JPH10287667A - Isocoumarin derivatives and their pharmaceutical composition - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain new isocoumarin derivatives capable of being utilized as an active ingredient exhibiting pharmacokinetics different from that of a conventional isocoumarin derivative in a pharmaceutical composition, e.g. a vascularization inhibitor. SOLUTION: The new isocoumarin compound is expressed by formula I [A is H, a lower alkyl; B is H, an OH-protecting group; R<1> is cyano, (substituted) carbamoyl, etc.; R<2> , R<3> are each H, a lower alkyl, a (substituted) aryl-lower alkyl, etc.], e.g. 2-(8-hydroxy-6-methoxy-1H-2-benzopyran-1-on-3-yl)-2- methylpropionic acid. The compound of formula I is obtained e.g. by introducing one or two lower alkyl groups, etc., into the methyl portion in the cyanomethyl group of a compound of formula II to produce a synthetic intermediate, and subsequently hydrolyzing the cyano group.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

[0001] The present invention relates to isocoumarin derivatives and pharmaceutical preparations containing them.

[0002]

BACKGROUND OF THE INVENTION Isocoumarin derivatives, especially formulas

[0003]

Embedded image

[0004] 3-hydroxymethyl-6-
Methoxy-8-hydroxy-1H-2-benzopyran-
1-one has been found for the first time as a compound produced by Streptoverticillium eurocidicum, and the antibiotic MI43 showing growth inhibitory activity against various animal cells and human cancer cells.
-37F11 (JP-A-3-217)
No. 7). Thereafter, the antibiotic MI43-37F11 was used.
(Hereinafter abbreviated as MI43), and an isocoumarin derivative having a leaving group-containing methyl group such as benzyloxymethyl or a methyl halide group at the 3-position of the isocoumarin skeleton is used as an intermediate. JP-A-4-112884), and the above-mentioned third position is represented by the formula

[0005]

Embedded image

Wherein R ₃ and R ₄ are each independently a substituted or unsubstituted alkyl group, alkoxy, alkanoyloxy, mono- or di-substituted amino, phenylthio,
Or an isocoumarin derivative having a group such as N ₃
Compounds having the same pharmacological activity as 43
No. 97441).

On the other hand, since MI43 significantly suppresses adjuvant arthritis in rats and collagen-induced joint salts in mice by oral administration, its use as an immunomodulator has also been proposed (JP-A-6-183966). Also,
As an isocoumarin derivative having more excellent efficacy against these diseases, a group represented by -C at the 3-position of the isocoumarin skeleton.
A compound to which HRCOOH (where R is a hydrogen atom or a lower alkyl group) is also proposed (JP-A-8-176138).

[0008]

SUMMARY OF THE INVENTION The above-mentioned Japanese Patent Application Laid-Open No. 8-176 is disclosed.
Although the compounds described in the '138 publication are particularly interesting in that they also have excellent bioavailability, there remains a need for compounds with even better potency. Therefore, an object of the present invention is to provide a compound other than the compounds described in the above-mentioned gazettes, which exhibits excellent efficacy for the above-mentioned diseases.

[0009]

Means for Solving the Problems The present inventors have, for example,
Some compounds described in JP-A-8-176138, in which the methine (-CH =) portion of the group -CHRCOOH (where R is a lower alkyl group) of an isocoumarin derivative has a more bulky substituent, Was found to exhibit pharmacological activity and a different biokinetics from known isocoumarin derivatives. In addition, a novel synthetic intermediate for efficiently producing such a compound has been found.

Thus, according to the present invention, there are provided isocoumarin derivatives represented by the following formula (I) and, where appropriate, salts thereof.

Formula (I):

[0012]

Embedded image

In the formula, A represents a hydrogen atom or a lower alkyl group; B represents a hydrogen atom or a protecting group for a hydroxyl group;
¹ represents a cyano group, a carbamoyl group or a carboxyl group which may be N-mono- or di-alkyl-substituted, and R ² and R ³ each independently represent hydrogen, lower alkyl or aryl-lower alkyl. Wherein the aryl moiety may be optionally substituted with one or more substituents, or R ² and R ³ together with the carbon atom to which they are attached, may have any of the three to seven members A C _2-6 alkylene group forming an alicyclic ring, provided that A
Is a methyl group and R ² and R ³ represent a hydrogen atom at the same time, or when one represents a hydrogen atom and the other represents a lower alkyl group, R ¹ is a group other than a cyano group or a carboxyl group.

