JP2967033B2 - Novel Kalihinol Derivatives and Their Production and Use - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a novel kalihinol derivative, 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. Therefore, bis (tri-n-)
Organotin compounds such as (butyltin) oxide (TBTO) have been used, but their use is extremely limited due to environmental pollution due to their toxicity. Therefore, there is an urgent need to develop antifouling agents to replace them. Some marine organisms inhibit attachment of attached organisms by chemical substances 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 ecosystem of marine life as well as its application.

Further, in addition to the above functions, antitumor activity,
Compounds having biological activities such as various enzyme inhibitory activities have been used 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 above-mentioned situation, the present inventors conducted a search for marine organisms, particularly active substances that inhibit the attachment and metamorphosis of larvae of barnacle barnacles, targeting marine organisms. Succeeded in isolating and inhibiting the structure of the larvae of the barnacle larvae from the sponge, and succeeded in determining the structure, thereby completing the present invention.

[0006]

The present invention includes the following inventions. (1) The following formula (I):

[0007]

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(Wherein, R ¹ represents a formamide group, an isocyano group or an amino group, R ² represents a hydrogen atom or an acyl group, and R ³ represents an isocyano group, an isocyanato group, an isothiocyanato group, a formamide group or an amino group. Represents, R
⁴ and R ⁵ are one of the other is a hydrogen atom with a chlorine atom. However, this excludes the case where both R ¹ and R ³ are isocyano groups. ). (2) The following formula (I ') characterized by being extracted and purified from a sponge:

[0009]

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(Wherein, R ¹ ′ represents a formamide group or an isocyano group, R ² ′ represents a hydrogen atom, R ³ ′ represents an isocyano group, an isocyanato group, or an isothiocyanato group;
One of R ⁴ ′ and R ⁵ ′ is a chlorine atom and the other is a hydrogen atom. )). (3) The following formula (I ″):

[0011]

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(Wherein R ¹ ″ represents a formamide group, an isocyano group or an amino group, R ² ″ represents a hydrogen atom or an acyl group, and R ³ ″ represents an isocyano group, an isocyanato group, an isothiocyanato group, a formamide group or Represents an amino group;
⁴ "and R ^5" is one of the other is a hydrogen atom with a chlorine atom. An antifouling agent containing a compound represented by the formula (1) as an active ingredient.

In the above formulas (I) and (I ″), R ²
Or an acyl group represented by R ² ", for example, an aliphatic acyl group having 1 to 5 carbon atoms such as a formyl group, an acetyl group, a propionyl group, a butyryl group, a valeryl group; a benzoyl group, a cinnamoyl group, a toluoyl group In the formula (I), R ¹ represents a formamide group or an isocyano group, R ² represents a hydrogen atom, R ³ represents an isocyano group, an isocyanato group, or an isothiocyanato group; A compound in which one of R ⁴ ′ and R ⁵ ′ is a chlorine atom and the other is a hydrogen atom, that is, the compound represented by the formula (I ′) can be obtained by extraction and purification from a sponge.

As a raw material animal used here, a sponge Vulgaris sp.
members of the genus hella, such as Acanthella cavernosa
, Acanthella klethra and the like. The extraction solvent generally includes an organic solvent, preferably a water-containing solvent such as methanol, ethanol, and acetone.

After the obtained extract is concentrated, it is preferably partitioned with water and a non-polar solvent such as hexane, benzene, carbon tetrachloride or the like, and the non-polar solvent phase is purified to efficiently produce the desired carihinol derivative. Obtainable. Purification can be performed by appropriately combining silica gel chromatography, reverse phase silica gel chromatography, high performance liquid chromatography and the like.

Further, among the above-mentioned kalihinol derivatives,
R in formula (I) or (I ″) ^ThreeOr R^Three"The Holm
Compounds that are amide groups are represented by R^ThreeOr R^Three"Is an isocyano group
Is hydrolyzed with acetic acid at room temperature.
Obtainable. Further, in the above formula (I) or (I ″),
R in¹, R^Three, R¹"Or R^Three"Is an amino group
Things are R¹, R^Three, R¹"Or R^ThreeIs a formamide group
Hydrolyzed with 6N hydrochloric acid or sodium borohydride
It can be obtained by reduction with lithium. Also,
R in the above formula (I) or (I ″)^TwoOr R^Two"Reed
Compounds that are^TwoOr R^TwoIs a hydrogen atom
The compound is converted to the corresponding acid chloride in a basic solvent such as pyridine.
Or by acting with an acid anhydride.
You.

