JPH09227492A - Novel sesquiterpene and its production and use thereof - Google Patents

Abstract

PROBLEM TO BE SOLVED: To obtain a novel sesquiterpene which has an activity of inhibiting the attaching and transformation of fouling organisms such as barnacles and is useful as an antifouling agent that can be used in an antifouling composition such as antifouling coating to be coated on ship bottoms and underwater structures. SOLUTION: This sesquiterpene is represented by formula I [R<1> and R<2> are each OH, an acyloxy, an alkoxy and they may be incorporated with each other to form an epoxy group; R<3> is dichloromethylenamino, isocyano, isothiocyanato, a (substituted)amino], typically 2chloro-1-dichloromethyleneamino-7,11-dimethyl-3- methylene-6-dodecene-10,11-epoxide. The compound of formula I is obtained by extracting a poriferan with a water-miscible solvent such as ethanol, partitioning the extract with water and a low polar solvent, concentrating the low polar solvent phase, anrtitioning the concentrate with a non-polar solvent and an ;tqueous alcohol, purifying both of the phases by a variety of' chromatography to collect a compound of formula II(R<1> ' and R<2> ' are OH or they may be incorporated with each other to form an epoxy group), when necessary, converting the product to another derivative in a usual manner.

Description

Detailed Description of the Invention

[0001]

[0001] The present invention relates to a novel sesquiterpene, a method for producing the same, and use of the compound as an antifouling agent.

[0002]

2. Description of the Related Art Conventionally, biofouling of marine structures, ships, fish nets, and the like due to attached organisms such as barnacles has caused enormous damage to ships and fisheries industries. Organotin compounds such as bis (tri-n-butyltin) oxide (TBTO) have been used as an antifouling agent against these biofoulings until recently, but their use is extremely limited due to environmental pollution due to their toxicity. I have. Therefore, there is an urgent need to develop antifouling agents to replace them. Some marine organisms inhibit the attachment of other organisms by chemicals produced by themselves. It is thought that a cleaner and stronger antifouling agent can be developed by using these biological compounds. It will also lead to basic research to elucidate the ecosystems of marine life, as well as to its application.

Further, in addition to the above functions, antitumor activity,
Compounds having biological activities such as various enzyme inhibitory activities are provided as lead compounds for the development of pharmaceuticals and research reagents. Therefore, research on bioactive substances obtained from marine organisms has attracted attention not only as antifouling agents but also from the viewpoint of effective use of marine biological resources in a wide range of fields.

In such a situation, the present inventors have:
We have been studying the mechanism of attachment and metamorphosis of larvae of marine organisms. In this area, there are still many unsolved areas such as chemicals that control the process of swimming larvae transforming into adults that are stuck to life, or changes in various organs accompanying metamorphosis. . There are some reports on active substances that inhibit the process of attachment and metamorphosis of larvae of marine organisms, but at present it has not been fully understood.

[0005]

In view of the situation described above, the present inventors conducted a search on marine organisms for an active substance that inhibits the attachment and metamorphosis of the barnacle larvae of the barnacle. -The metamorphosis inhibitor was isolated and the structure was determined, and the present invention was completed.

[0006]

The present invention includes the following inventions.

(1) The following equation (I):

[0008]

Embedded image

(Wherein R ¹ and R ² each independently represent a hydroxyl group, an acyloxy group or an alkoxy group, or jointly represent an epoxy group, and R ³ represents a dichloromethyleneamino group, an isocyano group, an isothiocyanato Or a substituted or unsubstituted amino group).

(2) The following formula (I ') characterized by being extracted and purified from a sponge:

[0011]

Embedded image

(Wherein R ¹ ′ and R ² ′ each represent a hydroxyl group or jointly represent an epoxy group).

(3) The following equation (I):

[0014]

[Chemical 6]

(Wherein R ¹ and R ² each independently represent a hydroxyl group, an acyloxy group or an alkoxy group, or jointly represent an epoxy group, and R ³ represents a dichloromethyleneamino group, an isocyano group, an isothiocyanato An antifouling agent containing, as an active ingredient, a compound represented by a group or a substituted or unsubstituted amino group.

In the above formula (I), the acyloxy group represented by R ¹ or R ² includes, for example, an aliphatic acyloxy having 1 to 5 carbon atoms such as a formyloxy group, an acetoxy group and a methylbutenoyloxy group. Groups; aromatic acyloxy groups such as a benzoyloxy group and a mono- or dihydroxybenzoyloxy group. The alkoxy group preferably includes an aliphatic alkoxy group having 1 to 3 carbon atoms, such as a methoxy group, an ethoxy group, and a propoxy group.

