JPH05331060A - Phthalide derivative and cytostatic agent containing the phthalide derivative as active component - Google Patents

Abstract

PURPOSE:To provide a new compound having cytostatic action and useful for the treatment of arteriosclerosis, etc. CONSTITUTION:The compound of formula (R1 to R4 are H, hydroxyl or methoxy; R5 is O or S; R5 is O, S or nitrogen atom having H or 1-7C alkyl; R7 and R6 are H, 1-7C alkyl or together form 1-7C alkylidene), e.g. (Z)-3- butylidene-5,6-dimethoxyphthalimidine. The compound of formula can be produced e.g. by subjecting 3,4-dimethoxybenzoic acid to an arbitrary reaction selected from nitrogen-substitution reaction, reducing reaction, demethylation reaction, sulfur-substitution reaction, alkylation reaction, etc., and their combination.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a phthalide derivative having a cell growth inhibitory action and useful as a drug such as an anti-atherosclerotic agent.

[0002]

2. Description of the Related Art Conventionally, antihyperlipidemic agents and antihypertensive agents, which are therapeutic agents for hyperlipidemia, hypertension, etc., have been widely used in order to clinically suppress the onset of arteriosclerotic diseases. ..
In addition, platelet anti-aggregating agents and the like are also used for the purpose of preventing the formation of thrombus.

However, at present, these drugs only prevent risk factors, and development of drugs having an epoch-making therapeutic effect on arteriosclerotic diseases has been desired.

[0004]

[Means for Solving the Problems] Arteriosclerosis is a complex lesion formed by various phenomena. Among them, proliferation of smooth muscle cells in the intima is also observed in the early stage of lesion formation, and the lesion progression It is one of the most important phenomena because it plays a central role. Therefore, it is considered that arteriosclerosis can be treated if the proliferation of smooth muscle cells can be suppressed.

Therefore, the inventors of the present invention have conducted extensive studies to develop a drug having an epoch-making therapeutic effect on arteriosclerosis, and as a result, the phthalide derivative represented by the following formula I has a cell growth inhibitory action. The present invention has been completed and the present invention has been completed.

That is, the present invention provides the following formula I: (Wherein R ₁ , R ₂ , R ₃ and R ₄ are a hydrogen atom, a hydroxyl group or a methoxyl group, R ₅ is an oxygen atom or a sulfur atom, and R ₆ is an oxygen atom, a sulfur atom or a hydrogen atom or the number of carbon atoms. A nitrogen atom having an alkyl group of 1 to 7, both R ₇ and R ₈ are hydrogen atoms, or one of them is a hydrogen atom, and the other is an alkyl group of 1 to 7 carbon atoms, or both Together, they are a phthalide derivative represented by an alkylidene group having 1 to 7 carbon atoms (hereinafter referred to as the compound of the present invention) and a cell growth inhibitor containing the phthalide derivative as an active ingredient.

The compound of the present invention can be obtained by appropriately selecting reactions such as nitrogen substitution reaction, reduction reaction, demethylation reaction, sulfur substitution reaction and alkylation reaction, and combining them as necessary.

The nitrogen substitution reaction is described in the literature [Pharmaceutical Journal, 57 ,
783 (1937)]. For example, hydrazine hydrate may be added and reacted in a solvent such as ethanol.

The reduction reaction can be easily carried out by a commonly used method such as adding a metal and an acid. For example, the target substance can be obtained by adding zinc using concentrated hydrochloric acid as a solvent.

The demethylation reaction may be carried out by a commonly used method, for example, in an organic solvent such as methylene chloride,
This is accomplished by adding boron tribromide. At this time,
It is preferable to carry out the reaction by substituting with an inert gas such as argon gas.

The sulfur substitution reaction is described in the literature [J. Med. Ch
em. , 29, 1996 (1986), tetrah.
edrom, 35, 1339 (1979), etc.]. For example, phosphorus pentasulfide may be reacted in an organic solvent such as xylene.

The alkylation reaction may be carried out by a commonly used method. For example, in the case of methylation, it can be easily carried out by adding methyl iodide in the presence of sodium hydroxide in an organic solvent such as dimethylsulfoxide. You can

Next, the aortic smooth muscle cell proliferation inhibitory activity of the compound of the present invention will be described with reference to experimental examples. Experimental Example Dulbecco's Modified Eagle (Dulbecco's Mo) containing 10% fetal bovine serum (FBS) and antibiotics
defined Eagle, DME) medium,
SD rat thoracic aorta was separated from vascular smooth muscle by an enzymatic method, and the thoracic aorta was isolated in an atmosphere containing 95% air and 5% carbon dioxide.
The cells were cultured at ° C and subcultured constantly. In the 4th generation, cells were trypsinized and adjusted to 21500 cells in 1 ml in DME medium containing 10% FBS and antibiotics, and then 24-well plate (Corning)
1 ml each, S at a density of 21500 per 2 cm ²
MC was seeded and cultured. After culturing for 3 days, the medium is DME medium containing only antibiotics, control medium or test medium 1
Replaced with ml. Control medium consisted of DME medium containing 10% FBS and antibiotics and ethanol at 0.1% (v / v) concentration. The compound of the present invention was previously dissolved in ethanol so that the final concentration of ethanol was 0.1% (v / v). DM containing only antibiotics
After replacing with E medium, control medium or test medium, the cells were further incubated for 2 days. At the end of the incubation period, cells were trypsinized and counted by counting the cell suspension in a Coulter counter.