The above-mentioned kalihinol derivative obtained as described above has an activity of inhibiting the attachment and transformation of harmful organisms in water such as barnacle barnacles and is useful as an antifouling agent. When the above-mentioned kalihinol derivative 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 a paint, a solution, an emulsion or 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 kalihinol derivative 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 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 kalihinol derivative is 0.1% based on the weight of the paint.
5050%, preferably 1-30%.

When the antifouling agent of the present invention is used as a solution, for example, the above-mentioned kalihinol derivative is mixed with a film-forming agent to form a solution dissolved in a solvent, which 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 a film forming agent, for example, natural resin,
Synthetic resins, artificial rubbers and the like are used, and toluene, xylene, cumene, ethyl acetate, methyl isobutyl ketone, methanol and the like are used as solvents. If necessary, additives such as a plasticizer can be added to this solution. When used as a solution, the kalihinol derivative is incorporated at a ratio of 0.1 to 100%, preferably 0.1 to 30% based on the weight of the solution.

When used as an emulsion, the above-mentioned kalihinol derivative 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 kalihinol derivative is 0.1 to 80 based on the weight of the emulsion.
%, Preferably 0.1 to 30%. Also,
The antifouling agent of the present invention can also be used by kneading it into a polymer resin of a material used in water such as aquaculture nets and fixed nets.

[0021]

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 900 g (frozen weight) of Acanthella sponge collected in Hachijojima was pulverized with a blender, extracted with 3 L of ethanol, and filtered. The residue was further extracted with 1 L of ethanol, filtered, combined with the above filtrate and concentrated. After that, concentrate the water 50
Distribute 3 times with 0 ml and 500 ml of hexane and extract 5 g of hexane extract
I got

The hexane extract was purified using the activity of inhibiting the attachment of the barnacle to the larvae of the barnacle as an indicator. First, rough separation was performed by silica gel column chromatography (chloroform, 3, 10, 20, 50% methanol / chloroform, methanol). The fraction eluted with chloroform is subjected to silica gel column chromatography (hexane, 50
% Hexane / benzene, benzene, 50% benzene / chloroform, chloroform, 10% methanol / chloroform, 50% methanol / chloroform, methanol). Next, the 50% benzene / chloroform eluted fraction was subjected to reverse phase high performance liquid column chromatography (ODS;
(Methanol) to give an active fraction. The active fraction was purified again by reversed-phase high performance liquid column chromatography (ODS; 80% methanol / water) to obtain 7.0 mg of the adhesion-inhibiting active substance 1.

Further, after removing the methanol-soluble portion of the 3% methanol / chloroform elution fraction in the silica gel column chromatography, purification by reverse phase high performance liquid column chromatography (ODS; 80% methanol / water) is performed. As a result, 3.1 mg of an adhesion-inhibiting active substance 2 was obtained. Similarly, the methanol-soluble portion of the 10% methanol / chloroform eluted fraction in the silica gel column chromatography was subjected to reverse phase high performance liquid column chromatography (OD
S; methanol) to obtain an active fraction. The active fraction was purified again by reversed-phase high-performance liquid column chromatography (ODS; 80% methanol / water) to obtain 5.8 mg of the adhesion-inhibiting active substance 3, 1.9 mg of the adhesion-inhibiting active substance 4, and 1.9 mg of the adhesion-inhibiting active substance. 2.2 mg of 5 were obtained. In addition, 10.0 mg of the aforementioned adhesion-inhibiting active substance 1 was also obtained.

(2) Determination of Structure (Adhesion Inhibiting Substance 1) Two (M + H) ⁺ ion peaks [m / z] having an intensity of about 3: 1 in FAB mass spectrum (positive ion mode; matrix: glycerol) 411, 413] show that there is one chlorine atom, and that ¹ H, ¹³ C
C ₂₂ H ₃₅ ClN ₂ O ₃
It was decided. By analyzing various two-dimensional NMR spectra and comparing with the chemical shift of ¹³ C NMR spectrum of kalihinol A which is a known substance [J. Am. Chem. Soc., 109 , 6119 (1987)], the structure of this compound is represented by the following formula. (A):

[0025]

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It was decided that: Since this compound has a formamide group, it has s-cis form and s-tran form of the formamide moiety.
It exists as an equilibrium mixture of the two isomers of the s-isomer under NMR measurements. Therefore, in the portion near the formamide group in the ¹ H, ¹³ C NMR spectrum, two pairs of signals appear in pairs. Its ratio in CDCl ₃ is ¹ HN
From the integrated value of the MR spectrum, it was found that a trans-isomer having a structural stability of about 2: 3 was the main component. This 2 below
The species structures and ¹ H and ¹³ C chemical shifts are shown.