Of these, A is a hydrogen atom or a lower alkyl group, B is a hydrogen atom, and R ¹ is a carboxyl group or a carbamoyl group optionally substituted with N-mono- or di-lower alkyl. The compounds of formula (I) have, for example, excellent anti-angiogenic activity. On the other hand, the compound of the formula (I) wherein A is a lower alkyl group, B is a protecting group for a hydroxyl group, and R ¹ is a cyano group is useful as an intermediate for synthesizing the compound exhibiting the above biological activity. .

[0015]

DETAILED DESCRIPTION OF THE INVENTION The lower alkyl group used for specifying the isocoumarin derivative of the present invention has 1 to 6 carbon atoms.
Means a straight or branched alkyl group, specifically, methyl, ethyl, n- or iso-
Examples include propyl, n-, iso-, sec- or t-butyl, n-pentyl, isoamyl, and n-hexyl groups.

Aryl-lower alkyl means a group wherein the lower alkyl is substituted by an aryl, preferably phenyl, optionally substituted by halogen, preferably chlorine, bromine, fluorine, One or more identical or different substituents selected from a hydroxyl group, a lower alkyl group, a carboxyl group, a nitro group, and an amino group;
Preferably, it may be substituted by one or two.

The protecting group for the hydroxyl group includes lower alkylsilyl (eg, t-butyldimethylsilyl, trimethylsilyl, etc.), acyl (eg, acetyl, propionyl, benzoyl, etc.), and optionally substituted alkoxymethyl (eg, methoxymethyl). , Methoxyethoxymethyl,
Benzyloxymethyl), and any group that can be used as a protecting group for a hydroxyl group in the field of synthetic organic chemistry.

The carbamoyl group (-CONH ₂ ) may be optionally substituted with N-mono- or di-lower alkyl. When substituted with di-lower alkyl, the lower alkyl groups may be the same or different. It may be.

Specific examples of the isocoumarin derivative according to the present invention in which the above groups are combined include, but are not limited to, those shown in Table I below.

[0020]

[Table 1]

[0021]

[Table 2]

[0022]

[Table 3]

[0023]

[Table 4]

[0024]

[Table 5]

[0025]

[Table 6]

[0026]

[Table 7]

[0027]

[Table 8]

[0028]

[Table 9]

The compound of the formula (I) can be provided in the form of a salt of a compound in which R ¹ represents a carboxyl group and a basic compound. These salts may be any salts as long as they meet the purpose of the present invention. However, preferably, a salt with a basic compound used for producing a pharmaceutically acceptable salt of a usual carboxyl group-containing drug is preferable.
Specific salts include lithium, alkali metals such as sodium and potassium, alkaline earth metals such as calcium and magnesium, and organic bases such as methylamine, ethylamine, dimethylamine, trimethylamine, triethylamine, pyridine, piperazine and piperidine. Salts.

The active compound represented by the above formula (I) can be obtained, for example, according to the method described in JP-A-8-176138 or a modification thereof.

[0031]

Embedded image

Using the starting material as a starting material, a typical example is described below or a modification thereof, in which one or two lower alkyl groups, one or two aryl-lower alkyl groups or an alkylene chain are added to the methyl portion of the cyanomethyl group. To produce a synthetic intermediate according to the present invention, followed by hydrolysis of the cyano group.

The above-mentioned compound according to the present invention or a pharmaceutically acceptable salt thereof exhibits, for example, an inhibitory effect on collagen-induced arthritis as described later in detail, and includes rheumatoid arthritis, systemic lupus erythematosus, Can be used mainly for the prevention or treatment of diseases associated with abnormal immunomodulatory effects, such as autoimmune diseases or malignant tumors such as sclerosis, periarteritis nodosa, ulcerative colitis and juvenile diabetes, and severe infections. I think that the.

For example, it shows an inhibitory effect on angiogenesis induced by tumor cells in the mouse subcutaneous subcutaneous method, and is mainly used for diseases associated with angiogenesis, such as growth and metastasis of malignant solid tumors, diabetic retinopathy, various chronic diseases, etc. It may be applicable to the prevention and treatment of inflammation, psoriasis, angiogenesis associated with corneal transplantation, and atherosclerosis.

