JPH0525092A - Physiologically active substance tan-1515, production thereof and its use - Google Patents

Abstract

PURPOSE:To obtain the new title substance useful as antiphlogistics because of phospholipase A2 inhibiting activity and useful as a synthetic intermediate for new derivative. CONSTITUTION:A physiologically active substance TAN-1515 having the following properties: Appearance, colorless crystal; melting point, 102-104 deg.C; molecular formula, C32H46O7; elementary analysis, C 70.82 H 8.54 (calculated value), C 70.99 H 8.91 (measured value); color reaction, positive to Barton reaction, iodine reaction, etc., and negative to ninhydrin reaction and Dragendorff reaction; classification of acidic, neutral or basic, acidic. The compound 12 is obtained by culturing a microorganism [e.g. Acremonium sp. FL-175559 strain (FERM BP-3487)] belonging to Acremonium and capable of producing a compound expressed by the formula and purifying the cultured mixture. Furthermore, temperature in the meddle period of culture is 20-30 deg.C and initial pH is preferably 6-8.

Description

Detailed Description of the Invention

[0001]

INDUSTRIAL APPLICABILITY The present invention produces a novel physiologically active substance TAN-1515 (hereinafter sometimes abbreviated as "TAN-1515") having a phospholipase A ₂ inhibitory activity which can be used as an anti-inflammatory agent. Regarding law and use.

[0002]

_2. Description of the Related Art Phospholipase A ₂ is considered to be a key enzyme for excising arachidonic acid from phospholipids and producing inflammatory mediators such as prostaglandins and leukotrienes. On the other hand, the action mechanism of a steroid drug having a strong anti-inflammatory effect has recently been investigated, and the protein produced by the administration of the steroid drug is phospholipase A.
The hypothesis of suppressing the activity of ₂ has been submitted. Therefore, a compound that effectively inhibits phospholipase A ₂ is
A strong anti-inflammatory effect can be expected without the side effects of steroids such as easy infection and adrenal atrophy.

[0003]

The compounds known to date as phospholipase A ₂ inhibitors include synthetic compounds such as mepacrine and p-bromophenacyl bromide.
In addition, natural products include sponge-derived manoalide, blue mold-derived plastatin, and bacterial-derived pripastatin.
However, these anti-inflammatory effects are far weaker than existing non-steroidal anti-inflammatory agents such as indomethacin and diclofenac, and they are not practical as anti-inflammatory agents. Therefore, there is a need for new phospholipase A ₂ inhibitors that can be used clinically as anti-inflammatory agents.

[0004]

Means for Solving the Problems Conventionally, phospholipase A
_Although research on ₂ inhibitors has been centered on exocrine enzymes such as pig pancreas as an enzyme source, the present inventors have found that they are derived from platelets, which are considered to act in the field of actual inflammation. As a result of searching phospholipase A ₂ as an enzyme source and searching for its inhibitor as a microbial metabolite, a compound having an inhibitory activity on platelet phospholipase A ₂ was found from a certain mold, and this was called a physiologically active substance TAN-1515. I decided. The present invention was completed as a result of purifying and isolating TAN-1515 from the culture solution, clarifying that it was a novel compound, and conducting further diligent research. That is, the present invention provides (1) a physiologically active substance TAN-1515 or a salt thereof,
(2) Belonging to the genus Acremonium, a physiologically active substance TAN-1
Culturing in a medium a microorganism capable of producing 515,
A physiologically active substance TAN-15, characterized in that the TAN-1515 is produced and accumulated in a culture, and the collected TAN-1515 is collected.
15 and a phospholipase A ₂ inhibitor comprising (3) a physiologically active substance TAN-1515. Any microorganism can be used in the present invention as long as it is a bacterium belonging to the genus Acremonium and producing the physiologically active substance TAN-1515. For example, Acremonium sp. FL-17559 isolated from the soil of Hyogo prefecture can be used as an example. The mycological symptoms of the FL-17559 strain are as follows.