In the above formula (I), examples of the substituted amino group represented by R ³ include an aliphatic acyl group having 1 to 5 carbon atoms such as a formyl group and an acetyl group, and an aromatic group such as a benzoyloxy group. Amino group substituted with an aromatic acyl group (eg, formamide group); amino group substituted with an alkoxycarbonyl group having 2 to 4 carbon atoms (eg, methoxycarbonylamino group); substituted with an alkyl group having 1 to 3 carbon atoms Examples include an amino group (for example, a methylamino group, a dimethylamino group) and the like.

In the above formula (I), R ¹ and R ² are
A compound which represents a hydroxyl group or jointly represents an epoxy group and R ³ represents a dichloromethyleneamino group, that is, a compound represented by the above formula (I ′), can be obtained by extraction and purification from a sponge. Can be.

The raw material animals used herein include the sponge (Demospongea) of the sponge (Hali)
chondrida), such as Halichondria, Axinyssa, Axinella, Acanthella, Pseuda
xinyssa and the like.

The extraction solvent generally includes an organic solvent, preferably a water-miscible solvent such as methanol, ethanol and acetone.

The obtained extract is concentrated, and preferably, the low-polar solvent phase obtained by partitioning with water and a low-polar solvent such as chloroform, dichloromethane, carbon tetrachloride, benzene, hexane and the like is concentrated. The desired sesquiterpene can be efficiently obtained by partitioning with a non-polar solvent such as benzene and hexane and an aqueous solution containing an alcohol such as methanol and ethanol, and purifying the non-polar solvent phase and the aqueous alcohol solution phase, respectively. . Purification can be performed by appropriately combining silica gel chromatography, reverse phase silica gel chromatography, high performance liquid chromatography and the like.

The formula (I ') obtained as described above
Can be converted to an isocyano group by reducing the compound with lithium aluminum hydride in tetrahydrofuran at -78 ° C, and hydrolyzing the isocyano compound in hydrous acetic acid. And a formamide compound, that is, a compound wherein R ³ in the formula (I) is a formamide group. Further, the isocyano compound can be converted to a compound in which R ³ is an isothiocyanato group in the above formula (I) by heating with excess sulfur. Further, a compound having a dichloromethyleneamino group can be converted into a compound in which R ³ is an amino group and a compound in which R ³ is an methoxycarbonylamino group in the above formula (I) by reacting 0.1 M phosphoric acid in methanol. . The compound wherein R ³ is an amino group is treated with an alkylating agent or an acylating agent by a conventional method,
Each can be converted into a compound in which R ³ is an amino group substituted with an alkyl group or an acyl group.

The compound in which R ¹ and / or R ² is a hydroxyl group in the above formula (I) is a compound in which R ¹ and / or R ² is an acyloxy group by esterification or acylation by a conventional method. Can be converted to. Further, by alkylation by a reaction with an alkyl halide by a conventional method, the compound can be converted to a compound in which R ¹ and / or R ² is an alkoxy group. The compound in which R ¹ and R ² together represent an epoxy group in the above formula (I) can be treated with a mineral acid to give a compound in which both R ¹ and R ² are hydroxyl groups, By treating with an alcohol, R ¹ and R ² can be converted to a compound in which one of them is a hydroxyl group and the other is an alkoxy group.

The above-mentioned sesquiterpene obtained as described above has an activity of inhibiting the attachment / transformation of harmful adhering organisms in water such as Taedima barnacles and is useful as an antifouling agent.

[0025]

DESCRIPTION OF THE PREFERRED EMBODIMENTS An example of a preferred embodiment of the present invention will be described below. In the above formula (I), R ¹ and R
Compounds in which ² represents a hydroxyl group or jointly represents an epoxy group and R ³ represents a dichloromethyleneamino group, that is, a compound represented by the above formula (I ′), is obtained by mixing a sponge with water or the like such as ethanol. After extraction with a non-polar solvent, the obtained extract is concentrated, and the low-polar solvent phase obtained by partitioning with water and a low-polar solvent such as chloroform is concentrated, and then contains a non-polar solvent such as hexane and an alcohol such as methanol. It can be obtained by partitioning with an aqueous solution and purifying the non-polar solvent phase and the alcohol aqueous solution phase by appropriately combining various types of chromatography. Furthermore, it can be converted to another derivative by a conventional method.