The results obtained were all expressed as an average value for 3 well cells. The growth inhibition rate (%) was calculated using the following formula.

[0015] (However, S in the formula is the we of the test medium when the test is completed
11 is the average number of cells per liter, C is the average number of cells per well of the control medium at the end of the test, and G is the average number of cells per well of the DME medium containing only the antibiotic at the end of the test. .. )

The results are shown as 50% inhibitory activity (IC ₅₀ , μ
g / ml) is shown in Table 1.

Table 1

From Table 1, it is clear that the compounds of the present invention have SMC growth inhibitory activity.

Next, the dose and formulation of the compound of the present invention will be explained.

The compound of the present invention can be administered to animals and humans as it is or together with a conventional pharmaceutical carrier. The dosage form is not particularly limited and may be appropriately selected and used as needed, and examples thereof include oral preparations such as tablets, capsules, granules, fine granules and powders, parenteral preparations such as injections and suppositories. Be done.

In order to exert a desired effect as an oral preparation, it varies depending on the age, body weight and degree of disease of the patient, but it is usually 50 mg to 5 g as the weight of the compound of the present invention in adults.
It is considered appropriate to take this in several divided doses a day.

Oral preparations include, for example, starch, lactose, sucrose,
Mannitol, carboxymethyl cellulose, corn starch, inorganic salts and the like are used according to a conventional method.

In this type of preparation, a binder, a disintegrating agent, a surfactant, a lubricant, a fluidity promoter, a flavoring agent, a coloring agent, a flavoring agent, etc. may be appropriately used in addition to the above-mentioned excipients. You can
Specific examples of each are as follows.

[Binder] Starch, dextrin, gum arabic powder, gelatin, hydroxypropyl starch,
Methyl cellulose, sodium carboxymethyl cellulose, hydroxypropyl cellulose, crystalline cellulose, ethyl cellulose, polyvinylpyrrolidone, macrogol.

[Disintegrant] Starch, hydroxypropyl starch, sodium carboxymethyl cellulose, calcium carboxymethyl cellulose, carboxymethyl cellulose, low-substituted hydroxypropyl cellulose.

[Surfactant] sodium lauryl sulfate,
Soy lecithin, sucrose fatty acid ester, polysorbate 80.

[Lubricant] Talc, waxes, hydrogenated vegetable oil, sucrose fatty acid ester, magnesium stearate, calcium stearate, aluminum stearate, polyethylene glycol.

[Fluidity promoter] Light anhydrous silicic acid, dried aluminum hydroxide gel, synthetic aluminum silicate, magnesium silicate.

The compounds of the present invention can also be administered as suspensions, emulsions, syrups and elixirs, and these various dosage forms may contain flavoring agents and coloring agents. Good.

In order to exert the desired effect as a parenteral agent, it depends on the age, body weight and degree of disease of the patient.
0.1 mg / day as the weight of the compound of the present invention in an adult
Intravenous infusion up to ~ 1 g, intravenous infusion, subcutaneous injection, and intramuscular injection seem appropriate.

This parenteral preparation is manufactured by a conventional method, and is generally used as a diluent in distilled water for injection, physiological saline, glucose solution, vegetable oil for injection, sesame oil, peanut oil, soybean oil, corn oil, propylene glycol, polyethylene glycol. Etc. can be used. Further, if necessary, a bactericidal agent, a preservative, and a stabilizer may be added. Further, from the viewpoint of stability, this parenteral preparation can be frozen after filling in a vial or the like, water is removed by a usual freeze-drying technique, and a liquid preparation can be re-prepared from the freeze-dried product immediately before use. Further, if necessary, a tonicity agent, a stabilizer, a preservative, a soothing agent and the like may be added appropriately.

Other parenteral agents include external preparations, coating agents such as ointments, and suppositories for rectal administration.
It is manufactured according to a conventional method.

Next, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

Example 1 4.4 ml of diisopropylamine was placed in a 500 ml two-necked flask equipped with a septum rubber, and the atmosphere was replaced with argon.
After adding and dissolving 50 ml of anhydrous tetrahydrofuran,
22 ml of 1.6Mn-butyllithium was added at -10 ° C, and the mixture was stirred at -10 ° C for 30 minutes.

Further, the reaction solution was heated at -40 ° C for 5,6
-Dimethoxyphthalide [200 ml with reflux condenser
Add 3,4-dimethoxybenzoic acid 5.0 to an eggplant-shaped flask.
g, 35% hydrochloric acid 50 ml, concentrated sulfuric acid 0.5 ml and dimethoxymethane 2.9 ml were added, and the mixture was stirred at an external temperature (90-90).
Heated at 100 ° C) for 7 hours. The reaction solution was returned to room temperature, extracted twice with chloroform (150 ml × 2), the chloroform layer was washed once with 100 ml of 5% aqueous sodium hydroxide solution and once with saturated saline solution, and dried (MgSO 4).
₄ ), filtered through Celite, and evaporated under reduced pressure to obtain a white solid. This was recrystallized from ethyl acetate-n-hexane to give 5,6-
1.36 g of dimethoxyphthalide was obtained as colorless scaly crystals. ] A suspension of 6.0 g of tetrahydrofuran in 130 ml was added over about 5 minutes, and the mixture was stirred at -40 ° C for 1 hour, and then-
Zinc chloride at 40 ° C (After finely crushing, stirring, under reduced pressure,
Heat and dry) 5.1 g of tetrahydrofuran 50
The ml solution was added, and the mixture was stirred at -40 ° C for 30 minutes.