(I) Morphological characteristics Mycelium: Mycelia are colorless and have partition walls, and the surface is smooth. The diameter is 1-3 μm and is not constant. Phialide: A large number of phialides with a septum at the base almost stand upright on hyphae. Almost no branching. It gradually becomes thinner from the base toward the tip. The length is 30-75 μm. The diameter of the base is 1.4-1.6 μm. Colorless and smooth surface. Conidia: Fiaro-type conidia. Spindle-shaped or banana-shaped. 1.2-1.5x
6.0-10.0 μm. Colorless and smooth surface. A spherical conidial mass is formed at the tip of the phialide, and its diameter is 10-30.
It exhibits μm and its size is not constant. (II) Growth condition on various media 1) Malt extract agar medium Growth was poor to moderate, colonies spread slowly at 24 ° C, and the diameter after 2 weeks was about 20-22 mm. The surface is flat and composed of thin mycelium, and the outer edge is slightly irregularly bordered. The central part is grayish white, and the middle part and the outer peripheral part are yellowish white. The development of aerial hyphae is poor, and a pale grayish white hyphae is slightly observed in the middle part. Conidia formation is good.
The back surface has a pale yellowish gray color. No formation of soluble pigment is observed. 2) The potato-glucose agar medium grew moderately at 24 ° C. and the colony diameter after 2 weeks was 3
It was 0-33 mm. The surface is flat and made of thin mycelium, and radial wrinkles are seen from the center. The outer edge is regularly bordered. The central part is grayish white, and the middle and outer parts are yellowish gray. The development of aerial hyphae is poor, but the middle part is occupied by rope-shaped hyphae. Conidia formation is good. The reverse side is yellowish gray to dark yellowish gray. No formation of soluble pigment is observed. 3) The growth of Czapek agar medium was poor, and the diameter of the colony after 2 weeks at 24 ° C was 40.
It was -42 mm. The surface is flat and very thin mycelium extends radially. The outer edge is regularly bordered. Both front and back are colorless. Conidia formation is good. No formation of soluble pigment is observed. 4) Oatmeal agar medium growth was moderate, and the diameter of colonies after 24 weeks was 5 ° C.
It was 0 mm. The surface is composed of thin mycelium and the outer edges are regularly bordered. Both front and back are colorless. Conidia formation is good. No formation of soluble pigment is observed.
From the above mycological properties, it is clear that this strain belongs to the genus Acremonium described in the following documents, and Acremonium sp. FL-175
59 (Acremonium sp. FL-17559). [WALTER GAMS; Cephalospori
um-artage Schimmelpilze (Hyphomycetes), Gustav F
ischer Stuttgart, (1971)] The above-mentioned Acremonium SP FL-17559 strain was deposited with the Fermentation Research Institute from June 26, 1991 under the deposit number IFO-32400.
This microorganism has been deposited under the Budapest Treaty with a deposit number FERM BP- from July 18, 1991, under the Budapest Treaty.
Deposited as 3487.

[0006] The genus Acremonium can be mutated naturally as a general property of microorganisms or by mutagens. For example, many mutant strains obtained by irradiation with radiation such as X-rays, gamma rays, and ultraviolet rays, separation of monospores, treatment with various drugs or culturing on a medium containing drugs, and mutating by other means, Alternatively, all naturally occurring mutants and the like having the property of producing TAN-1515 can be used in the method of the present invention.

The medium used for the culture of the present invention may be liquid or solid as long as it contains a nutrient source that can be utilized by the strain to be used, but it is more suitable to use a liquid medium when treating a large amount. A carbon source that can be assimilated, a nitrogen source that can be digested, an inorganic substance, and a micronutrient are appropriately added to the medium. Examples of carbon sources include glucose, lactose, sucrose, maltose, dextrin, starch, glycerin, mannitol, sorbitol, oils and fats (eg, soybean oil, olive oil, nuka oil, sesame oil, lard oil, chicken oil, etc.), nitrogen source Examples thereof include meat extract, yeast extract, dry yeast, soybean powder, corn steep liquor, peptone, cottonseed flour, urea, ammonium salts (eg, ammonium sulfate, ammonium acetate, etc.) and the like. Furthermore, salts containing sodium, potassium, calcium, magnesium, etc., metal salts such as iron, manganese, zinc, cobalt, nickel, etc., salts such as phosphoric acid, boric acid and acetic acid,
A salt of an organic acid such as propionic acid is appropriately used. In addition, amino acids (eg, glutamic acid, aspartic acid, alanine, lysine, valine, methionine, proline, etc.), peptides (eg, dipeptide, tripeptide, etc.), vitamins (eg, B ₁ , B ₂ , nicotinic acid, B ₁₂ ,
C, etc.), nucleic acids (eg, purines, pyridine and derivatives thereof) and the like may be contained. Of course, inorganic or organic acids, alkalis, buffers and the like are added for the purpose of adjusting the pH of the medium, or appropriate amounts of fats and oils, surfactants and the like are added for the purpose of defoaming.

