JP2002338554A - Bromotyrosine derivative and antimicrobial agent comprising the same as active ingredient - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a new bromotyrosine derivative, its derivative and an antimicrobial agent containing the derivative or its salt as an active ingredient, especially an antimicrobial agent having excellent antibacterial activity against periphytic bacteria and high safety. SOLUTION: This bromotyrosine derivative is represented by formula (1) or its salt.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

TECHNICAL FIELD The present invention relates to a novel bromotyrosine derivative or a salt thereof and an antibacterial agent containing the same as an active ingredient.

[0002]

2. Description of the Related Art Molds and bacteria cause various diseases in humans, and cause various pollutions in a wide range of industries and places of life such as agriculture, forestry and fisheries, industry, and the like. For example, trees, fruit trees, vegetables, etc. caused by phytopathogenic bacteria. Cause illness. Various antibacterial agents are used to protect against fungi and bacteria, but not only are they always needed to be more effective, safe and easily available, but new antibacterial agents are also required in relation to resistant species. Have been.

[0003] In recent years, various types of water-borne microorganisms have frequently caused obstacles, causing various adverse effects. For example, slime derived from bacteria and filamentous fungi occurs in heat exchangers and pipes of the circulating cooling system used in petrochemical plants, etc., and these slimes block pipes and reduce cooling efficiency, or In the ocean, these slimes form biofilms (the slimes are attached so as to cover objects) and promote the attachment of large organisms such as barnacles, increasing damage on ships, marine structures, fishing nets, etc. I have. Conventionally, as a measure to prevent adhesion of slime, biofilm, etc., antifouling paints containing an organotin compound, a copper compound, and the like have been applied to the bottom of a ship, a submerged portion of a marine structure, a fishing net, or the like (for example, Chemistry & In
dusty, 5 March 1990, 123-1
See page 27). However, seawater contamination due to long-term use of antifouling paints containing copper and tin compounds has become a problem, and there has been a demand for the development of highly safe antifouling paints that do not contain organic metals such as copper and tin compounds. I have.

[0004] By the way, sponges are a class of metazoan animals of which about 6000 species are known.
okadaic acid (J. oria okadai) (J.
Am. Chem. Soc. , 1981, 103, 246.
9. ) And halichondrins (J. Am. Chem. So
c. 1985, 107, 4796. Other compounds having various biological activities have been reported, including the sponge (Pseudoceratina pu) used in the present invention.
rpura) (for example, Tetrahedron Lett.)
1998, 4147. Tetrahedron, 1
996, 52, 8181. J .; Org. Che
m. , 1996, 61, 2936. Other) is.

Further, for example, the following formula (3) or (4)

[0006]

Embedded image

A bromotyrosine derivative represented by the following formula (for example, J. Chem. Soc. Chem.
Comm. , 1970, 752. ; Tetrahed
ronLett. 1979, 3921. J .; Na
Tural Products, 1989, 52, 75
3. J .; Natural Products, 19
92, 55, 822. J .; Natural Pro
ducts, 1994, 57, 705. ; Tetra
hedron, 1996, 52, 8181. other).

[0008] However, the sponge (Pseudoceratina purpurea) used in the present invention.
Has no report that the compound represented by the above formula (1) has been obtained. In addition, there has been no report of a bromotyrosine derivative which has been reported so far, such as the compound of the present invention represented by the above formula (1), which has an amine-type nitrogen and has a structure decarboxylated from bromotyrosine. .

Further, known bromotyrosine derivatives have antibacterial properties (for example, J. Natural Produ).
cts, 1990, 53, 615. Tetrahe;
dron, 1996, 52, 8181. Et al.), Cytotoxicity (eg, Tetrahedron Lett., 19).
79, 3921. J .; Natural Produ
cts, 1992, 55, 822. Et al.), Enzyme inhibitory activity (Tetrahedron Lett., 1985, 4).
517. Experientia, 1990, 4
6,548. ), Antiviral activity (J. Natural
Products, 1992, 55, 509. ) Etc., but there is no report showing that a small amount showed significant antibacterial activity like the compound represented by the formula (1).

