JP3046077B2 - Aliphatic tricarboxylic acid compound and method for producing the same - Google Patents

Description

Description: TECHNICAL FIELD The present invention relates to a novel aliphatic tricarboxylic acid compound, in particular, an unidentified fungus Z-285 (FERM BP-515)
No. 3 (deposited as No. 3). The present invention also relates to a method for producing the above-mentioned aliphatic tricarboxylic acid compound, and a pharmaceutical composition containing the above-mentioned compound, which is useful for treating hypercholesterolemia, fungal infection and the like.

BACKGROUND ART Hypercholesterolemia is known as one of the most dangerous factors for coronary arteriovenous heart disease such as arteriosclerosis. Bile acid sequestrant
Has been used for the treatment of this condition, and appears to be reasonably effective, but must be consumed in large quantities (ie, at a few grams per serving) and Bad taste.

An important approach to the reduction of low-density lipoprotein cholesterol is the inhibition of cholesterol biosynthesis. Currently available "lovastatin" is one of the effective anti-hypercholesterolemic agents, which inhibits HMG-CoA reductase, thereby
The level of mevalonic acid present is reduced. However, mevalonic acid is a non-steroidal isoprenoid such as dolichol, ubiquinone, and isopentenyl tR
It is also a genuine precursor for NA. In addition, mevalonic acid is required for the production of farnesyl diphosphate and geranylgeranyl diphosphate (ie, terpenoid substrates for protein prenylation).

Squalene synthetase (SQS) has an important branch point in the isoprenoid pathway and a new (de)
novo) catalyzes the first step involved in cholesterol biosynthesis. This enzyme catalyzes the reductive dimerization of two molecules of farnesyl diphosphate to form squalene via a presqualenediphosphate intermediate. Inhibition of SQS is ubiquinone, dolichol, and isopentenyl tR
It would provide a biosynthetic pathway without interference to NA.

Conventional efforts to find SQS inhibitors include Corey, E.
J. and R. Volante describe diphosphate-containing compounds as SQS inhibitors in "J. Am. Chem. Soc. 98: 1291,1976". Further, Biller, SA et al. Described farnesyl phosphonate as an SQS inhibitor in `` J. Am. Chem. Soc. 113: 85.
22,1991 ".

Recently, certain non-phosphorus containing SQS inhibitors, such as zaragozic acid C; Kenneth, FB et al., US
5,026,554, June 25, 1991) and viridiofungins (H
arris, GH et al., Tetrahedron Lett., 34: 5253, 1993)
Isolated from microbial sources.

BRIEF DISCLOSURE OF THE INVENTION The present invention provides a compound of formula (I): Wherein Y is an ethyl or ethenyl group; and
R ¹ , R ² and R ³ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms) or a pharmaceutically acceptable salt thereof. I will provide a.

Preferred compounds of the formula (I) are those wherein Y is an ethenyl group and R ¹ , R ² and R ³ are hydrogen atoms
CJ-13,981, and wherein Y is an ethyl group, and R ¹ ,
CJ-13,982 in which R ² and R ³ are hydrogen atoms are included.

The present invention also provides a FERM BP-5153 culture capable of producing an aliphatic tricarboxylic acid compound.

The present invention also provides a method for producing the aliphatic tricarboxylic acid compound, which comprises culturing a microorganism having the identification characteristics of FERM BP-5153, or a mutant or recombinant thereof. Further, the method can include a subsequent step of isolating the aliphatic tricarboxylic acid compound from the fermentation broth.

Furthermore, the present invention provides a fungus comprising a therapeutically effective amount of a compound of formula (I) or a pharmaceutically acceptable base addition salt thereof, and a pharmaceutically acceptable carrier. Compositions for treating infection and inhibiting SQS are provided.

The present invention also provides a method comprising administering to a subject in need of antifungal therapy a compound of formula (I) or a pharmaceutically acceptable base addition salt thereof in an antifungal amount. The present invention also relates to an antifungal treatment method.

Furthermore, the present invention provides a method for administering an effective amount of a compound represented by formula (I) or a pharmaceutically acceptable base addition salt thereof to a subject in need of SQS inhibition or reduction of serum cholesterol level. The method of inhibiting SQS or reducing serum cholesterol levels.

Therefore, the present invention is useful for treating or preventing hypercholesterolemia or fungal infection.

