JPH04134048A - Compound tan-1,415 derivative, production thereof and endothelin acceptor antagonist - Google Patents

Abstract

NEW MATERIAL:A compound shown by formula I (R1 and R4 are H or lower alkyl which may be substituted; R2 and R5 are H or CH3; R3 is H or Cl; with the proviso that when R3 is H, at least one of R1 and R4 is 2-6C alkyl which may be substituted and when R3 is Cl, a case where R1 is H or R4 is CH3 is omitted) or a salt thereof. USE:An endothelin acceptor antagonist useful for a preventive and remedy for myocardial infarction and renal failure. PREPARATION:Carboxyl groups of a compound shown by formula II are subjected to lower alkylation reaction to give a compound shown by formula I.

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to an endothelin receptor antagonist, a novel compound having the antagonistic action, and a method for producing the same.

BACKGROUND OF THE INVENTION Drugs commonly used as therapeutic agents for myocardial infarction include vasodilators and thrombolytic agents. These are applied to patients with mild symptoms, but surgery, i.e. coronary artery bypass grafting, is applied to patients with severe symptoms.
sgrafLing) and percutaneous transluminal coronary angioplasty (percutaneous transluminal coronary angioplasty)
nal coronary angioplasty)
etc. will be applied.

On the other hand, there are currently no effective drugs for acute renal failure, and only adenosine i-liphosphate (ATP) is administered as an energy booster.

Problems to be Solved by the Invention As mentioned above, the only treatments available for myocardial infarction or acute renal failure are symptomatic treatments, but recently endothelin (Endothelin), a vasoconstrictor present in vascular endothelial cells, has been developed.
oLhelin) was discovered [MYanagisa
wa et al., Nature, Volume 332, 4
11 pages (1988)]. E-endothelin is a peptide consisting of 21 amino acids, and exhibits strong vasoconstrictive activity, blood pressure increasing activity, and contractile force enhancing effect on cardiac muscle. Additionally, endothelin is associated with myocardial infarction [T, Watan
abe et al., Nature, vol. 344, p. 114 (1990)] or acute renal failure [YS
Hibouta et al., Life Science
5sciences), Volume 46, Page 1611 (19
90)], etc.).

Means for Solving the Problem The inventors have stated that, in view of the two situations, if it is possible to find a drug that selectively inhibits endothelin receptors,
Convinced that a fundamental preventive or therapeutic agent for the above-mentioned diseases could be obtained, he searched for its origin in microbial metabolites. As a result of intensive research using the method described below, compounds TAN-1,415A and B were isolated from the culture medium and found to exhibit strong endothelin receptor antagonism. Subsequently, as a result of further research, TAN-1415A was identified as astyl phosphoric acid and TAN-1415B was 4-chloroasterphosphoric acid, as described below, and new derivatives of these compounds were also synthesized, thereby completing the present invention.

That is, the present invention provides the following: 1) General formula (1) [wherein R and R4 are hydrogen atoms or optionally substituted lower alkyl groups, R3 and R5 are hydrogen atoms or methyl groups, and R3 is a hydrogen atom] Or indicates a chlorine atom. However, when R3 is a hydrogen atom, at least one of RI and R4 represents an optionally substituted lower alkyl group having 2 to 6 carbon atoms, and when R3 is a chlorine atom, R5 is hydrogen and R4 is Except when it is methyl. ] or a salt thereof, 2) General formula (2) [wherein R3 and R5 are hydrogen atoms or methyl groups,
R3 represents a hydrogen atom or a chlorine atom. ], and 3) a method for producing a compound of general formula (3), which comprises subjecting the carboxyl group of the compound to a lower alkylation reaction, and 3) general formula (3) [wherein R'1 and R'4 are R' and R'5 represent a hydrogen atom or a lower alkyl group which may be substituted; R'3 represents a hydrogen atom or a chlorine atom; ] It is an endothelin receptor antagonist containing a compound shown in the formula or a salt thereof as an active ingredient.