The means for culturing may be static culture, shaking culture, aeration stirring culture method, or the like. Needless to say, so-called deep aeration stirring culture is desirable for large-scale treatment. It is natural that the culture conditions are not constant depending on the condition of the medium, the composition, the strain type, and the means of culture, but they are usually at a temperature of about 15 ° C to 32 ° C.
It is recommended to select the initial pH around 5-9. Especially, the temperature in the middle stage of culture is about 20 ° C to 30 ° C, and the initial pH is about 6
A condition of -8 is desirable. The culturing period is not constant depending on the above-mentioned conditions, but it is preferable to cultivate until the concentration of various physiologically active substances becomes maximum. The time required for this is usually about 1-10 days in the case of shaking culture or aeration-agitation culture using a liquid medium. Generated physiologically active substance TAN-1515
Is present in the culture filtrate and cells, so the culture can be separated into a supernatant and cells by centrifugation or filtration, and can be purified from the supernatant and cells, respectively. In some cases, it may be advantageous to purify from an extract obtained by directly adding an organic solvent such as methanol, acetone or ethyl acetate to the product. In order to collect TAN-1515 from the culture solution, since the substance is a fat-soluble acidic substance, a separation / purification means usually used for collecting such microbial metabolites is appropriately used. For example, a method utilizing a difference in solubility with impurities, activated carbon, nonionic high-porous resin, chromatography using various carriers such as silica gel, alumina, dextran gel and the like are used alone or in combination.

The method for collecting TAN-1515 produced in the culture will be described in detail. First, the culture solution is extracted with an organic solvent that is acidic or neutral and is immiscible with water, such as ethyl acetate. Examples of the organic solvent used for extraction include esters such as ethyl acetate and butyl acetate, halogenated hydrocarbons such as chloroform and methylene chloride, and alcohols such as n-butanol and i-butanol. Alternatively, a water-miscible organic solvent such as methanol or acetone is added to the culture solution, and the mixture is stirred and extracted, and then the organic solvent of the extract is distilled off, and then the water-immiscible organic solvent (ethyl acetate Etc.) to extract the substance. The extract thus obtained is washed successively with acidic water and water, concentrated and the crude powder obtained is subjected to column chromatography on silica gel.
As the developing solvent, for example, a system in which an acid such as acetic acid or oxalic acid is added to a mixed solvent such as chloroform-methanol or hexane-ethyl acetate is used. By increasing the ratio of polar solvent, it is possible to separate and elute from other contaminants. The fraction containing TAN-1515 thus eluted is concentrated and n-hexane is added to obtain a powder of TAN-1515. This is redissolved in an organic solvent such as chloroform or ethyl acetate, washed successively with diluted hydrochloric acid and water, concentrated, and crystallized from water-containing methanol to give TAN-1515 as colorless crystals.

The physiologically active substance TA thus obtained
The physicochemical properties of N-1515 are as follows. 1) Shape: colorless crystal 2) Melting point: 102-104 ° C. 3) Mass spectrum: (SIMS): m / z 543 (M +)
H) 4) Molecular formula: C ₃₂ H ₄₆ O ₇ 5) Elemental analysis value: Calculated value: C, 70.82; H, 8.54 Actual value: C, 70.99; H, 8.91 6) Ultraviolet absorption spectrum: MeOH 1% λ (E): 210 (960), 269 (384), 302 (24
6) max 1cm 7) Infrared absorption spectrum (by KBr lock method): 332
5, 3075, 2920, 2850, 1665, 1610, 1595,1460, 1415,
1385, 1350, 1310, 1245, 1200, 1170, 1140, 1070, 105
5, 995, 850, 800, 720,685, 630,605 (cm ^-1 , see Fig. 2) 8) ¹ H nuclear magnetic resonance spectrum (300 MHz, d ₆ -DM
Chemical shift in SO, δppm): 0.85 (3H, br.t),
1.15-1.35 (28H, m), 1.53 (2H, m), 2.38 (3H, s), 2.61 (2H,
m), 6.18 (1H, d), 6.22 (1H, d), 6.52 (1H, br.d), 6.57 (1
H, d), 9.85 (1H, br.s), 10.13 (1H, s). 9) ¹³ C nuclear magnetic resonance spectrum (75 MHz, d ₆ -DM
Chemical shift in SO, δppm): 13.9 (q), 21.3
(q), 22.0 (t), 28.7 (t), 28.8 (t), 29.0 × 10 (t), 31.0
(t), 31.2 (t), 33.9 (t), 100.4 (d), 107.3 (d), 108.5
(d), 109.0 (s), 114.6 (d), 115.7 (s), 139.8 (s), 144.2
(s), 152.6 (s), 158.9 (s), 159.4 (s), 160.6 (s), 166.8
(s), 170.8 (s) However, s: singlet (d), d: doublet (doublet), t: triplet (triplet), q: quartet (quartet), br: broad (broad) are shown. 10) Color reaction positive: Burton, iodine, phosphomolybdate, potassium permanganate negative: ninhydrin, Dragendorff 11) Thin layer chromatography (carrier, silica gel glass plate 60F ₂₅₄ , 0.25mm, manufactured by Merck, Germany) : Developing solvent Rf Chloroform-methanol-acetic acid (100: 5: 1) 0.52 Toluene-methanol-acetic acid (20: 2: 1) 0.49 Ethyl acetate-acetic acid (100: 1) 0.5912) Acidic, Neutral or basic: From the physicochemical properties above acidic, TAN-1515 is a novel compound having the following structural formula.