In addition, the compounds of the present invention show higher stability in vivo than MI43, which is known to have an immunomodulatory effect, and in particular, significantly higher in vivo stability when administered orally. And excellent bioavailability. In addition, toxicity is hardly observed, and it can be used effectively and safely.

The compounds of the present invention can be used alone, for example, as pharmaceutical preparations for preventing or treating diseases associated with abnormal immunoregulatory effects or angiogenesis.
Preferably, it can be used in combination with a pharmaceutically acceptable auxiliary. Examples of such an auxiliary include fillers, extenders, binders, humectants, disintegrants, disintegration inhibitors, surfactants, diluents and excipients such as lubricants, which are commonly used in the art. Can be mentioned. Various dosage forms can be selected as the pharmaceutical preparation depending on the purpose of treatment. Representative examples are tablets, pills, powders, solutions, suspension syrups, emulsions, granules, capsules, suppositories. Injections (solutions, suspensions, etc.). In addition, since a particularly suitable administration route for exerting the action of the compound according to the present invention is recognized for oral administration, a dosage form for oral administration can be provided as a preferred dosage form among the above dosage forms.

For molding into tablets, known carriers can be widely used, for example, lactose, sucrose, sodium chloride, glucose, urea, starch, calcium carbonate, kaolin, crystalline cellulose, silica and the like. Agent,
Water, ethanol, propanol, simple syrup, glucose solution, starch solution, gelatin solution, binders such as carboxymethylcellulose, shellac, methylcellulose, potassium phosphate, polyvinylpyrrolidone, dried starch, sodium alginate, agar powder, laminaran powder, hydrogen carbonate Sodium, calcium carbonate, polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, stearic acid monoglyceride, starch, disintegrating agents such as lactose, sucrose,
Disintegration inhibitors such as cocoa butter, hydrogenated oil, quaternary ammonium bases, absorption promoters such as sodium lauryl sulfate, humectants such as glycerin and starch, adsorption of starch, lactose, kaolin, bentonite, colloidal silicic acid, etc. Agents, purified talc, stearates, boric acid powder, lubricants such as polyethylene glycol, and the like can be used.

Further, the tablets can be made into tablets coated with a usual coating, if necessary, such as sugar-coated tablets, gelatin-encapsulated tablets, film-coated tablets, double tablets and multilayer tablets. In molding into pill form, those conventionally known in the art can be widely used as carriers, for example, excipients such as glucose, lactose, starch, cocoa butter, hydrogenated vegetable oil, kaolin, talc, gum arabic powder, Binders such as tragacanth powder, gelatin and ethanol, and disintegrants such as laminaran and agar can be used.

For molding into a suppository form, as the carrier, those conventionally known in the art can be widely used, and examples thereof include polyethylene glycol, cocoa butter, higher alcohol esters of higher alcohol, gelatin, and semi-synthetic glyceride. Can be. When prepared as an injection, the liquid preparation and suspension are preferably sterilized and isotonic with blood. When preparing these liquid preparations and suspensions, they are commonly used as diluents in this field. Can be used, such as water, ethanol,
Propylene glycol, ethoxylated isostearyl alcohol, polyoxylated isostearyl alcohol, and polyoxyethylene sorbitan fatty acid esters can be used. In this case, sodium chloride, glucose or glycerin may be contained in the preparation in an amount sufficient to prepare an isotonic solution, and ordinary dissolution aids, buffers, soothing agents and the like may be used. May be. For oral administration, a coloring agent, a preservative, a flavoring agent, a flavoring agent, a sweetening agent, and other pharmaceuticals may be contained in the preparation as required.

The amount of the above-mentioned compound of the present invention contained in the pharmaceutical preparation according to the present invention is not particularly limited and may be selected from a wide range. In the case of solid preparations such as tablets, granules and capsules, the total composition is usually 1 to 70% by weight of the product, preferably 5 to
It is 50% by weight, and in the case of liquid preparations such as liquid preparations, injections and suspensions, it is 0.1 to 10% by weight.

The method of administering the compound of the present invention is not particularly limited, and the compound is administered according to various preparation forms, the age and sex of the patient, other conditions, the degree of the disease, and the like. For example, tablets, pills, solutions, suspension syrups, emulsions, granules and capsules are orally administered. As described above, oral administration is preferable, but in the case of an injection, intravenous administration alone or mixed with a normal replenisher such as glucose and amino acids, and further intramuscularly, intradermally, alone if necessary. It may be administered subcutaneously or intraperitoneally. In the case of suppositories, they are administered rectally.