As the alkyl group in the optionally substituted lower alkyl group represented by R'1 or R'4 in general formula (3), CI-8 alkyl (e.g., methylethyl, propyl, isopropyl, butyl, isobutyl, pentyl ,
impentyl, hexylisohexyl, etc.), and the substituents include, for example, C3-8 cycloalkyl (e.g., cyclopropyl cyclobutylcyclopentyl, cyclohexyl, etc.), phenyl which may have it[14, etc.
phenyl, 0-hydroxyphenyl, m-hydroxyphenyl, p-hydroxyphenyl, etc.), hydroxy, mercapto.

C1-3 alkylthio (eg, methylthio, ethylthiopropylthio, etc.), carboxy, guanidino, amino.

Examples include imidazolyl. Specific examples of the optionally substituted alkyl include methyl, ethyl, propyl, isopropyl, butyl, and isobutyl.

Cyclohexylmethyl, benzyl, p-hydroxybenzyl, hydroxymethyl, mercaptomethyl 1-hydroxyethyl, 2-methylthioethyl, carboxymethyl, 2-carboxyethyl, 3-guanidinopropyl, 4
-Aminobutyl, 4-imidazolylmethyl, and the like. - In murder (1), R and R4 also have the same meanings as the optionally substituted lower alkyl group, and the optionally substituted lower alkyl group having 2 to 6 carbon atoms is methyl. It has the same meaning as ``except for the base''.

The compound in which RZ+ R', R'3 and R'5 are hydrogen atoms and R/4 is a methyl group in the above-murder (3) is based on the literature [H, NiN15hika et al.; Society Japan (Bull, Agr, Chem, Soc,
Japan) + Vol. 18, p. 13, 1942], aster phosphoric acid
ic acid: 1). Also - murder (3)
where R' l + R' 2 and R' are hydrogen atoms, R
The compound in which '3 is a chlorine atom and R'4 is a methyl group is 4-chloroasterlic acid (2), which is a known compound described in Japanese Patent Publication No. 188084/1984.

Compounds 1 and 2 can also be produced by strain FL-8556, which was newly isolated from soil in Oita Prefecture.

This strain exhibits the following properties.

a) Morphological characteristics This strain FL-8556 grows well on malt extract agar, potato-glucose agar, synthetic mucor agar, etc., and also forms abundant conidia. Mycelia are transparent;
It has septa, and conidiophores arise directly from basal hyphae and aerial hyphae. Conidiophores are almost unbranched, 100μ
It has a length of more than m, and its surface is smooth. At its tip, 3 to 5 metula branches are often found at two points, or another metula is often found just below. At the tip of each metμ, there are several brush-like phialides (7 to 10μm) of almost uniform length.
phialides), several dozen chained conidia are observed in a broom-like shape, and a broom-like body (pen ic i l l
+) is formed. Conidia are almost spherical; 2.
It has a diameter of 0 to 3°0 μm and its surface is smooth.

Furthermore, no sclerotia or fungal bundles were observed on the agar medium examined.

b) Properties on agar medium ■Zupek agar medium: Growth at 24°C is very limited in the early stage, with a colony diameter of 25 m on the 7th day.
m, it does not grow in a circular shape and is amorphous, but it gradually grows vigorously and reaches 58 mm on the 144th day, growing in an almost circular shape. At the early stage of growth, there are almost no aerial hyphae and they are leather-like, but gradually relatively long aerial hyphae come to cover the entire colony.

Its color also changes from gray-white to gray-yellow-green to dark brown. The underside changes from light yellowish brown to dark brown.

After 10 days, a pale yellow pigment is observed on the agar medium.

■Malt extract agar medium: Growth is good at 24°C, reaching 35mm on the 7th day. The surface is velvety and changes from gray to gray-green to dark yellow-green on the 144th day, a color typical of the genus Penicillium.

