WO1996022996A1

WO1996022996A1 - Dihydrophenazine derivatives

Info

Publication number: WO1996022996A1
Application number: PCT/JP1996/000128
Authority: WO
Inventors: Haruo Seto; Kazuo Shin-Ya
Original assignee: Nippon Chemiphar Co., Ltd.
Priority date: 1995-01-25
Filing date: 1996-01-25
Publication date: 1996-08-01
Also published as: AU4495996A

Abstract

Dihydrophenazine derivatives represented by structural formulae (I, II), obtained from the strains of ray fungi of the genus Streptomyces deposited with an international depositary authority, and having an antioxidant effect and the effect of suppressing the toxicity of L-glutamic acid against mammalian neurons.

Description

Specification

Dihydrophenazine derivative

[Technical field]

TECHNICAL FIELD The present invention relates to a novel dihydrophenazine derivative, a strain capable of producing the same, and a drug using the same.

[Background technology]

L one Glutamate is a kind of Amino acid Natural constituting the evening protein, known to be force ^s' has a toxicity to neurons. If a substance having the effect of suppressing glutamate toxicity is obtained, it is expected that the substance can be used as a drug to activate and improve brain metabolism.

Diseases that may be related to reactive oxygen species include inflammation, rheumatoid arthritis, autoimmune diseases, radiation-induced skin disease, Parkinson's disease, and ischemic heart and brain disorders. If an antioxidant capable of appropriately removing active oxygen can be obtained, it is expected that the antioxidant can be used as a therapeutic agent for these diseases.

By the way, most of the many aquatic creatures known in advocacy survive in soil. Microorganisms that produce useful substances such as antibiotics, especially actinomycetes, are also largely derived from tsuji. Producing new and useful substances from soil ¾ϊ Work to find surnames has been ongoing. For example, JP-A-64-22861 discloses the following compound produced from a strain shown as Streptomyces 厲.

According to the description in JP-A-64-22861, the above compound has an action of removing superoxide.

In addition, Tetrahedron Letters, No. 7, 943-946 (1991) discloses the following compounds produced from a strain belonging to Streptomyces 厲.

Said R is H or CH _3.

According to the above-mentioned article, the above compound exhibits a function as a radical avenger of radicals such as vitamin E.

The present inventors have discovered a new actinomycete strain belonging to the genus Streptomyces from soil. This strain has been deposited on December 22, 1994 at the Institute of Biotechnology and Industrial Technology, 1-3-1 Higashi, Tsukuba, Ibaraki, Japan (Accession number: F ERM P-14717). Based on the Budapest Treaty on the International Recognition of the Deposit of Microorganisms for Patent Procedures, it was internationally deposited on January 27, 1995 with the Institute of Biotechnology and the Institute of Biotechnology. The accession number is FERM BP-5303

[Disclosure of the Invention]

The present inventors have conducted research on this strain and found that this strain is capable of producing a novel dihydrophenazine derivative represented by the following formula I or II.

Further studies on the dihydrophenazine derivatives I and II of the present invention revealed that each of these substances has a glutamate toxicity inhibitory action and an antioxidant action.

[Brief description of drawings]

FIG. 1 is a graph showing the UV-visible absorption β spectrum of dihydrophenazine derivative I.

FIG. 2 is a graph showing an infrared absorption spectrum of the dihydrophenazine derivative I.

FIG. 3 is a graph showing the 'H-NMR spectrum of the dihydrophenazine derivative I. FIG. 4 is a graph showing a ¹³ C-NMR spectrum of the dihydrophenazine derivative I.

FIG. 5 is a graph showing an ultraviolet-visible absorption spectrum of a dihydrophenazine derivative.

FIG. 6 is a graph showing an infrared absorption spectrum of the dihydrophenazine derivative.

FIG. 7 is a graph showing a Η-NMR spectrum of the dihydrophenazine derivative Π.

FIG. 8 is a graph showing a ¹³ C-NMR spectrum of a dihydrophenazine derivative.

FIG. 9 is a micrograph (× 100) of the strain.

FIG. 10 is a micrograph (approximately 120 × 0 ×) of the strain.

FIG. 11 is a graph showing the results of measuring the glutamate toxicity inhibitory action of dihydrophenazine derivative I.

FIG. 12 is a graph showing the results of measuring the glutamate toxicity inhibitory action of a dihydrophenazine derivative.

[Best Mode for Carrying Out the Invention]

First, the novel dihydrophenazine derivative I (that is, the compound represented by the formula I) will be described.

