellow tint.

(9) Glycerin / nitrate agar medium (cultured at 30 ° C) Aerial mycelia of white to pink white [5ba, Shell Pink] are formed on the growth of colorless to light yellow to light yellow tea [3le, Cinnamon]. The soluble pigment has a slight yellow tint.

(10) Starch agar medium (cultured at 30 ° C) Light yellow tea [2le, Mustard]-grayish yellow tea [3ni, Clove Bro
wn], on which white aerial hyphae grow slightly. No soluble dye is observed.

(11) Lime malate agar medium (cultured at 30 ° C) Colorless to light yellow growth, white to pink white [5ba, Shell
Pink]. No soluble dye is observed.

(12) Cellulose (Synthetic solution with filter paper strips, cultured at 30 ° C) The growth is colorless, and slightly forms white aerial hyphae. No soluble dye is observed.

(13) Gelatin stab culture In a simple gelatin medium (cultured at 20 ° C), the growth is colorless to pale yellow, no aerial mycelia form, and no soluble pigment is observed. In the case of a glucose / peptone / gelatin medium (cultured at 27 ° C.), the growth is pale yellow, no aerial hyphae are formed, and no soluble pigment is observed.

(14) Skimmed milk (cultured at 37 ° C) The growth is pale yellow to pale yellow tea, aerial mycelia do not form, and the soluble pigment is slightly brownish from around the 18th day after culture.

3. Physiological properties (1) Growth temperature range Using starch / inorganic salt agar medium (ISP-medium 4),
As a result of the test at each of 0 ° C., 24 ° C., 27 ° C., 30 ° C., 37 ° C., and 50 ° C., the cells grew at any temperature except 50 ° C. The optimum growth temperature is around 30 ° C.

(2) Liquefaction of gelatin (15% simple gelatin medium, cultivated at 20 ° C; glucose / peptone gelatin medium, cultivated at 27 ° C) In a simple gelatin medium, liquefaction starts around 11 days after culturing, but its effect is weak It is. In the case of the glucose / peptone / gelatin medium, liquefaction is observed from about 5 days after the culture, and the action is moderate to weak.

(3) Hydrolysis of starch (starch / inorganic salt agar medium, ISP-medium 4, and starch agar medium, all at 30 ° C)
Culture) Water degradability is observed in any of the media, and the action is moderate.

(4) Coagulation / peptone conversion of skim milk (defatted milk, cultivation at 37 ° C.) From about the fifth day after the culture, a coagulated state is exhibited, and after completion, peptone formation starts immediately. The progress of peptone formation was slow and incomplete after three weeks of observation.

(5) Melanin-like pigment formation (trypton yeast,
Broth, ISP-medium 1; peptone yeast / iron agar medium, I
SP-medium 6; tyrosine agar medium, ISP-medium 7; all at 30 ° C
Culture) It seems negative on tryptone yeast broth and tyrosine agar, and positive on peptone yeast iron agar.

(6) Utilization of carbon source (Pridham Gottlieb agar medium, ISP-medium at 9,30 ° C. culture) Growth using L-arabinose, D-xylose, D-glucose, D-fructose, rhamnose, D-mannitol However, sucrose, inositol, raffinose and lactose are not used.

(7) Dissolution of malate lime (maleate lime agar medium, 30
(Culturing at ℃) No dissolution of lime malate is observed.

(8) Nitrate reduction reaction (Peptone water containing 0.1% potassium nitrate, ISP-medium 8, cultured at 30 ° C.) Positive.

(9) Decomposition of cellulose (synthetic solution containing filter paper pieces, culture at 30 ° C.) Negative.

In summary, the MI215-NF3 strain forms a key or spiral spore chain on the aerial hyphae, with a spore linkage of 7
Count ~ 15. The spore surface is wart-like. Rotating branches and spores are not found. In addition, there was no division of the mycelium in the base. In various media, white-gray-white or pink-white basal hyphae grow on light yellow to light yellow to grayish yellow tea. The soluble pigment has a slight yellow tint.

The production of melanin-like pigment was negative on tryptone yeast broth and tyrosine agar, and
Shows positive on yeast / iron agar medium. Proteolytic power is moderate to weak, and starch has moderate water degradability. The nitrate reduction reaction is positive.

