JPH10139771A - New compound am6927 and its production - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound, produced by a specific microorganism belonging to the genus Metarhizium, capable of manifesting specifically inhibiting actions on the production of IgE antibodies and useful as a therapeutic and preventing agent for atopic diseases, pollinosis, allergic rhinits, etc. SOLUTION: This new compound is 7-oxa-3-oxo-1-hydroxy-1-(2,3-epoxy-7- hydroxy 6-methyl-5-hepten-2-yl)-2-methoxyspiro[5.2]octane represented by the formula. The compound represented by the formula is obtained by culturing a microorganism, having the ability to produce the compound and belonging to the genus Metarhizium, e.g. Metarhizium anisopliae var. anisopliae M6927 strain (FERM P-15942) and collecting the resultant compound from the cultured product. A medicine comprising the compound as an active ingredient can usually be administered to an adult in 20-1,000mg daily dose expressed in terms of the amount of the active ingredient.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel compound having an inhibitory effect on IgE antibody production, its production method and use, and a producing bacterium.

[0002]

2. Description of the Related Art Allergic reactions are classified into type I allergic reactions, type II allergic reactions, type III allergic reactions and type IV allergic reactions, depending on the mechanism of onset and the difference in symptoms. A typical allergic reaction is a type I allergic reaction which is also called an immediate allergic reaction. In the type I allergic reaction, IgE is generated by an antigen that has entered the living body, and the IgE antibody fixed to the cell membrane of mast cells in tissues or basophil cells in blood reacts with the antigen to produce the cells. Releases histamine, serotonin, and various other chemical mediators. The chemical mediators cause various tissue reactions such as contraction of smooth muscle and enhancement of vascular permeability. The pathological conditions of type I allergy include atopic bronchial asthma, atopy
Dermatitis, allergic rhinitis, hay fever, anaphylaxis, food allergies and the like.

[0003] At present, as agents for suppressing the type I allergic reaction, drugs for inhibiting the release of chemical messengers from mast cells and basophils and antagonists for chemical messengers have been developed. However, there are few known chemical mediators that specifically inhibit the release or production of chemical mediators or specific antagonists, and there are a wide variety of chemical mediators. At present, it is necessary to analyze the interaction of various chemical mediators in order to suppress the activity. Furthermore, the above agents do not suppress the production of IgE, the underlying cause of the allergic reaction, and cannot be a fundamental therapeutic agent in that sense.

Attempts have also been made to suppress IgE production with an immunosuppressant such as cyclosporin A to treat allergic diseases. However, non-specific immunosuppression is accompanied by the problem of side effects. Is difficult to overcome.

[0005]

There has been a demand for providing novel compounds useful as immunosuppressants.

[0006]

Means for Solving the Problems The present inventors have conducted intensive searches to find novel biologically active substances that specifically inhibit IgE antibody production from microbial metabolites. as a result,
Metarhiz isolated from forest soil in Okinawa
It has been found that a novel physiologically active substance having an inhibitory activity on IgE antibody production is produced from a culture solution of strain M6927 belonging to the genus ium), and thus the present invention has been completed.

That is, the present invention provides a compound of the formula (I)

[0008]

Embedded image

(Hereinafter, the compound of the present invention may be referred to as AM6927 or AM6927 compound). The properties of the AM6927 compound of the present invention are as follows. 1) Appearance: colorless needle-like crystal 2) Distinguishing between acidic, basic and neutral: neutral substance 3) Specific rotation: [α] _D ²⁴ -68.4 ° (C = 1.0,
(MeOH) 4) Molecular formula: C ₁₆ H ₂₄ O ₆ 5) Molecular weight: FAB-MS m / z 313 (M + H)
⁺ 6) Elemental analysis: Calculated (%) (C ₁₆ H ₂₄ to _{O 6) C, 61.52; H} , 7.74 Found (%) C, 61.61; H , 7.86 7) UV absorption spectrum: terminal absorption (MeOH) 8) Infrared absorption spectrum: (KBr, cm ^-1 ) As shown in FIG.