Further, to this reaction solution, at -40 ° C, 4.1 ml of n-butyraldehyde and 10 ml of tetrahydrofuran.
The solution was added and stirred at -40 ° C for 1 hour, then at room temperature for 2 hours.
Stir for hours. This reaction solution was ice-cooled, 5% hydrochloric acid 150 m
The mixture was poured into 1 l, extracted twice with ether (300 ml × 2), the ether layer was washed once with saturated brine, dried (magnesium sulfate), and the solvent was evaporated under reduced pressure to give a yellow oil. This was subjected to flash column chromatography (silica gel, 23
0-400 mesh, φ6.5 × 20, about 300 g; eluate, ethyl acetate: n-hexane = 1: 1, pressure 0.2
kg / cm ² , 1 fraction = 100 ml) was separated and purified, and 3- (1-hydroxybutyl) -5,6 represented by the above formula ◆ from the elution fraction (1900 to 6000 ml).
-5.74 g of dimethoxyphthalide was obtained as a colorless oil.

In a 300 ml round-bottomed flask equipped with a reflux condenser, 5.7 g of 3- (1-hydroxybutyl) -5,6-dimethoxyphthalide was placed and replaced with argon, and 20 ml of anhydrous benzene and 4.3 ml of anhydrous pyridine were added. After dissolving, 3.3 ml of methanesulfonyl chloride was added, and the mixture was heated under reflux for 1 hour. The reaction solution was poured into ice water and extracted twice with ether (200 ml × 2), and the ether layer was 5%.
Washed once with hydrochloric acid and saturated saline, and dried (Mg
SO _4), the solvent was distilled off under reduced pressure, to give 3- (1-methanesulfonyloxy-butyl) -5,6-dimethoxy-phthalide as a yellow oil. Next, this oily substance was mixed with anhydrous benzene 7
It was dissolved in 0 ml, DBU (4.8 ml) was added, and the mixture was heated under reflux for 1 hr. 200 ml of 2% hydrochloric acid obtained by ice-cooling this reaction solution
And extracted twice with ether (200 ml × 2), the ether layer was washed once with water and saturated brine, dried (MgSO ₄ ), and the solvent was evaporated under reduced pressure to give a yellow oil. This was subjected to flash column chromatography (silica gel, 230-400 mesh, φ6.5 × 30, about 450 g; eluent, chloroform, pressure 0.2 kg / c).
m ₂ , 1 fraction = 200 ml) was separated and purified to obtain 2.0 g of a white solid, which was recrystallized from ethyl acetate-n-hexane to give 3-butylidene-5,6-dimethoxyphthalide as colorless fine needles. Obtained as crystals.

In a 500 ml round-bottomed flask equipped with a reflux condenser, 5.0 g of 3-butylidene-5,6-dimethoxyphthalide, 50 ml of ethanol and 5 hydrazine hydrates were added.
0 ml was added, and the mixture was heated under reflux for 2 hours with stirring. The reaction mixture was poured into ice water, and the precipitated insoluble material was collected by filtration (washed well with water) to obtain 4.7 g of a white powder. This was recrystallized from chloroform to give 4-butyl-6,7-dimethoxyphthalazinone (4-butyl) having the following physicochemical properties.
-6,7-dimethyoxyphthalazino
ne) (4.3 g) was obtained as colorless fine needle crystals. Structural formula; C ₁₄ H ₁₈ O ₃ N ₂ molecular weight; 262.31 Melting point: 226-228 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3152, 2952, 1642, 1604, 1
504, 1402, 1304, 1266, 1216, 1
172, 1080 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 0.99 (3H,} t, J = 7.3Hz),
1.50 (2H, m), 1.80 (2H, m), 2.9
0 (2H, t, J = 7.8Hz), 4.05 (6H,
s), 7.10 (1H, s), 7.82 (1H, s),
10.26 (1H, br, D ₂ O exchange
ble) Mass spectrum (EI-MS) m / z (%): 262
(M ⁺ , 21), 220 (100)