[Chemical 3] Since TAN-1515 is an acidic substance, it can be prepared by a method known per se by a metal salt such as sodium salt, potassium salt or calcium salt, ammonium salt, or a salt with an organic amine such as triethylamine salt or ethanolamine salt. And the like, physiologically acceptable salts can be formed.

[0011]

[Function] The physiologically active substance TAN-
The rabbit platelet phospholipase A ₂ inhibitory activity of 1515 is shown together with the test method.

Experimental Example 1 Rabbit Platelet Phospholipase A
Preparation of ₂ crude enzyme solution Separation of platelets is carried out in accordance with a conventional method (information transmission and cell response, below, sequel biochemistry course 7), and the obtained platelets are crushed by the freeze-thaw method to prepare a phospholipase A ₂ crude enzyme solution. did. Blood obtained by whole blood collection from the carotid artery of a rabbit (white Japanese breed) using ethylenediaminetetraacetic acid as an anticoagulant was centrifuged at 140 xg for 15 minutes to obtain a platelet-rich fraction. . This fraction is centrifuged at 940 × g for 15 minutes to precipitate platelets, and then the supernatant plasma is removed. Platelets were loaded with 1.2 mM EDTA and 0.14 M Na.
Suspended in 10 mM Tris buffer (pH 7.4) containing Cl,
Centrifuge at 940 xg for 15 minutes to pellet the platelets and remove the supernatant. The platelets were then added to 0.14M NaC.
It is suspended in 10 mM Tris buffer (pH 7.4) containing 1 and the same operation is repeated twice. 5 × 10 ⁹ platelets / m 2
The suspension is suspended in 10 mM Tris buffer (pH 7.4) containing 0.14 M NaCl so as to be 1 and frozen at -80 ° C. After thawing frozen platelets at 37 ℃, 10,000 × g
After removing the precipitate by centrifugation for 10 minutes, the supernatant is further centrifuged at 105,000 × g for 60 minutes to obtain a phospholipase A ₂ -rich supernatant. This preparation was divided into aliquots, stored at -20 ° C, and diluted with 10 mM Tris buffer (pH 7.4) containing 0.1% bovine serum albumin and 0.14M NaCl before use.

