JPH10101666A - New benzoxacyclotridecyne compound and medicinal composition containing the same - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new compound exhibiting chymase inhibitory activity and antimicrobial activity, thus useful as a therapeutic agent for chronic cardiovascular disturbance or allergic diseases, or as an antimicrobial agent. SOLUTION: This new compound is shown by the formula [R<1> is OH, R<2> is CHO, and R<3> forms a double bond with the carbon atom to which R<1> is bound to, or R<1> is O and R<2> is joined, together with R<3> to form CH(OR<4> ) (R<4> is H or an acyl) or C(=0)], having the following physicochemical characteristics [provided that R<1> is 0; R<2> is joined together with R<3> to form CH(OH)]: appearance: pale yellow needle crystal; solubility: soluble to chloroform, diethyl ether, acetone, ethyl acetate, methanol; sparingly soluble to n-hexane; insoluble to water; melting point: 146-146.5 deg.C; elemental analysis (as C25 H26 O5): C = 73.83 %, H=6.55%; etc. This compound of the formula is obtained by culturing microorganisms belonging to the genus Stachybotrys (e.g. Stachybotrys cylindrospora).

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

The present invention relates to a novel compound and a medicine containing the same, and more particularly to a chymase inhibitor and / or an antibacterial agent containing the novel benzoxacyclotridesine compound.

[0002]

2. Description of the Related Art Human chymase is a neutral serine protease having a molecular weight of about 30,000, isolated in 1990, and is synthesized, stored and secreted mainly in mast cells, and is used in the heart, blood vessels,
It has been found to be present on the skin. Its main effect is the production of angiotensin II. Conventionally,
It was thought that angiotensin converting enzyme (hereinafter, abbreviated as ACE) acts on the production of angiotensin II, but recently, ACE acts on the production of angiotensin II in the human heart. It has been revealed that only about 10 to 15% is involved, and more than 80% is caused by the action of human chymase (Circulation Research).
Research) Vol. 66, p. 883, 1990
Year, Journal of Biological Chemistry (Journal of Biological Ch
mistry), vol. 266, p. 17173, 19
1991). For this reason, chymase inhibitors are attracting attention as a new type of therapeutic agent for cardiovascular disorders. Chymase has also been implicated in promoting histamine release from mast cells (see Journal of Biochemistry).
y) 103, 820-822, 1988),
The inhibitors hold promise as potential new types of anti-inflammatory and anti-allergic agents.

[0003] In addition, human chymase has various activities, such as promotion of macrophage foam cell formation in vitro, production of active collagenase from procollagenase, and limitation of extracellular matrix such as collagen, fibronectin and vitronectin. Degradation, thrombin and Ig
It has already been clarified that it has an action such as limited decomposition of G. It is also known pathologically that chymase activity is elevated in blood vessels after balloon injury and in hearts with cardiomyopathy. From such findings, it can be predicted that chymase is also involved in arteriosclerosis, inflammation, rheumatism and the like.

To date, International Patent Application Publication No. 93/2
5574, 95/27053, 95/2705
No. 5 discloses a peptidic chymase inhibitor. Further, as non-peptide chymase inhibitors, only imidazolidine derivatives are disclosed in WO 96/04248, and triazine derivatives are disclosed in EP 713876.

[0005]

At present, only a small number of non-peptidic chymase inhibitors have been found, and they are used for elucidating the physiological function of chymase and for developing clinically useful drugs. In addition, a non-peptidic chymase inhibitor having a strong activity has been demanded.

[0006]

Means for Solving the Problems In view of the above situation, the present inventors have conducted intensive studies and as a result, have found that one of the filamentous fungi, Stachybotry.
s. cylindrospora ) A compound having a strong human chymase inhibitory activity was found in the culture of the RF-5900 strain, and the compound was isolated and purified to complete the present invention. Furthermore, it has been found that the compound (I) of the present invention also has an antibacterial action.

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

Embedded image

Wherein R ¹ is hydroxy and R ² is-
CHO and R ³ forms a double bond with the carbon atom to which R ¹ is attached, or R ¹ is ＯO and R ² and R ³ together form —CH (OR ⁴⁾ - (wherein R ⁴ is hydrogen or acyl) or -C (= O) - there is provided a compound or a pharmaceutically acceptable salt thereof or a hydrate thereof represented by the form to) a. The present invention also provides a pharmaceutical composition containing compound (I) or a pharmaceutically acceptable salt thereof or a hydrate thereof, specifically a chymase inhibitor and / or an antibacterial agent. Further, a microorganism belonging to the genus Stachybotrys and capable of producing any of the compounds (I) is cultured, and the compound is collected from the obtained culture, and further subjected to ring opening, ring closing, acylation or oxidation, if desired. Features,
The present invention relates to a method for producing compound (I). Furthermore, the present invention relates to a microorganism belonging to Stachybotrys cylindrospora and capable of producing any of the compounds (I), more specifically, relates to Stachybotrys cylindrospora RF-5900.