In a 500 ml eggplant-shaped flask, 4-butyl-
4.0 g of 6,7-dimethoxyphthalazinone was added to concentrated hydrochloric acid 24
After suspending in 0 ml, 40 g of zinc was added little by little under stirring (because of the exothermic reaction, divided into about 8 times, taking care not to make the reaction extreme). The reaction solution was poured into ice water, extracted twice with ethyl acetate (200 × 2), the ethyl acetate layer was washed twice with water, once with saturated saline solution and dried (MgSO ₄ ), the solvent was distilled off under reduced pressure, and washed with light brown. A colored solid was obtained. This is flash column chromatography (silica gel, 230-40
0 mesh, φ4.5 × 20, about 160 g, eluate; ethyl acetate: n-hexane = 1: 1 to 2: 1, pressure 0.2
kg / cm ² , 1 fraction = 80 ml),
58 mg of a white solid was obtained from the first elution fraction (11-23). Further, this was recrystallized from chloroform-n-hexane to give (Z) -3-butylidene-5,6-dimethoxyphthalimidine [(Z) -3 having the following physicochemical properties.
-Butylidene-5,6-dimethoxy
[phthalimidine] was obtained as colorless needle crystals. Structural formula: C ₁₄ H ₁₇ O ₃ N Molecular weight: 247.30 Melting point: 188-190 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3176, 2960, 2928, 1678, 1
612, 1492, 1354, 1276, 1214 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 1.01 (3H,} t, J = 7.3Hz),
1.58 (2H, tq, J = 7.3, 7.3Hz),
2.27 (2H, dt, J = 7.8, 7.3Hz),
5.48 (1H, t, J = 7.8Hz), 7.06 (1
H, s), 7.27 (1H, s), 7.66 (1H, b)
r, D ₂ O exchangeable) mass spectrum (EI-MS) m / z (%): 247
(M ⁺ , 23), 218 (100)

From the second eluted fraction (28-45), 2.04 g of white solid was obtained. Further, it was recrystallized from chloroform-n-hexane to give 3-
Butyl-5,6-dimethoxyphthalimidine (3-Bu
tyl-5,6-dimethyoxyphthalim
Idine) was obtained as colorless fine needle crystals. Structural formula: C ₁₄ H ₁₉ O ₃ N Molecular weight: 249.31 Melting point: 177-179 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3184, 3068, 2928, 2864, 1
688, 1614, 1502, 1350, 1282, 1
216 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 0.91 (3H,} t, J = 6.8Hz),
1.35 (4H, m), 1.60 (1H, m), 1.9
5 (1H, m), 3.94 (3H, s), 3.96 (3
H, s), 4.53 (1H, dd, J = 7.3, 3.9).
Hz), 6.87 (1H, s), 7.30 (1H,
s), 6.60-7.10 (1H, br, D ₂ O ex
changeable) mass spectrum (EI-MS) m / z (%): 249
(M ⁺ , 16), 192 (100)

Example 2 (Z) -3-butylidene-in a 25 ml eggplant-shaped flask.
After adding 30 mg of 5,6-dimethoxyphthalimidine and substituting with argon gas and adding 0.36 ml of anhydrous methylene chloride, 0.36 ml of a 1M solution of boron tribromide in methylene chloride was added at -20 ° C. Stirred at 20 ° C for 2 hours. The reaction mixture was poured into ice water and extracted twice with ethyl acetate (1
0 × 2), the ethyl acetate layer was washed twice with saturated brine, dried (MgSO ₄ ) and the solvent was evaporated under reduced pressure to give a white solid.
This is flash column chromatography (silica gel, 230-400 mesh, φ3.5 × 5, about 40
g, eluent; methanol: chloroform = 1: 15, pressure 0.2 kg / cm ² , 1 fraction = 30 ml), and the following physicochemical properties were obtained from the first elution fraction (6) (Z). ) -3-Butylidene-5,6-dihydroxyphthalimidine [(Z) -3-Butylidene-
5,6-dihydroxyphythalimidin
e] 10 mg were obtained as a white solid. Structural formula; C ₁₂ H ₁₃ O ₃ N molecular weight; 219.24 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{D 3 OD): 0.99 (3H} , t, J = 7.3Hz),
1.54 (2H, tq, J = 7.3, 7.3Hz),
2.31 (2H, dt, J = 7.9, 7.3Hz),
5.48 (1H, t, J = 7.9Hz), 7.07 (1
H, s), 7.09 (1H, s) Mass spectrum (EI-MS) m / z (%): 219
(M ⁺ , 18), 190 (100)

Example 3 100 mg of 3-butyl-5,6-dimethoxyphthalimidine was placed in a 25 ml round-bottomed flask, the atmosphere was replaced with argon gas, and 1.2 ml of anhydrous methylene chloride was added.
1.2m methylene chloride solution of 1M boron tribromide at 0 ° C
1 was added, and the mixture was stirred at -20 ° C for 2 hours. The reaction solution was poured into ice water, extracted twice with ethyl acetate (containing 5% methanol) (40 × 2), and the ethyl acetate layer was washed once with saturated saline solution.
Drying (MgSO ₄ ) and distilling off the solvent under reduced pressure gave a white solid. This is flash column chromatography (silica gel, 230-400 mesh, φ3.5 × 5, about 4
0 g, eluate; methanol: chloroform = 1: 10,
Pressure 0.2 kg / cm ² , 1 fraction = 30 ml)
And 3-butyl-5,6-dihydroxyphthalimidine (3-Butyl-5,6-dihydroxox) having the following physicochemical properties from the first elution fraction (4-8).
67 mg of yphthalimidine) was obtained as a white solid. Structural formula: C ₁₂ H ₁₅ O ₃ N Molecular weight: 221.26 Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3416, 3316, 3080, 2928, 1
648, 1610, 1476, 1406, 1316, 1
188,1154 Proton nuclear magnetic resonance spectrum (δ ppm in C
D ₃ OD): 0.91 (3H, t, J = 7.0Hz),
1.30 (4H, m), 1.60 (1H, m), 1.8
5 (1H, m), 4.47 (1H, dd, J = 6.6,
4.6 Hz), 6.87 (1 H, s), 7.11 (1
H, s) Mass spectrum (EI-MS) m / z (%): 221
(M ⁺ , 9), 164 (100)