Conidia are formed in great abundance. Initially, the color of the back side is yellow-green at the periphery and dark green at the center, but gradually changes to dark green overall. Soluble pigments may not be observed at the initial stage, or pale yellow pigments may be observed after 140 hours. No sclerotia formation is observed.

■YpSs agar medium; growth is good at 24°C.
The colony size reaches 35 mm in the white of the eye.

The hyphae do not spread, their surface varies from felt-like to velvety-like, and the conidia are dark green to blue-green in color with a white margin. The underside is yellowish brown to bright orange. A yellow soluble pigment is observed.

C) Physiological properties The growth conditions of this strain were investigated on a potato-glucose agar medium. Optimum growth temperature is 20-28°C5 Optimum pH is 2-28°C
7, and the growth temperature is 9 to 30°C, and the pH is 2 to 8.
, 5.

As a result, mainly based on morphological features, A Manualof the Benicilia
he Pencillia by B, Rapp
er and C, Thom 1949), The Williams & Willikins Company (The
Williams and Witkins Comp
any ), and A Laboratory Guide to Common Penicillium Subesii (A Labo
ratoy Guide to Common Pen
iciliumSpecies, by Jhon
I, Pity, I 988), Commonwealth Scientific and Industrial Research Organization (Commonwealth Scientific and Industrial Research Organization)
5Ccientific and Industrial
real Research Organization
Penicillium citrinum (
It was identified as Penicillium citrinum).

This strain was acquired by Fermentation Research Institute (I) on September 3, 1990.
FO) with accession number IFO3232/l, and on September 10, 1990, with the Ministry of International Trade and Industry, Agency of Industrial Science and Technology, Microbial Research Institute (FRI), accession number FERM BP-3.
093, respectively.

The medium used for culturing the compound-producing bacteria may be either liquid or solid as long as it contains a nutrient source that can be used in a vacuum, but it is more appropriate to use a liquid medium when processing in large quantities. The medium is appropriately mixed with carbon sources, nitrogen sources, inorganic substances, and micronutrient sources that can be assimilated by the compound-producing bacteria. Examples of carbon sources include glucose, lactose, sucrose,
Maltose, dextrin, starch, glycerin, mannitol
Nitrogen sources include meat extracts, yeast extracts, dried yeast, soybean flour, corn staple liquor, etc. , peptone, cottonseed flour, blackstrap molasses, urea, ammonium salts (eg, ammonium sulfate, ammonium chloride, ammonium nitrate, ammonium acetate, etc.) and others are used. 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, etc., and salts of organic acids such as acetic acid, propionic acid, etc. are used as appropriate. . Other amino acids (
e.g., glutamic acid, aspartic acid, alanine, lysine, methionine, proline, etc.), peptides (e.g., dipeptides, tripeptides, etc.), vitamins (e.g., B1, B
2, nicotinic acid, B12, C, etc.), nucleic acids (eg, purines, pyrimidines, derivatives thereof, etc.). Of course, inorganic or organic acids or alkalis, buffers, etc. may be added for the purpose of adjusting the pH of the medium, or appropriate amounts of oils and fats, surfactants, etc. may be added for the purpose of defoaming. In liquid culture, the pH of the medium is preferably near neutral, particularly pH 5 to 8. Culture temperature is approximately 20'C
~30'C, and the culture time is preferably about 48 hours to 168 hours.

As a method for isolating active substances from culture fluid, is the generally known isolation method for fat-soluble substances applied, solvent extraction method as shown in the reference example, chromatography method using adsorbent resin, etc.? Used to advantage. Further, the desired compound can be obtained as a solid by concentrating, crystallizing, or freeze-drying the fractionated components.

Using the thus obtained compounds 1 and 2, the known literature [
R8P, CurLis et al., Journal of the Chemical Society (J, Chem, Soc, 196
0, page 4838], the methyl derivatives described below can be prepared.

