JPH07233066A - New antitumor agent - Google Patents

Abstract

PURPOSE:To obtain an antitumor agent showing excellent inhibitory action on multiplication of human ovarian cancer cell, comprising demethylpyronetin as an active ingredient. CONSTITUTION:This antitumor agent contains 0.1-100wt.% of a compound of the formula or its salt (NK10958P), (5R*, 6R*)-5-ethyl-5,6dihydro-6-[(E)-(2R*,3 S*,4R*,5S*)-2,4-dihydroxy-3,5-di-methyl-7-nonenyl]-2H-pyran-2-one as an active ingredient. In the case of an injection, 0.1-30wt.% of the compound of the formula is contained and pharmaceutically manufactured and in the case of an oral medicine, 2-100wt.% of the compound of the formula is contained and pharmaceutically manufactured. A dose is 1-100mg/kg/day in the case of parenteral administration and 5-500mg/kg/day in the case of oral administration.

Description

Detailed Description of the Invention

[0001]

The present invention relates to the following formula (1)

[0002]

[Chemical 2]

Demethylpyronetin (NK1 represented by
0958P) as an active ingredient.

[0004]

2. Description of the Related Art Alkylating agents, antimetabolites, antibiotics, steroids, antifolates, plant alkaloids and the like have been known as antitumor agents. On the other hand, demethylpyronetin (NK10958P) is produced by microorganisms, and is known to have agricultural and horticultural use, herbicidal action, plant growth regulating action, antibacterial activity, etc. (JP-A-5-1481).
17).

[0005]

The conventional antitumor agents are not yet sufficient, and there is a demand for agents that are more effective.

[0006]

Therefore, as a result of various investigations, the inventors of the present invention obtained the formula (1)

[0007]

[Chemical 3]

Demethylpyronetin (NK10 represented by
958P) was found to have an antitumor effect. The present invention has been completed based on the above findings.

That is, the present invention relates to demethylpyronetin (NK
10958P) or a pharmacologically acceptable salt thereof as an active ingredient.

The demethylpyronetin (NK10958P) represented by the formula (1) in the present invention is a strain of Streptomyces sp. (Storestomyces SP.) NK10958 (NK10958P) as described in Japanese Patent Laid-Open Publication No. 5-148117. It is a publicly known compound produced by Micro Inst. (Hereinafter, the compound represented by the above formula (1) of the present invention is referred to as "the present compound".)

This compound is the compound described in the above-mentioned JP-A-5-148117.
According to the method described in Japanese Patent Publication No. NK10958,
It can be advantageously produced using the strain.

[0012] In general, the radiobacterium is apt to mutate its properties naturally or artificially, and the antibiotic of the present invention can be produced by using a mutant strain of the NK10958 strain, and is not limited to the above. Alternatively, it can be produced by using various known bacteria belonging to the genus Streptomyces.

In order to produce the present compound, first, the strain is cultivated aerobically in a medium containing nutrients that can be utilized by the radiant bacterium. As the nutrient source, known ones that have been conventionally used for culturing a radiant bacterium can be used.For example, as the carbon source, glucose, fructose, glycerin, sucrose, dextrin, galactose, organic acid, etc., alone or in combination. Can be used.

As the inorganic and organic nitrogen sources, ammonium chloride, ammonium sulfate, urea, ammonium nitrate,
Sodium nitrate, peptone, meat extract, yeast extract, dry yeast, corn steep liquor, soybean powder, cottonseed oil residue, casamino acid, bactosoyton soluble vegetable protein, oatmeal and the like can be used alone or in combination.

If necessary, inorganic salts such as sodium chloride, calcium carbonate, magnesium sulfate, copper sulfate, iron sulfate, zinc sulfate, manganese chloride, and phosphate can be added, and organic substances such as amino acids, vitamins and nucleic acids can be added. It is possible to appropriately add a substance or an inorganic substance. As the culturing method, the liquid culturing method, particularly the deep agitation culturing method is most suitable.

It is desirable to carry out the culture at a culture temperature of 20 to 45 ° C. and a slightly acidic or slightly alkaline pH. In liquid culture, when the culture is generally performed for 1 to 5 days, the present compound is produced and accumulated in the culture medium. When the production amount in the culture solution reaches the maximum, the culture is stopped, and the target product is purified and isolated from the culture solution obtained by filtering the bacterial cells.

For the purification and isolation of the substance from the culture filtrate, the separation and purification method generally used for isolating microbial metabolites from the culture medium is used. For example, the culture is separated into a filtrate and a precipitate by a filtration method. The filtrate is adsorbed on the porous polymer resin and eluted with acetone. The precipitate is extracted with acetone.

The resulting acetone solution is further dissolved in a general organic solvent immiscible with water and concentrated under reduced pressure to obtain an oily substance containing the present compound. The crude substance is further purified by a known method usually used for lipophilic purification, that is, chromatography using silica gel, or recrystallization method alone or in combination.

Silica gel is used as a suitable example for purification,
A column chromatography method using n-hexane ethyl acetate or n-hexane-acetone as an eluent can be mentioned. A colorless crystal of the present compound can be obtained by concentrating and drying the active fraction obtained by these methods.

The compound forms a salt with an acid, and the acid for forming the salt may be any pharmacologically acceptable acid, and for example, hydrochloric acid, sulfuric acid, nitric acid, phosphoric acid and the like are preferable.