Example 4 (Z) -3-butylidene-in a 50 ml eggplant-shaped flask.
2.0 g of 4,5-dimethoxyphthalide, phosphorus pentasulfide 3.
After adding 36 g and substituting with argon, anhydrous xylene 20 was added.
ml was added, and the mixture was heated and stirred at an external temperature (120 ° C) for 4 hours. Water was added to the reaction solution and extracted twice with ethyl acetate (150 ×
2), the ethyl acetate layer was washed once with saturated saline and dried (M
gSO ₄ ) and the solvent were distilled off under reduced pressure to obtain a green oily substance. This is flash column chromatography (silica gel, 230-400 mesh, φ4.5 × 20, about 16
0 g, eluate; ethyl acetate: n-hexane = 1: 5, pressure 0.2 kg / cm ² , 1 fraction = 80 ml), and a yellow solid 1.5 from the first elution fraction (6-10).
4 g was obtained. This was recrystallized from chloroform-n-hexane to give (Z) -3-butylidene-4,5-dimethoxy-1 (3H) -isobenzofuranthion [(Z) -3-Butylidene-] having the following physicochemical properties.
4,5-dimethyloxy-1 (3H) -isobe
nzofuranthoione] was obtained as yellow needles. Structural formula: C ₁₄ H ₁₆ O ₃ S Molecular weight: 264.34 Melting point: 85-87 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 2932, 1606, 1504, 1450, 1
332, 1274, 1180, 1076, 1056, 1
004 Proton Nuclear Magnetic Resonance Spectra (δ ppm in C
_{DCl 3): 1.01 (3H,} t, J = 7.3Hz),
1.59 (2H, tq, J = 7.3, 7.3Hz),
2.55 (2H, dt, J = 8.1, 7.3Hz),
3.96 (3H, s), 3.98 (3H, s), 6.0
5 (1H, t, J = 8.1Hz), 7.04 (1H,
d, J = 8.6 Hz), 7.76 (1H, d, J = 8.
6 Hz) Mass spectrum (EI-MS) m / z (%): 264
(M ⁺ , 100), 235 (90), 222 (35),
207 (56), 189 (26)

Example 5 (Z) -3-butylidene-in a 25 ml eggplant-shaped flask
100 mg of 4,5-dimethoxy-1 (3H) -isobenzofuranthion was added, the atmosphere was replaced with argon gas, 1.1 ml of anhydrous methylene chloride was added, and then 1 M solution of boron tribromide in methylene chloride 1 was added at -20 ° C. Add 1 ml and -2
Stirred at 0 ° C for 2 hours. The reaction mixture was poured into ice water, extracted twice with ethyl acetate (40 × 2), the ethyl acetate layer was washed twice with saturated brine, dried (MgSO ₄ ), and the solvent was evaporated under reduced pressure to give a yellow solid. This is flash column chromatography (silica gel, 230-400 mesh, φ
3.5 × 5, about 40 g, eluate; methanol: chloroform = 1: 15, pressure 0.2 kg / cm ² , 1 fraction = 30 ml), and separated from the first elution fraction (3-4) (Z) -3-Butylidene-4,5-dihydroxy-1
(3H) -isobenzofuranthione [(Z) -3-Bu
tylidene-4,5-dihydroxy-1
(3H) -isobenzofuranthoion
e] 56 mg was obtained as a white solid. Structural formula: C ₁₂ H ₁₂ O ₃ S Molecular weight: 236.28 Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3170, 2956, 1632, 1608, 1
520, 1462, 1328, 1294, 1216, 1
180 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{D 3 OD): 1.02 (3H} , t, J = 7.3Hz),
1.60 (2H, tq, J = 7.3, 7.3Hz),
2.50 (2H, dt, J = 8.1, 7.3Hz).
6.02 (1H, t, J = 8.1Hz), 6.93 (1
H, d, J = 8.1 Hz), 7.38 (1H, d, J =
8.1 Hz) Mass spectrum (EI-MS) m / z (%): 236
(M ⁺ , 52), 207 (71), 194 (100)

Example 6 In a 500 ml eggplant-shaped flask equipped with a reflux condenser, a reference [Heterocycles, vol. 32,1737
(1991)] 3-butylidene-4,5-dimethoxyphthalide (5.0 g), ethanol (50 ml) and hydrazine hydrate (50 ml) were added, and the mixture was heated under reflux for 2 hours with stirring. The reaction mixture was poured into ice water, and the precipitated insoluble matter was collected by filtration (washed well with water) to obtain 5.3 g of a white powder. This was recrystallized from chloroform to give 4-butyl-5,6-dimethoxyphthalazinone (4
-Butyl-5,6-dimethyoxytha
Lazone) 4.5 g was obtained as colorless scaly crystals. Structural formula; C ₁₄ H ₁₈ O ₃ N ₂ molecular weight; 262.31 Melting point: 185-186 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3172, 2952, 1674, 1602, 1
298, 1280, 1026 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 0.96 (3H,} t), 2.46 (2H,
m), 2.70 (2H, m), 3.04 (2H, t, J
= 7.6 Hz), 3.94 (3H, s), 4.03 (3
H, s), 7.38 (1H, d, J = 8.8 Hz),
8.27 (1H, d, J = 8.8Hz), 9.86 (1
H, br, D ₂ O exchangeable mass spectrum (EI-MS) m / z (%): 262
(M ⁺ , 14), 231 (22), 220 (28), 2
05 (100)

