JP2661367B2 - WF11243 substance - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a WF11243 substance or a salt thereof, a production method thereof, and a use thereof. WF11243 substance is a conventionally unknown novel substance isolated and collected from microorganisms, especially mold cultures, and has excellent antifungal and antiprotozoal properties,
Reduction or prevention of various harmful effects derived from fungi and protozoa,
It is useful as various drugs such as agents for prevention and treatment of various diseases, and is also very useful for prevention and treatment of pneumocystis carinii.

Therefore, the present invention plays an important role in various technical fields such as pharmaceuticals, cosmetics, and foods and beverages.

2. Description of the Related Art Conventionally, various antifungal agents have been produced from cultures of microorganisms or by chemical synthesis. Although some of these antifungal agents are excellent, there are various problems such as the emergence of resistant bacteria, safety problems, etc., in addition to their medicinal properties. At present, there is no antifungal agent to be obtained.

In addition, recently, diseases caused by protozoa, particularly various diseases caused by Pneumocystis carinii, for example, carinii pneumonia, have been highlighted, and there is a strong demand in the art for the development of prevention and treatment methods therefor. .

Problems to be Solved by the Invention The present invention has been made in view of the above-mentioned state of the art, and inhibits the growth of Pneumocystis carini and other protozoa that are pathogenic microparticles that cause carinii pneumonia and other diseases, In addition to new antiprotozoal agents that are effective in killing,
The purpose of the present invention is to develop a new and unknown new antifungal agent which is effective in inhibiting the growth and killing of various fungi.

Means for solving the problems As a result of examining from various directions to achieve the above objectives,
The present inventors have focused on natural products from the aspect of safety, and have come to focus on fermentation products of microorganisms, and as a result of searching for various microorganisms, newly separated from defoliation samples collected in Kyoto Prefecture and Ayabe City It was discovered that the fungus No. 11243 accumulated the target substance in the culture solution. Further detailed studies of the physicochemical properties of this substance confirmed that it was a novel substance that was previously unknown, and newly added this substance to WF11243.
Substance, and the hydrochloride salt of WF11243 substance is FR901469
The substance was named and further research resulted in the establishment of an industrial process and completed the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS FIG. 1 is a drawing showing ¹³ C nuclear magnetic resonance spectrum of FR901469 substance.

FIG. 2 is a drawing showing ¹ H nuclear magnetic resonance spectrum of FR901469 substance.

The WF11243 substance according to the present invention is a novel substance whose hydrochloride FR901469 substance has the following physicochemical properties.

Physicochemical properties of FR901469 substance (1) (1) Color and state of substance White powder (2) Melting point 182 to 187 ° C (3) Specific rotation ▲ [α] ²³ _D ▼ + 29 ° (c = 1.5, methanol) ( 4) Molecular formula C ₇₁ H ₁₁₆ N ₁₄ O ₂₃ · HC1 (5) Elemental analysis Calculated value (%) (for C ₇₁ H ₁₁₆ N ₁₄ O ₂₃ · HC1 · 8H ₂ O):
C, 49.74; H, 7.82; N, 11.44 Observed value (%): C, 49.65; H, 7.72; N, 11.40 (6) Solubility in solvent Easily soluble: methanol, poorly soluble in water: acetone insoluble: n- Hexane (7) Color reaction Positive: iodine vapor reaction, cerium sulfate reaction, ninhydrin reaction, negative: Maurish reaction, Ehrlich reaction (8) Thin layer chromatography (9) Infrared absorption spectrum (FT-IR (KBr)) Significant peaks are as follows.

(10) High Performance Liquid Chromatography Column: YMC Pack ODS-AM (AM-303, S-5, 120A, 4.6m
mID × 250mm; YMC CO., Ltd) Detection: 210nm Developing solvent: 45% acetonitrile aqueous solution containing 0.5% NH ₄ H ₂ PO ₄ Flow rate: 1ml / min Retention time (RT): 10.9min (11) ¹³ C nucleus Magnetic resonance spectrum (100 MHz, CD ₃ OD) The chart is shown in FIG.