The MI215 to NF3 strains have meso-diaminopimelic acid as a cell component in all cells and glucose, galactose, majulose, and ribose as a sugar component, and have been described in Lechevalier et al., “Internationa
l Journal of Systematic Bacteriology, 20, 435, 1
970) Type III B, a major component of the cell wall
showed that. Phospholipids are PI type (excluding phosphatidylethanolamine, phosphatidylmethylethanolamine, phosphatidylcholine and unknown glucosamine-containing phospholipids, and include phosphatidylinositol and phosphatidylinositol mannoside). The composition of menaquinones _{MK-9 (H 6),} MK-9 (H 8) and a small amount of
It is MK-9 (H _4).

Based on the above, MI215-NF3 strain was obtained from Actinomadura (Ac
It is considered to be actinomycetes belonging to the genus tinomadura). When a species similar to the MI215-NF3 strain was searched for from known bacterial species of the genus Actinomadura, Actinomadura citrea (Actinomadura citrea, Reference 1 “An”
tibiotiki, Vol. 17, pp. 965-970, 1972; and Literature 2, "Sys
tematic & Applied Microbiology ", 6, 264-270, 198
5 years). That is, a spore chain exhibiting a key or spiral shape, a wart-like spore surface, a yellow soluble pigment in various media, and a menaquinone composition of the bacterial cell were MK-9.
(H ₆ ) and MK-9 (H ₈ ). However, there are many unclear points in the literature.
We plan to obtain a standard strain of Citrea and conduct comparative experiments. From these facts, at present, the MI215-NF3 strain was transformed into Actinomadura sp.
It is named. The MI215-NF3 strain was applied for deposit on November 22, 1988 at the Research Institute of Microorganisms and Industrial Technology of the Ministry of International Trade and Industry in Tsukuba, Ibaraki, Japan, and was deposited as Microtechnical Laboratory No. 10397. Currently, it is deposited under deposit number "FERM BP-2675" under the provisions of the Budapest Treaty.

In the method of the present invention, the production of the MI215-NF3 substance is performed as follows.

That is, the production of the MI215-NF3 substance is preferably performed by culturing the MI215-NF3 substance-producing bacterium in a nutrient medium at a temperature of 27 to 30 ° C while stirring under aerobic deep conditions. Nutrient media used for such purposes include sugars, starch, glycerin and assimilable carbon sources such as molasses, corn steep,
It can contain organic nitrogen sources such as liquor, soy flour, cottonseed flour, peptone, meat extract and hops meal, and inorganic nitrogen sources such as ammonium sulfate, ammonium nitrate and ammonium chloride. If excessive foaming occurs during the fermentation, an antifoam such as vegetable oil or silicone can be added to the fermentation medium. If the pH of the medium during fermentation changes, a buffer such as calcium carbonate may be added. Preferably, the aeration of the medium in the submerged tank maintains about 1/2 to 2 volumes of fresh air to 1 volume of broth per minute. Agitation is effected by means of agitation generally well known in the fermentation industry.

As a seed mother for the production of the MI215-NF3 substance, a growth obtained from a slope culture of the MI215-NF3 strain on an agar medium is used. The growth can be used to inoculate either the shake flask or the seed tank, or the seed tank may be inoculated with the growth from the shake flask.

Growth of this MI215-NF3 strain usually peaks in 5 to 7 days. However, the speed at which the bacteria used reach the maximum growth varies depending on the equipment used, the aeration speed, the stirring speed, and the like. Generally, fermentation is continued until sufficient antimicrobial activity has been imparted to the medium. The time-dependent change in the titer of the MI215-NF3 substance in this culture solution can be measured by a cylindrical plate evaluation method using Staphylococcus aureus smith as a test bacterium.