The production method of the compound (I) of the present invention by microorganism culture and the production method by chemical synthesis are described below. Production Method by Microbial Culture In the present microorganism culture method, a microorganism capable of producing the compound (I) of the present invention is cultured under the medium composition and medium conditions used for normal fermentation production, and the normal fermentation product is separated and collected. Compound (I) is isolated. Microorganisms that can produce the compound (I) of the present invention include microorganisms belonging to the genus Stachybotrys, for example, Stachybotrys cylindrospora RF
-5900 strains are exemplified. Stachybotrys cylindrospora RF-5900 strain had the following morphological properties.

[0010] The colony on the cone-meal agar medium grows a little slower, its surface becomes moist, and turns green-black as conidia are formed. The conidiophores are solitary, hardly branched, and stand upright from the medium. The vegetative mycelium is colorless and the width is 1.0
多 3.0 mm, composed of multicellular and branched. Each conidiophore is colorless and consists of 3-5 cells, 65-95 m high
m, the width of the base is 5.0 to 5.5 mm, and narrows toward the top, and becomes 3.0 to 3.5 mm at the phialide-adhered portion. The surface is smooth or granular, and the upper part is particularly strong. The phialide is a single cell having a height of 10.0 to 13.5 mm and a width of 6.0 to 7.5 mm, and has a club-like shape, the surface is smooth, and 4 to 7 conidiophores are formed at the tip. Conidia length 1
Cylindrical single cell of 1.0 to 13.0 mm in width and 4.0 to 4.5 mm in width. It has a pale green black color, has a ridge-like surface structure along its long axis, and has a spherical wet surface on phialide. It is formed as a glossy, conidial mass.

On a potato dextrose agar medium, it grows in the range of 10 ° C. to 32 ° C., and the optimum growth temperature is 2 ° C.
0 ° C to 28 ° C. Since this strain does not form a sexual reproductive organ and has the property of forming conidia as a spherical wet shiny conidia mass at the tip of phialide at the tip of the conidiophore, it is a strain belonging to the genus Stachybotrys Is determined.

[0012] These properties are described by Dermatias Theus
High fomisetes (Dermatiaceous H)
yphomycetes), Commonwealth Mycological Institute, Kew, Slay, England (Commonwealth Mycology)
cal Institute, Kew, Surrey,
England), pp. 542-544, 1971.
Year; More Dermatiaseus Hyphomisetes
mycetes) Commonwealth Mycological Institute, Kew, Slay, England (C
ommonwealth Mylogical I
nstate, Kew, Surrey, Engla
nd), pp. 463-464, 1976; Compendium of Soil Fungi reprint (Compendium of Soil Fungi).
Reprint 1993, pp. 742-743.
Page; Icones Microfungor a Matsushima Rectrum, Icones Microfungor
um a Matsushima Lectorum,
Kobe), 1975; and the like. As a result of comparison with known species of the genus Stachybotrys described in literatures such as Stachybotrys cylindrospora C. based on conidial shape, size, surface structure and other properties. W. Jensen (CW Je
nsen) 1912. This strain was obtained from the National Institute of Advanced Industrial Science and Technology (AIST) under the accession number "FERM P-
15742 "has been deposited on July 17, 1996.

As a medium for producing the compound (I) of the present invention, any of a synthetic medium and a natural medium can be suitably used as long as it contains a carbon source, a nitrogen source and an inorganic salt. If necessary, vitamins or other nutrients may be added as appropriate.

Examples of the carbon source include sugars such as glucose, maltose, fructose, sucrose and starch; alcohols such as glycerol and mannitol;
One or more kinds may be appropriately selected and used from general carbon sources such as amino acids such as glycine, alanine and asparagine, and fats and oils such as soybean oil and olive oil in consideration of the assimilation of microorganisms. Nitrogen sources include soy flour, corn steep liquor, beef extract, peptone, yeast extract,
Amino acid mixture, organic nitrogen-containing compounds such as fish meal or ammonium salts, inorganic nitrogen compounds such as nitrates and the like,
One or more kinds may be appropriately selected and used in consideration of the assimilation property of the microorganism. As the inorganic salt, for example, calcium carbonate, sodium chloride, potassium chloride, magnesium sulfate, copper sulfate, manganese chloride, zinc sulfate, cobalt chloride, and various phosphates may be added as needed. Further, an antifoaming agent, for example, vegetable oil, polypropylene glycol or the like can be added as needed.

