JPH0769872A - Anti-methicilin resistant staphylococcus agent - Google Patents

Abstract

PURPOSE:To obtain the subject atoxic, tasteless and odorless agent having antibacterial action on methicillin resistant staphylococcus auricularis(MRSA), comprising a specific anthraquinone derivative as an active ingredient. CONSTITUTION:This agent comprises an anthraquinone derivative of the formula (at least one of R2, R3, R5 and R7 at the beta-position is a functional group and is preferably OH group), in which at least one among four beta-positions contains a functional group, as an active ingredient. The anthraquinone derivative is preferably one or more of erythrolaccin, desoxyerythrolaccin, isoerythrolaccin, 1,3,5,6-tetrahydroxy-8-methyl-8(10H)-anthacenone and 1,3,6-trihydroxy-8-methyl-9(10 H)-anthacenone.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an anti-methicillin-resistant staphylococcal agent having antibacterial properties against methicillin-resistant staphylococcus (MRSA).

[0002]

2. Description of the Related Art Methicillin-resistant Staphylococcus aureus (MRSA) has been a problem in medicine because of its multidrug resistance. Gentamicin has been proposed as the antibacterial agent, but it was further desired to provide effective antibacterial agents of other species.

The present inventor discovered that certain anthraquinone derivatives have anti-methicillin-resistant staphylococcal properties and completed the present invention.

[0004]

Means, Actions and Effects for Solving the Problems That is, the anti-methicillin-resistant staphylococcal agent of the present invention comprises an anthraquinone derivative having a functional group in at least one of the four β-positions as an active ingredient.

Anthraquinone derivatives are represented by the general formula (I)

[0006]

[Chemical 1]

It is represented by Anthraquino in the present invention
The derivative of R is at the β-position. ₂Group, R₃Group, R₆
Group and R₇At least one of the groups is a functional group.
And are required. Functional group is reactive in the molecule
Means a rich group. In the present invention, the functional group is free
Those having a nucleophilic property in the state of
Is preferred.

In the present invention, the functional group is an ant.
Β-position of raquinone derivative [R₂, R₃, R₆Or R₇] To
It is important to be. α-position [R₁, R_Four, R_FiveOr
R₈], Even if a functional group such as a hydroxyl group is present,
The hydroxyl group is in a free state (-O ^-), The following equation
(II)

[0009]

[Chemical 2]

As shown in (1), it has an oxygen atom contained in the anthraquinone ring and a six-membered ring structure, and is in a stable state. Therefore, it is considered to be inactive. On the other hand, when a functional group such as a hydroxyl group is present at the β-position and is in a free state, the stable state like the α-position is not obtained.

The R ₁ , R ₄ , R ₅ or R ₈ group at the α-position may be a group consisting of any atomic group as long as it does not adversely affect the living body. Examples thereof include organic groups such as alkyl groups and functional groups such as hydroxyl groups. Further, among the R ₂ , R ₃ , R _6, and R ₇ groups at the β-position, any group other than the functional group can also be used as long as it does not adversely affect the living body. It may be a group composed of atomic groups.

Further, the anthraquinone derivative of the present invention may have at least one of the four β-positions of anthrone in which the oxygen atom at the 10th position is replaced by hydrogen, having a functional group. The mechanism by which the anthraquinone derivative of the present invention exhibits an anti-methicillin-resistant staphylococcal activity has not yet been clarified, but it is considered that the state in which the functional group is released is involved in the expression of the effect.

As the anthraquinone derivative in the present invention, for example, erythrolacsin, desoxyerythrolaccin, isoerythrolaccin, 1,3,5,6-tetrahydroxy-8-methyl-9 (10H) -anthracene and 1,3,3. 6-trihydroxy-8-methyl-9
(10H) -anthracene and the like.

Erythrolacin has the following formula (III)

[0015]

[Chemical 3]

It is represented by Desoxyerythrolacin has the following formula (IV)

[0017]

[Chemical 4]

It is represented by Isoerythrolacin has the following formula (V)

[0019]

[Chemical 5]

It is represented by 1,3,5,6-tetrahydroxy-8-methyl-9 (10H) -anthracenone has the following formula (VI)

[0021]

[Chemical 6]

It is represented by Further, 1,3,6-trihydroxy-8-methyl-9 (10H) -anthracene is represented by the following formula (VII)

[0023]

[Chemical 7]

It is represented by These compounds can be obtained, for example, by extracting and separating them from the resinous secretions of Lactophora lactus. Lack scales are about 0.6-long in body length
It is a 0.7 mm species of mosquito, belonging to the order Hemiptera Homoptera, along with the cicada, planthopper and Arabian insects.