As for the chemical structure, as shown in the above formula I, a hydroxyl group at the 1-position of α-L-rhamnose having a viranose structure is bonded to the carboxyl group of the carboxydihydrophenazine derivative by an s-ester bond. This ester bond can be hydrolyzed and recombined. Accordingly, this compound is decomposed into α-L-rhamnose and a carboxyl dihydrophenazine derivative, and the carboxyl dihydrophenazine derivative, which is a decomposition product, is ester-bonded to another sugar to form another type of dihydrophenazine. A nadine derivative can also be synthesized.

The physicochemical properties of dihydrophenazine derivative I are as follows.

Shape: orange powder Melting point: 63. 0-64. O

Specific rotation: [a] _D ^ao = — 139.8. (C-0.006, methanol) Molecular formula:

Οτ

Molecular weight: 612. 2841 (actual value, HRFAB-MS)

612. 2835 (calculated value)

Solubility: soluble in methanol, chloroform, dimethyl sulfoxide, and acetone, insoluble in hexane

The above molecular formula and molecular weight were determined by FAB mass spectrum using m-nitrobenzyl alcohol as a matrix. This compound showed a (M) ♦ peak of mZz612 in the FAB mass spectrum.

FIG. 1 shows an ultraviolet-visible absorption spectrum of the dihydrophenazine derivative I. The measurement was performed in methanol and the scanning speed was 120. OnmZ min. The band pass is 2. OOnm. As shown in FIG. 1, there is a 1β beak force at 232 nm (2690 0), 246 nm (25800), 298 nm (19000), 368 nm (3300), and 495 nm (9700). No change was observed in the absorption beak due to acid and alcohol.

FIG. 2 shows an infrared absorption spectrum of the dihydrophenazine derivative I. The measurement was performed by the KBr tablet method. In the infrared absorption spectrum, absorption powers derived from a hydroxyl group (3450 cm- ¹ ), a secondary amine (3330 cm- ¹ ), a ketone (1680 crrr) and an ester (1680 cm- ', 1260 cm-') were observed. The figure shows the 'H-NMR spectrum of the dihydrophenazine derivative I. The measurement was performed at 500 MHz in deuterated acetone.

FIG. 4 shows a ¹³ C-NMR spectrum of the dihydrophenazine derivative I. The measurements were performed at 500 MHz in heavy acetone.

The Rf value of the dihydrophenazine derivative I by thin-layer chromatography (cloform form Z methanol-10-1) was 0.42.

Next, the novel dihydrophenazine derivative Π (that is, the compound represented by the formula Π) will be described.

As shown in the above formula 式, the chemical structure of α-L-rhamnose having a viranose structure There is an ester bond between the hydroxyl group at position 1 and the carboxyl group of the carboxyl dihydrophenazine compound. In addition, this Estezo Yoshigo is capable of hydrolysis and recombination. Therefore, this compound is decomposed into α-L-rhamnose and a carboxyl dihydrophenazine compound, and the carboxyl dihydrophenazine compound, which is a decomposition product, is ester-bonded to another saccharide to form another type of dihydrophenazine. Penazine compounds can also be synthesized.

The physicochemical properties of the dihydrophenazine compound are as follows.

Shape: orange powder

Melting point: 59.0 ~ 61.0

Specific rotation: [α]. = 1 106.3 ° (C = 0.005, methanol) Molecular formula: C ₃₁ H ₃₂ N ₂ Οτ

Molecular weight: 544.221 1 (actual, HRFAB-MS)

544. 2209 (Calculated value)

The above molecular formula and molecular weight were determined by FAB mass spectrum using m-nitrobenzyl alcohol as a matrix. This compound showed (Μ) + beak of ΓηΖζδΖζ in the FAB mass spectrum.

FIG. 5 shows an ultraviolet-visible absorption spectrum of the dihydrophenazine compound. The measurement was performed in methanol and the scanning speed was 120. Onm / min. The band pass is 2.00 nm. As shown in FIG. 5, there are absorption peaks at 229 nm (28 900), 245 nm (27400), 296 nm (26000), 367 nm (4000) and 490 nm (1 1700). No change in the absorption beak due to acid and alkali was observed.

FIG. 6 shows the infrared absorption spectrum of the dihydrophenazine compound. The measurement was performed by the KBr tablet method. In the infrared absorption spectrum, a hydroxyl group (3450 cm- single, secondary Amin (3330Cm- absorption derived was observed in '), a ketone (1680Cnr one and esters ^{(1680crtT l, 1260 cnr')} . FIG. 7 Shows the 'H-NMR spectrum of dihydrophenazine compound 測定. Was performed in heavy acetone at 500 MHz.