The culturing temperature can be appropriately changed within a range in which the microorganism grows and produces the compound (I) of the present invention, but is preferably from 10 ° C to 32 ° C, more preferably from 18 ° C to 25 ° C.
° C. The pH is preferably around 6 to 8, and the culture time is usually about several days to several weeks. The culture may be terminated when the production of the compound (I) of the present invention reaches the maximum. Any culturing method can be suitably used as long as it is a commonly used method such as solid layer culturing or aeration and stirring culturing.

The method of separating and collecting the fermentation product from the culture includes filtration, centrifugation, various types of ion exchange resins and other active adsorption, chromatography, extraction with various organic solvents, and the like. A material separation and purification method can be used. For example, cells isolated and purified from a culture may be extracted with an organic solvent such as ethyl acetate, acetone, or methyl ethyl ketone, and then separated and purified by a combination of silica gel column chromatography and thin layer chromatography.

[0017] The process optionally by chemical synthesis, the present invention compound obtained in fermentation of (I), ring-opening reaction under appropriate conditions, may be subjected to acylation reaction or oxidation reaction. For example, R ¹ is ＯO, R ² and R
Compound (I) in which ³ together forms -CH (OH)-in a solvent such as acetonitrile, tetrahydrofuran, dioxane, ethyl ether, methanol, ethanol, cyclohexane, etc., under ice-cooling to heating, preferably around room temperature. For several minutes to several hours, if treated with an alkali such as sodium hydroxide, potassium hydroxide, sodium carbonate, potassium carbonate, R ¹ is hydroxy, R ² is —CHO, and R ³ is R ¹ The compound (I) which forms a double bond with the bonded carbon atom is obtained.

Further, the same R ¹ as above is ＝ O, and R ²
And R ³ together form -CH (OH)-in a solvent such as tetrahydrofuran, dioxane or ethyl ether in the presence of a base such as pyridine or triethylamine in the presence of a base such as acetyl chloride or propionyl chloride. When acylation is performed by reacting with an acid anhydride such as acid chloride or acetic anhydride under ice-cooling to heating, preferably at about room temperature for several hours to several tens of hours, R ¹ is ＯO, R ² and R ³ Can be combined to form -CH (acyloxy)-to obtain a compound (I). In this reaction, the base may be used in a solvent amount.

Further, R ¹ is と O, and R ² is the same as above.
And R ³ together form —CH (OH) — to form compound (I) in tetrahydrofuran, dioxane, ethyl ether, acetone, methyl chloride, dichloromethane,
Sarett oxidation (CrO ₃ -pyridine) in a suitable solvent such as 1,2-dichloroethane or Jones reagent (chromic anhydride + sulfuric acid), sodium hypochlorite, sodium chlorite, manganese dioxide Using a commonly used oxidizing agent such as, for example, under ice-cooling to room temperature, preferably under ice-cooling for several hours to several tens of hours, R ¹ =
O, and R ² and R ³ together form -C (= O)-
Can be obtained.

The compound (I) of the present invention also includes pharmaceutically acceptable salts thereof, for example, alkali metal salts such as sodium and potassium, and alkaline earth metal salts such as calcium and barium. These salts can be formed using commonly used reactions. The compound of the present invention also includes a hydrate thereof, and one molecule of the compound (I) of the present invention may be bonded to one or more water molecules.

Since the compound (I) of the present invention has a strong chymase inhibitory activity and / or an antibacterial activity, it can be administered as a medicament to animals including humans. Specific examples of chymase inhibitors include myocardial infarction, cardiac hypertrophy, heart failure, cardiomyopathy, hypertension, intimal thickening after PTCA (percutaneous coronary angioplasty), peripheral circulatory disorders, vasculitis, and atherosclerosis It is effective in treating chronic cardiovascular disorders such as diabetic and non-diabetic renal disorders and allergic diseases such as inflammation, allergy, rheumatism and atopy. As an antibacterial agent, it is effective for treatment of deep mycosis such as visceral candidiasis and pulmonary aspergillosis and dermatomycosis.

When the compound (I) of the present invention is administered as a medicament, it can be administered orally or parenterally. For oral administration, tablets, granules, powders,
What is necessary is just to prepare and administer in a normally used dosage form such as a capsule, pill, liquid, suspension, syrup, buccal or sublingual. For parenteral administration, any commonly used dosage form, such as injections such as intramuscular administration, suppositories, transdermal absorption agents, inhalants, etc., can be suitably administered, but intravenous administration is particularly preferred.