The dose of the compound of the present invention can be appropriately increased or decreased depending on the usage, the age and sex of the patient, other conditions, the degree of the disease, etc., but it is pharmacologically effective in the body of the animal to which it is administered. For example, 0.3 to 300 mg / kg body weight per day for oral administration and 0.03 to 30 mg / kg body weight for parenteral administration are suitable so as to achieve optimal levels.

Among the compounds according to the present invention, Compound 1 and Compound 36 were converted to DBA at 30 mg / kg body weight, respectively.
Administered orally to 7 / 1J male mice, no animals died and no animals showed any toxic symptoms. This suggests that the compounds used in the present invention do not show any acute toxicity. Similarly, Compound 1 and Compound 36 were orally administered to the above seven mice at 20 mg / kg / day for 3 weeks, but there were no dead individuals and no individual showing any toxic symptoms. Thus, the compounds of the present invention also show little subacute toxicity. In addition, the compound of the present invention represented by the formula (I) can be used extremely safely, so that it can be estimated that other compounds according to the present invention also show the same properties.

Furthermore, as specifically described below, the compounds according to the present invention show remarkable effects in tests using model animals for rheumatoid arthritis and model animals for angiogenesis test. In addition, as specifically described below, the compound according to the present invention shows a high blood concentration over time when administered orally to mice, and particularly has excellent properties as an orally administered drug.

[0045]

The present invention will be described below in more detail with reference to specific examples, but it is not intended that the present invention be limited to these embodiments.

Example 1 (Reference): 8-t-butyldimethylsilyloxy-3-cyanomethyl-6-methoxy-1H-
Production of 2-benzopyran-1-one

[0047]

Embedded image

3-cyanomethyl-8-hydroxy-6
Methoxy-1H-2-benzopyran-1-one 19.2
After dissolving 0 g (83.11 mmol) in 160 ml of N, N'-dimethylformamide and cooling to 0 ° C., 11.90 g (175.16 mmol) of imidazole, 21.14 g of t-butyldimethylsilyl chloride (140.13 mmol)
l) was added and the mixture was stirred as it was for 2 hours. 3 water in the reaction solution
00 ml, toluene 400 ml-ethyl acetate 200
The mixture was extracted twice with ml of the mixed solvent. The organic layer was washed with water (100m
1) and washing with saturated saline (200 ml), and then the solvent was distilled off under reduced pressure. 200 ml of ethanol was added to the residue, and the mixture was heated and dissolved. The mixture was stirred at room temperature for 13 hours, and the resulting crystals were separated by filtration to obtain 21.62 g of the title compound.

¹ H-NMR (CDCl ₃ ); δ 6.46 (1
H, s), 6.45 (2H, s), 3.87 (3H, s), 3.5
9 (2H, d, J = 1.4 Hz), 1.05 (9H, s), 0.2
7 (6H, s) Example 2 : 2- (8-hydroxy-6-methoxy-1H-2
-Benzopyran-1-one-3-yl) -2-methylpropionic acid

[0050]

Embedded image

8-t-butyldimethylsilyloxy-3
After dissolving 400 mg (1.16 mmol) of -cyanomethyl-6-methoxy-1H-2-benzopyran-1-one in 4 ml of tetrahydrofuran and cooling to -30 ° C,
116 mg of 60% oily sodium hydride (2.90 m
mol), and 440 μl (6.96 mmol) of methyl iodide.
l) Added and stirred for 1 hour. The reaction solution was diluted with 100 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by subjecting it to a silica gel column.
t-butyldimethylsilyloxy-3-cyanodimethylmethyl-6-methoxy-1H-2-benzopyran-1-
369 mg of ON were obtained.

The resulting 8-t-butyldimethylsilyloxy-3-cyanodimethylmethyl-6-methoxy-1H
369 mg of 2-benzopyran-1-one in 4 ml of acetic acid
And 4 ml of concentrated hydrochloric acid was added thereto, followed by stirring at 70 ° C. for 15 hours. 15 ml of water was added to the reaction solution, and the mixture was stirred at room temperature for 15 minutes, and the precipitated crystals were separated by filtration. 219 mg of the obtained crystals
Was dissolved in 10 ml of ethanol, 100 mg of activated carbon was added, and the mixture was stirred at 50 ° C. for 1 hour. The solvent was distilled off under reduced pressure except for the activated carbon. Dissolve the residue in 4 ml of ethanol,
9 ml of hexane was added and the mixture was stirred at room temperature, and the precipitated crystals were separated by filtration to obtain 126 mg of the title compound.