[0013] Measurement phospholipase A ₂ activity of Example 2 Phospholipase A ₂ activity, 1-palmitoyl-2
The (1- ¹⁴ C) arachidonyl -L-alpha-phosphatidylcholine used as a substrate to measure the release of free [1- ¹⁴ C] arachidonic acid. Substrate is 1-palmitoyl-2-
(1- ¹⁴ C) arachidonyl -L-α- phosphatidylcholine (New England Nuclear, Inc., manufactured by the United States)
Unlabeled 1-palmitoyl on [50 mCi / mmole]
It was diluted to 16 mCi / mmole by adding 2-arachidonyl-L-α-phosphatidylcholine (Avanchi Polar Lipids, USA). 1.5 ml
Add the following additives to a volume polypropylene tube: 0.9 nmo
le (15nCi) 1- palmitoyl -2- (1- ¹⁴ C) arachidonyl - phosphatidylcholine, dimethyl sulfoxide 12.5 [mu] l, 10 [mu] l of 500mM Tris buffer (pH9.0), 80mM CaCl ₂ of 2.5 [mu] l, 1
0 μl of phospholipase A ₂ crude enzyme solution (amount that gives 20-30% hydrolysis) and drug are added and the final volume is adjusted to 50 μl with water. Stir the mixture and mix in a constant temperature water bath for 3
Incubate at 0 ° C for 60 minutes. The reaction is stopped by adding 10 μl of acetic acid to each tube. Then, 100 mg of silica gel 60 (Merck, Germany) and 1 ml of ethyl acetate were added, and the mixture was stirred for 3 minutes using a vial mixer (Tayo Service Center, Japan). After stirring, centrifugation was performed at 10,000 × g for 1 minute, silica gel was allowed to settle, 700 μl of the supernatant was taken in a vial, 3 ml of scintillator A (Wako Pure Chemical Industries, Ltd., Japan) was added, and after stirring, a 2200 CA liquid scintillation counter ( Radioactivity was measured by Packard Co., Ltd. (USA). IC of phospholipase A ₂ inhibitory activity of the compound of the present invention and manolide used as standard drugs measured by this method
_{The 50} values are shown in [Table 1].

[Table 1] In the acute toxicity test using mice, the compound of the present invention had an LD ₅₀ value of 100 mg / kg or more after intraperitoneal administration.
The compound of the present invention has phospholipase A ₂ inhibitory activity, and various diseases caused by oxidative metabolites of arachidonic acid, such as allergic diseases, bronchial asthma, rheumatoid arthritis, osteoarthritis, pancreatitis, and various inflammatory diseases. In addition, it can be applied as a therapeutic and preventive agent for thrombosis and arteriosclerosis, and as an analgesic. The present compound, in addition to oral administration, is parenterally injected or used topically on the skin, mucous membranes, rectum, etc.,
The dose may vary depending on the disease to be administered, the administration route, etc., but when it is used for the treatment of inflammatory diseases, for example, it is usually 0.01-100 mg / kg per adult, especially 0.1.
-20 mg / kg is orally or parenterally administered about 1 to 3 times a day. For oral administration, capsules, tablets,
In the form of granules, syrups, powders, etc., the compound of the present invention and additives such as excipients, binders, disintegrants, surfactants, colorants, crosslinking agents, stabilizers, etc. are included. Is also good. Among them are starch, sucrose, fructose, lactose, glucose, mannitol, sorbitol precipitated calcium carbonate, crystalline cellulose, carboxymethyl cellulose,
Examples thereof include dextrin, gelatin, gum arabic, magnesium stearate, talc, hydroxypropyl methylcellulose and the like. For parenteral administration, the active ingredient can be dissolved or suspended in a conventional diluent (aqueous or non-aqueous carrier). As a diluent,
Examples include physiological saline, Ringer's solution, glucose aqueous solution, alcohols, fatty acid esters, glycols, glycerin fatty acid glyceride, oil sources derived from plants and animals, paraffins and the like. In addition, in pharmaceutical preparations, as additives, emulsifiers, suspending agents, solubilizers, stabilizers, preservatives, soothing agents, tonicity agents, buffers, pH adjusting agents, coloring agents, coating agents, etc. May be included. In addition, these preparations can be manufactured by a usual method.

[0014]

EXAMPLES The present invention will be described in more detail below with reference to examples. In the following examples, the composition percentage of the medium indicates weight / volume percentage.

Example 1 Bacto potato dextrose agar (manufactured by Difco,
A platinum loop of Acremonium sp. FL-17559 preliminarily grown sufficiently on a slant culture medium consisting of USA), glucose 2.0%, maltose 3.0%, raw soybean flour 1.5%, corn steep Inoculate a sterilized 200 ml Erlenmeyer flask with 40 ml of a seed culture medium consisting of 1.0% liquor, 0.5% polypeptone, 0.3% yeast extract and pH 6.0, and inoculate on a rotary shaker at 28 Cultivated at ℃ for 2 days. Glucose 1.0%, dextrin 4.0%, raw soybean flour 0.5%, malt extract 0.5
%, Polypeptone 0.5%, yeast extract 0.2%, iron sulfate heptahydrate 0.05%, magnesium sulfate heptahydrate 0.05%
Manganese sulfate heptahydrate 0.05%, monopotassium phosphate 0.5
%, Precipitated calcium carbonate 0.5%, pH 7.5, 40 ml of main culture medium was dispensed into a 200 ml Erlenmeyer flask in which the bacteria had been dispensed, and the mixture was cultured at 24 ° C. for 4 days on a rotary shaker, and then TAN-1515. Was generated and accumulated.