δc: 176.1 (s), 174.6 (s), 174.0 (s), 173.8
(S), 173.4 (s), 172.8 (s), 172.6 (s), 172.5 (s), 172.4
(S), 172.1 (s), 171.9 (s), 171.8 (s), 171.7 (s), 171.2
(S), 156.1 (s), 131.4 (d) x 2,128.6 (s), 117.9 (d) x 2,74.2
(D), 72.0 (d), 70.5 (d), 70.1 (d), 70.0 (d), 69.1 (d), 6
8.8 (d), 68.3 (d), 67.7 (d), 61.4 (d), 60.9 (d), 60.3 (d), 5
9.8 (d), 58.2 (d), 57.9 (t), 57.6 (d), 57.2 (d), 56.8 (d), 5
1.7 (d), 50.8 (d), 46.4 (t), 43.1 (t), 42.9 (t), 40.4 (t), 3
9.9 (t), 38.9 (t), 37.6 (t), 35.8 (t), 34.7 (t), 33.1 (t), 3
1.6 (d), 30.8 (t), 30.7 (t), 30.7 (t), 30.7 (t), 30.6 (t), 3
0.5 (t), 30.4 (t), 30.3 (t), 28.1 (t), 25.9 (t), 24.9 (t), 2
3.7 (t), 21.2 (q), 20.9 (q), 20.3 (q), 19.2 (q), 18.9 (q), 1
8.7 (q), 17.2 (q), 14.4 (q).

(12) ¹ H nuclear magnetic resonance spectrum (400 MHz, CD ₃ OD) The chart is shown in FIG.

δH: 7.08 (2H, d, J = 8 Hz), 6.98 (2H, d, J = 8 Hz), 5.18 to 5.09 (2H, m), 4.88 (1H, brs), 4.76 to 4.69
(3H, m), 4.65 (1H, m), 4.48 (1H, m), 4.46 (1H, d, J = 5H
z), 4.41 (1H, d, J = 9 Hz), 4.36 to 4.27 (3H, m), 4.24 to
4.13 (6H, m), 3.98 (1H, m), 3.92 to 3.78 (3H, m), 3.70 (1H, dd, J = 11 and 4Hz), 3.58 (1H, brd, J = 1
1Hz), 3.00-2.93 (3H, m), 2.80 (1H, dd, J = 13 and 11H
z), 2.56 (1H, dd, J = 13 and 11Hz), 2.45 (1H, dd, J = 14 and 9Hz), 2.37 (1H, br d, J = 1
3Hz), 2.30 (1H, m), 2.21 to 2.05 (3H, m), 2.03 to 1.93 (2
H, m), 1.89 (1H, m), 1.77 (1H, m), 1.62-1.50 (4H, m), 1.39 (3H, d, J = 6Hz), 1.35 (3H, d, J = 6.5Hz) , 1.35 (3H, d, J = 6.5 Hz), 1.30 (3H, d, J = 6 Hz), 1.28 (22 H, m), 1.13 (3H, d, J = 6 Hz), 0.89 (3H, t, J
= 7Hz), 0.81 (3H, d, J = 6.5Hz), 0.80 (3H, d, J = 6.5Hz).

(13) Molecular formula and FAB-MS C ₇₁ H ₁₁₆ N ₁₄ O ₂₃ · HC1 FAB-MS m / z 1533 (M + H) ⁺ ; HRFAB-MS m / z 1555.8240 (Calculated value of C ₇₁ H ₁₁₆ N ₁₄ O ₂₃ Na 1555.8235) (14) Amino acid analysis Thr (4), Gly (1), Ala (1), Val (1), Tyr
(1), Orn (1) (6N HC1, 110 ° C, hydrolysis for 20 hours).

(15) Ultraviolet absorption spectrum Significant peaks are as follows.

Further, the WF11243 substance according to the present invention shows the properties of the polypeptide in view of the above-mentioned physicochemical properties, but succeeded in determining its structure, and succeeded in determining the structure, and the putative chemical structure represented by the following formula [I] I got the formula.