The pharmaceutical composition of the present invention contains various pharmaceutical additives such as excipients, binders, wetting agents, disintegrants, lubricants and diluents suitable for the final dosage form in an effective amount of the active ingredient. Can be mixed and prepared as needed. In the case of an injection, a preparation may be prepared by sterilizing with an appropriate carrier.

Specifically, the excipients include lactose, sucrose,
Glucose, starch, calcium carbonate or crystalline cellulose, etc .; binders such as methylcellulose, carboxymethylcellulose, hydroxypropylcellulose, gelatin or polyvinylpyrrolidone; disintegrants: carboxymethylcellulose, sodium carboxymethylcellulose, starch, sodium alginate, agar powder or lauryl Lubricants such as sodium sulfate include talc, magnesium stearate and macrogol. Cocoa butter, macrogol, methylcellulose, or the like can be used as a suppository base. In addition, when prepared as a liquid or emulsion, suspension injection, a solubilizing agent usually used,
Suspending agents, emulsifiers, stabilizers, preservatives, isotonic agents and the like may be appropriately added, and in the case of oral administration, a flavoring agent, a fragrance and the like may be added.

The dose of the chymase inhibitor is preferably determined in consideration of the patient's age, body weight, administration route, type and degree of disease, and the like. Usually 1 μg to 1000 mg /
The dose may be administered once to several times per kg / day. Also,
When administered parenterally, 0.1 μg to 100 mg / kg / day usually varies depending on the administration route.
It may be administered in one to several doses. Similarly, it is desirable to set the dose as an antibacterial agent in consideration of the patient's age, body weight, administration route, type and degree of illness, etc. Normally 1μ
In the case of parenteral administration of g to 200 mg / kg / day, 0.1 μg to 20 mg / kg / day may be administered once to several times each.

[0026]

EXAMPLES The present invention will be described in more detail with reference to the following Examples, but it should not be construed that the present invention is limited thereto. Example 1 Production of Compound (I) by Fermentation (1) Fermentation Step Polypeptone 1.0%, glucose 2.0%, beef extract 0.3%, yeast extract 0.2%,
Stachybotrys cylindrospora RF-59 was placed in a 500 ml Mayer flask containing 100 ml of a medium consisting of 0.1% sodium chloride tap water (before pH 7.0 sterilization).
00 spores from the seed culture slant and 180
For 3 days on a rotary shaker for 3 minutes. Each 4 ml of this culture solution was inoculated into 200 500 ml Mayer flasks containing brown rice medium (25 g brown rice, 0.5 g glucose, 0.1 g difco yeast extract, 50 ml tap water) autoclaved at 121 ° C. for 30 minutes. The cells were cultured at 28 ° C. for 14 days.

(2) Separation Step The fermented products in the respective Meyer flasks obtained in the fermentation step were collected, added with 20 L of acetone and extracted with stirring. After suction filtration, the filtrate containing acetone was evaporated under reduced pressure to remove the solvent, the aqueous layer of the residue was extracted with ethyl acetate, and the ethyl acetate layer was concentrated to dryness to obtain a crude fraction (178.0 g). This crude fraction was divided into two equal parts, and the crude fraction (89.0 g) was subjected to silica gel column chromatography (Merck Kieselgel 60, 500 g,
Purified with 30-400 mesh, toluene: acetonitrile (9: 1). Elution with a total of 1800 ml of toluene: acetonitrile (9: 1)
The l was collected and fraction A was collected, and the next 1000 ml was collected to form the B fraction. The two batches were combined and the solvent was distilled off under reduced pressure to obtain the A fraction (5.39 g) and the B fraction (3.61 g). Got each.

Fraction A (5.39 g) was again subjected to silica gel column chromatography (Merck Kieselgel 60, 2).
30 g, 230-400 mesh, toluene: acetonitrile (95: 5)) to give crude compound (I-1) 9
24 mg were obtained. Furthermore, silica gel column chromatography (Merck Kieselgel 60, 100 g, 230-4
Purification was performed using 00 mesh, toluene: ethyl acetate: isopropanol (59: 8: 1), and fractions were collected in 10 g portions, and fractions 13 to 18 were collected and the solvent was distilled off under reduced pressure. The oily residue thus obtained is treated with n-hexane 5
After recrystallizing in ml, the crystals were collected by filtration to obtain 725 mg of compound (I-1) as pale yellow needles.