[0010]

SUMMARY OF THE INVENTION The present invention has been made in view of the above circumstances, and provides a novel bromotyrosine derivative and a salt thereof, as well as an antibacterial agent containing the bromotyrosine derivative as an active ingredient, especially against adherent bacteria. It is intended to provide a highly safe antibacterial agent having excellent antibacterial activity.

[0011]

Means for Solving the Problems The present inventors have made intensive studies to solve the above-mentioned problems, and have arrived at the present invention. The above-mentioned formula (1) separated from sponges collected near the Zamami Islands in Okinawa Prefecture, Japan. The bromotyrosine derivative represented by
Rhodospirillum salexigen, one of the marine adherent bacteria, is
gens) have been found to have growth inhibitory activity against SCRC-113, and thus have completed the present invention. In addition, this strain was sent to FERM P-
No. 13599 (JP-A-6-3274).
No. 95).

That is, the present invention relates to formula (1)

[0013]

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A first aspect of the present invention relates to a bromotyrosine derivative represented by the formula (1) or a salt thereof.

Further, in the first invention, the present invention relates to a bromotyrosine derivative or a salt thereof extracted and separated and purified from sponge Pseudoceratina purpurea (a second invention).

Also, the present invention relates to an antibacterial agent comprising a bromotyrosine derivative or a salt thereof according to any one of the first and second aspects as an active ingredient (third aspect). In the third invention, the present invention relates to the antibacterial agent according to the third invention, which has an antibacterial activity against adherent bacteria (fourth invention). Here, the adherent bacterium refers to a bacterium that forms a slime in a water system, for example, a water distribution pipe, or forms a biofilm prior to the attachment of large organisms such as barnacles in biofouling in the ocean. In the fourth invention, the adherent bacterium relates to an antibacterial agent belonging to the genus Rhodospirillum (fifth invention). In the fifth invention, a bacterium belonging to the genus Rhodospirillum is Rhodospirillum.
lum salexigens) SCRC-113 (F
ERMP-13599) (sixth invention).

Further, the present invention relates to a net coated or impregnated with the antibacterial agent according to any one of the third to sixth aspects (seventh aspect). The net is a net used in a water system such as an ocean or a river, and examples thereof include a fishing net and a shark avoiding net. The present invention relates to a rope coated or impregnated with the antibacterial agent according to any one of the third to sixth inventions (eighth invention). A rope means a rope used in a water system such as an ocean or a river. The present invention relates to a buoy coated or impregnated with the antibacterial agent according to any one of the third to sixth inventions (a ninth invention). The present invention relates to a paint containing the antibacterial agent according to any one of the third invention to the sixth invention (No. 10).
Invention). A pigment containing the antibacterial agent according to any one of the third to sixth inventions (an eleventh invention). The present invention relates to a rust inhibitor containing the antibacterial agent of any one of the third to sixth inventions (twelfth invention).

Equation (2)

[0019]

Embedded image

(Wherein R ¹ , R ² , R ^{1 ′} and R ^{2 ′} independently represent a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom or an iodine atom; R ³ , R ⁴ , R ^{3 ′} , R ^{4 ′} independently represents a hydrogen atom, a hydroxyl group which may have a protecting group, an amino group, a lower alkyl group, an alkenyl group, an alkoxy group or an alkylamino group, and R ⁵ and R ^{5 ′ each} independently represent a hydrogen atom. Or a protecting group for an amino group.
^' Independently represents an oxygen atom or a nitrogen atom, a sulfur atom or a phosphorus atom which may have a simple substituent. )
Or a salt thereof (a thirteenth invention).