The WF11243 substance according to the present invention is produced by, for example, a microorganism No. 11243 strain newly isolated from the defoliation sample collected by the present inventors in Ayabe City, Kyoto Prefecture, and is also produced by a chemical synthesis method such as peptide synthesis. be able to.

This No. 11243 strain spreads suppressively on various media,
It forms a pale orange settlement. The bacteriological properties of No. 11243 strain are shown below.

The properties of the culture on various media are shown in Table 1 below. When inoculated at the center of a malt extraction agar medium and cultured at 25 ° C for 14 days, the growth was extremely suppressed and spread to 0.5 to 1.0 cm in diameter.
The surface of the settlement was raised and pale orange. It also produced a colorless viscous leachate. The backside of the village was gray orange. An anamorph resulted. When the same culture was performed on a potato dextrose agar medium, growth was extremely suppressed. (0.5 / 1.0cm). The settlement surface was raised, producing radial grooves and pale orange. It also produced a colorless viscous leachate. The back of the settlement was pale yellow. An anamorph resulted.

The characteristics described above were observed after culturing at 25 ° C. for 14 days. The description of the color tone was made based on the Methuen Handbook of Color (Reference (1)).

-References- (1) A corner wrap and JH
Wonthshire, Metune Handbook of
Color, 3rd edition, Metune, London, 1983 (A.Ko
rnerup and JHWanscher: Methuen Handbook of Colou
r, Third ed., Methuen, London, 1983) No.11243 strain can grow at 7 ~ 29 ℃, and the optimal growth temperature is
22-26 ° C. (Measured on potato dextrose agar).

As a result of comprehensive examination of these properties, this No. 1124
The three strains were identified as belonging to fungi and named No. 11243 strain, which was deposited with the Research Institute of Microbial Industry and Technology, Ministry of International Trade and Industry (Accession No. FERM BP-3373, deposited on April 2, 1991).
3 days).

It is to be understood that the production of the WF11243 material is not limited to the use of the particular microorganisms described herein for illustrative purposes only. The present invention provides X-ray irradiation, ultraviolet irradiation, N-methyl-
It also encompasses the use of all mutants capable of producing WF11243, including artificial mutants and natural mutants, which can be obtained by mutation treatment of N'-nitro-N-nitrosoguanidine, 2-aminopurine, etc. .

The WF11243 substance according to the present invention is inoculated into a nutrient medium containing a carbon and nitrogen source capable of assimilating the fungus producing the substance belonging to the fungus (eg, strain No. 11243) and cultured under aerobic conditions (for example, , Shaking culture, aeration stirring culture, etc.).

As the carbon source, it is preferable to use glucose, sucrose, starch, modified starch, fructose, glycerin and other carbohydrates.

Nitrogen sources include oatmeal, enzyme extract, peptone, gluten meal, cottonseed flour, cottonseed oil cake, soybean flour, corn steep liquor, dried yeast, wheat germ, peanut flour,
It is preferable to use chicken bone meat meal and the like, but inorganic and organic nitrogen compounds such as ammonium salts (eg, ammonium nitrate, ammonium sulfate, ammonium phosphate, etc.), urea and amino acids can also be advantageously used.

These carbon and nitrogen sources are advantageously used together, but need not necessarily be pure. This is because impurities may include a growth factor or a trace element, and may be advantageous.

If necessary, for example, the following inorganic salts may be added to the medium: sodium carbonate, potassium carbonate, sodium phosphate, potassium phosphate, sodium chloride, potassium chloride, sodium iodide, potassium iodide, Magnesium salts, copper salts, cobalt salts, etc.

In particular, if the medium foams strongly, liquid paraffin, animal oil, vegetable oil, mineral oil, silicon, etc. may be added when necessary.

In order to industrially produce the target substance in large quantities, it is preferable to carry out aeration and stirring culture as in the case of other fermentation products. For small-scale production, shake culture using a flask is preferred.