The B fraction (3.61 g) was subjected to silica gel column chromatography (Merck Kieselgel 60, 170).
g, 230-400 mesh, n-hexane: acetone (2: 1)). Fractions were collected in 10 g portions, and fractions 49 to 89 were collected and the solvent was distilled off under reduced pressure to obtain 583 mg of compound (I-2).

Subsequently, fractions 32-36 were collected and the solvent was distilled off under reduced pressure to obtain 289 mg of a crude compound (I-3). This was subjected to thin layer chromatography (Merck Precoated TLC plate, silica gel F-254, 0.5 m
m, n-hexane: acetonitrile (2: 1)) and thin layer chromatography (Merck Precoated TLC)
The mixture was repeatedly purified on a plate, silica gel F-254, 0.5 mm, toluene: acetonitrile (2: 1)) to obtain 35 mg of compound (I-3).

Compound (I-1) R ¹ is ＝ O, and R ² and R ³ are taken together to form —CH (O
H)-formed (physicochemical properties) Properties: pale yellow needles Solubility: chloroform, diethyl ether, acetone,
Ethyl acetate, soluble in methanol, slightly soluble in n-hexane, insoluble in water Melting point: 146 to 146.5 ° C. [α] _D ²³ = −408.0 ± 8.7 ° (c = 0.514,
Chloroform) Elemental analysis: (C ₂₅ H ₂₆ O as ₅₎ Calculated; C: 73.87%, H: 6.45% Found; C: 73.83%, H: 6.55% L-SIMS, m / z: 407 (MH ⁺ )

IR, λ _max KBrcm ^-1 : 3560,3
436, 2979, 2966, 2933, 2912, 2
857, 1722, 1691, 1666, 1631, 1
598, 1579, 1470, 1446, 1401, 1
388, 1286, 1257, 1238, 1227, 1
187,1153,1114,1001,957,90
9,878,857,779 UV (methanol), nm (ε): 260 (10,60
0), 320 (3,700)

¹ H NMR (CDCl ₃ , 600 MHz)
δ: 1.04 (3H, s like), 1.52 (3
H, s like), 1.58 (3H, s like)
e), 2.14 (3H, d, J = 1.4 Hz), 2.2
3 (1H, dd, J = 7.0, 14.2 Hz), 2.4
4 (1H, m), 2.58 (1H, t, J = 12.7H
z), 2.86 (1H, dd, J = 9.0, 14.2H
z), 3.67 (1H, d, J = 12.5 Hz), 4.
62 (1H, m), 4.77 (1H, m), 4.95
(1H, d, J = 12.4 Hz), 5.18 (1H,
m), 5.88 (1H, ddd, J = 0.8, 1.0,
12.5 Hz), 6.68 (1H, q, J = 1.4H)
z), 6.96 (1H, ddd, J = 0.8, 1.2,
8.2 Hz), 7.34 (1H, ddd, J = 1.0,
1.2, 7.2 Hz), 7.58 (1H, m)

¹³ C NMR (CDCl ₃ , 150 MHz)
δ: 11.39 (q), 17.94 (q), 19.64
(Q), 25.84 (q), 35.56 (t), 36.
05 (t), 55.98 (s), 69.06 (d), 7
6.54 (t), 91.16 (s), 115.74
(D), 116.67 (d), 120.24 (d), 1
21.44 (s), 131.67 (d), 131.72
(D), 132.57 (s), 136.58 (d), 1
36.66 (s), 152.39 (s), 153.56
(S), 158.16 (s), 187.13 (s), 1
91.05 (s), 205.17 (s)

Compound (I-2) R ¹ is —OH, R ² is —CHO, and R ³ forms a double bond with the carbon atom to which R ¹ is bonded (7-trans form) (Physicochemical properties) ) Properties: pale yellow powder Solubility: chloroform, diethyl ether, acetone,
Ethyl acetate, soluble in methanol, slightly soluble in n-hexane, insoluble in water [α] _D ²³ = −425.2 ± 9.1 ° (c = 0.509,
Chloroform) L-SIMS, m / z: 407 (MH ⁺ ) HR-LSIMS, m / z (as C ₂₅ H ₂₆ O ₅ ) Calculated: 407.1857 Actual; 407.15959 (MH) ⁺

IR, λ _max CHCl ₃ cm ^-1 : 302
8,3016,2978,2920,2889,285
7,2737,1703,1657,1643 (s
h), 1563, 1465, 1417, 1386, 13
47,1259,1040,1009,955,92
5,868 UV (methanol), nm (ε): 265 (8,50
0), 278 (6,900)