When the sesquiterpene is used as an antifouling agent, the compound may be used alone or in combination with another antifouling agent. The antifouling agent of the present invention is prepared and used in the form of paints, solutions, emulsions, capsules and the like. These preparations can be carried out by employing a commonly used general formulation.

For example, when used as a paint, the above-mentioned sesquiterpene is blended with a paint preparation agent to prepare an antifouling paint, which can be applied to a ship bottom, an underwater structure, a cooling water intake channel, and the like. Examples of the film forming agent used at this time include oil varnish, synthetic resin, artificial rubber and the like. Solvents, extenders and the like can be further added to the antifouling paint as desired. In this case, the sesquiterpene is 0.1 to 50%, preferably 1 to 30% based on the weight of the coating.
%.

When the antifouling agent of the present invention is used as a solution, for example, the above-mentioned sesquiterpene is mixed with a film-forming agent to form a solution dissolved in a solvent, and this is used for the purpose of preventing the adhesion and propagation of organisms in water. It can be applied to aquaculture nets, fixed fishing nets and the like. As the coating film forming agent, for example, a natural resin, a synthetic resin, an artificial rubber, or the like is used, and as the solvent, toluene, xylene, cumene, ethyl acetate, methyl isobutyl ketone, methanol, or the like is used. If necessary, additives such as a plasticizer can be added to this solution. When used as a solution, the sesquiterpene is 0.1-100%, preferably 0.1-30%, based on the weight of the solution.
In the proportion of

When used as an emulsion, the above-mentioned sesquiterpene is dissolved in a solvent, and a surfactant is further added to prepare an emulsion by a conventional method. As the surfactant, an ordinary surfactant is generally used. When used as an emulsion, the sesquiterpene is incorporated at a ratio of 0.1 to 80%, preferably 0.1 to 30% based on the weight of the emulsion. When used as a capsule, the sesquiterpene in the order of mM is included in the capsule, and attached to a fishing net or the like so as to be gradually released and diffused. Further, the antifouling agent of the present invention can be used by kneading it into a polymer resin of a material used in water such as aquaculture nets and fixed nets.

[0030]

EXAMPLES The present invention will be described more specifically with reference to the following examples, but the scope of the present invention is not limited to these examples.

(Example 1) (1) Isolation operation 53 g of sponges belonging to the genus Axinyssa (Demospongea) (Halichondrida) collected from Hachijojima
Was extracted four times with 150 ml of ethanol using a homogenizer, filtered, and the filtrate was dried under reduced pressure. The operation of adding 150 ml of water and chloroform each and dividing into two phases was repeated three times. 100 mL of hexane and 90% methanol aqueous solution were added to each of the combined chloroform phases which were dried under reduced pressure. This two-phase distribution operation is repeated three times,
The hexane phases were combined and evaporated to dryness under reduced pressure to obtain 340 mg of a hexane fraction that inhibits the adhesion of larvae of Taedema barnacles. The operation of adding water to the 90% methanol phase to make 60% methanol, adding 100 ml of dichloromethane and partitioning the two phases was repeated three times.
The dichloromethane phases were combined and evaporated to dryness under reduced pressure to obtain 290 mg of a dichloromethane fraction which inhibits attachment of the larvae of the barnacle.

The hexane fraction was added to a column (3 × 15 cm) packed with Wako gel C-300 (silica gel), and hexane 100%, hexane: ethyl acetate (99: 1), hexane:
Ethyl acetate (98: 2), hexane: ethyl acetate (97: 3),
Hexane: ethyl acetate (95: 5), hexane: ethyl acetate
(9: 1), hexane: ethyl acetate (8: 2), hexane: ethyl acetate (1: 1), ethyl acetate 100%, and chloroform:
Elution was performed sequentially with methanol (1: 1). Hexane: ethyl acetate (97: 3) 31 mg fraction was concentrated under reduced pressure and then subjected to high performance liquid chromatography (Shodex SIL-5B 4.6 x 250 mm, solvent hexane: ethyl acetate 97: 3, flow rate 1.0 ml / min, detection RI) As a result, a fraction inhibiting the adhesion for a retention time of 14 to 15 minutes was obtained.
From this fraction, high performance liquid chromatography (Capcellp
1.2 mg of adhesion inhibitor-1 was isolated by ak silica AG 120 4.6 × 250 mm, solvent hexane: ethyl acetate 99: 1, flow rate 1.0 ml / min, detection RI) for 64 minutes. Also,
12 mg of the hexane: ethyl acetate (95: 5) fraction from the column packed with the above Wako gel C-300 (silica gel) was concentrated under reduced pressure and then subjected to high performance liquid chromatography (Shodex SIL-5).
B 4.6 × 250 mm, solvent hexane: ethyl acetate 97: 3, flow rate 1.0 ml / min, detection RI), and 0.6 mg of the adhesion inhibiting substance-2 having a retention time of 32 minutes was isolated.