When culturing is carried out in a large tank, in order to prevent the growth of the bacteria during the production process of the WF11243 substance, a relatively small amount of the medium is first inoculated with the produced bacteria, and then the culture is transferred to a large production tank. It is preferable to carry out production culture there. In this case, the composition of the medium used for the preculture and the composition of the medium used for the production culture may be the same or may be changed if necessary.

The cultivation is preferably performed under aeration and stirring conditions. For example, known methods such as stirring with a propeller or another machine, rotation or shaking of a fermenter, pumping, and blowing of air are used as appropriate. Use sterilized air for ventilation.

The culturing temperature can be appropriately changed within a range in which the present WF11243 substance-producing bacterium produces the present substance, but it is usually preferable to culture at 1 to 40 ° C, preferably 14 to 36 ° C. The culturing time varies depending on the culturing conditions and the culturing amount, but is usually about 1 day to 1 week.

After completion of the fermentation, recover the desired WF11243 substance from the culture. That is, the cells may be directly extracted with water and / or an organic solvent, or may be disrupted mechanically or using known means such as ultrasonic waves, and then extracted with water and / or an organic solvent, followed by a conventional method. Collect and purify according to In the case of a culture solution, it may be directly collected and purified according to a conventional method.

The method for recovery and purification includes, for example, solvent extraction with water, an organic solvent, and a mixed solvent thereof; chromatography;
Conventional methods such as recrystallization from a single solvent or a mixed solvent may be used alone or in combination as appropriate.

The recovery and purification of the WF11243 substance are appropriately performed using known methods as described above.
The culture was extracted with acetone water, adsorbed on a neutral and neutral adsorbent resin (eg, HP-20 (manufactured by Mitsubishi Kasei)), eluted with acidified acetone water, concentrated, and washed with ethyl acetate. , Butanol, and if necessary, desorption with a neutral adsorption resin is repeated for purification. The WF11243 substance is an amphoteric substance and can react with a base or acid to form a salt. WF11243 substance is free (WF11243 substance itself)
However, it can be recovered and purified, and also can be recovered and purified as a salt thereof. These can also be mutually converted by an ordinary method.

Salts of WF11243 substances with bases include sodium salts, alkali metal salts such as potassium salts, alkaline earth metal salts such as calcium salts and magnesium salts, salts with inorganic bases such as ammonium salts, methylamine salts, ethylamine salts, and propyl salts. Amine salts, isopropylamine salts, butylamine salts, t-butylamine salts, dimethylamine salts, diethylamine salts, trimethylamine salts, triethylamine salts,
Pyridine salt, picoline salt, dicyclohexylamine salt,
Salts with organic bases such as organic amine salts such as N, N'-dibenzylethyleneamine salt, amino acid salts such as arginine salt, asparagine salt and glutamate, and the like, and salts with acid include hydrochloride, odor Inorganic acid addition salts such as hydrochloride, hydroiodide, sulfate, nitrate, phosphate, etc., acetate, trifluoroacetate, maleate, tartrate,
Methanesulfonate, benzenesulfonate, formate,
Acid addition salts such as organic acid addition salts such as toluenesulfonate, and amino acid addition salts such as aspartate and glutamate.

The preparation of the present invention is characterized by the rectal administration of the WF11243 substance and / or salts thereof as an active ingredient, pulmonary (nasal or buccal inhalation), nasal, ophthalmic, topical (topical), oral or parenteral (subcutaneous, intravenous and (Including intramuscular) and solid, semi-solid or liquid preparations containing organic or inorganic carriers or excipients suitable for administration or inhalation. Active ingredients include, for example, tablets, pellets,
Lozenges, capsules, suppositories, creams, ointments, aerosols, powders for inhalation, solutions, emulsions, suspensions, and other conventional non-toxic pharmaceutically acceptable dosage forms suitable for use It can be formulated with a carrier. In addition, auxiliary, stabilizing, thickening, coloring and flavoring agents can be used as needed. The WF11243 substance and / or salts thereof may be contained in the preparation in an amount sufficient to produce a desired therapeutic effect on the course or condition of the disease.