¹ H-NMR (DMSO-d ₆ ); δ 10.87
(1H, s), 6.74 (1H, s), 6.71 (1H, d, J =
2.2Hz), 6.55 (1H, d, J = 2.2Hz), 3.8
6 (3H, s), 1.45 (6H, s) Example 3 : 2- (8-hydroxy-6-methoxy-1H-2
Preparation of -benzopyran-1-one-3-yl) -3-phenylpropionic acid

[0054]

Embedded image

8-t-butyldimethylsilyloxy-3
-Cyanomethyl-6-methoxy-1H-2-benzopyran-1-one (500 mg, 1.45 mmol) was dissolved in dichloromethane (2 ml), 1N aqueous sodium hydroxide solution (2 ml) was added, the mixture was cooled to 0 ° C., and n-tetrabutylammonium was added. 117 mg of bromide (0.36 mmol
l), 180 μl (1.45 mmol) of benzyl bromide
l) Added and stirred vigorously for 15 minutes. The reaction solution was diluted with 50 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by a silica gel column to give 124 mg of 3-benzylcyanomethyl-8-t-butyldimethylsilyloxy-6-methoxy-1H-2-benzopyran-1-one.

The 3-benzylcyanomethyl-8-t-butyldimethylsilyloxy-6 obtained by the above method
Methoxy-1H-2-benzopyran-1-one 292m
g (0.67 mmol) was dissolved in 2 ml of acetic acid, 2 ml of concentrated hydrochloric acid was added, and the mixture was stirred at 70 ° C. for 16 hours. 10 ml of water was added to the reaction solution, the mixture was stirred at room temperature, and the precipitated crystals were separated by filtration. 203 mg of the obtained crystals were dissolved in 10 ml of ethanol, 100 mg of activated carbon was added, and the mixture was stirred at 50 ° C. for 1 hour. After filtering off the activated carbon, the solvent was distilled off under reduced pressure to obtain 191 mg of the title compound.

¹ H-NMR (DMSO-d ₆ ); δ 11.22
(1H, s), 7.12-7.27 (5H, m), 6.59 (1H,
d, J = 2.2 Hz), 6.58 (1H, s), 6.54 (1H,
d, J = 2.2 Hz), 3.93 (1 H, t, J = 7.3 Hz,
8.1 Hz), 3.83 (3H, s), 3.26 (1H, dd, J
= 6.6 Hz, 14.0 Hz), 3.08 (1H, dd, J =
8.8 Hz, 14.0 Hz) Example 4 : 2- (8-hydroxy-6-methoxy-1H-2)
-Benzopyran-1-one-3-yl) propionamide

[0058]

Embedded image

8-t-butyldimethylsilyloxy-3
-Cyanomethyl-6-methoxy-1H-2-benzopyran-1-one (2.64 g, 7.65 mmol) was dissolved in dichloromethane (30 ml), 1N aqueous sodium hydroxide solution (30 ml) was added, and the mixture was cooled to 0 ° C. 635 mg (1.91 mmol) of butyl ammonium bromide
l), 2.1 ml (30.60 mmol) of methyl iodide
In addition, the mixture was vigorously stirred for 1 hour. After the dichloromethane layer was dehydrated and dried with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by subjecting it to a silica gel column to obtain 1.06 g of 8-t-butyldimethylsilyloxy-3-cyanomethylmethyl-6-methoxy-1H-2-benzopyran-1-one.

8-t-butyldimethylsilyloxy-3
531 mg (1.48 mmol) of -cyanomethylmethyl-6-methoxy-1H-2-benzopyran-1-one were dissolved in 10 ml of dioxane, and 10 ml of concentrated hydrochloric acid was added.
Stirred at 50 ° C. for 2.5 hours. Ethyl acetate 10
It was diluted to 0 ml and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified on a silica gel column to give 334 mg of the title compound.