Furthermore, the following method is generally known as an example of introducing an alkyl group into an acidic group such as a carboxylic acid or phenol, and can be fully applied to the method for producing the compound of the present invention.

1) The raw material compound is a diazoalkane (e.g. diazomethane)
and a solvent (eg, ethyl ether, tetrahydrofuran, dioxane, methanol, etc.) at about 0°C to reflux temperature for about 2 minutes to 10 hours.

2) Activate the raw material compound with an alkyl halide (e.g.
methyl iodide, n-butyl chloride, etc.). Appropriate reaction conditions include using a solvent (for example, dimethylformamide, dimethylacetamide, etc.) and allowing the reaction to occur at about 0° C. to 60° C. for about 2 minutes to 20 hours. Even if an alkali metal salt (eg, sodium carbonate, potassium carbonate), ammonia, triethylamine, etc. are present in this reaction solution, there is no problem in the progress of the reaction.

3) Reacting the raw material compound with an alcohol such as methanol. In this reaction, a mineral acid such as hydrochloric acid, sulfuric acid or p-toluenesulfonic acid is used as a condensing agent. The reaction conditions are about 0'C to reflux temperature for 15 minutes to 20 hours, and the solvent used is the corresponding alcohol or a mixture thereof with chloroform, dichloromethane, dioxane, etc.

Pharmaceutically acceptable salts of the compounds represented by general formulas (1) and (3) include sodium salts, potassium salts,
Examples include calcium salts or ammonium salts,
These salts can be prepared by methods known per se.

As the compound represented by the general formula (3), the following compound examples can be specifically mentioned.

(Margins below) The above compounds 1 to 4 were obtained in the reference examples described below, IO and 11 were obtained in the examples described below, and compounds 5 to 9 were obtained in the above literature [J, Chem, Soc.

1960, page 4838].

For production Next, the biological activity of the compound of general formula (3) will be described.

(a) Radio receptor measurement Pig hearts were used for endothelin radio receptor measurements, with the aorta, atrium and fat removed.
Ventricles (200-250 g) were used. After the ventricles are washed,
Shred and add 10 volumes of C solution (20mM NaHCO
a, 0.1mM PMSF (Phenylmethyl
lsulfonyl fluoride), 1mM
E D T A (Ethylenediaminet
The mixture was homogenized using a Polytron homogenizer in aqueous solution (etraacetic acid). After filtration with gauze, centrifuge at 1,500 g for 15 minutes, and remove the supernatant (5
00ml) was further centrifuged at 40,000g for 20 minutes. The precipitate was dissolved in TPE buffer (50mM Tris buffer (pH 7).
, 4), O, 1mM PMSF, 1mM EDTA) and stored at 180 degrees as a crude receptor membrane fraction, and used by suspending in TPE buffer as necessary.

5I-endothelin as a radioligand] (8
1,4T Bq/mmol, manufactured by DuPont, USA) at 3
70 Bq of the crude membrane fraction, 1.4 μg of protein, and the sample were added.

The amount of non-specific binding was measured by adding XIO-'M endothelin-1. The reaction was carried out in 0.2ml of TPE buffer at 37°C for 190 minutes. After the reaction, a glass filter (CF/B,
Whatman, USA) to stop the reaction.
The filter was washed three times with 10 mM NaK-phosphate buffer (pH 7.4), and the radioactivity remaining on the filter was measured using a gamma counter.

The results are shown in Table 1.

Table 1 Radio receptor binding inhibitory activity Compounds that inhibit endothelin-1 binding I C5o (μg/ml
) (b) Measurement of guinea pig ileal contraction inhibitory effect on guinea pigs (S td 1lartley male,
After concussion, blood was exsanguinated from the carotid artery and the ileum was removed from a 250 g (Japan Nippon Nippon Nippon Nippon Nippon Nippon Nihon Nishin Nihon Nishin Nihon Nishin Nippon Nippon Nihon Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishi Nishi Nishin Nishi Nishi Nishi Nishi Nishin Nishi Nishi Nishi Nishi Nishin Nishi Nishi Nishi Nishi Nishin Nishi Nishi Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi Nishin Nishi) concussion. 2 pieces of approximately 3cm
In a Magnus tube filled with 0 ml of Tyrode's buffer,
It was suspended under a tension of 0.5 g.