On the other hand, the dichloromethane fraction was added to Wako gel C-
Add to a column (2 x 10 cm) packed with 300 (silica gel) and add chloroform 100%, chloroform: methanol
(99: 1), chloroform: methanol (98: 2), chloroform: methanol (95: 5), chloroform: methanol (9: 1), chloroform: methanol (8: 2), chloroform: methanol (7: 3) , And chloroform: methanol (1: 1). Chloroform 100% fraction 3.5
After concentrating mg under reduced pressure, high performance liquid chromatography (Develos
il 60-5 4.6 × 250 mm, solvent hexane: ethyl acetate 8:
2, retention time 7 ~ 8 depending on flow rate 1.0 ml / min, detection RI)
Was isolated in an amount of 0.3 mg.

(2) Determination of Structure and Activity of Adhesion Inhibitor-1 One-dimensional and two-dimensional NMR spectra and mass spectrum of the obtained adhesion inhibitor-1 were analyzed, and the structure of the compound was represented by the following formula (A). It was decided.

[0035]

Embedded image

The physicochemical constants of the adhesion inhibitor-1 are as follows.

Molecular formula C ₁₆ H ₂₄ Cl ₃ NO [α] _D = + 1.7 ° (c = 0.060, CHCl ₃ ) EIMS [M-Cl] ⁺ = [C ₁₆ H ₂₄ ³⁵ Cl ₂ NO] ⁺ = 316.1223 ( Δ -1.2 mm
u), [C ₁₆ H ₂₄ ³⁵ Cl ³⁷ ClNO] ⁺ = 318.1205 (Δ 0.0 mmu), [C
^{_{16 H 24 37 Cl 2 NO]}} + = 320.1197 (Δ +2.1 mmu) IR (neat) 1654 cm -1 1 H NMR (CDCl 3) 3.82 (H 2 -1), 4.60 (H-2), 2.10, 2.25
(H ₂ -4), 2.22 (H ₂ -5), 5.17 (H-6), 2.08, 2.16 (H _2-
8), 1.62 (H ₂ -9), 2.68 (H-10), 1.28 (H ₃ -12), 1.25 (H
₃ -13), 1.63 (H ₃ -14), 5.04, 5.18 (H ₂ -15) ppm ¹³ C NMR (CDCl ₃ ) 59.5 (C-1), 62.4 (C-2), 145.8 (C-
3), 31.6 (C-4), 26.2 (C-5), 123.9 (C-6), 135.2 (C-
7), 36.3 (C-8), 27.4 (C-9), 64.1 (C-10), 58.3 (C-1
1), 24.9 (C-12), 18.8 (C-13), 16.1 (C-14), 114.5
(C-15), 127.0 (C-16) ppm Attachment inhibitor-1 inhibited the attachment of Cypris larvae to the barnacle at a concentration of 1.0 μg / ml by 90% or more, but the mortality was 5% or less. Was.

(3) Determination of Structure and Activity of Adhesion Inhibitor-2 One-dimensional and two-dimensional NMR spectra and mass spectrum of the obtained adhesion inhibitor-2 were analyzed, and the structure of the compound was represented by the following formula (B). It was decided.