When this formulation is applied to humans, it is preferably by intravenous, intramuscular or oral administration. The therapeutically effective amount of the active ingredient varies depending on the age and conditions of each patient to be treated.In general, the active ingredient is administered intravenously at a dose of 0.01 to 100 mg / kg human body weight per day, and intramuscularly. In this case, the daily dose is 0.1 to 100 mg per kg of human body weight, and in the case of oral administration, the daily dose is 0.5 to 100 mg per kg of human body weight, for the treatment or prevention of infectious diseases.

The WF11243 substance and / or salt thereof according to the present invention is widely effective in preventing and / or treating various fungal infectious diseases, but is also widely effective in preventing and / or treating various protozoal diseases. , Pneumocystis pneumonia and other Pneumocystis carinii infections are particularly useful, but the following points should be particularly noted in the treatment or prevention of Pneumocystis carini infections.

For administration by inhalation, the compounds of the present invention are conveniently delivered in the form of an aerosol spray from pressure vessel or nebulizer. The compound can also be supplied as a powder that can be formulated and the powder composition can be inhaled by a gas-inhaled powder inhaler. A preferred delivery system for inhalation is a metered dose inhalation aerosol,
It can be formulated as a suspension or solution of the compound in a suitable propellant, for example a fluorocarbon or hydrocarbon.

Aerosol administration is the preferred method of administration because it is desirable to treat the lungs and bronchi directly. Insufflation is also a desirable method, especially if the infection has spread to the ears and other body cavities.

Parenteral administration can also be used using intravenous infusion.

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

Example 1 ((1) Fermentative production of WF11243 substance) Sucrose 4%, cottonseed oil cake 2%, dried yeast 1%,
Peptone 1%, KH ₂ PO ₄ 0.2%, CaCO ₃ 0.2%, Tween (T
Ween) A pre-culture medium consisting of 800.1% was dispensed 160 ml at a time into a 500 ml Erlenmeyer flask and sterilized at 121 ° C for 30 minutes. No. 11243 strain (FERM BP-3373) was added to each medium.
Of the slant culture was inoculated one loop at a time and shake-cultured at 25 ° C. for 4 days.

Next, modified starch 2%, glucose 0.5%, cottonseed oil cake 1
%, Gluten meal _{1%, KH 2 PO 4 2} %, Na 2 HPO 4 · 12H 2 O
_{1.5%, ZnSO 4 · 7H 2} O0.001%, Adecanol LG-109
A main culture medium consisting of 0.025% (manufactured by Asahi Denka Co., Ltd.) and 0.025% of silicon KM70 (manufactured by Shin-Etsu Chemical Co., Ltd.) was prepared, and 20 l of the main culture medium was injected into a 30-liter jar fermenter. After sterilizing this at 121 ° C. for 30 minutes, the preculture obtained above was inoculated at 2% and cultured at 25 ° C. for 4 days. Stirring is 200rp
m, the air flow was 20 l / min. The amount of the WF11243 substance in the culture was determined by HPLC [column: Hibar LiChrosper 100RP18 (manufactured by Merck); solvent: 50% acetonitrile water containing 0.5% NH ₄ H ₂ PO ₄ ; detection: UV 210 nm; flow rate: 1 ml / min] Quantified. As an assay sample, an equal amount of acetone was added to the culture solution, filtered, and then appropriately concentrated.

((2) Extraction and purification of WF11243 substance) To 75 l of the culture obtained by the above culture method, add an equal volume of acetone, leave the mixture at room temperature overnight with occasional stirring, and filter to obtain a culture extract. Was. To the extract was added 65 l of water, and the pH was adjusted to pH 6.5 with 6 N NaOH, followed by 6.5 l of HP-20.
(Mitsubishi Kasei). 35l water, 40% acetone water 2
After washing the column with 7 l, a final concentration of 0.002 N HC1
The target substance was eluted with 48 liters of 80% acetone water.