During this time, the temperature was kept at 37°C, and the mixed gas (95%02.5
%CO,) was bubbled through. Add sample 5
After stabilization for minutes, the ileum was contracted by adding 1X10-8 M of endothelin-1, and the degree of inhibition of contractions without sample addition was expressed as a percentage. The contraction was performed using an isotonic quadrangle juicer (ME-/1013, manufactured by Suruga Electronics Co., Ltd.) and a recorder (Rectiphon)-8K.

(manufactured by JEOL SANEISHA). The activity of the sample was expressed as the ED value, which is the concentration showing 50% inhibition of maximal contraction.

The results are shown in Table 2.

]　　　　Xl0-' 2　　　Xl0-5 1 × 10 o, sxi.

O,8X]0 (c) Antagonistic effect of endothelin 1 on rat pressor response Male Wistar rats (body weight 250-300 g) were used under pendoparbital anesthesia. Blood pressure was measured using a cannula inserted into the right carotid artery. Endothelin-1 0
, 3 mmol/kg was administered intravenously to examine the effect on gastric pressure. One hour later, Compound 3 (50 mg/kg) was administered intravenously, and 10 minutes later, the same dose of endothelin-1 was administered intravenously to examine the effect on blood pressure.

The results are shown in Table 3.

Table 3 Test drug numbers for the pressor response of endothelin-1 are % changes from before endothelin-1 administration (n = 3
) Compound 3 suppresses the pressor reaction of endothelin-1 by 47%, and it is clear that it has an antagonistic effect.

- Compound (3), such as Compound 1, showed no abnormal response when administered intraperitoneally at 400 mg/kg in an acute toxicity test using mice.

The above results demonstrate that the compound (3) or its salt was effective in vitro and in vivo (1n
It has been clearly demonstrated that the action of endothelin was inhibited via the endothelin receptor in both systems (vivo). These facts indicate that the compound of general formula (3) or a salt thereof is an endothelin receptor antagonist.

An endothelin receptor antagonist containing the compound of general formula (3) or a salt thereof as an active ingredient is useful as a therapeutic or preventive agent for myocardial infarction or acute renal failure in mammals, and It can be obtained by mixing with a commercially acceptable carrier. The present invention can be provided in, for example, a drip, an injection, a capsule, a tablet, a waste, a solution, a suspension, and other pharmaceutically appropriate dosage forms.

When manufacturing parenteral preparations, such as injection preparations, tonicity agents (
Examples: glucose, D-sorbitol, D-mannitol,
(sodium chloride, etc.), preservatives (e.g., benzyl alcohol, chlorobutanol, methyl parahydroxybenzoate,
parahydroxybenzoic acid, propyl, etc.), anticoagulants (e.g., dextran sulfate, heparin, etc.) and buffering agents (e.g.,
phosphate buffer, sodium acetate buffer, etc.) may be used.

Oral preparations are in the form of capsules, tablets, syrups, powders, granules, etc., and contain the compound of (3) or its salt as well as additives such as excipients, binders, disintegrants, and lubricants. Agents, colorants, flavoring agents, stabilizers, etc. may also be included.

Regarding the administration method, for example, the compound of (3) or a salt thereof is administered parenterally, subcutaneously, intravenously or intramuscularly as an injection at a dose of about 0.1 to 100 mg/kg/day, preferably 0.5 to 100 mg/kg/day, preferably 0.5 to 100 mg/kg/day. Administer 50 mg/kg/day.