Example 2 Bacto potato dextrose agar (manufactured by Difco,
5 platinum loops of Acremonium sp. FL-17559 preliminarily grown sufficiently on a slant medium composed of USA, glucose 2.0%, maltose 3.0%, raw soybean flour 1.5%, corn steep -Inoculate a 2 liter Sakaguchi flask sterilized by dispensing with 500 ml of a seed culture medium consisting of 1.0% liquor, 0.5% polypeptone, 0.3% yeast extract, and pH 6.0, and put on a reciprocating shaker. It was cultured at 28 ° C for 2 days. Glucose 1.
0%, dextrin 4.0%, raw soybean powder 0.5%, malt extract 0.5%, polypeptone 0.5%, yeast extract 0.2
%, Iron sulfate heptahydrate 0.05%, magnesium sulfate heptahydrate 0.05%, manganese sulphate heptahydrate 0.05%, monopotassium phosphate 0.5% and precipitated calcium carbonate 0.5%, pH 7 .
120 liters of main culture medium consisting of 5 was sterilized by injection
It was transplanted to a 00-liter fermentor. This main fermentation is 24
℃, aeration (120 l / min), stirring (140 r.p.
m.) and cultivated for 114 hours under an internal pressure of 1.0 kg / cm ² .
TAN-1515 was produced and accumulated.

Example 3 The culture solution (2.2 liters) obtained in Example 1 was subjected to centrifugation to separate it into a supernatant and cells. The supernatant (1.8 liters) was extracted twice with ethyl acetate (0.9 liters) at pH 3.0. The ethyl acetate layer (1.7 liter) was washed with water (1 liter) and concentrated to give an oily residue (1.25 g). The cells were extracted with 80% acetone water (1.8 liter) with stirring for 30 minutes and then filtered using Hyflo Super Cell. The resulting filtrate (1.5 liters) was concentrated to 200 ml, and water (8 liters) was added.
After diluting with (00 ml), the mixture was extracted twice with ethyl acetate (0.5 liter) to a pH of 3.0. Ethyl acetate layer (0.
9 liters) is washed with water (1 liter) and concentrated,
An oily residue (1.13 g) was obtained. The supernatant and the oily residue obtained from the cells were combined (2.38 g), dissolved in chloroform (10 ml) and subjected to column chromatography on silica gel (50 g). The column was chloroform (200 ml), chloroform-methanol (49:
1,600 ml), chloroform-methanol (19:
It was developed sequentially with 1,400 ml). Fractions No. 5 to 11 were collected and concentrated, and hexane was added to obtain 570 mg of TAN-1515 powder. Powdered TAN-1515 (150 mg) was dissolved in ethyl acetate (30 ml) and 0.5N hydrochloric acid (10 ml) was added.
And washed with water (10 ml). The ethyl acetate layer was dried over anhydrous sodium sulfate, concentrated, and crystallized from water-containing methanol to give colorless crystals of TAN-1515 (88 m
g) was obtained.

Example 4 100 L of the culture broth of Example 2 was treated in the same manner as in Example 3 to obtain 4 g of colorless crystals of TAN-1515.

[0019]

The novel physiologically active substance TAN-15 of the present invention
15 has a strong phospholipase A ₂ inhibitory activity, can be used as an anti-inflammatory agent, and is also useful as an intermediate for the synthesis of new derivatives.

[0020]

[Brief description of drawings]

FIG. 1 UV spectrum of TAN-1515

FIG. 2 IR spectrum of TAN-1515

FIG. 3 ¹ H NMR spectrum of TAN-1515

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification code Internal reference number FI technical display location C12R 1: 645)

Claims (3)

[Claims] 1. The formula: Or a salt thereof. 2. A compound belonging to the genus Acremonium and represented by the formula: A method for producing a compound represented by the above formula, which comprises culturing in a medium a microorganism having an ability to produce the compound represented by, allowing the compound to accumulate in the culture, and collecting the compound. 3. A phospholipase A ₂ inhibitor comprising the compound according to claim 1 or a salt thereof.