[0021]

BEST MODE FOR CARRYING OUT THE INVENTION The bromotyrosine derivative represented by the above formula (1) of the present invention is a marine organism collected near the Zamami Islands in Okinawa Prefecture, for example, sponge (Pseudocerat).
Ina purpura) can be obtained by extracting with an organic solvent, followed by separation and purification.

As the solvent used for the extraction, methanol,
Examples include ethanol, acetone, ethyl acetate, chloroform, benzene, and hexane. Known methods can be used for separation and purification. Chromatography can be used alone or in combination with column chromatography, thin-layer chromatography, high performance liquid chromatography, and the like. In addition to silica gel, Sephadex LH20, reverse Phase-related RP-
18 or the like can be used. As thin-layer chromatography and high-performance liquid chromatography, in addition to silica gel,
RP-18 or the like is used.

The bromotyrosine derivative of the present invention represented by the above formula (1) has excellent antibacterial properties and is expected to have various pharmacological activities.

When the bromotyrosine derivative of the present invention represented by the above formula (1) is used as an antibacterial agent, it may be used as it is, or may be used as a formulation. In the case of formulation, conventional diluents, carriers, extenders, additives and the like can be used. The antibacterial agent of the present invention can be used for protection against slime formation by filamentous fungi and marine bacteria.
Specifically, it can be used for disinfecting and disinfecting clothing, tableware, medical equipment, etc., as well as disinfecting and deodorizing agents for industrial manufacturing processes or livestock sheds, plant antibacterial agents, disinfectants for water treatment such as pools and cooling water, slime It is useful as a control agent, an aqueous pollution control agent, and the like.

The adherent bacterium which the antibacterial agent of the present invention has an antibacterial activity may be any as long as it has an ability to adhere to an object to form a slime or a biofilm. Bacteria belonging to the genus Aum, Hyphomicrobium, Hyphomonas, Pedimicrobium, Prosececomicrobium, Artecomonas, Delaea, Pseudomonas, Marinomonas, etc. can be exemplified.

The antibacterial agent of the present invention can be used by being applied or impregnated on nets such as fishing nets, ropes used in oceans and rivers, and the like. In addition, it can be used by applying or impregnating it to members used in oceans and rivers such as buoys, preventing filamentous fungi and attached bacteria from forming slime and biofilms on nets, ropes or buoys etc. it can. Further, the antibacterial agent can be used by being contained in a paint, a pigment, a rust inhibitor or the like.

The antibacterial agent of the present invention can also be used as a salt of a bromotyrosine derivative represented by the formula (1). Such salts include inorganic or organic acid addition salts, for example, hydrochloride, Phosphate, sulfate, citrate, tartrate, maleate, fumarate, oxalate, benzoate and the like, but are not limited thereto, and conventionally known methods Can form a salt.