DETAILED DESCRIPTION OF THE INVENTION The microorganism used in the present invention is strain Z-285, which is from Texas Technical University (USA, USA).
It was isolated from the roots of plants in Chihuahuan Desert by a group of Dr. John Zak of Lubbock, Texas). In accordance with the Budapest Treaty, it was commissioned on July 4, 1995 to the Institute of Biotechnology and Industrial Technology, Ministry of International Trade and Industry (address: zip code: 305; 1-3-1, Higashi 1-3-1, Tsukuba, Ibaraki, Japan). Deposit number FERM BP-5153.

From a slant of yeast extract-malt extract agar, inoculate FERM BP-5153 onto a plate of confirmation media and incubate the plate under fluorescence or invisible light for 12 hours.
And incubated at 25 ° C under alternating conditions for 12 hours in the dark for 1-5 weeks. The results were read at various times, but most commonly on day 11. Hue is
`` Color Standards and Color Nomenclature, Ridgw
ay, R., 1912 ".

The media used to describe the cultures and references for their preparation are as follows.

1. Cornmeal agar: Carmichael,
JW, Mycologia49: 820-830,1957.

2. Czapek-sucrose agar: Raper, KBand DI
Fennell, 1965. The Genus Aspergillus, p.36.

3. Malt extract agar: ibid, p.38.

4. Oatmeal agar medium: Quaker oats 30g, agar
15g, 1 liter of tap water.

5. Potato dextrose agar: peeled potatoes
100g, dextrose 10g, coconut milk 50ml, agar 20
g, 1 liter of tap water.

6. V-8 juice agar: ATCC medium343, ATCC M
edia Handbook, p. 17, 1984.

7. Potato carrot agar: Lechevalier, MP, J. La
b. Clin. Med. 71: 934-944, 1968, using only 30 g of potato, 2.5 g of carrot and 20 g of agar.

8. Yeast extract soluble starch agar: Emerson,
R., Mycologia 50: 589-621,1958.

 The FERM BP-5153 culture exhibited the following characteristics:

Cornmeal agar: reaches 8.3 cm in diameter in 11 days; white to cream (XVI); smooth, sunken to fluffy; reverse cream (XVI) to light ) Olive-gray (LI); no soluble pigment.

Czapek-sucrose agar: reaches 5.5 cm in diameter in 11 days; pale flesh color to seashell pink (XIV); smooth, sunken to fluffy state; reverse safrano Pink (I)
I) to light salmon (XIV); no soluble pigment.

Malt extract agar: 8.5 cm in diameter in 11 days; pale pink buff to pinkish buff (XXIX); smooth, wool-like to fine funiculose ; Obi reverse pink buff (XXI
X), olive gray to oliveaceous
Black (LI); no soluble pigment.

Oatmeal agar: 8.5 cm in diameter in 11 days; ochraceous-buff to light buff (X
V); Smooth, coarse wool-like state to fine funiculose state; Light aucrasus-buff (XV) to light salmon color (XIV), concentric mouse gray (LI) Yes; no soluble pigment.

Potato dextrose agar: reaching 8.5 cm in diameter in 11 days; pale aucrasus-salmon (XV); smooth, wool-like to funiculose state; reverse pale aucrasus-salmon to light aukurasus-salmon (X
V), with concentric light olive-grey to Oliveas black (LI) areas; no soluble pigment.

V-8 juice agar: reaching 8.5 cm in diameter in 11 days;
Pale cinnamon-pink to pale pink cinnamon (XXIX); smooth, wool-like to funiculose, reverse salmon to flesh-color (XIV), concentric light rat to rat (LI) Areas may be present; no soluble pigment.

Potato carrot agar: 8.0 cm in diameter in 11 days; white to off-white; smooth, submerged to fluffy; reverse colorless to pale auraceus bath (XV); no soluble pigment.

Yeast extract soluble starch agar: 8.5 cm diameter in 11 days; white to cartridge buff (XXX); smooth, wool-like to funiculose; reverse pale aucrasus-salmon to light aukurasus- Salmon (XV); no soluble pigment.

Morphological characteristics: Morphological characteristics were observed after 2, 3 and 5 weeks of incubation. The following observations were obtained after 3 weeks of incubation on a malt extract agar medium under alternating conditions of 12 hours of fluorescence and 12 hours of darkness. Branched hyphae; septat
e); hyaline brown to black; smooth to rough; diameter 1.5-8.0 μm. Thick-walled spores are single or chain; terminal, lateral or internodal; hyaline is brown to black; spherical, subspherical, oval, oval, or irregular; 4-13 μm or 5-17 × in diameter 4-11 μm. After 5 weeks of incubation, no spores or conidia were produced in any of the media used.