The main absorptions are as follows. 3475, 2
971, 2935, 2875, 1727, 1432, 1
384, 1288, 1218, 1103, 1027, 9
60,927 9) ¹ H-NMR spectrum: (400 MHz, CDC
l ₃ ) As shown in FIG.

10) ¹³ C-NMR spectrum: (100
MHz, CDCl ₃ ) δ (ppm) 207.2 (s), 138.9 (s), 119.6 (s)
(D), 86.7 (d), 79.2 (s), 68.8
(T), 61.2 (s), 61.0 (s), 59.9
(Q), 57.1 (d), 52.0 (t), 37.2
(T), 30.8 (t), 27.1 (t), 15.0
(Q), 14.6 (q) 11) Solubility: chloroform, ethyl acetate, acetone,
It is soluble in methanol and insoluble in water.

12) Color test: Positive for iodine vapor reaction and potassium permanganate decolorization reaction, negative for ferric chloride reaction and ninhydrin reaction. 13) Thin layer chromatography: stationary phase developing solvent Rf value silica gel f S201 Hexane: acetone 0.58 (manufactured by Tokyo Kasei) (1: 1) 14) High performance liquid chromatography: separation column: Shodex C18-5A (column size) φ4.5 × 150 mm, manufactured by Showa Denko KK Solvent: 18% acetonitrile Flow rate: 1.0 ml / min Detection: UV 205 nm Retention time: 11.6 minutes Obaricin having an immunosuppressive action as having a similar structure to the compound of the present invention [ Helv. Chim. Acta, 51: 1395
(1968)] and a PF1131 compound [JP-A-7-1795]
No. 7], fumagillin having antiprotozoal activity [Science,
113: 202 (1951)], FR-65 having an immunosuppressive action
814 [JP-A-61-33181], 1771A and B having antitumor activity [JP-A-1-233275]
No. WF2015A and B having angiogenesis inhibitory action [JP-A-2-233610], FR125
No. 035 (JP-A-3-47120) and derivatives of fumagillol, which is a hydrolyzate of fumagillin (JP-A-3-7222 and JP-A-3-279376), are known. However, the compounds of the present invention differ in chemical structure from these known compounds, and the present invention is considered to be novel compounds.

[0013] The present invention is also a method for producing AM6927, which comprises culturing a microorganism belonging to the genus Metallidium having the ability to produce an AM6927 compound, and collecting the AM6927 compound from the culture. The microorganism belonging to the genus Metallidium having the ability to produce the AM6927 compound of the present invention is not particularly limited as long as it has the ability to produce the compound of the present invention. . anisopliae) M
The 6927 strain (FERM P-15942) is exemplified.

The M6927 strain is a filamentous fungus isolated from forest soil in Okinawa Prefecture, and its bacteriological properties are as follows. 1. Growth state on each medium 1) East-phosphate-soluble starch (YpSs) agar medium The colony reached a diameter of 14 mm or more after culturing at 25 ° C for 7 days, and was white (1A1) or yellow on the medium. White (1A
2) It becomes a felt-like colony. The back of the colony is light yellow (2A5). After culturing at 25 ° C. for 20 days, the diameter of the colony reached 32 mm, the surface was felt-like, and the color was white (1A1) to yellow-white (1A).
2). The back of the colony is light yellow (2A
5). No growth of colonies at 37 ° C. is observed.

2) Potato-glucose agar medium Cultured at 25 ° C. for 7 days, the diameter of the colony reached 17 mm or more, and the surface was partially raised in a felt-like shape with greenish yellow (1A7) or vivid. Yellow (3A8). The back surface of the colony is (2A3) to (2A4). When cultured at 25 ° C. for 20 days, the colonies reached a diameter of 50 mm or more, and were colored greenish yellow (1A7) to vivid yellow (3A).
8). The color of the back surface is (2A3) to (2A4). No growth of colonies at 37 ° C. is observed.