In the method of the present invention, the MI215-NF3 substance is collected from the culture obtained as described above. As a collection method, a means normally used for collecting a metabolite produced by a microorganism is appropriately used. Consisting of For example, a method using the difference in solubility with impurities, a method using a difference in adsorption affinity to various adsorbents, an extraction with a solvent that does not mix with water, and the like can be used alone or in combination with MI215.
-Can collect NF3 substances. In principle, the following method is preferred. That is, the cells are separated from the culture by filtration or centrifugation, and the resulting culture filtrate is extracted with an organic solvent immiscible with water, such as n-butyl acetate, ethyl acetate, and the like, and then washed with water to remove the organic solvent. Evaporate to dryness. The dried product usually contains the MI215-NF3 substance in an amount of about 10%. Furthermore, the crude MI21 obtained as a dried product
In order to increase the purity of the 5-NF3 substance, purification by silica gel column chromatography
MI215-NF3 substance can be obtained. The MI215-NF3 substance obtained by the above method is in the form of a salt such as sodium or potassium, or a free carboxylic acid, but can be made into a single salt by a known method. For example, the present MI215-NF3 substance is dissolved in ethyl acetate, washed sequentially with 1M aqueous hydrochloric acid and a saturated sodium carbonate solution, dried over an organic solvent layer over anhydrous sodium sulfate, concentrated to dryness, and crystallized from ethyl acetate-hexane. The sodium salt of the MI215-NF3 substance is obtained as colorless plate crystals.

The MI215-NF3 substance (sodium salt) obtained by purification as described above is dissolved, for example, in an organic solvent such as ethyl acetate or methanol, and a strongly acidic ion exchange resin (H ⁺ type) is added to the solution. Upon treatment, the MI215-NF3 material in the form of the free carboxylic acid can be formed. This free acid form
When the MI215-NF3 material is treated with, for example, an aqueous solution of a basic alkali metal inorganic salt, for example, sodium carbonate or potassium carbonate, or a basic water-soluble alkaline earth metal compound, for example, calcium hydroxide or magnesium hydroxide in an organic solvent. MI215-NF3 material in the form of the corresponding alkali metal salt or alkaline earth metal salt can be produced. Similarly, the corresponding aluminum salt or iron salt can be prepared by treating with another metal salt such as aluminum hydroxide or ferric hydroxide.

Further, according to the third invention, MI21 of the above formula (I)
An agent for controlling chicken coccidiosis comprising a 5-NF3 substance or a salt thereof as an active ingredient is provided.

In this coccidiosis controlling agent, the MI215-NF3 substance or a salt thereof as an active ingredient compound is, as in known drugs for similar uses, a known animal medically acceptable carrier, such as starch, cellulose powder, wood flour, Alternatively, it can be mixed with ethanol or the like or a food material. The amount of the MI215-NF3 substance or salt thereof in the composition obtained by mixing with the carrier can be at a concentration of 50 to 100 ppm which is effective for treating coccidiosis. The coccidiosis controlling agent of the present invention can also be in the form of a solution in which the MI215-NF3 substance or a salt thereof is dissolved in a suitable organic solvent, or a concentrated stock solution or dispersion in which the substance is suspended in a water-containing organic solvent. By adding such a solution or dispersion to the feed, the concentration of the active ingredient substance in the feed is reduced.
It should be 50-100ppm.

According to another aspect of the present invention, Actinomadura sp. MI215-NF3 is provided as a useful and novel microorganism capable of producing a useful antibiotic of the formula (I) as described above.

 Hereinafter, the present invention will be described in more detail with reference to examples.

Example 1 Actinomadura sp. MI
Cells of a slant culture of strain 215-NF3 (FERM BP-2675) were
ml seed mother medium (galactose 2%, dextrin 2 for chemistry)
%, Soy Peptone 1%, Corn Steep Liquor 0.5
%, Ammonium sulfate 0.2% and calcium carbonate 0.2%, and inoculated into a 500 ml rotary flask containing pH 7.4) and cultured on a rotary shaker at 30 ° C. for 144 hours. The obtained culture solution was divided into 110 ml each of a main culture medium (1.5% glycerin, 1.5% soluble starch, 0.5% soybean meal, 1.5% fish meal, 0.2% calcium carbonate, pH 7.4) in 500 ml portions. Volumetric rotary flasks were inoculated at 3% by weight. The inoculated medium was cultured at 27 ° C. for 144 hours on a rotary shaker at 180 rpm. The cells were collected from the obtained culture by filtration, and about 4 of the obtained culture filtrate was extracted with ethyl acetate 3
After extraction with, the extract is washed with water and dried over anhydrous sodium sulfate. The insoluble part was filtered off from the dried ethyl acetate solution,
The filtrate is evaporated to dryness. The residue was dissolved in 100 ml of ethyl acetate, the insoluble portion was filtered off again, and the filtrate was evaporated to dryness to give MI215-NF3.
2.4 g of crude material were obtained.