FIG. 8 shows a ' ³ C-NMR spectrum of the dihydrophenazine compound. The measurement was performed at 500 MHz in deuterated acetone.

The Rf value of the dihydrophenazine compound 薄 by thin-layer chromatography (chloroform / methanol = 10 = 1) was 0.42.

The dihydro off We phenazine derivatives I and Π are all at the moment, no I connexion Shi force is only in the culture of the present invention have force ⁵ discovered strains.

Next, this strain will be described.

Deposit No. FE RM BP-5303, deposited on January 27, 1995 at the Institute of Life Science and Engineering, was isolated from soil samples collected in Bunkyo-ku, Tokyo. Things. This strain is an actinomycete belonging to Streptomyces II.

First, the basic hypha does not break. Aerial hyphae form a short main axis, and irregularly long branches, straight or curved, to form 10 to 50 or more spore chains. The spores are non-motile, cylindrical or oblong and have a width of 0.5 to 0.8 "m. The spore surface is smooth. Spores, spores, and other special forms No cell wall chemical type is type I.

Micrographs of this strain are shown in FIGS. 9 (100 ×) and 10 (about 12000 ×). The length of the white bar seen in Fig. 10 is 500nm.

Next, the strains were (1) sucrose-nitrate agar, (2) glucose-asparagine agar, (3) glycerin-asparagine agar, (4) inorganic salt-starch agar, (5) tyrosine agar. The results of culturing on a medium, (6) nutrient agar medium, (7) yeast 'malt agar medium, and (8) oatmeal agar medium are shown. Table 1 shows the culture characteristics on the above various media. Table 1 Medium Color of the flora on the colony surface Color of the lining of the colony Diffusible dye

(1) Gray series (d) Light brown taste gray (5cb) None

(2) Gray series (3fe) Dark brown (6pl-6pn) Light orange (5ca) (3) Gray series (d) Dark brown to dark brown (6pl-6pn) Light orange (4ea)

(4) Gray series (3fe-5fe) Dark brown (6nl) Light red (61 / 2gc)

(5) Gray series (5) Dark reddish orange (61e) Pinky white (5cb)

(6) No aerial mycelium Light brown taste gray (3ec-3dc) None

(7) Gray series (5fe) brown to dark brown ho ni ~ 5pn) dark orange ho ic)

(8) Gray series (3fe ~ 5fe) Ashy reddish brown (61 / 2ni) Pale red (7gc) Note: Colors in parentheses are colors of color, harmony manual (container, corporation, ob. USA, 1950) Indicates the mark code.

As shown in Table 1, the flora color on the colony surface is a gray series. The back color is dark reddish brown and dark brown. Diffusible dyes are pale orange to pale red or dark orange. The hues of these dyes did not change with changes in PH. Table 2 below shows the physiological properties and the assimilation of carbon sources.

Table 2 Physiological properties and assimilation of carbon sources Growth temperature range 20 to 37 'C Optimum 30 to 37 Melanin-like pigment production: Tyrosine agar +

Peptone, yeast iron agar + Tribton, yeast, broth + starch hydrolysis +

Liquefaction of gelatin

Conversion of skim milk to milk + coagulation of skim milk

Reduction of nitrate + assimilation of carbon source: D—glucose +

L-arabinose +

D-xylose +

D—Fructose

Sucrose

L-Ramnose

Lahu North

i-inositol

As described above, this strain is mesophilic, and uses glucose, arabinose, xylose, and mannitol as carbon sources assimilating ability. Based on the morphological characteristics and cell wall chemical type of this strain, this strain is located in Streptomyces II.

Based on the above-mentioned properties (spore chain morphology, spore surface, fungal mold color, lining color, diffusible dye, assimilation ability of carbon source, etc.), the "Bacterial name approval list, 1990 J Effectiveness A search was made for Streptomyces species listed in the name list, and nearby species were selected.

When the diagnostic properties of Streptomyces purophofuscus (Str-marked tomyces purpeofuscus) are selected, as shown in Table 3 below, the properties of this strain and Streptomyces * parvofuscus are based on the assimilation ability of the carbon source ( Except for the mannits).