Temperature relationship: Growth is moderate at 20 ° C, moderate to good at 28 ° C, good to excellent at 37 ° C, and 45 ° C
And at 50 ° C.

FERM BP-5153 is characterized by its pale yellow (yellowis
h), pale orange-pink, pale pink buff, light buff to tan colony; its pale yellow, pale pink orange, pale orange, pale orange-buff, pale buff, pale Gray, gray to black colony reverse; and lack of soluble pigment. The mycelium had a septum, was branched, and was initially hyaline but changed to a yellow-brown, pink buff or Aucrasus buff.

They will turn brown, grey or black after 4-5 weeks of prolonged incubation or upon exposure to fluorescence or dark light. Various forms of chlamydospores were produced on some media. No spores or conidia were produced in any of the media used.

Based on the above results, FERM BP-5153 could not be identified as a fungal genus because there are no spores or conidia required for identification as a fungal genus, and thus are considered to be unidentified fungi. This was deposited as an unidentified fungus under the accession number FERM BP-5153 with the Institute of Biotechnology and Industrial Technology of the Ministry of International Trade and Industry of Japan.

In the present invention, FERM BP-5153 having the ability to produce the aliphatic tricarboxylic acid compound represented by the formula (I).
Mutants or recombinants from can also be used. Mutants or recombinants may be accidental mutations, or may be artificial mutations due to ultraviolet irradiation or treatment with a mutagen (eg, N-methyl-N'-nitro-N-nitrosoguanidine or ethyl methanesulfonate), or It can be obtained according to a well-known method by a cell technical method (for example, protoplast fusion or genetic manipulation).

According to the present invention, the aliphatic tricarboxylic acid compound of the formula (I) can be prepared under the same conditions as those generally used for the production of bioactive compounds by fermentation under conditions similar to those of FERM.
It can be produced by aerobically fermenting BP-5153, or a mutant or recombinant thereof.

FERM BP-5153, or a mutant or recombinant thereof,
Usually, fermentation is performed under aerobic conditions in the liquid for 1 to 10 days while stirring at a temperature of 20 to 40 ° C, and these conditions can be changed according to the fermentation conditions. The cultivation of FERM BP-5153 for producing the aliphatic tricarboxylic acid compound is preferably performed in an aqueous nutrient medium at a temperature of 25 ° C to 35 ° C for 1 to 3 days. The pH of the medium can be adjusted in the range of 4.0 to 9.0, preferably 5.5 to 7.0.

Suitable nutrient media useful for fermentation include assimilable carbon sources (eg, sugars, starch and glycerol); organic nitrogen sources [eg, casein, oxygen digest of casein, soybean meal, groundnut meal, peanuts. Meal, wheat gluten, soy flour (flower), meat extract, and fish meal]. Growth material sources (eg, inorganic salts,
Sodium chloride, and calcium carbonate); and trace elements (eg, iron, magnesium, copper, zinc, cobalt, and manganese) can also be utilized with advantageous results. If excessive foaming occurs during the fermentation, an antifoaming agent (eg, polypropylene glycols or silicones) can also be added to the fermentation medium.

The aeration of the culture medium in the fermenter for submerged growth is carried out at a rate of 3 to 200% by volume of sterilized air per minute / volume of the medium, preferably 5%.
Maintained at 0-150% volume / medium volume. The stirring speed depends on the type of stirrer used. Fermenters are usually 300-2,0
Operate at 00rpm, but shake flasks usually have 150-250r
Drive at pm. Of course, sterility must be maintained during the transplantation of the organism and throughout its growth.

The aliphatic tricarboxylic acid compound represented by the formula (I) is
It can be isolated by standard techniques, such as extraction and various chromatographic techniques.

CJ-13,981 which is the aliphatic tricarboxylic acid compound
And CJ-13,982 were isolated from the fermentation mixture in substantially pure form and identified by a variety of spectroscopic techniques such as UV spectrophotometry, NMR and mass spectroscopy. According to the above analysis, these compounds have the following chemical formula: It is considered to be represented by

Compounds of formula (I) wherein R ¹ , R ² , and R ³ are groups other than hydrogen atoms can be prepared using suitable alkylating agents under suitable conditions known to those skilled in the art using CJ- It can be prepared by alkylating 13,981 or CJ-13,982.

SQS of the aliphatic tricarboxylic acid compound produced by the method of the present invention, i.e., CJ-13,981 and CJ-13,982.
Inhibition and antifungal activity was measured by the standard in vitro protocol described below.