[0027]

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Also, the ¹ H spectrum of the compound and ¹³ CN
The MR spectra are shown in FIGS. 1 and 2, respectively. Other physicochemical constants are as follows. [Α] _D = + 13 ° (c = 0.1, CHCl ₃ ) IR spectrum (cm ^-1 ): 3300, 2150, 1670, 700 (Adhesion inhibitory substance 2) ¹ H NMR, ¹³ C NMR and FABMS data are literatures Since it was consistent with that described in [J. Am. Chem. Soc., 109 , 6119 (1987)], the following formula (B):

[0029]

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Kalihinol E was identified. (Adhesion-inhibiting active substance 3) A chlorine atom is formed by two (M + H) ⁺ ion peaks [m / z 411, 413] having an intensity of about 3: 1 in the FAB mass spectrum (positive ion mode; matrix: glycerol). it was shown to exist one, also, ¹ H, ¹³ C
C ₂₂ H ₃₅ ClN ₂ O ₃
It was decided. By analyzing various two-dimensional NMR spectra and comparing it with the chemical shift of the ¹³ C NMR spectrum of the adhesion inhibitory active substance 2 (calihinol E), the structure of the compound was represented by the following formula (C)

[0031]

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Was determined. Since this compound has a formamide group, it has s-cis form and s-tran form of the formamide moiety.
It exists as an equilibrium mixture of the two isomers of the s-isomer under NMR measurements. Therefore, in the portion near the formamide group in the ¹ H, ¹³ C NMR spectrum, two pairs of signals appear in pairs. Its ratio in CDCl ₃ is ¹ HN
From the integrated value of the MR spectrum, it was found that a trans-isomer having a structural stability of about 2: 3 was the main component. This 2 below
The species structures and ¹ H and ¹³ C chemical shifts are shown.

[0033]

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Further, the ¹ H spectrum of the compound and ¹³ CN
The MR spectra are shown in FIGS. 3 and 4, respectively. Other physicochemical constants are as follows. IR spectrum (cm ^-1 ): 3300, 2100, 1670, 750 (Adhesion-inhibiting active substance 4) Two (M + H) of about 3: 1 intensity in FAB mass spectrum (positive ion mode; matrix: glycerol) ⁺ chlorine atom is shown to be present one by ion peak [m / z 427, 429] , also, ¹ H, ¹³ C
Combine the molecular formula with C ₂₂ H ₃₅ ClN ₂ O ₄
It was decided. By analyzing various two-dimensional NMR spectra and IR spectra and comparing the chemical shift of the ¹³ C NMR spectrum of the adhesion-inhibiting active substance 1, the structure of the present compound was represented by the following formula (D):

[0035]

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Was determined. This compound is different from the adhesion-inhibiting active substance 1 in the functional group at the C-5 position, whereas that of the adhesion-inhibiting active substance 1 is an isocyanato group, whereas that of the present compound is an isocyanato group. Since this compound also has a formamide group, the s-cis form and s-trans formamide moiety
It exists as an equilibrium mixture of the two isomers under NMR measurements. Therefore, in the portion near the formamide group in the ¹ H, ¹³ C NMR spectrum, two pairs of signals appear in pairs. Its ratio of in CDCl ₃ ¹ H NMR
From the integrated value of the spectrum, it was found that a trans-isomer having a structural stability of about 2: 3 was the main component. The two structures and the chemical shifts of ¹ H and ¹³ C are shown below.