[0039]

Embedded image

The physicochemical constants of the adhesion inhibitory substance-2 are as follows. Molecular formula C ₁₆ H ₂₆ Cl ₃ NO ₂ [α] _D = −22.5 ° (c = 0.020, CHCl ₃ ) EIMS [M-Cl-H ₂ O] ⁺ = [C ₁₆ H ₂₄ ³⁵ Cl ₂ NO] ⁺ = 316.1233 (Δ -0.
2 mmu) IR (neat) 1649 cm ^{-1 1} H NMR (CDCl ₃ ) 3.82 (H ₂ -1), 4.60 (H-2), 2.07, 2.22
(H ₂ -4), 2.21 (H ₂ -5), 5.18 (H-6), 2.05, 2.28 (H _2-
8), 1.44, 1.70 (H ₂ -9), 3.47 (H-10), 1.58 (H ₃ -12),
1.53 (H ₃ -13), 1.62 (H ₃ -14), 5.05, 5.19 (H ₂ -15) ppm ¹³ C NMR (CDCl ₃ ) 59.5 (C-1), 62.4 (C-2), 145.8 (C -
3), 31.7 (C-4), 26.2 (C-5), 124.1 (C-6), 135.5 (C-
7), 36.4 (C-8), 29.7 (C-9), 78.4 (C-10), 76.2 (C-1
1), 29.2 (C-12), 27.2 (C-13), 16.1 (C-14), 114.5
(C-15), 127.0 (C-16) ppm Attachment inhibitor-2 inhibited adhesion of cypris larvae to barnacles at a concentration of 1.0 μg / ml by 50% or more, but the mortality was 5% or less. Was.

(4) Determination of Structure and Activity of Adhesion Inhibitor-3 One- and two-dimensional NMR spectra and mass spectrum of the obtained adhesion inhibitor-3 were analyzed, and the structure of the compound was represented by the following formula (C). It was decided. This is a diastereomer having the same planar structure as the adhesion inhibitory substance-2 represented by the formula (B), but having a different configuration at the 10-position.

[0042]

Embedded image

The physicochemical constants of the adhesion inhibitory substance-3 are as follows. Molecular formula C ₁₆ H ₂₆ Cl ₃ NO ₂ [α] _D = + 7.3 ° (c = 0.015, CHCl ₃ ) EIMS [M-Cl-H ₂ O] ⁺ = [C ₁₆ H ₂₄ ³⁵ Cl ₂ NO] ⁺ = 316.1221 (Δ -1.
4 mmu) IR (neat) 1649 cm ^{-1 1} H NMR (CDCl ₃ ) 3.82 (H ₂ -1), 4.59 (H-2), 2.09, 2.24
(H ₂ -4), 2.21 (H ₂ -5), 5.20 (H-6), 2.09, 2.30 (H _2-
8), 1.66, 1.95 (H ₂ -9), 3.78 (H-10), 1.28 (H ₃ -12),
1.27 (H ₃ -13), 1.61 (H ₃ -14), 5.03, 5.19 (H ₂ -15) ppm ¹³ C NMR (CDCl ₃ ) 59.5 (C-1), 62.4 (C-2), 145.7 (C -
3), 31.6 (C-4), 26.2 (C-5), 124.8 (C-6), 134.4 (C-
7), 37.1 (C-8), 31.2 (C-9), 73.6 (C-10), 72.8 (C-1
1), 26.5 (C-12), 25.2 (C-13), 16.0 (C-14), 114.5
(C-15), 127.0 (C-16) ppm Attachment inhibitor-3 at a concentration of 0.5 μg / ml inhibited the attachment of cypris larvae of the barnacle to the barnacle, but the mortality was less than 5%. Was.

[0044]

INDUSTRIAL APPLICABILITY The present invention provides a sesquiterpene which has an activity of inhibiting the attachment / transformation of harmful adhering organisms in water, such as Tagejima barnacle, and is useful as an antifouling agent.

─────────────────────────────────────────────────── ─── Continuation of the front page (51) Int.Cl. ⁶ Identification code Office reference number FI Technical indication location A01N 47/46 A01N 47/46 C07C 269/08 9451-4H C07C 269/08 335/32 7106-4H 335/32

Claims (3)

[Claims] (1) The following formula (I): (Wherein, R ¹ and R ² each independently represent a hydroxyl group, an acyloxy group or an alkoxy group, or jointly represent an epoxy group, and R ³ represents a dichloromethyleneamino group, an isocyano group, an isothiocyanato group or a substituted Or an unsubstituted amino group.). 2. The following formula (I ′) characterized by being extracted and purified from a sponge: (Wherein R ¹ ′ and R ² ′ represent a hydroxyl group, or
Together they represent an epoxy group. )). 3. The following formula (I): (Wherein, R ¹ and R ² each independently represent a hydroxyl group, an acyloxy group or an alkoxy group, or jointly represent an epoxy group, and R ³ represents a dichloromethyleneamino group, an isocyano group, an isothiocyanato group or a substituted Or an unsubstituted amino group) as an active ingredient.