The growth conditions are limited to a part of the subtropical region, India and Thailand are the two major producing areas, and they are also produced in China, Myanmar, Taiwan, Indochina, Vietnam, etc. Lactophyllidae parasitize various plants of different strains as mother trees, but the main ones are legumes and mulberry plants. Among them, it often parasitizes the Japanese soybeans, the jackfruit tree, and the American alder tree.

In aquaculture, these plants are seeded with twigs with eggs of the larvae of the larvae. The eggs of the larvae of Lacquer worms hatch, walk to new branches rather than twigs, pierce the sucker tube into the skin of the mother tree, absorb the sap, nourish it, and grow. Then, it secretes a resinous substance, forms a resin layer, and lives in it. Adults are shed several times in a few months. The secretion of resin is mainly performed by females, and the lifespan is longer than that of males and they survive for about 6 months. There are three tubes that secrete resin,
The resin has a function of wrapping and protecting the insect body. The resin is secreted thickly by the branches. A collection of this is called a stick rack. The components of the stick rack are composed of a resin component, a wax component, a pigment component, and water, although they vary depending on the origin and type of the raw material.

A stick rack is obtained by crushing and washing the stick rack to remove impurities. The resin component extracted from the seed rack is called shellac, which has long been used as a coating agent for medicines and foods, and has been recognized as safe worldwide. It is also used in various fields industrially.

Erythrolacsin, desoxyerythrolacsin, isoerythrolaccin, 1,3,5,6-tetrahydroxy-8-methyl-9 (10H) -anthracenone and 1,3,6-trihydroxy-8-methyl-9
(10H) -anthracene is a yellow rack dye contained in the dye component of the stick rack, and is about 0.12%, about 0.03%, and about 0.1% in the stick rack resin, respectively.
15%, about 0.70%, about 0.35%.

Erythrolacine from stick rack,
Desoxyerythrolacsin, Isoerythrolacsin,
1,3,5,6-tetrahydroxy-8-methyl-9
(10H) -anthracenone and 1,3,6-trihydroxy-8-methyl-9 (10H) -anthracene are extracted, separated and purified by extracting the components with a solvent,
It can be obtained by a known method of separating and purifying using chromatography.

The anthraquinone derivative in the present invention may be obtained by synthesis. The effective dose of the anthraquinone derivative in the present invention is 0.25 g to 1 g per day for intravenous injection, and about 1 to 3 g per day for oral administration, and it is advisable to take it about 3 times a day depending on the symptoms. To be

The anthraquinone derivative in the present invention is
If desired, a conventional pharmaceutical carrier, solvent, or excipient can be used to prepare a medical preparation according to a conventional method for manufacturing a preparation. Next, the fact that the anthraquinone derivative of the present invention has an antibacterial action will be described with reference to Examples.

[0032]

【Example】

A. Extraction and Purification of Test Substance First, as test substances, erythrolacsin and desoxyerythrolacsin were extracted from stick rack and purified into respective components.

(1) Extraction 1 part of stick rack is immersed in 2 parts of ethyl acetate for extraction. (2) Concentration The extract is concentrated.

(3) Preparation of sample The concentrated extract is dissolved in a 10-fold amount of the following alkaline liquid to prepare a sample. Alkaline liquid: Methanol: 1% aqueous sodium hydrogen carbonate solution = 7: 3 (4) Column chromatography A column chromatography is used to separate a mixture of erythrolacsin and desoxyerythrolacsin.

For column chromatography, the following reverse phase carrier is used. The column is pre-filled with the following acidic liquid. Carrier ・ ・ ・ Chromatorex ・ Chromatography ・
Silica gel (Chromatorex Chromatography Silica Ge
l) ODS 100-200 mesh (Fuji Devison Chemical Co., Ltd.) Acid solution: Methanol: 1% acetic acid aqueous solution = 7: 3 In a column, develop the above sample with the packed acid solution.

(5) Precipitation The precipitate deposited from the yellow band fraction obtained by column chromatography is collected by filtration. This precipitate is a mixture of erythrolacsin and desoxyerythrolacsin.

(6) High Performance Liquid Chromatography The above mixture is separated into erythrolacsin and desoxyerythrolacsin by high performance liquid chromatography. High performance liquid chromatography is performed under the following conditions using a reversed phase column.