Oral dosage: about 0.1 to 400 mg/kg/day;
Preferably, the dose is 0.5 to 200 mg/kg/day.

The present invention will be explained in more detail with reference to Reference Examples and Examples below, but the present invention is not limited thereto. Note that percentages (%) in the culture medium are expressed as weight/volume percentages unless otherwise specified.

Reference example 1 Potato Dextrose Broth (Difco).

Penicillium citrinum FL-8556 (I FO32324, FERM
BP-3093) with 2% glucose, 3% maltose, 1.5
% Raw Soybean Flour, 1.0% Corn Steep Liquor (C
3L), 0.5% peptone, 0.3% yeast extract, 0.3L), 0.5% peptone, 0.3% yeast extract, 0.
40 ml of seed medium (pH, 6,5) containing 3% NaC (!
) in a 200 ml Erlenmeyer flask at 28°C.
A seed culture was obtained by culturing on a rotary shaker for 148 hours. 1 ml of the obtained seed culture solution was placed in a 200 ml flask.
0ml of 1.0% glucose, 3.0% maltose, 2.0%
Cottonseed flour, 1.0% C8L, 0.5% dry yeast, 0.5%
The cells were transferred to a fermentation medium containing calcium carbonate and cultured on a rotary shaker at 28°C for 5 days.

Reference Example 2 The culture solution (4,1) obtained in Reference Example 1 was adjusted to pH with 3N hydrochloric acid.
3, extracted with ethyl acetate (3,5C), and filtered with Hyflo Supercel (John Manpil Co., USA).
An ethyl acetate extract (3,2f2) was obtained. This 2
% sodium bicarbonate aqueous solution, and transfer solution (4Q)
After adjusting the pH to 3 with 3N hydrochloric acid, redissolving with ethyl acetate,
The obtained reconverted solution (3Q) was washed with water, dried over anhydrous sodium sulfate, and then evaporated under reduced pressure to obtain a crude oily substance (3 g). This was carried out using silica gel column chromatography (
130g) and solvent system toluene-acetone (10~
5:l.

1.5Q-fraction I), (3-1:1.21 and (l
.

Elution fraction 1 (930 mg) in 5.700 m1-fraction ■)
and fraction 11 (1.15 g) were obtained. Fraction I was crystallized from methanol to yield colorless needle-shaped crystals of Compound 1 (310 mg). Mother liquor concentrate (620mg
) was subjected to silica gel column chromatography (30 g), and the solvent system was hexane-ethyl acetate (5-3:1).
Compound 1 (375 mg) was obtained by eluting the active fraction by adding acetic acid to a concentration of 1%.

Fraction ■ was subjected to silica gel column chromatography (50 g
) and the solvent system, hexane-ethyl acetate (5-3:
1.1.1), acetic acid was added to give a concentration of 1% and elution was performed to obtain Compound 1 (147 mg), a mixture of Compounds 1 and 2 (145 mg), and Compound 2 (123 mg). Compound 2 was recrystallized from acetone to give colorless needles (38 mg).