¹ H-NMR (CDCl ₃ ); δ 10.93 (1
H, s), 6.50 (1H, d, J = 2.2 Hz), 6.40 (1
H, s), 6.38 (1H, d, J = 2.2 Hz), 5.90 (1
H, s), 5.56 (1H, s), 3.87 (3H, s), 3.4
7 (1H, q, J = 6.6 Hz), 1.54 (3H, d, J = 6.
6 Hz) Example 5 : 2- (8-hydroxy-6-methoxy-1H-2)
-Benzopyran-1-one-3-yl) -2-dipropylpentanamide

[0062]

Embedded image

8-t-butyldimethylsilyloxy-3
After dissolving 500 mg (1.45 mmol) of -cyanomethyl-6-methoxy-1H-2-benzopyran-1-one in 3 ml of tetrahydrofuran and cooling to −30 ° C.,
174 mg (4.35 m) of 60% oily sodium hydride
mol), and 1.5 ml (11.60) of n-propyl iodide.
mmol) and stirred for 1.5 hours. The reaction solution was diluted with 100 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified on a silica gel column to give 189 mg of 8-t-butyldimethylsilyloxy-3-dipropylcyanomethyl-6-methoxy-1H-2-benzopyran-1-one.

The 8-t-butyldimethylsilyloxy-3-dipropylcyanomethyl-6 obtained by the above method
-Methoxy-1H-2-benzopyran-1-one 346
mg (0.71 mmol) was dissolved in acetic acid (2 ml), concentrated sulfuric acid (2 ml) was added, and the mixture was stirred at 70 ° C. for 16 hours. The reaction solution was diluted with 50 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by a silica gel column to give 201 mg of the title compound.

¹ H-NMR (CDCl ₃ ); δ 6.50 (1
H, d, J = 2.2 Hz), 6.43 (1H, s), 6.39 (1
H, d, J = 2.2 Hz), 5.73 (1H, brs), 5.4
8 (1H, brs), 3.87 (3H, s), 1.87-1.99
(4H, m), 1.18-1.33 (4H, m), 0.97 (3H,
t, J = 6.0 Hz) Example 6 : 2- (8-hydroxy-6-methoxy-1H-2)
-Benzopyran-1-one-3-yl) -3-diphenylmethylpropionamide

[0066]

Embedded image

8-t-butyldimethylsilyloxy-3
After dissolving 300 mg (0.87 mmol) of -cyanomethyl-6-methoxy-1H-2-benzopyran-1-one in 2 ml of tetrahydrofuran and cooling to −30 ° C.,
87 mg (2.17 mm) of 60% oily sodium hydride
ol) and 850 ml (11.60 m) of benzyl bromide.
mol), and the mixture was stirred as it was for 0.5 hour. The reaction solution was diluted with 50 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by a silica gel column to give 395 mg of 8-t-butyldimethylsilyloxy-3-cyanodibenzylmethyl-6-methoxy-1H-2-benzopyran-1-one.

The resulting 8-t-butyldimethylsilyloxy-3-cyanodibenzylmethyl-6-methoxy-1
395 mg of H-2-benzopyran-1-one (0.71
mmol) was dissolved in 2 ml of acetic acid, 2 ml of concentrated sulfuric acid was added, and the mixture was stirred at 70 ° C. for 16 hours. Reaction solution ヲ ethyl acetate 5
It was diluted to 0 ml and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure.
The residue was purified on a silica gel column to give 258 mg of the title compound.

¹ H-NMR (CDCl ₃ ); δ 10.97 (1
H, s), 7.14-7.24 (10H, m), 6.51 (1H,
d, J = 2.2 Hz), 6.25 (1H, s), 6.23 (1H,
d, J = 2.2 Hz), 5.60 (2H, brs), 3.84
(3H, s), 3.34 (2H, d, J = 14.0 Hz), 3.2
9 (2H, d, J = 13.2 Hz) Example 7 : 2- (8-hydroxy-6-methoxy-1H-2)
-Benzopyran-1-one-3-yl) -2-cyclopropaneacetamide

[0070]

Embedded image

8-t-butyldimethylsilyloxy-3
After dissolving 100 mg (0.29 mmol) of -cyanomethyl-6-methoxy-1H-2-benzopyran-1-one in 1 ml of tetrahydrofuran and cooling to -30 ° C,
29 mg (0.73 mm) of 60% oily sodium hydride
ol) and then add 1,3-diiodopropane for 40 m
1 (0.35 mmol) was added and the mixture was stirred for 1 hour. The reaction solution was diluted with 50 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate,
The solvent was distilled off under reduced pressure. The residue was purified by subjecting it to a silica gel column to give 8-t-butyldimethylsilyloxy-
3-cyanocyclopropylmethyl-6-methoxy-1H
15.7 mg of -2-benzopyran-1-one was obtained.