The purification was carried out by antibacterial activity against Candida albicans and HPLC [column: YMC Paked Column AM-303 (S-5,1
20A, ODS) YMC Co., LTD]; mobile phase: containing 0.5% NH ₄ H ₂ PO ₄
45% acetonitrile water; detection: UV 210 nm; flow rate: 1 ml / min;
Retention time (RT): 10.9 minutes].

The eluate obtained above was concentrated under reduced pressure to 1.9 l, and the pH was adjusted.
After correcting to 6.0, it was washed with twice the volume of ethyl acetate. Next, the active ingredient was extracted with 1.9 l of butanol, and concentrated under reduced pressure.
Was replaced. After adjusting the pH of the aqueous solution to 3.0 with 1N HCl,
The aqueous solution was washed again with ethyl acetate 1 and then the active ingredient was extracted with butanol 1. The organic solvent layer was washed with 1% aqueous sodium bicarbonate 1 and aqueous hydrochloric acid 1 at pH 4, and then washed.
The organic solvent layer was concentrated under reduced pressure. The residue was then dissolved in 3.0 l of 50% acetonitrile water and applied to a 1.3 l HP-20 column. Column is 3.5 l water, 50 l methanol water 3.5 l, 80%
After washing with 3.5 l of methanol water and 3.8 l of methanol, the active ingredient was eluted with 2.7 l of 80% acetone water containing 0.002N HCl as a final concentration. Next, the eluate was concentrated under reduced pressure, and the residue was dissolved in 5 L of 20% methanol water.
-Gel (ODS-AM, 120-S50, manufactured by YMC Co., LTD) column (5
00 ml). Column contains 0.5% NH ₄ H ₂ PO ₄ in advance 2
Equilibrate with 0% acetonitrile water, charge the solution containing the target substance, wash with 30% acetonitrile water containing 0.5% NH ₄ H ₂ PO ₄ , 35% acetonitrile water 1 and 40% acetonitrile water 1 The target substance was eluted with 45% acetonitrile and 2 l of water.

After diluting 280 ml of the eluted active fraction with an equal volume of water, the column (180 ml) was reverse-phased (YMC-Gel, ODS-AM, 120-S50) again.
30% acetonitrile water containing 0.5% NH ₄ H ₂ PO ₄
The column was washed with 0.4 l of 35% acetonitrile water and 0.4 l of 40% acetonitrile water, and developed with 43% acetonitrile water to elute the target substance. 55 ml of this eluted active fraction was diluted with an equal amount of water, and applied to a 40 ml HP-20 column. The column is diluted with 180 ml of water with an equal volume of water and 40 ml of HP-20
Column. After washing the column with 180 ml of water, 100 ml of 80% acetone water containing 0.002N HCl as a final concentration is used.
The target substance was eluted. The eluate was concentrated under reduced pressure, acetone was removed, and then lyophilized to obtain 72 mg of a white powder of hydrochloride of WF11243 substance (referred to as FR901469 substance).

((3) Physicochemical properties of FR901469 substance) The physicochemical properties of the FR901469 substance thus obtained are as shown in Tables 2 to 5. The amino acid analysis was performed as follows. First, FR901469 substance 1m
g was hydrolyzed with 6N HCl (1 ml) in a sealed tube at 110 ° C. for 20 hours. After completion of the hydrolysis, the reaction mixture was evaporated to dryness,
Use this with an amino acid analyzer (Hitachi 835 Automatic Amino
-Acid Analyzer). The result is Thr
(4), Gly (1), Ala (1), Val (1), Tyr
(1) and Orn (1). And its chemical structure is
It was determined as in the following equation (I).

Example 2 (Biological properties of FR901469 substance) ((1) Antibacterial activity) The antibacterial action of the FR901469 substance was measured by a microbroth dilution method using a 96-well multi-tray described below.