The compound of the formula (2) or a salt thereof is expected to have antibacterial activity or various pharmacological activities like the bromotyrosine derivative of the formula (1). The compound of the formula (2) or a salt thereof can be formulated and used in the same manner as the boromotyrosine derivative of the formula (1), and can be suitably used as an antibacterial agent, particularly for adherent bacteria. Further, the compound of the formula (2) or a salt thereof may be applied and impregnated on a rope or buoy,
It can be used by being contained in a paint, a pigment or a rust preventive. In the formula (2), as a protecting group for a hydroxyl group or an amino group, an acyl group such as an acetyl group, a propionyl group, a pivaloyl group and a benzoyl group; an alkoxycarbonyl group such as a methoxycarbonyl group and a t-butoxycarbonyl group; An aralkyloxycarbonyl group such as a group; an arylmethyl group such as a benzyl group and a naphthylmethyl group; a silyl group such as a trimethylsilyl group, a triethylsilyl group, a benzyldimethylsilyl group and a t-butyldiphenylsilyl group; an acetonide group; Examples thereof include a lower alkoxymethyl group such as a methoxymethyl group. The lower alkyl group means a linear, branched or cyclic alkyl group having 1 to 6 carbon atoms, and specific examples thereof include a methyl group, an ethyl group, a propyl group, an isopropyl group, a butyl group and an isobutyl group. Group, sec-butyl group, pentyl group, hexyl group, cyclopropyl group, cyclopentyl group, cyclohexyl group and the like. The alkenyl group is particularly preferably a straight-chain or branched one having 1 to 6 carbon atoms, and includes ethenyl, propenyl, butenyl, isobutenyl, sec-butenyl, pentenyl, hexenyl and the like. Can be. The alkoxy group means a linear or branched alkoxy group having 1 to 6 carbon atoms, and specific examples thereof include methoxy, ethoxy, propoxy, isopropoxy, butoxy, isobutoxy, and pentyloxy. And a hexyloxy group. Examples of the cycloalkoxy group include a cyclopropyloxy group and a cyclopentyloxy group. The alkylamino group means an amino group in which one or both of the hydrogen atoms is substituted by a lower alkyl group, and specific examples thereof include an N-methylamino group, an N-ethylamino group, an N-propylamino group, -Isopropylamino group, N, N-dimethylamino group, N, N-diethylamino group, N-ethyl-N-methylamino group, N, N-dipropylamino group and the like. Simple substituents refer to a hydrogen atom, an oxygen atom or a lower alkyl group. The salt of the compound of the formula (2) can be used as an inorganic or organic acid addition salt in the same manner as the salt of the bromotyrosine derivative of the formula (1).

The bromotyrosine derivative of the present invention represented by the above formula (1) can be easily modified by a known method, so that it can be converted into a derivative having more excellent properties as an antibacterial agent and the like. It can also be used.

[0030]

EXAMPLES Next, the present invention will be described with reference to examples, but the present invention is not limited to the above-described embodiments and the following examples.

(Separation and Purification of Bromotyrosine Derivative (1)) Sponge (Pse) collected near the Zamami Islands in Okinawa Prefecture
240 g of udoceratinapurpura) was added to 1 L of acetone and pulverized using a Waring blender. After allowing to stand at room temperature for 3 days, the mixture was filtered, and the filtrate was heated and the organic solvent was distilled off under reduced pressure. The aqueous residue was washed with ethyl acetate (0.
5L), and the solvent was distilled off and concentrated to obtain 1.98 g of a crude extract.

The above crude extract was mixed with 100 mL of hexane and 90 mL
Then, the 90% methanol layer was further washed twice with 100 mL of hexane, and the organic solvent was distilled off under reduced pressure with heating to obtain a 90% methanol-soluble fraction.
36 g were obtained.

A glass column was filled with 50 g of silica gel suspended in chloroform / methanol = 50/1, and the above 90% methanol-soluble fraction dissolved in a small amount of chloroform / methanol = 50/1 was adsorbed thereto. . Chloroform / methanol = 50/1, chloroform / methanol = 20/1, chloroform / methanol = 9/1, and methanol were sequentially flown through this column by four column volumes. Chloroform / methanol = latter half of 50/1 and 20
The first half of the / 1 eluted fraction was collected and concentrated to a yellow oily substance (263 m).
g was obtained.

In a glass column, benzene / acetone = 4 /
20 g of silica gel suspended in 1 was filled, and the yellow oil dissolved in a small amount of benzene / acetone = 4/1 was adsorbed thereto. To this column, benzene / acetone = 4/1, benzene / acetone = 2/1, benzene / acetone = 1/1, acetone, and methanol were respectively flown in three column volumes. The first about 分 の of the benzene / acetone = 4/1 elution fraction was combined and concentrated to 84.2 mg of a yellow oil.
I got

A glass column was filled with 7 g of silica gel suspended in chloroform / ethyl acetate = 4/1, and the yellow oil dissolved in a small amount of chloroform / ethyl acetate = 4/1 was adsorbed thereto. In this column, chloroform / ethyl acetate = 4/1, chloroform / ethyl acetate = 2/1, chloroform / ethyl acetate = 1/1, chloroform / ethyl acetate = 1/2, ethyl acetate, and methanol were each quadrupled in column capacity. Shed. The fraction eluted with chloroform / ethyl acetate = 2/1 was collected and concentrated to give a yellow oily substance of 19.0 m
g was obtained.