Table 3 Comparison items This strain Streptomyces

Perovskus spore chain form: straight or curved + +

Spore surface: smooth + +

Flora color: gray + +

Back color: red Z orange + +

PH sensitivity

Diffusible dye: production + +

PH sensitivity

Melanin pigment + +

Starch hydrolysis + +

Nitrate reduction + +

Growth temperature: 1 0

3 7 V + +

4 ++

Carbon source assimilation ability: arabinose + +

Xylose + inositol one

Mannite +

Rhamnose one Rough Ino One One

Sucrose

Fructose

Galactose ++ As described above, this strain is the closest approximation to Streptomyces parovuskaska, and was deposited under that species name. The designation of the deposited strain (label for identification by the depositor) is 2887—SVS2.

The dihydrophenazine derivatives I and の of the present invention can be produced by aerobically culturing a production bacterium such as the present strain in a suitable medium and collecting the target substance from the culture. The culture medium is composed of nutrient sources available for the dihydrophenazine derivative producing bacteria. Glycerol is preferred as the carbon source. As a nitrogen source, molasses, casein and polypeptone force can be preferably used. In addition, inorganic salts can be added according to the amount of water. An appropriate antifoaming agent (eg, silicone) may be added to suppress foaming during acidification.

For large production of dihydrophenazine derivatives I and II, it is preferable to employ aerobic submerged culture conditions. These conditions are the same as the culture conditions for mass production of other Η¾δ-like bioactive substances. For small-scale production, shaking culture in a flask can be employed.

Alternatively, the cells may be cultured using a jar fermenter. In that case, it is preferable to use vegetative cells of the microorganism for inoculation in the acid fermentation tank in order to avoid growth delay in the production process. For this purpose, it is preferable to inoculate a relatively small amount of medium with the cells of the substance, culture the inoculated medium to produce vegetative cells of the microorganism, and then transfer the cultured vegetative cells to a fermentation tank. The culture medium for producing vegetative cells may be different from the culture medium for producing the dihydrophenazine derivatives I and II.

Agitation and aeration of the culture mixture can be performed in a conventional manner. For stirring, a blower or a similar mechanical stirring device can be used. You may search by rotating or shaking the acid fermentation tank. Various bomb devices can be used for ventilation. Alternatively, aeration may be performed by passing sterilized air through the medium. The acid fermentation temperature is generally from 10 to 40 ° C, preferably from 20 to 30 ° C. The fermentation time is generally 50 to 200 hours. The acid fermentation time varies depending on the acid fermentation conditions and scale.

After the completion of the fermentation, dihydric phenazine derivatives I and から can be recovered from the culture solution by various conventional recovery and purification methods. As the recovery method, solvent extraction, chromatography, or recrystallization can be employed. For solvent extraction and recrystallization, an appropriate solvent is used for each of the dihydrophenazine derivatives I and Π. Two or more solvents may be used in combination.

The dihydrophenazine derivatives I and Π are generally found in cultured bacterial cell components. Thus, extracts and whether we microbial cells obtained by the culture solution was centrifuged or墟過a suitable solvent, it forces ^s preferred to implement the purification treatment dihydro Hue phenazine derivatives. As the solvent, acetone or methanol can be used.

Purification of the dihydrophenazine derivatives I and II can be performed by treating the extract in accordance with a conventional method. For example, removing the solvent from the extract by evaporation or distillation, re-extracting with an appropriate solvent (eg, ethyl acetate), and repeatedly drying to obtain a residue containing the dihydrophenazine derivatives I and Π Can be. Using the crude product as a crude sample, a distilling method such as silica gel chromatography or HPLC separation can be performed to purify the dihydrophenazine derivatives I and それぞれ, respectively.

The ability to isolate the dihydrophenazine derivative I and II producing strains from the natural world by the usual method is also important for nonproprietary products other than this strain. Specifically, it can be carried out in the same manner as the method for isolating antibiotic-producing bacteria. The strain may be subjected to irradiation or other mutation-inducing treatment to improve the productivity of dihydrophenazine derivatives I and II.

It is presumed that many genes are involved in the biosynthesis of dihydrophenazine derivatives I and II, as well as the antibiotics produced by actinomycetes. With the development of genetic recombination technology, biosynthesis of such substances has become genetically manipulable. Therefore, a gene involved in the biosynthesis of dihydrofunazine derivatives I and の of this strain is introduced into another strain, and the resulting transformant is transformed with the dihydrophenazine derivatives I and Π. It can also be produced.