Preparation of microsomes Freshly obtained livers from Wistar rats were rinsed in ice-cold PBS and buffer A containing protease inhibitors [PMSF (500 μM), aprotinin (10 μM), leupeptin (10 μM) and chymostatin (10 μM)] [ MOPS-NaOH
(50 mM); pH 7.4, MgCl ₂ (10 mM), EDTA (1 mM) and DTT (1 mM).
0 mM)]. The homogenate was centrifuged at 3,000 xg for 20 minutes at 4 ° C.
The supernatant was re-centrifuged at 20,000 xg for 30 minutes at 4 ° C.
The supernatant was centrifuged again at 4 ° C., 100,000 × g for 60 minutes. After centrifugation, the supernatant was removed and the pellet (microsome-related fraction) was suspended in buffer A. The protein concentration of this microsome-related fraction is approximately
13.2 mg / ml. The microsome-related suspension was stored at -70 C until use. Under these conditions, SQ
S activity is stable for at least several months.

Squalene synthetase analysis The analysis was performed in a 96-well microtiter plate containing 50 μl reaction mixture per well. The reaction mixture was composed of MOPS-NaOH (50 mM) (pH 7.4), KF (10 mM), MgCl ₂
(10mM), CHAPS (2mM), DTT (10mM), NADPH (0.5m
M), ascorbate (50 mM), ascorbate oxidase (20 units / ml), glucose-6-phosphate (2 m
M), glucose-6-phosphate dehydrogenase (20 units / ml), rat liver microsome-related protein and [ ³ H] farnesyl diphosphate (6 μM). After incubation at 37 ° C. for 30 minutes, 1-propanol (20%) containing unlabeled squalene (1%) was added.
The reaction was terminated by adding μl). The reaction mixture (30 μl) was applied to a polyester backed silica gel TLC sheet (Sigma, 10 × 10 cm) and developed with n-hexane-ethyl acetate (7: 3). After drying, the radioactivity in the area containing squalene (Rf = 0.8) was scraped and measured by a scintillation counter. The SQS inhibitory activity was calculated by the following equation.

Antifungal Assay The antifungal activity of the compound of the present invention was measured using an 8 mm paper disc (ADVANTEC, trademark), an agar plate medium (Antibiotic Medium).
11, Difco) and test organisms (Candida albicans; C)
andida albicans). The aliphatic tricarboxylic acid compound, CJ-13,981
And CJ-13,982 showed antifungal activity.

Pharmaceutically acceptable base addition salts of CJ-13,981 and CJ-13,982 are obtained by preparing a solution or suspension of the free acid with about 1 chemical equivalent of a pharmaceutically acceptable base. Prepared in the usual way of processing. For the isolation of the salt, a conventional concentration technique and a recrystallization technique are used. Examples of suitable bases include alkali metal hydroxides (eg, sodium or potassium hydroxide), alkaline earth metal hydroxides (eg, magnesium or calcium hydroxide),
And ammonium or organic amines (eg, diethanolamine or N-methylglucamine).

The aliphatic tricarboxylic acid compounds of formula (I) and pharmaceutically acceptable base addition salts thereof are useful for treating hypercholesterolemia, fungal infection, and the like. The aliphatic tricarboxylic acid compound represented by the formula (I) can be administered alone or in combination with a pharmaceutically acceptable carrier in single or multiple doses. Suitable pharmaceutical carriers include inert solid diluents or fillers, sterile aqueous solution and various organic solvents. Accordingly, pharmaceutical compositions formed by combining an aliphatic tricarboxylic acid compound of Formula (I) with a pharmaceutically acceptable carrier can be used in a variety of dosage forms, such as tablets, powders, It can easily be administered in the form of lozenges, syrups and injectable solutions. These pharmaceutical compositions can, if desired, contain additional ingredients such as flavorings, binders, and excipients. Thus, for oral administration, tablets containing a variety of excipients, such as sodium citrate, calcium carbonate, and calcium phosphate, may be combined with various disintegrants, such as
It can be used with starch, alginic acid, and certain complex silicates, and binders such as polyvinylpyrrolidone, sucrose, gelatin, and gum arabic. In addition, lubricants such as magnesium stearate, sodium lauryl sulfate, and talc are often useful for tableting purposes. Solid compositions of a similar type can also be used as fillers in soft-filled and hard-filled gelatin capsules. Preferred materials in this connection include lactose (or lactose) and high molecular weight polyethylene glycols. If an aqueous suspension or elixir is desired for oral administration, the essential active ingredient and various sweetening or flavoring agents, coloring or dyes, and, if desired, emulsifying or suspending agents, are combined with a diluent, For example, it can be combined with water, ethanol, propylene glycol, glycerin, and combinations thereof.