2. Physiological properties When cultivated in a potato-glucose liquid medium, the range of pH at which the cells can grow is 2.0 to 12.0.
H is 5.0 to 8.0. When cultivated on a potato glucose agar medium, the temperature range at which the cells can grow is 14 to
39 ° C, and the optimal growth temperature is 20 to 25 ° C.

3. Morphological characteristics under the microscope The bacterium does not form migratory spores or sporangia. Only the asexual generation is formed, not the sexual generation. The fungus forms an olive-colored (1F6) densely converged columnar conidial mass. The mode of conidium formation is of the phiaro type. The phialide is cylindrical and has a narrowed tip. Conidiophores are complex and densely branched. Conidia are single cells and cylindrical, colorless or slightly colored, and are formed in a chain from phialides.

4. Identification This bacterium belongs to the incomplete fungi because it forms only the asexual generation and not the sexual generation. In addition, formation of an olive-colored (1F6) densely converged columnar conidium mass is observed in the fungus. Due to these characteristics, this bacterium is known as Metahizhi
It belongs to the genus um. The genus Metahizium is currently M.
flavoviride and M.A. anisopliae is known ^{1) In} particular, M. anisopliae depends on the size of conidia. anisopliae var.
major and M.M. anisopliae var. a
nispriae. ²⁾ The conidia of this bacterium are cylindrical and have a size of 2-3 × 6-7 μm. Good agreement with the characteristics of anisopliae. Therefore, this strain was transformed into Metarhidium anisoliae varaetas anisopriae (Metatarhizium).
anisopriavar. anisoplia
e) Called M6927. In addition, this strain is FERM P-15 by the National Institute of Biotechnology and Industrial Technology.
No. 942.

The color of the growth state in each medium is indicated by K
ornerup A. and Wanscher J .;
H. 1978 [Methuen Handbook
ofColor 3rd ed. ] Eye Meth
uen, London. References 1). Hawksworth, D.W. L. , Kirk,
P. M. Sutton, B .; C. and Pegle
r, D. N. 1995. “Ainsworth & Bi
sby's Dictionary of the F
ungi ", 8th ed., pp. 377. CAB
International, UK. 2). Tulloch, M .; 1976. Thegen
us Metalhizium. Trans. Br.
Mycol. Soc. 66: 407-411. A typical and preferable method for producing AM6927 according to the present invention is a method for producing the compound belonging to the genus Metallidium (for example, Metalhidium anisoploiae v.).
ar. anisopliae M6927) is cultured in a suitable medium, and the isolated culture is used as an example.

The medium used for producing the substance of the present invention is most preferably, but not limited to, shaking culture or aeration-agitation culture using a liquid medium. The medium is AM692
No particular limitation is imposed as long as the 7 producing bacteria grow and accumulate the compound in the medium. Examples of carbon sources include glucose, sucrose, dextrin, starch, glycerin, molasses, and organic acids. Etc. can be used. Examples of the nitrogen source include yeast extract, peptone, meat extract, soybean powder, cottonseed powder, gluten meal, wheat germ, corn plica, amino acids, ammonium salts, nitrates, and various other organic or inorganic substances. Nitrogen compounds are used. As the inorganic salt, sodium chloride, potassium chloride, calcium carbonate, various phosphates, magnesium salts, copper salts, cobalt salts and the like may be added. In addition, growth of bacteria and AM692
7 Vitamins, coenzymes, and the like that promote production may be added. In particular, if the medium foams strongly, liquid paraffin, animal oil, vegetable oil, mineral oil, silicon, etc. may be added when necessary.