A small amount of the above yellow oil (chloroform /
Ethyl acetate = 10/1) solution, and high performance liquid chromatography (column: Develosil 30-3, 2)
0 mm × 250 mm; solvent: chloroform / ethyl acetate = 10/1; flow rate: 5.0 mL / min. ; Detection: UV
254 nm), and the mixture was collected for a retention time of 75 minutes to 130 minutes and concentrated to give 13.2 mg of a colorless oil.

The above colorless oil was converted into a small amount of a 40% methanol solution and subjected to high performance liquid chromatography (column: YM
C-Pack ODS-AQ, 10 mm x 250 mm;
Solvent: 40% methanol-water; flow rate: 2.0 mL / mi
n. ; Detection: UV 254 nm) and retention time 13
From 26 minutes to 26 minutes and concentrated to give a colorless oil
1.9 mg were obtained.

The above colorless oil was converted into a small amount of a 10% methanol solution and subjected to high performance liquid chromatography (column: YM
C-Pack ODS-AQ, 4.6 mm x 150 m
m; solvent: 10% methanol-water; flow rate: 1.0 mL /
min. Detection: UV 254 nm), fractionated for a retention time of 25 to 27 minutes, and concentrated to give 0.82 mg of the bromotyrosine derivative (1) as a colorless oil.

Molecular formula: C ₁₈ H ₁₈ Br ₄ N _{2
^{O 6 1 H-NMR (800MHz}} , CDCl 3): δ2.4
5 (1H, d, J = 5.1 Hz, 1-OH), 2.52
(1H, s, 8-NH), 2.81 (1H, d, J = 1
6Hz, 7a-H), 2.85 (1H, d, J = 16H)
z, 7b-H), 3.77 (3H, s, 3-OMe),
5.44 (1H, d, J = 5.1 Hz, 1-H);
^{42 (1H, s, 5-} H) 1 H-NMR (800MHz, CD 3 OD): δ2.8
1 (1H, d, J = 17 Hz, 7a-H), 2.85
(1H, d, J = 17 Hz, 7b-H), 3.73 (3
H, s, 3-OMe), 4.11 (1H, d, J = 1.
1 Hz, 1-H), 6.34 (1 H, d, J = 1.1 H)
z, 5-H) ¹³ C-NMR (200 MHz, CDCl ₃ ): δ2
5.5 (t, 7-C), 60.3 (q, 3-OMe),
74.3 (s, 6-C), 77.4 (d, 1-C), 1
12.3 (s, 2-C), 116.6 (s, 8-C),
121.2 (s, 4-C), 131.6 (d, 5-
_{C), 148.1 (s, 3} -C) IR (CHCl 3): 3580,3350 (br), 1
580 cm ^-1 [α] ²⁹ _D + 248 ° (c 0.012, CHCl
₃ ) ESIMS (m / z): 696.7766 (Δ2.9)
mmu) [M + Na] ⁺ .

