KR101149382B1 - Anti-scuticociliatic composition containing Polysiphonia morrowii Harvey extract, or synthesized or isolated compounds from the same - Google Patents

Abstract

The present invention relates to a composition for preventing, treating or inhibiting Scutika infection of fish containing Polysiphonia morrowii Harvey extract or a nonpolar solvent soluble fraction thereof, or a compound or derivative thereof isolated from the extract. Morrow red yarn extract or fractions thereof exhibits significant anti-skutica activity, and compounds and derivatives thereof isolated from the fractions have an effect of significantly reducing the mortality of fish by Scutika insects.

Description

Anti-scuticociliatic composition containing Polysiphonia morrowii Harvey extract, or synthesized or isolated compounds from the same}

FIELD OF THE INVENTION The present invention relates to an anti-suctica composition comprising an extract of Polysiphonia morrowii Harvey or a compound isolated or synthesized therefrom. More specifically, the present invention relates to an extract of Morrow red thread or a nonpolar solvent soluble fraction thereof. The present invention relates to a composition for preventing, treating or inhibiting Scutika infection of fish containing one compound or a derivative thereof.

Scutika insects frequently occur in flounder, the representative fish of Korea, and the mortality rate is very high when it occurs in childhood, and the mortality rate is high when there is a lot of organic matter in the aquaculture environment or when other diseases and complex infections occur. high. Scutika are parasitic in gills, body surface and fins, and tissues collapse. As symptoms progress, the muscles are exposed and penetrate the brain and various organs, causing necrosis of tissues.

Currently, formalin, which is approved as a medicinal product, is mainly used for the control of Scutika insects in Korea, but the concentration that can control more than 200 ppm of Scutica insects causes severe stress on the flounder, and the infiltrating body is impossible to remedy. to be.

Regarding the control of the conventional Skutika insects, Republic of Korea Patent No. 10-0279281 Ae is disclosed a fish parasite Skutika insecticide using an alginate oligosaccharide derivative as an active ingredient, and in Korea Patent Application No. 10-2005-0048037 A composition for the control of parasitic diseases, including Scutica ciliated insects of cultured fish, which has a pesticidal effect on parasites such as Scutica ciliates, which are harmless to the human body and do not adversely affect the environment by mixing sodium percarbonate at a predetermined ratio. Is disclosed.

In addition, Korean Patent No. 10-0735038 discloses 38kDa recombinant antigenic protein of Scutica's ciliated protein, which effectively immunizes the flounder as well as prevents and treats Scutica. It is disclosed that there is an excellent effect, the Republic of Korea Patent No. 10-0768143 No. 0.5ppm or more and 10ppm or less, 0.015ppm or more and 0.3ppm or less of the organic carboxylic acid stabilized chlorine dioxide or chlorite in a water-blocked water tank It is disclosed that the fish suffering from Scutica infection can be treated by bathing the fish by adding 5 ppm or more to 30 ppm or less, respectively.

In the prior domestic patent literature on the control of Scutica insects as described above, there are chlorine dioxide, butylaldehyde, ketoconazole, antifungal agents, ketoconazole and terbinapine hydrochloride, but these may be problematic in case of their toxicity or remaining in the fish body. There is this. In addition, chitosan, alginic acid oligosaccharides, and truncated pine extracts are registered as Scutica insect remedies.

Polysiphonia morrowii Harvey is a red algae in seawater, belongs to the Rhodomelaceae. (Algaebase: http: //www.algaebase.org). To date, several bromophenol compounds have been isolated and reported from Morrow red yarn, and their physiological activities include α-glucosidase inhibitory activity [Fisheries Science (1999), 65 (2), 300-303]. Antimicrobial Activity against Fish Bacteria Presented by the Inventors' Prior Research [J. Fish Pathol., 18 (2): 147-156 (2005)], a domestic patent relating to antiviral activity against fish disease virus [Food Sci. Biotechnol. 17 (5): 1074-1078 (2008); Korean Patent No. 10-0838621] and antioxidant activity [Food Sci. Biotechnol. 18 (1): 124-129 (2009). However, there has been no report on the control effect of Scutica insects of the Morrow red thread and the compound separated therefrom.

Accordingly, the present inventors have made intensive studies to improve the above-mentioned various problems in the control of Scutica insects, and as a result, extracts of Polysiphonia morrowii Harvey, fractions thereof, or bromine-containing separated or synthesized therefrom The compounds and compounds of this family were found to be effective in controlling Scutica insects and completed the present invention.