¹ H NMR (CDCl ₃ , 600 MHz)
δ: 1.32 (3H, s like), 1.54 (3
H, s like), 1.59 (3H, s like)
e), 2.09 (3H, d, J = 1.4 Hz), 2.2
7 (1H, dd, J = 8.0, 14.8 Hz), 2.2
9 (1H, m), 2.38 (1H, t, J = 12.5H
z), 2.90 (1H, m), 4.41 (1H, dl)
ike, J = 12.1 Hz), 4.55 (1H, m),
4.58 (1H, d, J = 12.1 Hz), 5.44
(1H, m), 6.37 (1H, q, J = 1.4H
z), 7.47 (1H, m), 7.50 (1H, m),
7.60 (1H, m), 9.96 (1H, s), 16.
44 (1H, s)

¹³ C NMR (CDCl ₃ , 150 MHz)
δ: 18.10 (q), 19.90 (q), 20.28
(Q), 25.74 (q), 34.26 (t), 38.
41 (t), 58.46 (s), 76.71 (t), 1
13.17 (s), 118.38 (d), 122.35
(D), 125.12 (d), 126.28 (s), 1
29.11 (d), 129.50 (d), 133.17
(S), 134.36 (s), 134.86 (s), 1
35.72 (s), 153.70 (s), 164.85
(S), 182.08 (s), 190.56 (d), 1
97.02 (s), 198.12 (s)

Compound (I-3) R ¹ forms —OH, R ² forms —CHO, and R ³ forms a double bond with the carbon atom to which R ¹ is bonded (7-cis form) (physicochemical properties) ) Properties: pale yellow powder Solubility: chloroform, diethyl ether, acetone,
Ethyl acetate, soluble in methanol, slightly soluble in n-hexane, insoluble in water [α] _D ²³ = −407.6 ± 8.9 ° (c = 0.50)
3, chloroform) L-SIMS, m / z: 407 (MH ⁺ ) HR-LSIMS, m / z (as C ₂₅ H ₂₆ O ₅ ) Calculated; 407.1857 Actual; 407.15959 (MH) ⁺

IR, λ _max CHCl ₃ cm ^-1 : 3012
2974, 2919, 2878, 1695, 1654,
1638, 1561, 1459, 1415, 1378,
1306,1261,1167,1101,957,9
27,880,865 UV (MeOH), nm (ε): 330 (7,700)

¹ H NMR (CDCl ₃ , 600 MHz)
δ: 1.50 (3H, br.s), 1.56 (3H, b.
r. s), 1.83 (3H, s), 2.10 (3H,
d, J = 1.4 Hz), 2.23 (1H, m), 2.2
4 (1H, m), 2.67 (1H, dd, J = 11.
0, 13.3 Hz), 2.83 (1H, m), 4.13
(1H, br.d, J = 8.4 Hz), 4.41 (1
H, d, J = 8.4 Hz), 4.51 (1H, m),
5.28 (1H, m), 6.34 (1H, q, J = 1.
4Hz), 7.21 (1H, dd, J = 0.8, 8.2)
Hz), 7.54 (1H, t, J = 8.2 Hz), 7.5.
68 (1H, dd, J = 0.8, 8.2 Hz);
02 (1H, s), 16.03 (1H, s)

¹³ C NMR (CDCl ₃ , 150 MHz)
δ: 18.12 (q), 20.25 (q), 22.99
(Q), 25.73 (q), 36.81 (t), 39.
09 (t), 59.17 (s), 70.55 (t), 1
13.07 (s), 117.72 (d), 121.88
(D), 122.25 (d), 123.46 (d), 1
23.96 (d), 130.72 (d), 134.13
(S), 134.40 (s), 134.50 (s), 1
35.66 (s), 156.07 (s), 164.70
(S), 181.72 (s), 190.91 (d), 1
96.73 (s), 198.56 (s)

Example 2 Compound (I-1) to Compound
(I-2) Derivation A 1N aqueous sodium hydroxide solution (0.02 ml) was added to a solution of compound (I-1) (5.2 mg) in acetonitrile (0.2 ml), and the mixture was stirred at room temperature for 30 minutes. After adding 1N hydrochloric acid (0.02 ml) to the reaction solution, water (1 m
1), and separated with ethyl acetate (1 ml). The ethyl acetate layer was dried over sodium sulfate, and the solvent was distilled off under reduced pressure to give compound (I-
2) 4.8 mg was obtained.