(Antibacterial activity test) The antibacterial activity test was performed by the disk diffusion method as follows. Rhodospirillum salexigens cultured in a liquid medium (0.5% peptone, 0.1% yeast extract, 1/2 concentration artificial seawater) for 3 days
ns) 100 μL of the SCRC-113 culture solution was applied to a diameter of 85
agar plate for antibacterial test (peptone 0.5%, yeast extract 0.1%, agar 1.5%,
(1/2 concentration artificial seawater). 15 μL of a methanol solution containing a predetermined concentration of the bromotyrosine derivative (1) obtained in Example 1 was placed on autoclave-sterilized circular filter paper (6 mm in diameter), and gently placed on the flat plate. 2
After culturing at 5 ° C. for 3 days, the diameter of the inhibition circle was measured. As a result, when 1.6 μg of the bromotyrosine derivative (1) was loaded per disk (1.6 μg / disk), the diameter of the blocking circle was 21 mm, and it was confirmed that the bromotyrosine derivative had excellent antibacterial activity against adherent bacteria. Was. For example, among the known bromotyrosine derivatives, (2c) and (3c), which are considered to have relatively strong antibacterial properties, 10 μg / d
It has been reported that an obstruction circle of about 10 mm was observed on an isk (J. Natural Products, 199).
0,53,615. Tetrahedron, 19;
96, 52, 8181. ).

[0041]

The present invention has the following effects because it is configured as described above in detail. The bromotyrosine derivative and the salt thereof of the present invention have an antibacterial activity and are useful as an antibacterial agent, and particularly have an excellent antibacterial activity against adherent bacteria,
Since the formation of slime and biofilm by the adherent bacteria is inhibited, damage to ships, marine structures, fishing nets, and the like due to these can be prevented. In addition, development of an antibacterial agent having more excellent activity can be expected because of easy chemical modification and easy synthesis of a derivative.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification symbol FI Theme coat ゛ (Reference) C09D 201/00 C09D 201/00 // A61K 31/423 A61K 31/423 (72) Inventor Reiko Watanabe Nishi, Aichi 2-10-3 Nami-Biwajima-cho, Kasugai-gun Castle F-term (reference) 4C056 AA01 AB01 AC01 AD09 AE02 AF08 4C086 AA01 AA02 AA03 BC68 GA09 MA01 MA04 NA14 ZB35 4C087 AA01 AA02 BB02 MA01 NA14 ZB35 4H011 AA01 DC10 DD07 4J038 EA011 JB38 KA02 KA05

Claims (13)

[Claims] (1) The following formula (1): Or a salt thereof. 2. A sponge (Pseudoceratin)
A bromotyrosine derivative or a salt thereof according to claim 1, which has been extracted, separated and purified from a purpura). 3. An antimicrobial agent comprising the bromotyrosine derivative or a salt thereof according to claim 1 as an active ingredient. 4. The antibacterial agent according to claim 3, which has an antibacterial activity against adherent bacteria. 5. The adherent bacterium is Rhodospirillum (Rhodo).
The antibacterial agent according to claim 4, which belongs to the genus spirlum). 6. Rhodospiril (Rhodospiril)
lum) bacteria is Rhodospirillum salexigens (Rhodospirillum salexigen)
s) The antibacterial agent according to claim 5, which is SCRC-113 (FERMP-13599). 7. A net coated or impregnated with the antibacterial agent according to any one of claims 3 to 6. 8. A rope coated or impregnated with the antibacterial agent according to any one of claims 3 to 6. 9. A buoy coated or impregnated with the antibacterial agent according to any one of claims 3 to 6. 10. A paint containing the antibacterial agent according to any one of claims 3 to 6. 11. A pigment containing the antibacterial agent according to any one of claims 3 to 6. 12. A rust preventive containing the antibacterial agent according to any one of claims 3 to 6. 13. The following general formula (2): (Wherein, R ¹ , R ² , R ^{1 ′} and R ^{2 ′} independently represent a hydrogen atom, a fluorine atom, a chlorine atom, a bromine atom or an iodine atom. R ³ , R ⁴ , R ^{3 ′} , R ^{4 ′)} Independently represents a hydrogen atom, a hydroxyl group or an amino group which may have a protecting group, a lower alkyl group, an alkenyl group, an alkoxy group or an alkylamino group, and R ⁵ and R ^{5 ′} independently represent a hydrogen atom or an amino group. X and X ^′ independently represent an oxygen atom or a nitrogen atom, a sulfur atom or a phosphorus atom which may have a simple substituent.) Or a salt thereof.