8-t-butyldimethylsilyloxy-3-cyanocyclopropylmethyl-6-methoxy-1H-2-benzopyran-1-one 1 obtained by the above method
9 mg was dissolved in acetic acid (1.5 ml), sulfuric acid (0.5 ml) was added, and the mixture was stirred at 70 ° C. for 16 hours. Ethyl acetate 2
It was diluted to 5 ml and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure.
The residue was purified by preparative TLC to give the title compound in 10%.
mg.

¹ H-NMR (CDCl ₃ ) δ: 10.94 (1
H, s), 6.50 (1H, d, J = 2.2 Hz), 6.45 (1
H, s), 6.39 (1H, d, J = 2.2 Hz), 5.83 (1
H, s), 5.58 (1H, s), 3.88 (3H, s), 2.7
2-2.79 (2H, m), 2.48-2.55 (2H, m), 1.
98-2.09 (2H, m) Example 8 : Preparation of 2- (6,8-hydroxy-1H-2-benzopyran-1-one-3-yl) propionic acid

[0074]

Embedded image

8-t-butyldimethylsilyloxy-3-cyanomethylmethyl-6 obtained according to the method of Example 4.
-Methoxy-1H-2-benzopyran-1-one 280
mg (0.78 mmol) in anhydrous dichloromethane 16m
After cooling to 0 ° C., add 1N boron tribromide solution to 4
Then, the mixture was stirred at room temperature for 48 hours. The reaction solution was diluted with 50 ml of dichloromethane and washed with water and saturated saline.
After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified on a silica gel column,
-Cyanomethylmethyl-6,8-hydroxy-1H-2
116 mg of -benzopyran-1-one were obtained.

186 mg (0.81 mmol) of 3-cyanomethylmethyl-6,8-hydroxy-1H-2-benzopyran-1-one obtained by the above method was added to 2 m of acetic acid.
and concentrated hydrochloric acid (2 ml) was added thereto, followed by stirring at 70 ° C. for 16 hours. The reaction solution was diluted with 50 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure. The residue was purified by silica gel column to give 54 mg of the title compound.

¹ H-NMR (DMSO-d ₆ ); δ 10.88
(2H, s), 6.62 (1H, s), 6.43 (1H, d, J =
2.2Hz), 6.35 (1H, d, J = 1.5Hz), 3.6
5 (1H, q, J = 7.3 Hz), 1.38 (3H, d, J = 7.
3 Hz) Example 9 : Preparation of 2- (6,8-dihydroxy-1H-2-benzopyran-1-one-3-yl) -2-methylpropionic acid

[0078]

Embedded image

8-t-butyldimethylsilyloxy-3-cyanodimethylmethyl obtained according to the method of Example 2
6-methoxy-1H-2-benzopyran-1-one 17
8 mg (0.48 mmol) of anhydrous dichloromethane 8 m
After cooling to 0 ° C, 1N boron tribromide solution was added to 2
Then, the mixture was stirred at room temperature for 19 hours. 19 hours later, 1N
2 ml of boron tribromide solution was added, and the mixture was further stirred for 24 hours. The reaction solution was diluted with 100 ml of methylene chloride, washed with water and saturated saline, dried over anhydrous sodium sulfate and dried, and the solvent was distilled off under reduced pressure. The residue was purified by subjecting it to a silica gel column to give 3-cyanodimethylmethyl-6,8.
-Hydroxy-1H-2-benzopyran-1-one is 9
3 mg were obtained.

The obtained 3-cyanodimethylmethyl-6,
8-hydroxy-1H-2-benzopyran-1-one 9
3 mg (0.38 mmol) was dissolved in 2 ml of acetic acid, 2 ml of concentrated hydrochloric acid was added, and the mixture was stirred at 70 ° C. for 16 hours. The reaction solution was diluted with 50 ml of ethyl acetate and washed with water and saturated saline. After dehydration and drying with anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to obtain 90 mg of the title compound.