Example 3 Preparation of Compound (I-4) A solution of compound (I-1) (4 mg, 0.01 mmol) in pyridine (0.15 ml) was added with acetic anhydride (0.05 ml).
Was added and left for 14 hours. Toluene was added to the reaction solution, and the solvent was distilled off under reduced pressure to obtain 4 mg of compound (I-4). Compound (I-4) R ¹ is ＯO, R ² and R ³ are taken together to form —CH (OA
c)-formed EIMS, m / z: 448 (M) ⁺ IR, λ _max CHCl ₃ cm ^-1 : 2933,1744,1
705,1676,1627,1603,1583,1
474,1452,1377,1296,1261,1
233, 1222, 1210, 1190, 1165, 1
100,1078,1049,1017,936,88
6,828

¹ H NMR (CDCl ₃ , 300 MHz)
δ: 1.06 (3H, s), 1.57 (6H, s),
2.12 (3H, s), 2.13 (3H, d, J = 1.
5 Hz), 2.26 (1H, dd, J = 6.6,14.
0 Hz), 2.40 (1H, m), 2.58 (1H,
t, J = 12.6 Hz), 2.85 (1H, dd, J =
8.5, 14.0 Hz), 4.65 (1H, m), 4.
76 (1H, m), 4.97 (1H, d, J = 12.6)
Hz), 5.15 (1H, m), 6.65 (1H,
m), 6.87 (1H, m), 6.96 (1H, m),
7.10 (1H, m), 7.54 (1H, m)

Example 4 Preparation of Compound (I-5) A Jones reagent (0.02 ml) was added to a solution of compound (I-1) (8 mg, 0.02 mmol) in acetone (0.47 ml).
5 mmol) and stirred for 1.5 hours under ice-cooling. Further, a Jones reagent (0.1 mmol) was added and the mixture was stirred under ice cooling for 5 hours. After the reaction was stopped by adding 2 drops of isopropanol to the reaction solution, the mixture was neutralized with a 10% aqueous sodium hydrogen carbonate solution, and the solvent was distilled off under reduced pressure. The residue was treated with ethyl acetate (20 m
l), partition extraction with water (20 ml) and concentration of the ethyl acetate layer, followed by thin layer chromatography (Merck Precoated TLC plate, silica gel F-254, 0.5 mm,
The product was purified with n-hexane: acetone (1: 1) to obtain 3.9 mg of compound (I-5).

Compound (I-5) R ¹ is ＯO, R ² and R ³ are taken together to form —C (（O)
L-SIMS, m / z: 405 (MH) ⁺ IR, λ _max CHCl ₃ cm ^-1 : 2976, 2931, ¹
772, 1756, 1716, 1703, 1674, 1
621, 1598, 1576, 1474, 1451, 1
379,1280,1263,1238,1221,1
210, 1198, 1140, 1109, 1063, 1
045,1020,970,819

¹ H NMR (CDCl ₃ , 300 MHz)
δ: 1.08 (3H, s), 1.50 (3H, s),
1.62 (3H, s), 2.14 (3H, d, J = 1.
2Hz), 2.23 (1H, dd, J = 7.2, 14.
1Hz), 2.46 (1H, m), 2.58 (1H,
t, J = 12.9 Hz), 2.89 (1H, dd, J =
8.7, 14.1 Hz), 3.67 (1H, d, J = 1)
2.5 Hz), 4.65 (1H, m), 4.87 (1
H, m), 5.04 (1H, d, J = 12.6 Hz),
5.26 (1H, m), 6.64 (1H, m), 7.3
1 (1H, m), 7.62 (1H, m), 7.68 (1
H, m)

¹³ C NMR (CDCl ₃ , 75 MHz)
δ: 11.20, 17.87, 19.64, 25.9
0, 35.40, 36.13, 57.60, 88.9
8, 115.50, 116.60, 126.86, 12
8.55, 131.92, 132.20, 133.8
6,136.38,137.06,144.66,15
4.40, 158.54, 185.47, 188.1
3,188.32, 201.19

Test Example 1 Chymase Enzyme Inhibitory Activity (1) Preparation of Compound (I) Compound (I) was all dimethyl sulfoxide (DM
(SO) to a concentration of 10 mg / ml.
The DMSO concentration brought in for the activity measurement was 1%. (2) Measurement of chymase inhibitory activity Buffer solution (0.1 M Tris-HCl, 1.8 M NaCl
Compound (I) dissolved in DMSO in pH 8.0)
And Urata et al.'S method (Journal of Biological
Chemistry (Journal of Biology)
cal Chemistry) Vol. 265, Vol. 2234
8 to 22357), and treated at 37 ° C. for 30 minutes.
Ala-Ala-Pro-Phe-pNA (BACHEM Feinchemikalien A
G) was added to a concentration of 0.5 mM, and an enzyme reaction was carried out at 37 ° C. After the reaction, the absorbance of the solution (405 n
m) was measured and the inhibition rate was calculated. (3) Results Regarding the above compound, 5% of its human chymase inhibitory activity
0% inhibition concentration (hereinafter referred to as IC ₅₀₎ shown in Table 1.