[0037]

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Further, the ¹ H spectrum of the compound and ¹³ CN
The MR spectra are shown in FIGS. 5 and 6, respectively. Other physicochemical constants are as follows. IR spectrum (cm ^-1 ): 3300, 2250, 1670, 700 (Adhesion inhibitory substance 5) Two (M + H) s of about 3: 1 intensity in FAB mass spectrum (positive ion mode; matrix: glycerol) ⁺ chlorine atom is shown to be present one by ion peak [m / z 443, 445] , also, ¹ H, ¹³ C
C ₂₂ H ₃₅ ClN ₂ O ₃
S was decided. By analyzing various two-dimensional NMR spectra and IR spectra and comparing the chemical shifts of the ¹³ C NMR spectrum of the adhesion-inhibiting active substance 1, the structure of the compound was represented by the following formula (E):

[0039]

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Was determined. This compound is different from the adhesion-inhibiting active substance 1 in the functional group at the C-5 position, and that of the adhesion-inhibiting active substance 1 is an isocyano group, whereas that of the present compound is an isothiocyanato group. Since this compound also has a formamide group, the s-cis form and s-tr of the formamide moiety
It exists as an equilibrium mixture of the two isomers of the ans isomer under NMR measurement. Therefore, in the portion near the formamide group in the ¹ H, ¹³ C NMR spectrum, two pairs of signals appear in pairs. Its ratio in CDCl ₃ is ¹ H
From the integrated value of the NMR spectrum, it was found that the trans-isomer having a structural stability of about 2: 3 was the main component. The two structures and the chemical shifts of ¹ H and ¹³ C are shown below.

[0041]

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Further, the ¹ H spectrum of the compound and ¹³ CN
The MR spectra are shown in FIGS. 7 and 8, respectively. Other physicochemical constants are as follows. IR spectrum (cm ^-1 ): 3300, 2100, 1630, 1250, 770 (3) Adhesion-inhibiting activity By using the adhesion-inhibiting active substances 1, 2, 3, 4, and 5, each of 5 μg / ml of Cyprididae was found. Larval adhesion was inhibited by 100%, and a significant difference was observed even at 0.05 μg / ml from the sample-free group.

[0043]

Industrial Applicability According to the present invention, there is provided a calihinol derivative which has an activity of inhibiting the attachment and transformation of harmful inhabiting organisms in water such as barnacle barnacles and is useful as an antifouling agent.

[Brief description of the drawings]

FIG. 1 is a diagram showing a ¹ H NMR spectrum of an adhesion inhibitory active substance 1.

FIG. 2 is a view showing a ¹³ C NMR spectrum of an adhesion-inhibiting active substance 1.

FIG. 3 is a diagram showing a ¹ H NMR spectrum of an adhesion inhibitory active substance 3.

FIG. 4 is a view showing a ¹³ C NMR spectrum of an adhesion inhibitory active substance 3.

FIG. 5 is a view showing a ¹ H NMR spectrum of an adhesion inhibitory active substance 4.

FIG. 6 is a view showing a ¹³ C NMR spectrum of an adhesion inhibitory active substance 4.

FIG. 7 is a view showing a ¹ H NMR spectrum of an adhesion inhibitory active substance 5.

FIG. 8 is a view showing a ¹³ C NMR spectrum of an adhesion inhibitory active substance 5.

──────────────────────────────────────────────────続 き Continued on the front page (58) Field surveyed (Int. Cl. ⁶ , DB name) C07D 309/00-309/08 CA (STN) REGISTRY (STN)

Claims (3)

(57) [Claims] (1) The following formula (I): (Wherein, R ¹ represents a formamide group, an isocyano group or an amino group, R ² represents a hydrogen atom or an acyl group, and R ³
Represents an isocyano group, an isocyanato group, an isothiocyanato group, a formamide group or an amino group, and R ⁴ and R ⁵
Has one chlorine atom and the other a hydrogen atom. Where R
Except when ¹ and R ³ are both isocyano groups. ). 2. The following formula (I ′), which is extracted and purified from a sponge: (Wherein, R ¹ ′ represents a formamide group or an isocyano group, R ² ′ represents a hydrogen atom, R ³ ′ represents an isocyano group, an isocyanato group, or an isothiocyanato group, and R ⁴ ′ and R ⁴ ′
^{In 5} ′, one is a chlorine atom and the other is a hydrogen atom. )). 3. The following formula (I ″): (Wherein, R ¹ "represents a formamide group, an isocyano group or an amino group, R ² " represents a hydrogen atom or an acyl group, and R ³ "
Represents an isocyano group, an isocyanato group, an isothiocyanato group, a formamide group or an amino group, and R ⁴ "and R ⁵ "
Has one chlorine atom and the other a hydrogen atom. An antifouling agent containing a compound represented by the formula (1) as an active ingredient.