JPH06234645A - Vasculatization inhibitor - Google Patents

Abstract

PURPOSE:To obtain a compound, having marked vascularization-inhibitory activity, thus having excellent effect as a vascularization inhibitor. CONSTITUTION:This vascularization inhibitor consists of a compound selected from compounds of formulas I to IV and salts thereof. The compound of the formula I can be obtained by culturing a strain belonging to Bacillus thuringiensis var. gerehiae. The compound of the formula II can obtained by reacting a formic acid solution with the compound of the formula I, and the compound of the formula III can be obtained by reacting sodium nitrite with the compound of formula I in the presence of acetic acid. The compound of the formula IV can be obtained by dissolving the compound of the formula II and sodium nitrite in distilled water followed by dripping acetip acid to the resultant aqueous solution to conduct reaction. This vascularization inhibitor can be used for the treatment of diseases caused by vasohyperplasia such as diabetic retinopathy, rheumatic arthritis, immature baby retinopathy or senile xanthosis degeneration.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel angiogenesis inhibitor, which is effectively used in the pharmaceutical industry.

[0002]

BACKGROUND OF THE INVENTION Angiogenesis inhibitors are expected to be effective against angiogenic diseases such as diabetic retinopathy, rheumatoid arthritis, retinopathy of prematurity, and senile macular degeneration. Some substances are known as
At present, it cannot be said that all of them have the properties that are satisfactory as pharmaceuticals.
Many researchers have reported that research is being conducted to find better angiogenesis inhibitors.

[0003]

In view of such a situation, the inventors of the present invention have made earnest studies to develop a new type of useful angiogenesis inhibitor.

[0004]

Means for Solving the Problems The present inventors have examined various compounds for the presence or absence of angiogenesis-inhibitory action, and found that the specific compounds described below have the action, and completed the present invention. Of.

That is, the present invention provides the following chemical formulas 5, 6, and 7
And an angiogenesis inhibitor characterized by comprising a compound selected from the compounds represented by Chemical Formula 8 and salts of these compounds.

[0006]

[Chemical 5]

[0007]

[Chemical 6]

[0008]

[Chemical 7]

[0009]

[Chemical 8]

Compounds represented by the above chemical formulas 5, 6, 6, and 8 (hereinafter, compound (A), compound (B),
The production method of compound (C) and compound (D)) is
Coll.Czechoslov.Chem. Commun.vol.34,891 (1968), same vo
l.41,3837 (1976), Biochim.Biophys.Acta vol.209,357 (1
970). And the like. Of these compounds, compounds (A), (B) and (C) have been reported to have an RNA polymerase inhibitory action, and compounds (A) and (B) are known to have a carcinostatic action. (Japanese Patent Laid-Open No. 56-2
9518, JP-A-55-31043, JP-A-55-10.
4214), and other physiological activities are not known at all. In the present invention, the compounds (A), (B), (C) and (D) and salts thereof are used as angiogenesis inhibitors. Examples of the salt include alkali metal salts, alkaline earth metal salts and the like. Preferred for the present invention are alkali metal salts such as sodium and potassium.

[0011]

The compound of the present invention has an excellent angiogenesis-inhibiting effect as will be shown in the following examples.

[0012]

EXAMPLES Next, the angiogenesis-inhibiting effect of the compound of the present invention will be described in detail with reference to Examples.

Method for producing compound (A): Bacillus thuringin Bacillus thuring
giensis var gelechiae) is bactocasitone 3.0%, sodium citrate 0.3%, monosodium phosphate 0.18%, disodium phosphate 0.38%, monoammonium phosphate 0.15%, sulfuric acid. Magnesium heptahydrate 0.005%, calcium chloride dihydrate 0.005
%, Ferrous sulfate heptahydrate 0.001%, vitamin B ₁₃
pH 7.0 medium containing ppm and 0.1% antifoam agent 60
It was cultured in a liter for 30 hours under the conditions of 30 ° C., 300 rpm, and aeration rate of 0.5 vvm. During the culture, the pH of the culture broth was maintained at 7.0 using 30% sulfuric acid. 12 of the obtained culture solution
After continuous centrifugation at 000 rpm, the supernatant was passed through an anion exchange resin, washed with water, and developed with a 0.2 M aqueous formic acid solution. Fractions containing compound (A) were collected and concentrated under reduced pressure to obtain a crude product of compound (A). The crude product was purified by liquid chromatography (anion exchange resin, 0.2M formic acid aqueous solution) to obtain a white solid compound (A).

Method for producing compound (B): 100 mg of compound (A) was added to 5 ml of 1M formic acid solution, stirred at 40 ° C. for 3 hours, concentrated under reduced pressure, and the residue was subjected to liquid chromatography (anion exchange resin). , 0.2M formic acid aqueous solution),
A white solid compound (B) was obtained.

Method for producing compound (C): 1 under acidic acetic acid
350 mg of sodium nitrite was added to 00 mg of the compound (A), and the mixture was stirred at room temperature for 4 hours. The reaction solution was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (anion exchange resin, 0.1 M formic acid aqueous solution). Fractions containing compound (C) were collected and freeze-dried to obtain 78 mg of white powdery solid.

Method for producing compound (D): 70 mg of compound (B) and 300 mg of sodium nitrite were dissolved in 4 ml of distilled water, and 0.4 ml of acetic acid was added dropwise with stirring at room temperature.
After stirring at room temperature for 4 hours, the mixture was concentrated under reduced pressure, and the residue was purified by high performance liquid chromatography (anion exchange resin, 0.1M formic acid aqueous solution). Fractions containing compound (D) were collected and lyophilized to give 51 mg of white powdery solid. The maximum absorption position λ of the ultraviolet-visible spectrum of the compound
_{The max} was 248 nm, and the IR spectrum measured by the KBr method is shown in FIG.

Angiogenesis-inhibiting effect on bovine pulmonary artery endothelial cells: E-MEM medium containing 10% fetal bovine serum was added to a 96-well microplate, 2000 bovine pulmonary artery endothelial cells were inoculated per well, and compound (A) was added. , PBS containing compound (B), compound (C) and compound (D)
Add a predetermined amount of the solution, and add 37% in a CO ₂ incubator.
Culturing was performed at 72 ° C for 72 hours. After culturing, the degree of cell proliferation was measured by the MTT method, and the compound (A) for vascular endothelial cells,
The growth inhibition rates of compound (B), compound (C) and compound (D) were calculated. The results are shown in Table 1.

[0018]

[Table 1]

[0019]

As is apparent from the above examples, the compound (A), the compound (B), the compound (C), the compound (D)
And these salts have a significant angiogenesis inhibitory effect, for the treatment of diseases caused by abnormal growth of blood vessels, such as diabetic retinopathy, rheumatoid arthritis, retinopathy of prematurity, and senile macular degeneration. Can be used.

[Brief description of drawings]

FIG. 1 is an IR spectrum of compound (D) produced in Example.

Front page continuation (72) Inventor Toshiaki Segawa 2nd Okubo, Tsukuba City, Ibaraki Toagosei Chemical Industry Co., Ltd.

Claims (1)

[Claims] 1. An angiogenesis inhibitor comprising a compound selected from the compounds represented by the following chemical formulas 1, 2, 3, and 4, and salts of these compounds. [Chemical 1] [Chemical 2] [Chemical 3] [Chemical 4]