¹ H-NMR (DMSO-d ₆ ); δ 10.90
(1H, s), 10.86 (1H, s), 6.68 (1H, s),
6.48 (1H, d, J = 2.2 Hz), 6.35 (1H, d, J
= 2.2 Hz), 1.44 (6H, s) Example 10 : Inhibition of angiogenesis using the mouse subcutaneous subcutaneous method As part of the demonstration that the compounds according to the invention have excellent pharmacological activity, mice are used below. Describe the angiogenesis inhibition test.

As a test method, the inhibitory effects of compounds 1 and 36 of the present invention on tumor angiogenesis induced by the mouse transplanted tumor S180 were examined by the mouse subcutaneous subcutaneous method. That is, 1 × 10 ⁷ S180 cells were injected into a chamber prepared by attaching a 0.45 μm pore size millipore filter to both sides of the millipore ring.
After closing the injection port, this chamber was transplanted into an air sack made under the back of an ICR female mouse (9 to 10 weeks old). A 0.5% carboxymethylcellulose solution of a test compound and a solvent control was orally administered for 5 days from the day of transplantation. On the 5th day after the transplantation, the skin was peeled off, an O-ring having the same shape as the ring and having an inner diameter of 10 mm was placed on the skin-contacting portion of the skin, observed under a stereoscopic microscope, and photographed. From the obtained photographs, the intensity of angiogenesis was divided into four stages of 0, 1, 2, and 3 based on the number of bent blood vessels having a length of 3 mm or more unique to tumor blood vessels, and scored based on the following criteria.

Score 0: number of tumor blood vessels is 0 1: number of tumor blood vessels is one 2: number of tumor blood vessels is two 3: number of tumor blood vessels is three or more The results are shown in Table II below.

[0084]

[Table 10]

According to the above table, Compound 36 was 10 mg / kg.
And 30 mg / kg orally, and Compound 1 contained 1
It can be seen that oral administration of 0 mg / kg significantly inhibits angiogenesis induced by tumor cell S180 in the mouse subcutaneous subcutaneous method.

[0086]

Industrial Applicability According to the present invention, there is provided a novel isocoumarin derivative which can be used as an active ingredient exhibiting in vivo kinetics different from a conventional isocoumarin derivative of a pharmaceutical composition such as an angiogenesis inhibitor. Also provided are novel synthetic intermediates for efficiently producing these derivatives.

 ──────────────────────────────────────────────────の Continued on the front page (72) Inventor Hiroshi Iguchi 1-37-10 Sakuragaoka, Hodogaya-ku, Yokohama-shi, Kanagawa Prefecture (72) Inventor Takeo Yoshioka 1782-10, Yoshioka, Ayase-shi, Kanagawa Prefecture (72) Inventor Masaaki Ishizuka Shizuoka Prefecture 6-7, Nishiwakacho, Mishima-shi (72) Inventor Tomio Takeuchi 5-1-11 701-A, Higashi-Gotanda, Shinagawa-ku, Tokyo

Claims (4)

[Claims] 1. An isocoumarin derivative represented by the following formula (I) or a salt thereof as appropriate. Formula (I): In the formula, A represents a hydrogen atom or a lower alkyl group, B represents a hydrogen atom or a protecting group for a hydroxyl group, R ¹ represents a cyano group, an N-mono- or di-alkyl-substituted carbamoyl group or a carboxyl group. A group represented by R ²
And R ^3, independently of one another, hydrogen, lower alkyl or aryl - represents a lower alkyl group, wherein the aryl moiety is optionally be substituted by one or more substituents or R ^2, And R ³ represent a C _2-6 alkylene group which, together with the carbon atom to which they are attached, forms a ^3- to 7-membered alicyclic ring, provided that A is a lower alkyl group , R ² and R ³ simultaneously represent a hydrogen atom, or when one is a hydrogen atom and the other is a lower alkyl group, then R ¹ is a group other than a cyano group or a carboxyl group. 2. A is a hydrogen atom or a lower alkyl group, B is a hydrogen atom, and R ¹ is a carboxyl group or a carbamoyl group optionally substituted with N-mono- or di-lower alkyl. 2. The compound according to 1. 3. The compound according to claim ¹ , wherein A is a lower alkyl group, B is a hydroxyl-protecting group, and R ¹ is a cyano group. 4. A pharmaceutical preparation comprising the compound according to claim 2 as an active ingredient.