For parenteral administration, solutions of an aliphatic tricarboxylic acid compound of formula (I) in sesame or peanut oil, aqueous propylene glycol or sterile aqueous solution may be employed. The aqueous solutions should be suitably buffered if necessary, and the liquid diluent first rendered isotonic with sufficient saline or glucose. These particular aqueous solutions are particularly suitable for intravenous, intramuscular, subcutaneous and intraperitoneal administration purposes. In this connection, the sterile aqueous media used are all readily available by standard techniques known to those skilled in the art.

In addition, when treating skin conditions, the aliphatic tricarboxylic acid compounds of formula (I) may be administered topically, which may be in accordance with standard pharmaceutical practices, creams, jellies, gels, pastes and the like. And ointment.

In general, the tricarboxylic acid compounds of the formula (I) are present in the above-mentioned administration forms at a concentration level in the range from 5 to 70% by weight, preferably from 10 to 50% by weight.

Generally, a therapeutically effective daily dose for the active compound will range from 0.01 to 100 mg / kg, generally from about 1 to about 5 mg / kg, of body weight for a patient to be treated for a fungal infection; For inhibiting SQS and reducing serum cholesterol levels, it is 0.2-100 mg / kg, for example 0.2-1 mg / kg. As is generally known, the effective dose of the active compound depends on the intended route of administration and other factors, such as the age and weight of the patient, as is generally known to the physician. . The dose also depends on the disease to be treated.

Examples Hereinafter, the present invention will be described specifically with reference to Examples.
These examples do not limit the scope of the invention. Spectral and physicochemical data were obtained with the following equipment. UV, JASCO Ubest-30; NMR, JEOL JNM-GX2
70 (updated with LSI-11 / 73 host computer, TH-5 adjustable probe, and version 1.6 software); and LRFAB-MS and HRFAB-MS, JEOL J
MS-700 Mstation. Unless otherwise noted, all NMR spectra were measured in CD ₃ OD and peak positions were ¹ H-NMR
3.35 ppm (parts per million) and ¹³ C
Expressed in ppm based on the internal standard peak CH ₃ OH in 49.0ppm respect -NMR. The shape of the peak is shown as follows. s = singlet; d = doublet; t = triplet; q = quadruple; m
= Multiple lines; and br = broad. All FAB-MS spectra were measured using a glycerol-matrix.

Example 1 Fermentation of FERM BP-5153 Medium 1 (2.4% potato dextrose broth, 0.5% yeast extract and 0.1% agar) in a 500 ml flask
The ml was inoculated with a vegetative cell suspension from a FERM BP-5153 slant culture. The flask was shaken on a rotary shaker (210 cm, 7 cm stroke) at 26 ° C. for 4 days to obtain a first inoculum culture.

5 ml of the first inoculum culture was inoculated into a 500 ml flask containing medium 1 (150 ml). The flask was shaken on a rotary shaker at 26 ° C. for 3 days to obtain a second inoculum culture.

The second inoculum culture was used to inoculate a 6 liter fermenter containing 3 liters of sterile medium [medium 2: potato dextrose broth 2.4%]. Aeration was performed at 26 ° C. for 9 days at 1700 rpm, 3 liter / min.

Extraction and isolation Add 2 liters of ethanol, then add fermentation broth (3
Liters). The filtrate was concentrated to an aqueous solution (1 liter) and adjusted to pH 3 with 1N HCl. Subsequently, extraction was performed three times with ethyl acetate (1 liter). The extract was dried over anhydrous sodium sulfate and evaporated. This extract (3.5 g) was added to YMC-pack O
The sample was placed on a DS AM-343 column (20 × 250 mm, Yamamura trademark) and eluted with methanol-0.1% TFA (in water) (85:15) at a flow rate of 10 ml / min. Detection was by UV absorbance at 220 mm. The eluting peak was collected and CJ-13,981
(198 mg) and CJ-13,982 (508 mg).

HPLC analysis aliphatic tricarboxylic acid compounds CJ-13,981 and CJ-13,9
Analytical HPLC of the sample containing 82 was performed on a YMC-Pack ODS-AM
Using a −312 column (6.0 × 250 mm, Yamamura trademark) at a flow rate of 0.8 ml / min, methanol-0.1% TFA (in water) (85: 1)
Performed by eluting in 5). CJ-13,981 and C
The retention times of J-13,982 were 7.0 minutes and 8.7 minutes, respectively.

