JPH05117278A - New compound rgt-1, its production and antitumor agent comprising the same compound as effective ingredient - Google Patents

Abstract

PURPOSE:To obtain new compound RGT-1 useful as an antitumor agent. CONSTITUTION:A compound of the formula having the following physical and chemical properties. Shape: white powder. Molecular weight: m/Z 757 (M+H)<+>. Solubility: soluble in methanol, dichloromethane and pyridine and insoluble in water. The compound of the formula is obtained by subjecting Gambierdiscns toxicns of Dinoflagellate to stationary culture at 25 deg.C for 21 days.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a novel compound RGT-1 produced by the dinoflagellate Gambierdiscus toxicus , a method for producing the same, and an antitumor agent containing the compound as an active ingredient.

[0002]

It has been reported that many seaweeds and unicellular algae contain substances exhibiting antibacterial or antifungal and antitumor actions. These substances generally include halogen compounds, cholines, phenol compounds, tannins, organic acids, macrolides, etc.
(Fenical.W.et.al .: "Food-Drugs from the Sea, Proce
edings 1972 "MTS (1973), pp199-202) and goniodomin from dinoflagellates-A (Murakami.M.et.al .: Tetrahedron
Lett. 29: 1149-1152 (1988)) and other antitumor substances have been obtained. In addition, amphidinolide derived from dinoflagellate
(J.Kobayashi.et.al.:Teterahedron Lett. 27. 5755 (1
986)) is known to have antitumor activity. However, these studies are still developing and the development of antitumor substances derived from marine products has not yet been put to practical use.

[0003]

An object of the present invention is to find a substance having excellent physiological activity from various marine products and provide it as an antitumor substance. And this is extremely important in expanding the industrial application field of marine products. From such a viewpoint, the present inventors conducted various searches for useful physiologically active substances from marine plankton, and found that the compound extracted and isolated from the culture of the dinoflagellate Gambierdiscus toxicus ( Gambierdiscus toxicus ) is They found that they have a tumor action and completed the present invention.

[0004]

The present invention has the following formula:

[0005]

[Chemical 2]

Is a novel compound RGT-1 Furthermore, the present invention cultivates a dinoflagellate, Gambierdiscus toxicus ( Gambierdiscus toxicus ), which is a type of algae, in a medium, and collects the novel compound RGT-1 from the culture. This is the manufacturing method of 1.

Further, the present invention provides a novel compound RGT-1.
Is an antitumor agent containing as an active ingredient. Hereinafter, the present invention will be described in detail. The dinoflagellate Gambierdiscus toxicus of the present invention is a strain that has been collected and continuously cultured on a Rangiroa coral reef in French Polynesia. Culture is ES-1 medium
(Prova-soli.L. (1966) in Proceeding of US Japan
Conference Held at Hkone, Sep-tember 12-15 (Watanab
eA & Hattori.A., eds.) pp63-75. Japan Society of
Using Plant Physiology, Tokyo), static culture was performed at 25 ° C. for 21 days in a 3-liter turnip type flask. See again 18
Illumination of 4000-8000 lux was applied at an interval of 6 hours in the dark. However, it goes without saying that the culture conditions such as the medium composition and the culture temperature are appropriately adjusted and selected so as to obtain preferable results.

In order to collect the novel compound RGT-1 of the present invention from a culture, separation and purification usually used for collecting microbial metabolites are taken into consideration in consideration of the physicochemical properties of the substance as described later. Means are used as appropriate. For example, extraction with a solvent, a method utilizing the difference in solubility with impurities,
Alternatively, chromatography or the like is used alone or in combination. As the carrier used in chromatography, conventional inorganic and organic carriers such as silica gel, polyvinyl resin, polystyrene resin and the like can be used.

In order to collect the RGT-1 produced and accumulated in the culture, an extract obtained by extracting the cultured algal cells obtained by centrifugation or filtration in advance with a solvent such as methanol is used. After concentration, extract with an organic solvent such as dichloromethane. The obtained extract is concentrated and then applied to a silica gel column. The active ingredient is eluted with a silica gel column such as acetone-methanol (9: 1).
The active ingredient is then subjected to column chromatography on polyvinyl resin. The column is developed with methanol, hydrous methanol, etc., the active ingredients are collected and concentrated, and then purified with hydrous acetonitrile as a solvent using a reversed-phase polystyrene, silica gel column or the like to obtain RGT-1 as a white powder.