Conditions such as culturing temperature, culturing time, stirring speed, aeration rate, and pH of the culture solution in culturing AM6927-producing bacteria are appropriately selected and adjusted so that the accumulated amount of AM6927 is maximized. For example, in the case of ordinary aeration and stirring culture, culture is preferably performed at a culture temperature of 27 to 30 ° C. for 2 to 5 days, and the pH of the culture solution is preferably adjusted to pH 5.0 to 8.0.

The time-dependent change in the amount of AM6927 accumulated in the culture solution over the course of the culture can be measured by the below-described production inhibitory activity or reverse phase HPLC. Usually, the production reaches its maximum in culture for 48 to 96 hours. After completion of the culturing, AM6927 accumulated mainly in the liquid portion of the cells is preferably removed by filtration or centrifugation of the cells and other solid parts and separated from the filtrate or supernatant. The compound can be separated from the culture solution without removing the compound. Various methods based on its physicochemical properties can be used for separating and purifying AM6927 from the culture solution. For example, AM692 present in the filtrate or supernatant
7 can be extracted and purified by an organic solvent immiscible with water under neutral pH conditions, for example, ethyl acetate, butyl acetate, butanol, chloroform, dichloromethane, methylene chloride, ethylene chloride, etc., alone or in combination. . Alternatively, for example, Diaion HP-20 (manufactured by Mitsubishi Kasei) or the like is used as an adsorbent. AM69
27 is passed through the adsorbent layer to remove impurities by adsorbing impurities, or after adsorbing AM6927, eluting with methanol water, acetone water, or the like to elute the compound. Can be obtained. Further, adsorption column chromatography using a carrier such as silica gel, alumina, and florisil, Sephadex LH-
Purification of AM6927 by partition column chromatography using No. 20 (manufactured by Pharmacia), Toyopar HW-40 (manufactured by Tohso), and high performance liquid chromatography using normal phase and reverse phase columns. Can be.

[0023] The present invention is also a medicament comprising an AM6927 compound as an active ingredient. The medicament of the present invention is usually used for AM
In addition to the 6927 compounds, it can be comprised of a pharmaceutically acceptable carrier. As the carrier, known carriers can be used, and depending on their properties, they can be classified as excipients, binders, disintegrants, lubricants, flavoring agents, solubilizing agents, suspending agents, coating agents, and the like. Among them, for example, lactose, corn starch, magnesium stearate and the like can be mentioned.

The medicament of the present invention is, for example, an immunosuppressant, and more specifically, an IgE production inhibitor or I
a preventive or therapeutic agent for an immune disease based on abnormal production of gE,
Examples thereof include anti-atopic bronchial asthma agents, anti-atopic dermatitis agents, anti-allergic rhinitis agents, anti-hay fever agents, anti-anaphylaxis agents or anti-food allergic agents. Although the medicament of the present invention can be administered in various forms, for example, oral administration by tablets, capsules, granules, syrups and the like or injections (intravenous, intramuscular, subcutaneous), eye drops, suppositories, ointments, Parenteral administration by spray, lotion and the like can be mentioned.

These medicaments vary depending on symptoms, age, body weight, administration method, dosage form and the like, but usually 20 mg to 1000 mg per day can be administered to an adult.

[0026]

Next, the present invention will be described more specifically with reference to examples, but the present invention is not limited to these examples. Example 1 (1) Fermentative production of AM6927 compound Glucose 2%, soluble starch 2%, soybean powder 2%, yeast extract 0.5%, NaCl 0.25% and CaCO 2
₃ Aqueous medium (pH 6.5) 100 containing 0.35%
was dispensed into a 500 ml Erlenmeyer flask and sterilized at 120 ° C. for 20 minutes. The M6927 strain (FERM P
-15942) was inoculated with one platinum loop and cultured at 28 ° C. for 3 days on a rotary shaker (200 rpm / min) to obtain a primary seed culture. The resulting primary seed culture was inoculated into 20 l of the same sterile medium described above in a 30 l jar fermenter. The fermenter was aerated at 20 l / min and aerated with stirring at 200 rpm and 28 ° C. for 48 hours to obtain a secondary seed culture solution.