Compound 1 m, p, 208-210°C (decomposed) eOH UV λ (ε) + 310 nm (7, Too)
, 245nm (13,600) ax 212nm (54,600) I R (cm', KBr) + 3300. 17
10. 1660 16201595, 1190
1050 EI-MS: m/z 348 (M+) Elemental analysis: Calculated value C,, H,80, C58,62,
H4, 63, 036, 75 measurement value C58, 29, H4
,92'HNMR (300MHz, heavy acetone, δppm)7
, 05 (IH, d, J= 2.811z), 6.9
2 (IH, d, J = 2.811z) 6, 45 (II
I, s), 5.90 (IH, s), 3.81 (3
11, s), 3.74 (3II, s), 2.16
(3H,s)”CNMR (75MHz, DMSOdo,
δppm) 170.6 (Q), 165.0 (Q),
160.5 (Q), 158.7 (Q) 155.1 (Q
), 153.2(Q), 143.5(Q), 13
4.0 (Q) 125.2 (Q), 109.9 (C
I), 107.6 (C11), 104.9
(CI+) 104.7 (CI+), 103.3 (Q)
, 56.0(CH3), 51.9(C1+3)21
, 4 (C1l,) (However, Q is a quaternary carbon, CH is methine, CH is methylene, and CH8 is a methyl group) Compound 2゜m, p, 233-235°C (decomposition) MeOI + UV λ ( e): 315 nm (1
3,800), 243 nm (22,900) ax, 213 nm (86,300) IR (cm-', KBr): 3430. 32
80. 1720. 1690°1600, 1190
．． 1070. 1060E I-MS: m/
z 382 (M+) elemental analysis, calculated value CI7Hlf
C53,35,H3,95033,4 as i○60ρ
4Cf! 9.26 Measured value C53,29,H3,9+ HNMR (300MH2, CDCl23-heavy methanol, δppm): 7.00 (111,d, J=2.8H
z), 6.79 (IILd.

J=2.8Hz), 5.92(IH,d, J=0.4
Hz), 3.79 (3H, s).

3,78(3H,s), 2.26(311,d,
J = 0.4Hz) I3CNMR (75MHz, DM
SO-do, δppm) 170.9 (Q), 164.
8(Q), 157.2(Q), 156.6(Q)
155.2 (Q), 153.1 (Q), 141.5
(Q), 133.6(Q).

125.1(Q), 113.3(Q), 107.6
(CH), 106.1 (CI) 105.0 (C) I)
, 104.1 (Q), 56.0 (CH3), 52
．． 0 (CIl!1) 20, 5 (CI+3) Reference Example 3 Compound 1 (], 55 g was dissolved in ethyl acetate (150 ml), 2% hydrogen carbonate) IJium aqueous solution (150 ml)
) was extracted. After concentrating the extract to 100 ml, Amberlite XAD-11 (200 ml.
The sample was passed through a column packed with HAAS (manufactured by Haas, USA). After washing with water (600 ml), it was eluted with 50% methanol water (1).The eluate was concentrated and lyophilized to yield compound 3 (1).
, 24g) was obtained.

II, O UV λ (ε): 316 nm (8,140)
, 251 nm (14,100) ax , 212 nm (48, too) IR (am-', KBr): 3400. 1
720. 1580. 1430 elemental analysis: Calculated value C,7H,,08Na-HtO as C53,58,H
4,41,037,08°Na 5.92 Measured value C52,68,H4,22,Na 6.4 Reference example 4 Sodium hydrogen carbonate (17mg) was added to compound 2 (77mg).
) was added and dissolved in water (10 ml). The obtained solution was concentrated and lyophilized to obtain Compound 4 (8iI mg).

uvλH, 12FA. x(e): 315 nm (7,
300), 250nm (10,500) 212nm (
48,500) I R (cm-', KBr): 3400. 17
10. 1580. 1440° elemental analysis: calculated value CI
? H1408CI2N a ・t, 5H, O as C1,29,H3,97,O35,20CQ 8.2
1 Na5.32 Measured value C47,59, H4,26 Example 1 Compound 2 (50 mg) was suspended in ether (3 ml), and an ether solution of diazomethane was added until the solid was dissolved. After stirring the resulting solution at room temperature for 10 minutes, the solvent was distilled off under reduced pressure. The residue was chromatographed on silica gel [silica gel (
2.5 g), solvent, hexane-ethyl acetate (5:1→3
:1)] to obtain compound IO (45 mg).

, 208nm (46,000) IR (cm""', KBr): 3400. 1
700. 1650. 16001430, 1200
1090 1060 elemental analysis, calculated value C+eH1
70ac Q as C54,49H4,3203226 (18,94 Measured value C54,24,H4,62'HNMR (300MHz, CDCl2+, δppm)
: 6.98 (IH, d, J=2.9Hz), 8.
72 (IH, d, J = 2.911z), 5.88 (I
H,s), 3.97(311,s), 3.75(3
11,s), 3.71 (311,s).