The dihydrophenazine derivatives I and 得 obtained as described above exhibit an inhibitory effect on glutamate toxicity. In addition, the dihydrophenazine derivatives I and π also have an antioxidant effect. Therefore, the dihydrophenazine derivatives I and の of the present invention are useful as inhibitors of glutamic acid toxicity or as antioxidants.

The inhibitory action of the dihydrophenazine derivative of the present invention on glutamate toxicity is stronger than any of the conventionally known compounds, and the dihydrophenazine derivative I can suppress glutamate toxicity by about 50% at several ηΜ. Glutamic acid toxicity can be suppressed by about 50% with dihydrophenazine derivative ヱ with more than 10 ηΜ. Known compounds having an inhibitory action on glutamate toxicity suppress nerve cell death due to mild cerebral ischemia and activate brain metabolism. Therefore, the dihydrofunadine derivatives I and II of the present invention are also effective as therapeutic agents for cerebral ischemic disorders such as cerebral infarction and cerebrovascular dementia.

Glutamic acid has been suggested to be toxic by various mechanisms, but it has been implicated in the expression of free radicals such as active oxygen in all systems. For this reason, it is known that antioxidants such as vitamin 抑制する suppress glutamate toxicity. The dihydrophenazine derivative of the present invention also exhibits a similar antioxidant effect. Therefore, the dihydrophenazine derivative of the present invention is also effective as a therapeutic agent for diseases involving active oxygen, such as inflammation, rheumatoid arthritis, and autoimmune diseases.

[Example 1]

(1) Fermentation

培地 Streptomyces parovuscus 2887-SVS 2 containing 15 mL of starch (1%), bolipeptone (1%) and meat extract (1%) in a 15 mL dispensed CI medium (ρ 前 7.0 before sterilization) Slant culture of the strain One platinum loop was inoculated and subjected to shake culture at 27 * for 3 days. Next, 2 mL of this preculture was added to a medium containing glycerin (2%), molasses (1%), casein (0.596), bolipeptone (0.1%) and calcium carbonate (0.4%). Six 500 mL Erlenmeyer flasks with knobs into which 1 OO mL was dispensed were inoculated, and cultured for 2 weeks at 27 days. 600 mL of seeds prepared as above are glycerin (2%), molasses (1%), A 60 liter production medium containing casein (0.5%), polypeptone (0.1%) and calcium carbonate (0.4%) was inoculated. The fermentation was carried out at 27 liters for 3 days with aeration in a jar fermenter at 30 liters per minute and agitation at 400 rpm.

(2) Isolation and purification

60 liters of the culture solution was centrifuged, and the obtained cells were extracted with an equal amount of acetone. After the acetone was concentrated and removed, the mixture was extracted with ethyl acetate under conditions of about 3.0. After the organic phase was concentrated under reduced pressure and washed with hexane, the insoluble fraction was subjected to silica gel chromatography (KOGEL C-100, 50 OmL). Wash the column with hexane Z ethyl acetate = 2 liters (1.5 liters).

(1.5 liters). The active fractions were collected, concentrated under reduced pressure, applied to a reversed-phase silica gel column (Senshu ODS-SS-1020-T, 50 OmL), and eluted with 90% methanol. The active fraction obtained is concentrated under reduced pressure, and then reverse-phase silica gel column.

(Senshu Pak. PEGASIL ODS, Naoki 2 OmmX 25 Omm) was used for HPLC fractionation in a solvent system of 90% methanol. The active beak was concentrated under reduced pressure to obtain 300 mg of the dihydrophenazine derivative I of the present invention as a red powder.

[Example 2]

30 L of the culture solution prepared by the same method as described in Example 1 was centrifuged, and the obtained cells were extracted with an equal volume of acetone. After the acetone was concentrated and removed, the mixture was extracted with ethyl acetate at pH 3.0. The organic phase was compressed under reduced pressure and washed with hexane, and the insoluble fraction was subjected to silica gel chromatography (WAKOGEL C-100, 50 OmL). The column was washed with hexane Z ethyl acetate = 2/1 (1.5 liters), and eluted with black form methanol = 20: 1 (1.5 liters). The active fractions were collected and concentrated under reduced pressure, and then applied to a reverse phase silica gel column (Senshu 0DS-SS_1020-T, 50 OmL), and eluted with 90% methanol. After the obtained active fraction was reduced under reduced pressure, HPLC fractionation was performed using a reversed phase silica gel column (Senshu Pak. PEGASIL ODS, diameter 2 Omm × 25 Omm) in a solvent system of 85% methanol. The activated beak was concentrated under reduced pressure to obtain 3 Omg of the dihydrofunazine derivative of the present invention as a red powder. [Example 3]

"Suppressive effect of glutamate toxicity on nerve cells"

For glutamine inhibition, a hybridoma (N18-RE-105 cells) of mouse neuroblast JfiM and rat retinal neurons was used.