From the X-ray crystallographic analysis and the like, the three-dimensional structure of the present compound obtained as described above is (5R ^* , 6R ^* )-5-
Ethyl-5,6-dihydro-6-[(E)-(2R ^* ,
3S ^* , 4R ^* , 5S ^* )-2,4-dihydroxy-
3,5-Dimethyl-7-nonenyl] -2H-pyran-2
-Confirmed to be on. In this compound, the hydroxyl groups at the 2- and 4-positions of the side chain nonenyl group may form an ester with an acid such as carboxylic acid or phosphoric acid, and further may form an ether.

When the present compound is used as an antitumor agent, it may be used alone or in admixture with an excipient or carrier for injection.
It is administered as an oral agent or suppository. As the excipient and the carrier, those which are pharmaceutically acceptable are selected, and the type and composition thereof depend on the administration route and administration method. For example, water, alcohol or soybean oil as a liquid carrier,
Animal and vegetable oils such as peanut oil, sesame oil and mineral oil, or sugars such as maltose and sucrose, amino acids, cellulose derivatives such as hydroxypropylcellulose and organic acid salts such as magnesium stearate are used as solid carriers in which synthetic oils are used. To be done.

In the case of injections, physiological saline, various buffer solutions, saccharide solutions such as glucose, inositol and mannitol, ethylene glycol, propylene glycol, etc.
Glycols such as polyethylene glycol are desirable.
Inositol, mannitol, glucose, mannose, maltose, sugars such as sucrose, and a freeze-dried preparation together with excipients such as amino acids such as phenylalanine, a suitable solvent for injection at the time of administration, such as sterile water, It can also be administered by dissolving it in a liquid for intravenous administration such as physiological saline, glucose solution, electrolyte solution and amino acid solution.

The content of the present compound in the preparation varies depending on the preparation, but is usually 0.1 to 100% by weight, preferably 1
Is about 98% by weight. For example, in the case of an injectable solution, it is preferable to contain the active ingredient in an amount of usually 0.1 to 30% by weight, preferably 1 to 10% by weight. In the case of oral administration, it is used in the form of tablets, capsules, powders, granules, liquids, dry syrups and the like together with the solid carrier or liquid carrier. Capsules, tablets, granules and powders generally contain 2-100% by weight, preferably 5-98% by weight, of the active ingredient.

The dose is determined according to the age, body weight, condition of the patient, purpose of treatment, etc., but the dose is generally 1 to 100 mg / kg · day for parenteral administration and 5 to 500 for oral administration.
mg / kg · day.

(Action) Next, the pharmacological action of the present invention will be shown by test examples.

Test Example 1 Growth inhibitory effect on human ovarian cancer cells RPMI164 supplemented with 10% fetal bovine serum and antibiotics
96-we of A2780 human ovarian cancer cells using 0 culture solution.
ll plate (Costar) was plated and cultured at 37 ° C. under 5% CO ₂ . After pre-culturing for 1 day, each sample serially diluted with physiological saline was added in triplicate at 25 μl / well.
The drug was treated for a day. The growth-suppressing effect of drugs is MTT
It was evaluated by assay. That is, MTT (3- (4,5-
Dimethylthiazol -2-yl) -2,5-diphenyltetraz
olium bromide) 1 mg / ml solution was added at 50 μl / well and cultured for about 4 hours. After sucking the culture solution, 100 μl
DMSO was added to dissolve purple formazan produced from MTT by mitochondrial dehydrogenase. 5
The absorbance at 70 nm (OD ₅₇₀ ) was measured by microplate reader (Dyn
atech), and the growth inhibition rate was calculated from the following equation. Growth inhibition rate (%) = (1- (OD _{570 of} drug treatment group) /
(OD _{570 of} control group) × 100 Further, the 50% growth inhibitory concentration with respect to the control group is shown as an IC ₅₀ value.

The results are shown in Table 1.

[Table 1]

As shown in Table 1, the compound of the present invention has a 50% growth inhibitory concentration (IC ₅₀ ) of 13 ng / human ovarian cancer cells.
It showed a strong antiproliferative action, showing ml.

[0030]

EXAMPLES The present invention will be specifically described below with reference to examples.

Example 1 1 g of the test drug, 1 g of polysorbate, Macrogol 40
0.1 g of the product was dispersed and dissolved in 100 g of distilled water for injection, filtered through a membrane filter, dispensed into ampoules, and freeze-dried by a conventional method. An injectable formulation containing 50 mg of the compound of the invention per ampoule was obtained.

[0032]

EFFECTS OF THE INVENTION As described above, the present compound is expected to be an antitumor agent because it exhibits a growth inhibitory action on human ovarian cancer cells.

Claims (2)

[Claims] 1. The following formula (1): A novel antitumor agent comprising a compound represented by: or a pharmacologically acceptable salt thereof as an active ingredient. 2. A compound of formula (1) is (5R ^* , 6R ^* )-
5-Ethyl-5,6-dihydro-6-[(E)-(2R
^* , 3S ^* , 4R ^* , 5S ^* )-2,4-dihydroxy-3,5-dimethyl-7-nonenyl] -2H-pyran-
The antitumor agent according to claim 1, which is a compound that is 2-on or a pharmacologically acceptable salt thereof.