Example 2 2.4 g of a crude product of the MI215-NF3 substance obtained in Example 1 was dissolved in chloroform, and Kieselgel silica gel manufactured by Merck & Co. was used.
Column chromatography packed with 60 (150 g) in chloroform, chloroform / acetone (5: 1)
Is eluted as a developing solvent. The MI215-NF3 material elutes in the eluate fraction 1600 ml to 3700 ml. After evaporating the solvent under reduced pressure from the active fraction, the residue was dissolved in 90 ml of ethyl acetate,
The organic layer was washed successively with 0.1M aqueous hydrochloric acid, 0.1M aqueous sodium hydroxide solution and saturated saline, and then dried over anhydrous sodium sulfate. Then, the solution is distilled under reduced pressure to remove the solvent.
255 mg of an oily substance were obtained. The obtained oil was crystallized from hexane and ethyl acetate to obtain the sodium salt of MI215-NF3 substance as colorless plate crystals (232 mg).

Example 3 200 mg of sodium salt of MI215-NF3 substance was added to chloroform 1
Dissolve in 100 ml, add 10 ml of water, adjust to pH 3.0 with hydrochloric acid,
Shaking extraction was performed to obtain a chloroform layer. The chloroform layer was washed with water, dehydrated with anhydrous sodium sulfate, and concentrated to dryness to obtain 196 mg of a MI215-NF3 substance in the form of a free carboxylic acid.

Example 4 20 mg of MI215-NF3 substance was dissolved in 1 ml of acetone. To this was added 0.3 ml of a 2.5% suspension of aluminum hydroxide, and after reaction at room temperature for 18 hours, 100 ml of water was added, and the mixture was extracted twice with 100 ml of chloroform. The chloroform layer is washed with water,
After the dehydration operation, the solution was concentrated to dryness to obtain 19.8 mg of an aluminum salt of MI215-NF3 substance.

(Effect of the Invention) The novel antibiotic MI215-NF3 substance of the present invention or a salt thereof is useful as an agent for controlling coccidiosis in chickens. And the types of chickens that can be treated are not limited to specific ones. Further, the antibiotic MI215-NF3 substance or a salt thereof according to the present invention is more effective at a small dose compared to conventionally used antibiotics for treating coccidium such as monensin or salinomycin.

[Brief description of the drawings]

FIG. 1 is an infrared absorption spectrum of the sodium salt of MI215-NF3 substance measured by the KBr tablet method. Fig. 2 shows MI215
-It is a proton nuclear magnetic resonance spectrum at 400 MHz measured at room temperature in deuterated chloroform of sodium salt of NF3 substance.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI C12R 1:03) (C12P 17/18 C12R 1:03) (72) Inventor Masaru Hamada 26-1 Naitocho, Shinjuku-ku, Tokyo Hidekazu, Tokyo Residence No. 405 (72) Inventor Hiroshi Naganawa 3-17, Denon Chofuhoncho, Ota-ku, Tokyo (72) Inventor Seiji Sato 200-96 Higashi-Tahara, Hadano-shi, Kanagawa Prefecture (58) Fields investigated (Int. Cl. ⁶ , DB name) C07D 493/10 A61K 31/35 C12P 17/00-17/18 C12N 1/20 WPI (DIALOG) REGISTRY (STN) CA (STN)

Claims (5)

(57) [Claims] 1. The following formula (I) (Wherein R is a hydrogen atom)
-NF3 substances and salts of MI215-NF3 substances wherein R in formula (I) is a metal or ammonium group. 2. The salt of the MI215-NF3 substance according to claim 1, wherein R in the formula (I) is an alkali metal, an alkaline earth metal or another metal or an ammonium group. 3. A method for culturing a microorganism producing the MI215-NF3 substance of the formula (I) or a salt thereof according to claim 1, which belongs to Actinomadura, and collecting the MI215-NF3 substance or a salt thereof from the culture. A method for producing the antibiotic MI215-NF3 or a salt thereof. 4. An agent for controlling coccidiosis in chickens, comprising the MI215-NF3 substance of formula (I) or a salt thereof according to claim 1 as an active ingredient. 5. An actinomadura sp. MI215-NF having a property of producing the MI215-NF3 substance of the formula (I) according to claim 1.
3 shares.