In a 500 ml eggplant-shaped flask, 4-butyl-
5,6-dimethoxyphthalazinone 4.0 g was added with concentrated hydrochloric acid 16
After suspending in 0 ml, 40 g of zinc was added little by little under stirring (because of the exothermic reaction, divided into about 8 times, taking care not to make the reaction extreme). The reaction mixture was poured into ice water, extracted twice with ethyl acetate (200 × 2), the ethyl acetate layer was washed twice with water, washed with saturated saline once and dried (MgSO4), the solvent was evaporated under reduced pressure, and the residue was light brown. To give a solid. This is flash column chromatography (silica gel, 230-40
0 mesh, φ4.5 × 20, about 160 g, eluate; ethyl acetate: n-hexane = 1: 1 to 3: 2, pressure 0.2
kg / cm ² , 1 fraction = 80 ml),
0.87 g of a white solid was obtained from the first elution fraction (6 to 13). Further, this was recrystallized from chloroform-n-hexane to give (Z) -3-butylidene-4,5-dimethoxyphthalimidine [(Z) -3 having the following physicochemical properties.
-Butylidene-4,5-dimethoxy
[phthalimidine] was obtained as colorless needle crystals. Structure; C ₁₄ H ₁₇ O ₃ N molecular weight; 247.30 mp: 130-132 ° C Infrared absorption spectrum ^{(IR, ν max cm -1,} K
Br): 3176, 3048, 2956, 1710, 1
620, 1500, 1368, 1268, 1096, 1
030 proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 1.00 (3H,} t, J = 7.3Hz),
1.59 (2H, tq, J = 7.3, 7.3Hz),
2.33 (2H, dt, J = 8.1, 7.3Hz),
3.93 (3H, s), 3.95 (3H, s), 6.0
7 (1H, t, J = 8.1Hz), 7.00 (1H,
d, J = 8.3 Hz), 7.58 (1H, d, J = 8.
3Hz), 8.61 (1H, br, D ₂ O excha
massable) mass spectrum (EI-MS) m / z (%): 247
(M ⁺ , 27), 218 (100), 175 (29)

From the second eluted fraction (21-30), 1.46 g of white solid was obtained. Further, it was recrystallized from chloroform-n-hexane and had the following physicochemical properties-3
-Butyl-4,5-dimethoxyphthalimidine (3-B
utyl-4,5-dimethyloxyphthali
to obtain colorless fine needle crystals. Structural formula: C ₁₄ H ₁₉ O ₃ N Molecular weight: 249.31 Melting point: 119-121 ° C Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3196, 3080, 2952, 2860, 1
710, 1620, 1496, 1372, 1268, 1
074,1028 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 0.88 (3H,} t, J = 7.0Hz),
1.30 (4H, m), 1.65 (1H, m), 2.1
5 (1H, m), 3.92 (3H, s), 3.94 (3
H, s), 4.70 (1H, dd, J = 7.3, 2.9).
Hz), 7.02 (1H, d, J = 8.3 Hz), 7.
21 (1H, br, D ₂ O exchangeable
e), 7.56 (1H, d, J = 8.3 Hz) Mass spectrum (EI-MS) m / z (%): 249
(M ⁺ , 9), 207 (10), 192 (100)

Example 7 (Z) -3-butylidene- was added to a 25 ml eggplant-shaped flask.
After adding 100 mg of 4,5-dimethoxyphthalazinone and replacing with argon gas, 1.2 ml of anhydrous methylene chloride was added, and then 1.2 ml of 1M boron tribromide in methylene chloride solution was added at -20 ° C. Stirred at 20 ° C for 2 hours.
The reaction solution was poured into ice water and extracted twice with ethyl acetate (40 ×
2), wash the ethyl acetate layer once with water and dry (MgS
O ₄ ) and the solvent were distilled off under reduced pressure to obtain a white solid. This is flash column chromatography (silica gel, 23
0-400 mesh, φ3.5 × 5, about 40 g, eluate; methanol: chloroform = 1: 15, pressure 0.2
kg / cm ² , 1 fraction = 30 ml)
From the first elution fractions (1 to 3), (Z) -3-butylidene-4,5-dihydroxyphthalimidine [(Z) -3-Butylidene-4, which has the following physicochemical properties,
5-dihydroxyphthalimidine]
84 mg was obtained as a white solid. Structural formula: C ₁₂ H ₁₃ O ₃ N Molecular weight: 219.24 Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3468, 3240, 2960, 1650, 1
626, 1604, 1286 Proton nuclear magnetic resonance spectrum (δ ppm in C
D ₃ OD): 1.00 (3H, t, J = 7.3Hz),
1.56 (2H, tq, J = 7.3, 7.3Hz),
2.33 (2H, dt, J = 7.9, 7.3Hz),
6.09 (1H, t, J = 7.9Hz), 6.86 (1
H, d, J = 8.1 Hz), 7.13 (1H, d, J =
8.1 Hz). Mass spectrum (EI-MS) m / z (%): 219
(M ⁺ , 20), 190 (100)

