JP2010064973A - Sponge-derived starfish repellent - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a starfish repellent using a compound having degradability under a marine environment. <P>SOLUTION: The starfish repellent is discovered by virtue of homarine, β-cholestanol, cholesterol, L-phenylalanine or a thymine compound alone or a plurality of their combinations. The extract of sponge including Kinubariitokaimen (a kind of sponge) adhering to seashells is found to have starfish repellent activity. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

The present invention relates to a sponge-derived compound or sponge extract having starfish repellent activity.

The damage caused by starfishes of shellfish such as scallop, scallop and abalone is an important issue in the fishery industry. Mixing of starfish into the aquaculture net and predation by starfish contribute to a high depletion rate when shellfish are released.

In order to prevent starfish from being damaged, starfish are being exterminated. However, when they are brought back to land, they become industrial waste, and it is very expensive to dispose of them. Yes.

The present inventors have discovered that shellfish to which sponge is attached are not preyed by starfish during diving. We also observed that the starfish prey immediately when the shellfish with sponges were removed from the shell.

Provided is a starfish repellent using a compound having degradability in a marine environment.

As a result of intensive studies on the above problems, the present inventors have found starfish repellent activity in a sponge extract, a sponge-derived compound alone or a mixture thereof, and completed the present invention.

Since the sponge extract of the present invention or the sponge-derived compound alone or a mixture thereof has starfish repellent activity, it can be used as a starfish control agent.

That is, the present invention relates to the following (1) to (3).
(1) Sponge extract containing homarine represented by Chemical Formula 1, β-cholestanol, cholesterol, L-phenylalanine or thymine represented by Chemical Formula 2, or a pharmacologically acceptable salt thereof.

(2) Sponge extract adhering to shells including quinuburite sponge.

(3) A method for producing a starfish repellent by using a compound containing homarin, β-cholestanol, cholesterol, L-phenylalanine or thymine, or a pharmacologically acceptable salt thereof, or a combination thereof.

Sponge adhering to the shellfish of the present invention can be collected by diving, and after collecting spongy adhering to the shell, it returns to the place of collection and grows again. Examples include kinuvalite sponges that attach to snails that live in the Tohoku region, and sponges that attach to bivalve red clams.

Sponge extract is obtained by the method used for the collection of substances usually performed. For example, the sponge is cut as it is or is cut into small pieces, extracted with a water-soluble organic solvent such as methanol, ethanol, or propanol. After concentration, the solution is made using a water-containing organic solvent such as water-containing methanol, and if necessary, degreased. Then, extraction is performed with an organic solvent such as water-saturated 1-butanol.

Concentration of the residue obtained by concentrating the adsorption chromatography, gel filtration chromatography, thin layer chromatography, high performance liquid chromatography, etc., as appropriate, or repeating them as necessary, leads to homarin, β-cholestanol, cholesterol Alternatively, L-phenylalanine and thymine can be isolated.

In the present invention, the pharmacologically acceptable salt is not particularly limited as long as it is a salt that can be usually taken. For example, a salt with a mineral acid or an organic acid, that is, a sodium salt, potassium salt, hydrochloride, sulfate, Acetate and the like are preferred.

The present invention also includes a starfish repellent containing as an active ingredient a compound of the above-described homarine, β-cholestanol, cholesterol, L-phenylalanine or thymine, or a pharmacologically acceptable salt thereof, or a mixture of combinations thereof. .

All five compounds are components contained in the living body of sponge. Homarine is a compound that exists in common in marine invertebrates, and β-cholestanol, cholesterol, thymine, and L-phenylalanine are compounds that exist in living organisms. It is speculated that a compound isolated from quinuvarite sponge that also lives in the inhabiting environment of a starfish to be a repellent, or a sponge extract is a compound having a degradation pathway in a marine environment.

Test Examples Extracts of sponges adhering to shells including quinuvalite sponges or compounds of homarin, β-cholestanol, cholesterol, L-phenylalanine or thymine, or pharmacologically acceptable salts thereof, or a plurality thereof The starfish repellent activity by the mixture of these is described.

Experiment 1
Kinuburite sponges adhering to snails were collected, chopped, extracted with ethanol, and concentrated to obtain an extract. Twelve were prepared by cutting 1 x 1 x 1 cm each of agar using clam soup containing 1 ml of this ethanol extract and agar prepared only with clam soup. Each agar was fixed on the bottom of a 30 × 30 × 60 cm flowing water tank with an acrylic rod fixed on an acrylic plate. The number of agar remaining after the passage of one night was counted after adding five starfishes.

Results The agar made from clam soup had 0 residues and the agar with ethanol extract had 10 residues.

Experiment 2
Sponge adhering to the bivalve red clam was collected, chopped, extracted with ethanol, and concentrated to obtain an extract. Twelve pieces of agar prepared using red cabbage broth containing 1 ml of this ethanol extract and agar prepared only with red cabbage broth were cut into 1 × 1 × 1 cm each. Each agar was fixed on the bottom of the same aquarium on a 40 × 50 × 5 cm depth of flowing water tank with an acrylic rod fixed on an acrylic plate. Five starfish and three starfish starfish were added and the number of agar remaining after 24 hours was counted.

Results The agar prepared from the scallop broth was 0 remaining, and the agar added with the ethanol extract had 8 remaining. The agar made only with red scallop broth was 0 remaining 6 hours after the start.