The physicochemical properties of RGT-1 are as follows. 1) Shape: White powder 2) Molecular weight measurement: m / z 757 (M + H) ⁺ 3) Solubility: Soluble in methanol, dichloromethane and pyridine, insoluble in water 4) ¹ H-NMR spectrum (400MHZ, deuterated methanol) Middle): As shown in FIG.

Next, the antitumor effect of the novel compound RGT-1 will be described. One DBA / 2 mouse was inoculated intraperitoneally with 5 × 10 ⁵ cells of mouse lymphoma cells L5178Y. Three mice were used per group. As a test substance, 0.1 mg / kg of L5178Y cells was intraperitoneally administered on the day after inoculation and every 5 days.
The average survival time of each administration group (T) and control group (C) was calculated,
From this, the T / C (%) life extension rate was calculated. Table-1 Antitumor effect of RGT-1 Drug Dose (mg / kg) Survival rate Life extension rate (T / C%) RGT-1 0 22 100 0.1 28 127 As is clear from Table 1, RGT-1 is a mouse. Shows antitumor activity against lymphoma. As is clear from these data, the crude product or purified product containing RGT-1 can be used as a therapeutic composition for human cancer patients. For example, it can be used for treating cancer by orally, parenterally or externally administering to human.

As is clear from the properties described above, RG
T-1 is a novel antitumor substance.

[0013]

INDUSTRIAL APPLICABILITY According to the present invention, a novel compound RGT-1 having an antitumor action is provided, and a method for producing the substance from the dinoflagellate Gambierdiscus toxius. And this compound is effective as an anti-cancer substance for cancer patients.

[0014]

【Example】

(1) Cultivation of Gambierdiscus toxicus ES-1 medium (pH 7.5) in a turnip flask of 3 liter container
After injecting 2000 ml, sterilized and sterilized, and further filtered 0.5 ml vitamin mixture (vitamin B ₁₂ 4 μg, thiamine 0.2 μg,
Biotin (containing 2 μg) was added. Add 200 ml (about 200,000 cells) of Gambierdiscus toxicas culture to this 25
Culturing was carried out for 21 days at an illumination cycle of 6000 lux of light, 16 hours of light and 8 hours of darkness.

(2) Extraction from culture The culture broth (600 liters) obtained by culturing 50 3 liter capacity turnip flasks by the method of (1) is filtered.
(Advantech Co., Ltd.), and the resulting filtration residue containing algae is mixed with methanol (3 liters) and ultrasonically disrupted for 30 minutes. After crushing, the methanol extract obtained by filtration was concentrated, the solvent was distilled off, and the residue was partitioned with 60% methanol and dichloromethane. Dichloromethane soluble fraction is concentrated and coarse powder
(47.4 mg) was obtained.

(3) Purification The obtained crude powder was dissolved in hexane-acetone (4: 1) and applied to a Florisil column. Hexane-acetone
After (4: 1), acetone-methanol (9: 1) was developed as a solvent to obtain an active ingredient in the acetone-methanol section. Toyo Pearl HW-40 (Tosoh)
Was purified by gel filtration chromatography (methanol). The active fraction was further added to Asahipak ODP-50 (2
× 25cm, 75-100% acetonitrile), Capcell Pak C-
8 (2 x 25 cm, 65-100% acetonitrile) was purified by reversed phase chromatography and finally Asahipak
RGT-1 white powder with ODP-50 (45% acetonitrile) 1.
I got 2 milligrams.

The structure was determined mainly by 1 H-NMR, 13 C-NMR and high resolution mass spectrum.

[Brief description of drawings]

FIG. 1 ¹ H nuclear magnetic resonance spectrum of RGT-1 (400 MHz
It is a figure which shows Z, heavy methanol-in heavy pyridine 1: 1).

Continuation of the front page (51) Int.Cl. ⁵ Identification code Office reference number FI Technical display location C07D 493/22 7329-4C A61K 31/35 ADU 7252-4C 35/80 Z 7180-4C C12P 17/18 D 2104 -4B // (C12P 17/18 C12R 1:89) 7804-4B

Claims (3)

[Claims] 1. A novel compound represented by the following formula (RGT-
1). [Chemical 1] 2. A method for producing compound RGT-1, which comprises culturing a dinoflagellate Gambierdiscus toxicus and collecting the compound RGT-1 according to claim 1 from the culture. 3. An antitumor agent comprising the compound RGT-1 according to claim 1 as an active ingredient.