Example 8 100 mg of 3-butyl-4,5-dimethoxyphthalimidine was placed in a 25 ml round-bottomed flask, the atmosphere was replaced with argon gas, 1.2 ml of anhydrous methylene chloride was added, and then -2.
1.2m methylene chloride solution of 1M boron tribromide at 0 ° C
1 was added, and the mixture was stirred at -20 ° C for 2 hours. The reaction solution was poured into ice water, extracted twice with ethyl acetate (containing 5% methanol) (40 × 2), and the ethyl acetate layer was washed once with saturated saline solution.
Drying (MgSO ₄ ) and distilling off the solvent under reduced pressure gave a white solid. This is flash column chromatography (silica gel, 230-400 mesh, φ3.5 × 5, about 4
0 g, eluate; methanol: chloroform = 1: 10,
Pressure 0.2 kg / cm ² , 1 fraction = 30 ml)
And 3-butyl-4,5-dihydroxyphthalimidine (3-Butyl-4,5-dihydro) having the following physicochemical properties from the first elution fraction (7-14).
67 mg of xyphthalimidine) was obtained as a white solid. Structural formula: C ₁₂ H ₁₅ O ₃ N Molecular weight: 221.26 Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3392, 2956, 2928, 1658, 1
628, 1416, 1292 Proton nuclear magnetic resonance spectrum (δ ppm in C
D ₃ OD): 0.89 (3H, t, J = 7.1Hz),
1.25 (4H, m), 1.70 (1H, m), 2.1
0 (1H, m), 4.66 (1H, dd, J = 6.8,
3.2 Hz), 6.88 (1H, d, J = 8.1H
z), 7.12 (1H, d, J = 8.1 Hz) Mass spectrum (EI-MS) m / z (%): 221
(6, M ⁺ ), 164 (100)

Example 9 In a 50 ml eggplant-shaped flask, (Z) -3-butylidene-
After adding 300 mg of 4,5-dimethoxyphthalimidine and 136 mg of potassium hydroxide and replacing with argon gas, 3 ml of anhydrous dimethylsulfoxide and 0.23 ml of methyl iodide were added, and the mixture was stirred at room temperature for 3 hours. This reaction solution was added with water, extracted twice with ethyl acetate (100 × 2), and the ethyl acetate layer was washed twice with saturated brine and dried (MgS
O ₄ ) and the solvent were distilled off under reduced pressure to obtain a colorless oily substance. This is flash column chromatography (silica gel, 2
30-400 mesh, φ4.5 × 20, about 160g,
Eluent; ethyl acetate: n-hexane = 1: 1, pressure 0.
2 kg / cm ² , 1 fraction = 80 ml), and 310 m of colorless solid from the first elution fraction (11-15)
g was obtained. Further, it was recrystallized from chloroform-n-hexane and had the following physicochemical properties (Z) -3-
Butylidene-4,5-dimethoxy-N-methylphthalimidine [(Z) -3-Butylidene-4,5-
dimethyl-N-methylphthali
to obtain needle-like crystals. Structure; C ₁₅ H ₁₉ O ₃ N molecular weight; 261.32 mp: 77-79 ° C IR absorption spectrum ^{(IR, ν max cm -1,} K
Br): 2952, 1694, 1648, 1498, 1
428, 1384, 1266, 1256, 1082, 1
056 Proton nuclear magnetic resonance spectrum (δ ppm in C
_{DCl 3): 1.03 (3H,} t, J = 7.3Hz),
1.60 (2H, tq, J = 7.3, 7.3Hz),
2.61 (2H, dt, J = 8.1, 7.3Hz),
3.49 (3H, s), 3.90 (3H, s), 3.9
3 (3H, s), 6.29 (1H, t, J = 8.1H
z), 6.97 (1H, d, J = 8.3 Hz), 7.5
5 (1H, d, J = 8.3 Hz). Mass spectrum (EI-MS) m / z (%): 261
(M ⁺ , 25), 232 (100)

Example 10 (Z) -3-butylidene-in a 25 ml eggplant-shaped flask
4,5-dimethoxy-N-methylphthalimidine 100
mg was added, the atmosphere was replaced with argon gas, 1.1 ml of anhydrous methylene chloride was added, 1.1 ml of a 1M solution of boron tribromide in methylene chloride was added at -20 ° C, and the mixture was stirred at -20 ° C for 2 hours. The reaction solution is poured into ice water and extracted with ethyl acetate 2
(40 × 2) times, the ethyl acetate layer was washed twice with saturated brine, dried (MgSO ₄ ) and the solvent was evaporated under reduced pressure to give a white solid. This is flash column chromatography
(Silica gel, 230-400 mesh, φ3.5 ×
5, about 40 g, eluate; methanol: chloroform =
1:15, pressure 0.2 kg / cm ² , 1 fraction =
(Z) -3-butylidene-4, which has the following physicochemical properties, from the first elution fraction (2-3).
5-dihydroxy-N-methylphthalimidine [(Z)
-3-Butylidene-4,5-dihydro
xy-N-methylphthalimidine]
Obtained 49 mg as a white solid. Structural formula: C ₁₃ H ₁₅ O ₃ N Molecular weight: 233.27 Infrared absorption spectrum (IR, ν max cm ^-1 , K
Br): 3464, 2956, 2924, 1646, 1
434, 1282, 1068 Proton nuclear magnetic resonance spectrum (δ ppm in C
D ₃ OD): 1.04 (3H, t, J = 7.3Hz),
1.60 (2H, tq, J = 7.3, 7.3Hz),
2.65 (2H, dt, J = 7.9, 7.3Hz),
3.47 (3H, s), 6.40 (1H, t, J = 7.
9Hz), 6.85 (1H, d, J = 8.1Hz),
7.12 (1H, d, J = 8.1Hz). Mass spectrum (EI-MS) m / z (%): 233
(M ⁺ , 19), 204 (100)