Properties The physicochemical properties of CJ-13,981 and CJ-13,982 are as follows.

CJ-13,981 was isolated as a white powder and showed the following spectral data: ¹ H NMR (270 MHz, CD ₃ OD) δ (pp
m) 1.14-1.40 (16H, m), 1.47 (1H, m), 1.79 (1H, m),
2.03 (2H, m), 2.64 (1H, dd, 2.9,11.7), 2.68 (1H, d, 16.
5), 3.07 (1H, d, 16.5), 4.87-5.01 (2H, m) and 5.79
(1H, ddt, 10.1,17.0,6.6); ¹³ C NMR (67.5 MHz, CD ₃ OD)
δ (ppm) 28.89, 29.61, 30.91, 31.01, 31.24 (2C), 31.40
(2C), 31.46,35.69,43.08,55.59,77.60,115.49,140.9
4,174.65,176.97 and 177.41; FAB-MS (m / z) 357 (M-
H) ^- ; UV max (MeOH) 215 nm; [α] ²⁶ _D (c 0.81, acetone)-18.52.

CJ-13,982 was isolated as a white powder and showed the following spectral data: ¹ H NMR (270 MHz, CD ₃ OD) δ (pp
m) 0.89 (3H, t, 6.5), 1.27 (20H, m), 1.49 (1H, m), 1.7
9 (1H, m), 2.64 (1H, m), 2.68 (1H, d, 16.5) and 3.08
(1H, d, 16.5); ¹³ C NMR (67.5 MHz, CD ₃ OD) δ (ppm) 1
5.26,24.52,28.87,29.60,31.26 (3C), 31.44 (2C), 31.
54 (3C), 33.86, 43.08, 55.57, 77.59, 174.68, 176.97 and
177.41; FAB-MS (m / z) 359 (MH) ^- ; HRFAB-MS (m / z
z) Found: 359.2073 (calculated theoretical values for C ₁₈ H ₃₁ O _7: 35
9.2067); UV max (MeOH) 215 nm; [α] ²⁶ _D (c2.6, acetone) -18.34.

Example 2 Preparation of Trimethyl Ester of CJ-13,981 A solution of CJ-13,981 (2 mg) in 0.5 ml of acetonitrile was added at room temperature in n-hexane to give (trimethylsilyl)
Treated with 10 equivalents of diazomethane. After 2 hours, the reaction mixture was applied to a YMC-Pack ODS-AM-343 column (20 × 250 m
m, Yamamura trademark) and eluted with aqueous methanol (85:15) at a flow rate of 10 ml / min. Detection at 220 nm
Performed by UV absorbance. The eluting peak was collected and CJ-1
3,981 trimethyl ester was obtained.

Example 3 Preparation of Trimethyl Ester of CJ-13,982 A solution of CJ-13,982 (2 mg) in 0.5 ml of acetonitrile at room temperature in n-hexane (trimethylsilyl)
Treated with 10 equivalents of diazomethane. After 2 hours, the reaction mixture was applied to a YMC-Pack ODS-AM-343 column (20 × 250 m
m, Yamamura ™) and eluted with methanol-water (85:15) at a flow rate of 10 ml / min. Detection was by UV absorbance at 220 nm. The eluting peak was collected and CJ-
13,982 trimethyl esters were obtained.

──────────────────────────────────────────────────続 き Continuation of front page (56) References JP-A-7-112954 (JP, A) JP-A-58-172341 (JP, A) US Patent 5,254,727 (US, A) US Patent 3,625,826 (US, A) (58) Fields investigated (Int. Cl. ⁷ , DB name) C07C 59/245 C07C 59/42 C07C 69/675 C07C 69/732 CAPLUS (STN) REGISTRY (STN) WPIDS (STN)

Claims (3)

(57) [Claims] (1) Formula (I): Wherein Y is an ethyl or ethenyl group; and
R ¹ , R ² and R ³ are each independently a hydrogen atom or an alkyl group having 1 to 5 carbon atoms) or a pharmaceutically acceptable salt thereof. 2. Y is an ethenyl group, and R ¹ , R ² and
The compound according to claim 1, wherein R ³ is a hydrogen atom. 3. Y is an ethyl group and R ¹ , R ² and R ³
The compound according to claim 1, wherein is a hydrogen atom.