4 l of the secondary seed culture thus obtained was further added to
Glucose 2%, peptone 1%, C.I. S. L. 1%, K
H ₂ PO ₄ 0.2%, MgSO ₄ .7H ₂ O 0.1% and antiform FS-028 (Dow Corni
ng K. 200 l of sterile medium in a 300 l stainless steel fermenter containing 0.03% (K company). The culture was performed at 28 ° C. for 72 hours under aeration at 280 l / min and stirring at 300 rpm.

(2) Purification of AM6927 Compound 200 ml of broth obtained by the above culture method was added to diatomaceous earth (5
kg) and filtered to obtain a filtrate (180 l). After the obtained filtrate was adjusted to pH 7.0 with hydrochloric acid, ethyl acetate (90 l) was added to perform an extraction operation. The ethyl acetate layer was concentrated under reduced pressure to obtain about 100 ml of a syrup. About 1 l of benzene was added to this syrup-like substance, insolubles were removed by filtration, and the filtrate was concentrated under reduced pressure. The concentrate was applied to a silica gel column (1900 ml) previously prepared with a hexane-acetone (10: 6) mixed solvent, eluted with the same mixed solvent, and fractionated by 150 ml. For detection of the active substance in the purification process, silica gel thin layer chromatography [TLC: silica gel f, manufactured by Tokyo Chemical Industry Co., Ltd.]
S201; developing system: benzene-ethyl acetate = 2: 10
The substance having an Rf value of 0.50 was used.

Fraction No. 31-No. At 49, an AM6927 component mainly showing an Rf value of 0.50 was eluted. Collect the fraction containing the AM6927 component,
It was concentrated under reduced pressure. The concentrate containing the AM6927 component was further applied to a silica gel column (330 ml) and eluted with chloroform-methanol (10: 0.2) to give 18 m
Each fraction was separated. Fraction No. 52-No. 57
Eluted the AM6927 component. This fraction was collected, concentrated under reduced pressure, and dissolved in about 1 ml of chloroform.
When about 10 ml of ethyl ether is added and left at room temperature, AM
6927 was obtained as needle-like crystals. The crystals were collected on a glass filter by filtration and dried to obtain purified colorless needle-shaped crystals AM6927 (248 mg). Example 2 (1) IgE Antibody Production Inhibitory Activity of AM6927 IgE antibody production inhibitory activity of AM6927 obtained in Example 1 was measured as follows, and the activity was confirmed.

The spleen of a 6-week-old female BALB / c mouse was aseptically removed and passed through a stainless steel mesh to remove large cell clumps, and then 10% fetal bovine serum (Hyclon).
e), 2 mM glutamine, 5 × 10 ⁻⁵ M2-mercaptoethanol, 0.2% NaHCO ₃ , penicillin G
(50 IU / ml) and streptomycin (50 μg)
/ Ml) containing RPMI-1640 medium (GIBCO
(BRL) to obtain a spleen cell suspension. After centrifuging this solution at 1200 rpm for 10 minutes, 140 mM N
Erythrocytes are destroyed with H ₄ Cl-15 mM Tris solution, and PB
S (-) (2.7 mM KCl, 1.5 mM KH ₂ P
O ₄ , 0.14 M NaCl, 8.1 mM Na ₂ HPO ₄
・ After washing twice with a solution (containing 7H ₂ O), the anti-T
T cells were killed using the hy-1 antibody and rabbit complement and used as a B cell source. This B cell source was washed twice with a PBS (−) solution, and then washed with the above RPMI-1640 medium at 5 × 10 6 cells.
^It was adjusted to ⁶ cells / ml to obtain a B cell suspension. LPS (Difco) was added to this B cell suspension as a B cell stimulant.
Company; (derived from E. coli) at a concentration of 50 μg / ml, and precultured for 24 hours at 37 ° C. under 5% CO ₂ . Thereafter, the cells were collected and washed twice with a PBS (-) solution. The cells were again cultured in the above RPMI-1640 medium for 5 minutes.
x10 ⁶ cells / ml, and 100 μl of this cell suspension
was placed in each well of a 96-well plate (5 × 10 ⁵ cells / well), and 5 μg of the above LPS and 100 U of commercially available mouse IL-4 (Genzyme) were added thereto. Then, 50 μl of AM6927 compound at various concentrations was added and 2
After culturing at 37 ° C. for 7 days using a 5% CO ₂ incubator at 00 μl / well, the amount of IgE in the medium was measured. Since the AM6927 compound was insoluble in water, it was dissolved in DMSO and then diluted with the above medium. The above operation was repeated except that no AM6927 compound was added as a control.