[0051]

[Table 1] Table 1 shows that the compound (I) of the present invention has chymase inhibitory activity.

Test Example 2 Antifungal Activity (1) Method for Preparing Specimens The compounds of the present invention were all dimethyl sulfoxide (DMS)
O) to give a concentration of 10 mg / ml. (2) Measurement of activity Candida albicans Ca-15-3 ( Candi
da albicans ) (Sabouraud dextrose broth (S) supplemented with 10% fetal bovine serum.
and suspended in about 2 × 10 ⁸ cfu / ml.
C.) stored in a yeast nitrogen base (YN Nitrogen Base; YN).
B, manufactured by Difco) diluted to 1 × 10 ⁵ cfu / ml with a medium, and 100 μl was added to each well of a 96-well microplate.
1 was dispensed. The compound of the present invention is diluted with this bacterial solution to give 2
A two-fold dilution series was prepared to examine growth inhibition.

(3) Determination of culture conditions and results The determination of growth inhibition was determined by culturing at 30 ° C. for 24 hours and then measuring the absorbance at 595 nm using an autoreader (BIO-RAD). 50% of the absorbance of
The concentration that inhibits the growth of bacteria is defined below as the minimum effective concentration (M
(IC ₅₀ ). Table 2 shows the results.

[0054]

[Table 2] Table 2 shows that the compound (I) of the present invention has an antibacterial activity.

Formulation Example 1 Compound (I) of the present invention 3 mg Gelatin 150 mg Phenol 4 mg The above components were dissolved in an appropriate amount of purified water for injection to prepare an injection solution.

[0056]

As apparent from the above test examples, the compound (I) of the present invention exhibits a chymase inhibitory activity and / or an antibacterial activity. Therefore, the compound (I) of the present invention is very useful as a therapeutic agent for chronic cardiovascular disorders, a therapeutic agent for allergic diseases and / or an antibacterial agent.

[Brief description of the drawings]

FIG. 1 is a graph showing an IR spectrum of compound I-1.

FIG. 2 is a graph showing ¹ HNMR spectrum of compound I-1.

FIG. 3 is a graph showing a ¹³ C NMR spectrum of compound I-1.

FIG. 4 is a graph showing an IR spectrum of compound I-2.

FIG. 5 is a graph showing ¹ HNMR spectrum of compound I-2.

FIG. 6 is a graph showing a ¹³ C NMR spectrum of compound I-2.

FIG. 7 is a graph showing an IR spectrum of compound I-3.

FIG. 8 is a graph showing ¹ HNMR spectrum of compound I-3.

FIG. 9 is a graph showing a ¹³ C NMR spectrum of compound I-3.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁶ Identification code FI A61K 31/335 ADZ A61K 31/335 ADZ AED AED C12P 17/08 C12P 17/08 // (C12P 17/08 C12R 1: 645)

Claims (7)

[Claims] 1. A compound of formula (I): Wherein R ¹ is hydroxy, R ² is —CHO, and R ³ forms a double bond with the carbon atom to which R ¹ is attached, or R ¹ is ＯO; And R ² and R
³ together -CH (OR ⁴⁾ - (wherein R ⁴ is hydrogen or acyl) or -C (= O) - to form) a compound or a pharmaceutically acceptable salt or thereof that shown Hydrate. 2. A pharmaceutical composition comprising the compound according to claim 1 or a pharmaceutically acceptable salt thereof or a hydrate thereof. 3. A chymase inhibitor comprising the compound according to claim 1, a pharmaceutically acceptable salt thereof, or a hydrate thereof. 4. An antibacterial agent comprising the compound according to claim 1, or a pharmaceutically acceptable salt thereof, or a hydrate thereof. 5. Stachybotr ( Stachybotr)
ys ) culturing a microorganism belonging to the genus and capable of producing any of the compounds according to claim 1, collecting the compound from the resulting culture, and optionally further opening, closing, acylating or oxidizing the compound. The method for producing a compound according to claim 1, characterized in that: 6. Stachybotrys cylindrospora ( S
tachybotrysylindrospora )
And a microorganism which produces any of the compounds according to claim 1. 7. Stachybotrys cylindrospora R
F-5900 ( Stachybotrys cylin
The microorganism according to claim 6, wherein the microorganism is Drospora RF-5900).