Experiment 3
Asari-simmered soup made of β-cholestanol, cholesterol, thymine, L-phenylalanine, which is a commercially synthesized product of a compound isolated from Sorbus, and a homarin synthesized by methylating aα-picolinic acid, which is a commercial product, with methyl iodide In the agar prepared in step 2, 2 mg each was added and cut into 1 × 1 × 1 cm to make 12 pieces. Each agar was fixed on the bottom of a 30 × 30 × 60 cm flowing water tank with an acrylic rod fixed on an acrylic plate. The number of agar remaining after the passage of one night was counted after adding five starfishes.

Results The agar made from clam soup was 0 remaining, and the remaining agar was adjusted by adding 12 β-cholestanol, 8 cholesterol, 6 thymine, 8 L-phenylalanine and 12 homarin. .

Experiment 4
Nittoru CL (Nitto Kasei Co., Ltd., Osaka), a commercially available fishnet antifouling agent, added with 10 mg of an extract of quinuvalite sponge adhering to a sea urchin shell, β-cholestanol, cholesterol, thymine, L-phenylalanine, A mixture of 5 mg each of homarin was added to gauze 3 × 6 cm and allowed to stand overnight at room temperature. A 40 × 50 × 5 cm deep flowing water tank was partitioned into a central portion from the flowing water port to 2/3 with an acrylic plate to make a Y-shaped water tank. Since starfish usually have the property of flowing, the inlet of running water is set to one side of the Y shape, and gauze is fixed near it. Five starfishes and five starfish starfish were added and the progress from the start of the experiment to the end of 120 hours was observed.

Results In the ethanol extract, individuals approaching gauze appeared after 48 hours from the start, and half approached gauze at 72 hours after the start. In the case where 5 mg each of β-cholestanol, cholesterol, thymine, L-phenylalanine, and homarin were mixed and added, there was a single starfish that approached gauze one hour after the start. No other individuals approached gauze until 96 hours passed, and individuals approaching gauze appeared from around 104 hours.

Although the Example of this invention is shown below, this is only an illustration, Comprising: This invention is not limited, Various modifications are possible. HPLC is high performance liquid chromatography.
Preparation method and separation of compounds of quinuvalite sponge adhering to Sorbus.
(A) Collected aspergillus in Mutsu Bay, Aomori Prefecture, and collected adhering kinuburite sponge with a spatula. Kinuvalite sponge 610 g was immediately treated with ethanol. After collecting kinuvalite sponges, the rodents returned to the sea. Ethanol extraction was performed 4 times to obtain 32.81 g of concentrated extract.

(B) Of the concentrated extract of (a), 5.24 g was subjected to liquid-liquid partition with n-hexane. The aqueous layer was further partitioned with ethyl acetate. When the n-hexane layer (85.6 mg) was subjected to silica gel column chromatography with ethyl acetate / n-hexane, β-cholestanol and Rf values similar to those of TLC were shown in ethyl acetate 10% and 15% fractions. A fraction containing a compound showing a spot was obtained. Since the GC / MS spectrum, various NMR spectra, and the retention time of HPLC of the commercial product were matched, β-cholestanol and cholesterol were determined.

(C) The ethyl acetate layer (189.6 mg) was obtained by the ethyl acetate partition of (b). The aqueous layer was further partitioned with 1-butanol to obtain 193 mg of butanol layer and 594 mg of aqueous layer. A 500 mg butanol-distributed aqueous layer was applied to an ODS cartridge column, and the 3% methanol eluate was subjected to reverse phase HPLC (C18, Capcel Pak 10φ x 250 mm, Shiseido, 0.05% TFA-containing water flow rate 1 mL / min, UV 210 nm monitor). Separation gave one compound. Analyzing mass spectra and various NMR spectra to estimate homarine, and comparing the retention time of HPLC and various NMR spectra of the homarin prepared by methylation of a-picolinic acid with methyl iodide. It was decided from Homarine.

(D) The 10% methanol eluate was fractionated by reverse phase HPLC (C18, Capcel Pak 10φ x 250 mm, Shiseido, 5% methanol flow rate 2 mL / min, UV 210 nm monitor) to isolate two compounds. Since the various NMR spectra and HPLC retention times of the commercial products matched, thymine and phenylalanine were determined.

(E) The phenylalanine of (d) was made into agar of clam soup with 2 mg each of commercially available synthetic products D-phenylalanine and L-phenylalanine, each cut into 1 × 1 × 1 cm and made into 12 pieces. Each agar was fixed on the bottom of a 30 × 30 × 60 cm flowing water tank attached with an acrylic rod fixed to an acrylic plate. The number of agar remaining after the passage of one night was counted after adding five starfishes. As a result, the number of D-phenylalanine was 3 and the number of L-phenylalanine was 8, and the D-phenylalanine agar was found to have contacted traces and was out of shape. did.

All of the five compounds are components contained in the body of sponge. Homarine is a compound that exists in common in marine invertebrates, and β-cholestanol, cholesterol, thymine, and L-phenylalanine are compounds that exist in living organisms. It is speculated that a compound or sponge extract isolated from quinuburite sponge that also inhabits the environment of starfish, which is the target of repellent, has a degradation pathway in the marine environment.

Claims (3)

A sponge extract comprising a homarine represented by Chemical Formula 1, β-cholestanol represented by Chemical Formula 2, cholesterol, L-phenylalanine, or thymine or a pharmacologically acceptable salt thereof.

Extracts of sponges attached to shells such as quinuvalite sponges. A method for producing a starfish repellent by using a compound containing homarine, β-cholestanol, cholesterol, L-phenylalanine, thymine, or a pharmacologically acceptable salt thereof, or a combination thereof.