The amount of IgE in the culture supernatant was measured by the following method. Commercially available rat anti-mouse IgE as primary antibody
2 µg / m of monoclonal antibody (Yamasa Shoyu Co., Ltd.)
and dissolved in a PBS (-) solution to obtain a 96-well EL.
The mixture was dispensed into an ISA plate (Dynatech) at a rate of 50 μl / well and kept at room temperature for 1.5 hours. Thereafter, the PBS (−) solution was discarded, and each well was washed by adding a PBS (−) solution containing 0.05% Tween 20 at a rate of 300 μl / well. This washing operation is performed 4 times.
Repeated times. After washing, a blocking solution [1% BSA (Miles; fr;
actionV) -containing PBS (-) solution]
For one day and night to perform blocking. The blocking solution was then discarded and the previously described 0.05% Tween
The washing operation with a PBS (-) solution containing 20 was repeated four times. Next, a series of sample diluents for activity measurement prepared in advance are added at a rate of 50 μl / well, and 1
Hold for hours. PBS containing 0.05% Tween 20
(−) The washing operation with the solution was repeated four times. Next, a commercially available biotin-labeled rat anti-mouse IgE monoclonal antibody (Yamasa Shoyu Co., Ltd.) was used as a secondary antibody at 2 μg / ml.
Was dissolved in a PBS (-) solution, added at 50 μl / well, and kept at room temperature for 1 hour. 0.05% Tween
After the washing operation with the n (20) -containing PBS (−) solution was repeated four times, the Horderadish Peroxidase diluted 2000-fold with the PBS (−) solution was used.
5 Avidin D (Vector, A2004)
0 μl / well was added and kept at room temperature for 1 hour. 0.05
After the washing operation with a PBS (-) solution containing 20% Tween 20 was repeated four times, an ABTS solution [0.129 g citric acid; 0.276 g Na ₂ HPO ₄ .12H ₂ O;
% H ₂ O ₂ 1 μl; 3 mg 2,2-Azino-bis
(3-ethylbenzothiazoline-6
-Sulfonic acid) diamonium
salt (Wako Pure Chemical Industries, Ltd .; 10 ml distilled water)
Add 0 μl / well, keep at room temperature for 10 minutes, and add 5% (w
(V) 50 μl / well of oxalic acid was added to stop the reaction. Then, the IgE amount was measured by measuring the absorbance at 415 nm.