A test bacterial suspension (viable cell count: 2 × 10 ⁵ cells / ml) was prepared from the yeast cultured on the slant medium in yeast nitrogen-based dextrose (YNBD) medium. A serial two-fold dilution series of the FR901469 substance in YNBD was made and 100 μl was added to each well.
Further, after adding 100 μl of the test bacterial suspension to each well, the cells were incubated at 37 ° C. for 24 hours (Candida and Aspergillus sp.) Or 48 hours.
Culturing for a time (genus Cryptococcus). After the completion of the culture, the turbidity of each well was measured, and the drug concentration of the well exhibiting half the turbidity of the well without drug was expressed as a 50% growth inhibitory concentration (IC ₅₀ ). The results are shown in Table 2 below.

((3) Infection Protective Effect of FR901469 Substance against Candida albicans in Experimental Mouse Infection) The efficacy of FR901469 substance as an antifungal agent in a living body was clarified by the following method.

As test animals, ICR female mice 4 weeks old and weighing 18 to 21 g were used, and one group consisted of 5 mice. Suspended in saline
Mice were infected with Candida albicans FP633 by injecting into the lateral tail vein of mice at 2 × 10 ⁶ viable cells per animal. One hour after the infection, a saline solution of FR901469 substance was administered to the mice by subcutaneous injection. This subcutaneous injection was repeated once a day for three days from the day after the infection. The number of surviving mice on day 14 after infection was observed, and the results obtained are shown in Table 3 below.

Table 3 FR901469 vivo infection protective effect FR901469 dose of substance (mg / kg) Survival animals Number 10 5 1 5 0 0 ICR strain female mice Five ((4) Toxicity Test) 5 weeks old, the FR901469 substance 1
Intraperitoneal injection was performed once daily for 3 days at a dose of 00 mg / kg, but there were no deaths, and the weight gain was exactly the same as in the non-administered mouse group, confirming the high safety of the WF11243 substance.

Example 3 (Production of Injection) (1) 5 g of FR901469 substance produced in Example 1 (2) 9 g of sodium chloride (3) 1 g of sodium bicarbonate All components of (1) to (4) were dissolved in 100 ml of distilled water. rear,
One ml was dispensed into each ampule to produce 1,000 injections.

Example 4 (Production of inhalable aerosol) 2% by weight of the compound represented by the formula (I), 33% by weight of ethanol, and 65% by weight of a propellant (propane: isobutane = 70: 30 mixture) were charged into a pressure-resistant container equipped with a valve. To produce an inhalable aerosol.

Effects of the Invention The present invention provides a WF11243 substance, which is a novel pharmacologically active substance which has been unknown so far, has excellent antifungal and antiprotozoal activities, and is used for pharmaceuticals, cosmetics, industrial chemicals, food and drink It is extremely useful for preventing or suppressing the growth and killing of fungi and protozoa in various technical fields of products. The substance according to the present invention is characterized in that it has such an excellent antifungal action and also has an antiprotozoal action, and the latter is, for example, effective in preventing and treating carinii pneumonia. Especially characteristic.

Also, since the estimated structural formula of the active ingredient according to the present invention has been clarified, it is possible to produce not only industrial compounds using microorganisms but also organic synthesis, and as a result, it becomes possible to produce various derivatives. And the development of new applications can be greatly expected.

Reference to the deposited microorganism in Rule 13bis of Rule 1 1.No. 11243 b. Name and address of the depository institution that deposited the microorganism Name Address to Institute of Microbial Engineering, Institute of Industrial Science and Technology, Ministry of International Trade and Industry Address: Ibaraki, Japan 305 1-3-3 Higashi, Tsukuba City Date of deposit at loyal depository April 23, 1991 Accession number FERM BP-3373 assigned by the depositary of Hiy for deposit

──────────────────────────────────────────────────続 き Continuation of the front page (51) Int.Cl. ⁶ Identification code Agency reference number FI Technical display location C12R 1: 645) (C12P 21/02 C12R 1: 645) (72) Inventor Masakuni Okuhara Ibaraki 2-14-10, Umezono, Tsukuba

Claims (3)

(57) [Claims] 1. A WF having the following structural formula:
11243 substances or salts thereof. 2. The WF11243 substance producing bacterium No. 11243 according to claim 1. 3. An antifungal agent comprising the WF11243 substance or a salt thereof according to claim 1 as an active ingredient.