When a high concentration of glutamate (1-1 OmM) is added to this hybridoma, cell death due to oxidative stress due to inhibition of cystine uptake is observed (Neuron 2, 1547 (1989), and J. Pharmacol. Exp. Ther, 250, 1132 (1989)). The effects of the dihydrophenazine derivatives I and の of the present invention on the glutamate-induced cell death were examined.

6.25 x 10 ³ cells / cm in a 96-well microplate containing Dulbecco's modified MEM medium containing 10% FCS and HAT (hypoxanthine 0,1 mM, aminopterin 40ηM, thymidine 0.14 mM, Sigma) Hybridoma cells were inoculated so that the number of cells became ³ . Twenty-four hours later, 1 OmM glutamic acid and the dihydrophenazine derivative of the present invention were added. After the addition of glutamic acid, the cells were further cultured for 24 hours, and then the lactate dehydrogen lactate (LDH) activity contained in the cells and the medium was measured. Glutamate toxicity was evaluated by calculating the LDH release rate according to the following equation.

01 01 release rate =

LDH activity in culture medium Z (intracellular + LDH activity in culture medium)} X 100 The measurement results of dihydrophenazine derivatives I and Π are shown in FIGS. 11 and 12, respectively. These figures are graphs obtained by blotting the measurement results, where the horizontal axis represents the concentration of the dihydrophenazine compound of the present invention and the vertical axis represents the LDH release rate.

Example 4]

"Lipid peroxidation inhibitory effect"

Microsomes were prepared from male Wistar rat livers according to the method of Ernster, Shimi et al. (Above Cell Biol. 15, 541 (1962)).

Liver microsomes (protein amount 0.5 mg), 0.2 M Tris-HCl buffer (pH 7.4), 2 mg / mLADP, DMS0 (dimethyl) of test substance (dihydrophenazine derivative I of the present invention) Sulfoxide) solution, and 0.5 mL of the reaction solution containing N ADPH in SmgZmL were incubated at 37 ° C for 1 hour. reaction Transfer the container to ice water, immediately add 0.05 mL of 0.5% butylhydroxytoluene-ethanol solution to stop the reaction, and further add 0.1% SDS (sodium dodecyl sulfate) 0.25 mL, 20% 1.75 mL of diacid and 1.5 mL of 0.8% thiobarbituric acid solution (pH 3.5) were added. Transfer the reaction vessel into boiling water,

Heat for 1 hour to develop color. After cooling, add 4 mL of n-butanol, centrifuge (3000 rpm, 10 minutes), measure the absorbance of the supernatant at 530 nm, and observe the lipid peroxidation reaction of the test substance. Was determined.

The results are shown in Table 4.

Table 4 Concentration suppression rate I Cso (M)

10- ⁷ M 0.5%

10 " ^β Μ 31.5% 1.5 x 10 1 '10" ^Β Μ 95.0%

10 " ⁴ Μ 99.1%

[Industrial applicability]

The dihydrophenazine derivative of the present invention exhibits a glutamate toxicity inhibitory action and an antioxidant action. Therefore, the dihydrophenazine derivative of the present invention has an effect as a glutamate toxicity inhibitor or an antioxidant.

Claims

The scope of the claims

1. A dihydrophenazine derivative represented by the following formula.

2 · An inhibitor of gluminamine activity comprising a dihydrophenazine derivative represented by the following formula.

3. An antioxidant comprising any one of the dihydrophenazine-induced derivatives described in claims 1 and 2.

4. An inhibitor of glutamic acid toxicity comprising the dihydrophenazine derivative according to any one of claims 1 and 2.

5. A strain of actinomycetes which belongs to Streptomyces I and has an ability to produce the dihydrophenazine derivative described in claim 1, Z or 2.

6. A strain of actinomycetes deposited in Streptomyces にて under the accession number FERM BP-5303 and deposited with the National Institute of Bioscience and Human Technology.

7. Belongs to Streptomyces 、 and has accession number FERM BP-5303! ^ Life Engineering: L-glutamic acid, which is derived from a strain of actinomycetes deposited at urns Sci. A substance that has the effect of suppressing toxicity.

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