Column to be used ・ ・ ・ Develosil
sil) ODS 10/20 20 × 250 Developing solvent: Acetonitrile: 1% acetic acid solution =
3: 7 Flow rate ··· 10 ml / min Measurement wavelength ··· 280 nm Under these conditions, desoxyerythrolacin is 100
After minutes, erythrolacsin is separated after 120 minutes. B. Antibacterial test (1) Preparation of test substance solution 2 mg of the test substance was collected in a sterilized vial bottle of 10 ml volume, and 0.25 ml of dimethyl sulfoxide (DM) was added.
SO), 0.5 ml absolute alcohol and 9.25 ml
Add sterile deionized water from and dissolve. Here, the concentration of the test substance in the solution is 200 μg / ml.

As the test substance, erythrolacsin and desoxyerythrolaccin extracted and purified from the stick rack according to the method A described above, and, for comparison, antimethicillin-resistant staphylococcal agents conventionally used were compared. Some gentamicin was used.

(2) Methicillin-resistant Staphylococcus aureus (MRS
Culture of A) Single methicillin-resistant Staphylococcus aureus (MRSA) cells were cultured under constant culture conditions as described below. Strains used: Staphylococcus aureus (methicillin resistant) ATC
C33591 medium: Nutrient liquid medium (Nutrient broth GIBCO) Culture time: 18 to 24 hours Culture temperature: 37 ° C. (3) Test method Methicillin-resistant Staphylococcus aureus (MRSA) -containing medium 1.9 ml of the medium obtained in the above (2) Into the test tube, 0.1 m of the test substance solution obtained in (1) above
Add l. Here, the concentration of the test substance is 10 μg / m
1 (ie initial test substance concentration).

After 4 days, the growth of cells of methicillin-resistant Staphylococcus aureus (MRSA) is measured by visual observation. If it is cloudy (+), if it is not growing or if there is a lack of cloudiness, it is recorded as (-). If positive, it is determined to be inactive and the test is stopped at that point.

If there is activity (ie, determined as negative in the above), the test substance is further diluted to determine the minimum effective concentration (ED100). That is, 1 ml of the medium prepared in was added to methicillin-resistant Staphylococcus aureus (MRS).
The concentration of the test substance is 3 μg in addition to 2 ml of the medium containing A)
/ Ml. Then, the activity is determined in the same manner as above.

Further, in the next dilution step, 1 ml is taken from the medium having the above-mentioned 3 μg / ml, and it is added to 2 ml of the medium newly containing methicillin-resistant Staphylococcus aureus (MRSA) so that the test substance becomes 1 μg / ml. To prepare. Then, the activity is determined in the same manner as above.

Methicillin-resistant Staphylococcus aureus (MRSA)
As a blank experiment, cell growth was also carried out on the cells to which the test substance was not added. The final number of cells in the medium is 10
It becomes ⁴ to 10 ³ pieces / ml. The minimum effective concentration for methicillin-resistant Staphylococcus aureus (MRSA) is determined by the above-mentioned method at which the growth inhibition can be visually recognized. Visual observation is determined by comparison with the blank experiment above.

(4) Results Both erythrolacsin and desoxyerythrolacsin showed activity against methicillin-resistant Staphylococcus aureus (MRSA). Minimum effective concentration is 10 μg / ml for both
Met. The fact that it was active against methicillin-resistant Staphylococcus aureus (MRSA), which is a multidrug-resistant bacterium, is a very important fact in medicine. It is expected to be utilized in the future.

On the other hand, the minimum effective concentration of gentamicin, which is a conventionally known anti-methicillin-resistant staphylococcal agent, against methicillin-resistant staphylococcus (MRSA) (ED10
0) was 1 μg / ml. Here, the effective dose of erythrolacsin or desoxyerythrolacsin is
It is advisable that the dose is 0.25 g to 1 g per day by intravenous injection, and about 1 to 3 g per day by oral administration. It is advisable to take 3 times a day depending on the symptoms.

Further, erythrolacsin or desoxyerythrolacsin can be prepared into a medical preparation by using a conventional pharmaceutical carrier, solvent or excipient, if desired, according to a conventional method for manufacturing a preparation. . The above-mentioned erythrolacsin and desoxyerythrolacsin are both nontoxic, tasteless and odorless and highly safe, and do not adversely affect the living body.

Claims (3)

[Claims] 1. An anti-methicillin-resistant staphylococcal agent comprising an anthraquinone derivative having at least one of four β-positions having a functional group as an active ingredient. 2. The anti-methicillin-resistant staphylococcal agent according to claim 1, wherein the functional group is a hydroxyl group. 3. The anthraquinone derivative is erythrolacsin, desoxyerythrolaccin, isoerythrolaccin, 1,3,5,6-tetrahydroxy-8-methyl-9 (10H) -anthracene or 1,3,6-tritrione. The anti-methicillin-resistant staphylococcal agent according to claim 1 or 2, which is any one or more of hydroxy-8-methyl-9 (10H) -anthracenone.