The IgE antibody production inhibitory activity is determined by measuring the concentration at which the amount of IgE antibody produced is reduced to 50% of the control by the IC.
The activity of AM6927 was calculated as ₅₀ (μg / ml). (2) Cytotoxicity of AM6927 compound In this study, mouse lymphoma P388D1 cells (AT
CC CCL-46) was used. Cell viability, Mic
hael C, Alley et al. [Cancer R
es. , 48 , 589-601 (1988)].
MTT [3- (4,5-dimethylthiazole
l-2-yl) -2,5-diphenyltetra
[Zolium bromide] reagent. That is, 10% fetal calf serum (Hyclone)
2 mM glutamine, 5 μM 2-mercaptoethane-l, 0.2% sodium bicarbonate, penicillin G (5
0 IU / ml) and streptomycin (50 μg / m
l) containing RPMI 1640 medium (GIBCO
BRL) to convert mouse lymphoma P388D1 cells to 1x
It was prepared in 10 ⁵ cells / ml. This cell suspension 100μ
1 was placed in each well of a 96-well plate (1 × 10 ⁴ cells / well). 100 μl / well of various concentrations of AM6927 compound solutions prepared using the above medium were added thereto, and the cells were cultured at 37 ° C. for 3 days using a 5% CO ₂ incubator. Thereafter, the above MTT reagent was dissolved in a PBS (-) solution to a concentration of 1 mg / ml, 50 µl was added to each well, and the cells were cultured at 37 ° C for 4 hours using a 5% CO ₂ incubator. After completion of the culture, the blue dye produced by the living cells was measured at 570 nm. In addition, AM692
Since 7 compounds are insoluble in water, after dissolving in DMSO,
Diluted with the above medium. AM692 as control
The above operation was repeated except that 7 compounds were not added. The concentration at which the cell number was reduced to 50% of the control was
The IC ₅₀ value of AM6927 was measured as C ₅₀ (μg / ml). Table 1 shows the above results.

[0033]

[Table 1]

As is apparent from the above results, AM69
27 compounds were compounds with low cytotoxicity, and were confirmed to have IgE antibody production inhibitory action at low concentrations. Therefore, the compound of the present invention is an immunosuppressant,
It is a prophylactic / therapeutic agent for an immune disease based on abnormal production of IgE, for example, an anti-atopic bronchial asthma agent, an anti-atopic dermatitis agent, an anti-allergic rhinitis agent, an anti-hay fever agent, an anti-anaphylaxis agent or It was confirmed that it could be used as an anti-food allergic agent.

(3) Acute toxicity of AM6927 compound After dissolving AM6927 compound in DMSO, it was diluted with a physiological saline solution and 30 mg / kg was administered once a day for 5 days.
Five CR mice (female, 5 weeks old) were intraperitoneally administered.
As a result, no abnormal symptoms were observed in the mice, and it was confirmed that the present compound was highly safe. Example 3 Oral Capsules 30 mg of AM6927 produced in Example 1 was mixed well with 170 mg of lactose, 140 mg of Cornstarch, and 2 mg of magnesium stearate.
mg was placed in a gelatin capsule to make a capsule.

[0036]

Industrial Applicability The present invention provides novel compounds, and these compounds do not show general immunosuppressive action but show excellent IgE antibody production inhibitory action. Therefore, in particular, diseases caused by abnormal production of IgE, such as atopic bronchial asthma, atopic dermatitis, allergic disease
It is very useful as a therapeutic or prophylactic agent for rhinitis, hay fever, anaphylaxis, food allergies and the like, or a material for conversion to them.

[Brief description of the drawings]

FIG. 1 shows the infrared absorption spectrum of AM6927 in a potassium bromide tablet.

FIG. 2 shows a 400 MHz ¹ H-NMR spectrum of AM6927 in a heavy chloroform solution.

──────────────────────────────────────────────────続 き Continued on the front page (51) Int.Cl. ⁶ Identification symbol FI // (C12N 1/14 C12R 1: 645) (C12P 17/16 C12R 1: 645)

Claims (5)

[Claims] 1. A compound of the formula (I) A compound represented by the formula: 2. A method for producing a compound according to claim 1, wherein a microorganism belonging to the genus Metallidium having the ability to produce the compound according to claim 1 is cultured, and the compound is collected from the culture. 3. A pharmaceutical comprising the compound according to claim 1 as an active ingredient. 4. The medicament according to claim 3, wherein the medicament is an immunosuppressant, an IgE production inhibitor, or an agent for preventing or treating an immune disease based on abnormal production of IgE. 5. The strain of metallisium anisopriae varaetus anisopriae M6927 (FERM P-1)
5942).