2, 23 (3B, s) Example 2 Compound 3 (103 mg) was dissolved in dimethylformamide (2 m
1), ethyl iodide (45 μQ) was added, and the mixture was stirred at room temperature for 8 hours. Add ethyl iodide (45μQ) to the reaction solution.
was added and further stirred for 16 hours. The solvent of the reaction solution was distilled off, water was added to the remaining solution, and the mixture was extracted with ethyl acetate. The organic layer was washed with saturated brine, dehydrated with sodium sulfate, concentrated to dryness, and the combined material (1
10 mg) was obtained. The obtained compound was purified by silica gel column chromatography [silica gel (5 g), solution 1: hexane-ethyl acetate (5:1 → 3:1)] to obtain compound 11 (
89 mg) was obtained.

eOH UV λ (ε): 315 nm (7,20
0), 250nm (12,800) ThaX, 210nm (47,400) I R(am"", KBr) + 3430. 29
80. 1710. 16201430, 1200.
1060 Elemental analysis: Calculated value C, 8H. 08・0.5H,O as C59,22,H5,49035,29 measurement value C59
,53,H5,33'HNMR (300MH2, CDCl23.δppm)
: 6.98 (IH, d, J=2.811z),
6.71 (IH, d, J=2.8Hz), 6.41
(it(, d, J=0.611z), 5.77(I
I(,d,J=0.6Hz), 4.41(2H,q
, J=7.1Hz), 3.73(38,s),
3.69 (3H, s), 213 (3H, s),
1.34 (3H, t, J = 7.1Hz) Example 3 Compound 3 (100g) was dissolved in distilled water 11, mannitol (25g) was added and dissolved, sterilized and filtered, and 5ml each was added to ampoules. Dispense. This is put into a freeze dryer and after drying is combined to obtain an ampoule for dissolution before use. For use, the ampoule is opened and dissolved in 5 ml of physiological saline to prepare an injection for subcutaneous, intravenous or intramuscular administration.

Example 4 Compound 3 (]OOmg) was added to cornstarch (50mg)
and lactose (50 mg) and granulated according to a conventional method. This is encapsulated in gelatin capsules according to the usual method,
Capsules and Sur.

Effects of the Invention The compound of general formula (3) has endothelin receptor antagonistic activity and is effective as a prophylactic or therapeutic agent for myocardial infarction or renal failure.

Claims (1)

[Claims] 1) General formula ▲ Numerical formulas, chemical formulas, tables, etc. ▼ [In the formula, R_1 and R_4 are hydrogen atoms or optionally substituted lower alkyl groups, R_2 and R_5 are hydrogen atoms or methyl group, R_3 represents a hydrogen atom or a chlorine atom. However, when R_3 is a hydrogen atom, R_
1 and R_4 represent an optionally substituted lower alkyl group having 2 to 6 carbon atoms, and R_4
Except when 3 is a chlorine atom, R_1 is hydrogen and R_4 is methyl. ] or its salt. 2) General formula▲There are mathematical formulas, chemical formulas, tables, etc.▼ [In the formula, R_2 and R_5 represent a hydrogen atom or a methyl group, and R_3 represents a hydrogen atom or a chlorine atom. 2. The method for producing the compound according to claim 1, which comprises subjecting the carboxyl group of the compound represented by the above formula to a lower alkylation reaction. 3) General formula▲ Numerical formula, chemical formula, table, etc.▼ [In the formula, R'_1 and R'_4 are hydrogen atoms or optionally substituted lower alkyl groups, R'_2 and R
'_5 represents a hydrogen atom or a methyl group, and R'_3 represents a hydrogen atom or a chlorine atom. ] An endothelin receptor antagonist containing a compound represented by or a salt thereof as an active ingredient.