Next, the formulation examples of the present invention will be described. [Formulation Example 1] Corn starch 44 g Crystalline cellulose 40 g Carboxymethyl cellulose calcium 5 g Light anhydrous silicic acid 0.5 g Magnesium stearate 0.5 g Compound obtained in Example 2 10 g Total 100 g According to the above formulation, It was compression molded with a tableting machine to obtain 200 mg tablets. 20 mg of the compound obtained in Example 2 is contained in one tablet, and 3 to 10 tablets for an adult are taken in several divided doses per day.

[Formulation Example 2] Crystalline cellulose 84.5 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Compound obtained in Example 3 10 g Total 100 g According to the above-mentioned prescription, a part of and was uniformly mixed, After compression molding, the mixture was crushed, and the remaining amount was added and mixed, and compression molding was carried out using a tablet machine to obtain 200 mg tablets. 20 mg of the compound obtained in Example 3 is contained in one tablet, and 3 to 10 tablets for an adult are taken in several divided doses per day.

[Formulation Example 3] Crystalline cellulose 79.5 g 10% Hydroxypropyl cellulose ethanol solution 50 g Carboxymethyl cellulose calcium 5 g Magnesium stearate 0.5 g Compound obtained in Example 5 10 g Total 145 g According to the above formulation, and After uniformly mixing, blending by a conventional method, granulating by an extrusion granulator, drying and crushing, and are mixed, compression molding is carried out by a tableting machine to obtain a tablet of 200 mg. It was 20 mg of the compound obtained in Example 5 is contained in one tablet, and 3 to 10 tablets for an adult are taken in several divided doses per day.

[Formulation Example 4] Corn starch 84 g Magnesium stearate 0.5 g Carboxymethyl cellulose calcium 5 g Light anhydrous silicic acid 0.5 g Compound 10 g obtained in Example 7 10 g Total 100 g After compression molding with a compression molding machine, it was crushed with a crusher and sieved to obtain granules. 1 g of this granule contains 100 mg of the compound obtained in Example 7, and 0.6 to 2 g per day for an adult
Take in several divided doses.

[Formulation Example 5] Crystalline cellulose 86.5 g 10% Hydroxypropyl cellulose ethanol solution 35 g Compound obtained in Example 8 10 g Total 131.5 g According to the above-mentioned prescription, the ingredients (1) to (4) were uniformly mixed and mixed. After granulating with an extrusion granulator, it was dried and sieved to obtain granules. 1 g of this granule contains 100 mg of the compound obtained in Example 8, and an adult daily dose of 0.6-
Take 2g in several divided doses.

Formulation Example 6 Cornstarch 89.5 g Light anhydrous silicic acid 0.5 g Compound obtained in Example 10 10 g Total 100 g
No. 2 capsule was filled. 20 mg of the compound obtained in Example 10 is contained in 1 capsule of this capsule, and 3 to 10 capsules for an adult are to be taken in several divided doses per day.

[Formulation Example 7] Distilled water for injection 89.5 g Soybean oil 5 g Soybean phospholipid 2.5 g Glycerin 2 g Compound 1 g obtained in Example 1 1 g Total amount 100 g The solution was added and emulsified to obtain an injection.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁵ Identification number Reference number within the agency FI Technical display location C07D 307/87 307/88 333/72 (72) Inventor Riki Oyama 3586 Yoshihara, Ami-cho, Inashiki-gun, Ibaraki Prefecture Tsumura Co., Ltd.

Claims (2)

[Claims] 1. The following formula I (Wherein R ₁ , R ₂ , R ₃ and R ₄ are a hydrogen atom, a hydroxyl group or a methoxyl group, R ₅ is an oxygen atom or a sulfur atom, and R ₆ is an oxygen atom, a sulfur atom or a hydrogen atom or the number of carbon atoms. A nitrogen atom having an alkyl group of 1 to 7, both R ₇ and R ₈ are hydrogen atoms, or one of them is a hydrogen atom, and the other is an alkyl group of 1 to 7 carbon atoms, or both Together, they are an alkylidene group having 1 to 7 carbon atoms). 2. The following formula I (Wherein R ₁ , R ₂ , R ₃ and R ₄ are a hydrogen atom, a hydroxyl group or a methoxyl group, R ₅ is an oxygen atom or a sulfur atom, and R ₆ is an oxygen atom, a sulfur atom or a hydrogen atom or the number of carbon atoms. A nitrogen atom having an alkyl group of 1 to 7, both R ₇ and R ₈ are hydrogen atoms, or one of them is a hydrogen atom, and the other is an alkyl group of 1 to 7 carbon atoms, or both And a phthalide derivative represented by C1 to C7 alkylidene group) as an active ingredient.