Accordingly, an object of the present invention is to prevent, treat or inhibit the infection of Scutica insects containing as an active ingredient extracts, fractions or bromine-containing compounds isolated or synthesized from Polysiphonia morrowii Harvey It is to provide a composition.

An object of the present invention as described above is to prepare a methanol extract from the Morrow red yarn, from which CH ₂ Cl ₂ , methanol, n -hexane fractions are prepared, and then the compounds are separated from the methanol fractions and antiscutica for each of them. While confirming the insecticidal activity, it was achieved by identifying the anti-skutica insect activity against derivatives similar to these compounds.

The present invention provides a composition for preventing, treating or inhibiting Scutika infection containing an extract of Polysiphonia morrowii Harvey as an active ingredient.

In the present invention, the term "active ingredient" refers to a substance or a group of substances (a pharmacologically active ingredient or the like, which is expected to express the efficacy or effect of the drug directly or indirectly by intrinsic pharmacological action). It means containing a main component).

In the present invention, the Morrow red thread extract is characterized in that any one selected from the group consisting of non-polar solvent soluble extract, preferably MeOH, n -hexane or CH ₂ Cl ₂ extract.

In another aspect, the present invention comprises 3-bromo-4,5-dihydroxybenzyl methyl ether of the formula (1) or derivatives thereof as the active ingredient Provided is a composition for preventing, treating or inhibiting Scutika infection of fish.

<Formula 1>

In the present invention, the derivative of the compound of Formula 1 is characterized in that 2-bromo-4-methoxyphenol of the formula (5).

<Formula 5>

Morrow red thread extract according to the present invention exhibits significant anti-Sucutica activity, the compounds and derivatives thereof isolated from the fractions have an effect of significantly reducing the mortality of fish caused by Scutika insects, the present invention It is a very useful invention for aquaculture.

1 is a diagram illustrating a process of extracting and fractionating from the Morrow red yarn according to the present invention.
2 is a view showing a process of separating the active material after the fractionation process from the Morrow red yarn according to the present invention.
FIG. 3 is a diagram showing a Scotica worm observed under a microscope 20 hours after treatment with the Morrow red thread extract and fractions therefrom.
FIG. 4 is a diagram showing a Scotic worm observed under a microscope 20 hours after treatment with two bromine-containing compounds separated from a Morrow red thread.

Hereinafter, preferred embodiments of the present invention will be described in more detail with reference to Examples. However, the scope of the present invention is not limited to these examples.

Example 1 Preparation of Morrow Red Thread Extract

In this example, the extraction of the Mororul red yarn was prepared using methanol (MeOH) as the extraction solvent. As shown in FIG. 1, the lyophilized Morrow red thread (133.5 g) was extracted with hot water (80 ° C.) for 1 h with 80% MeOH 3 L, and the total extract was collected and concentrated under reduced pressure with a vacuum evaporator. .

The 80% MeOH extract (33.54 g) obtained above was suspended in distilled water and fractionated with methylene chloride (CH ₂ Cl ₂ ). The methylene chloride soluble fraction was concentrated under reduced pressure, dissolved in 90% aqueous methanol solution, and fractionated with n-hexane to obtain a total of three fractions.

Example 2 Isolation of Active Compound from Morrow Red Thread Fraction

In this example, the active compound was separated from the 90% aqueous methanol fraction obtained in Example 1. As shown in FIG. 2, the 90% methanol aqueous solution fraction was first divided into nine small fractions by performing ODS column chromatography (MeOH: Water = 5: 95-100: 0). Small fraction fr. The second subfraction (180 mg) of the five subfractions obtained by Sephadex LH-20 column chromatography on II was subjected to Sephadex LH-20 column chromatography (MeOH: Actone = 1: 9), respectively. 1 (24.9 mg) and compound 2 (46.9 mg) were obtained. In addition, HPLC purification of the small fraction fr.VI was carried out to obtain compound 3.

Example 3 Chemical Structure Analysis and Identification of Compounds 1, 2 and 3

Chemical structures of the compounds 1 to 3 obtained in Example 2 were analyzed. In order to analyze the chemical structure of the compound 1 , spectroscopic analysis was performed using EIMS, ¹ H-NMR and ¹³ C-NMR.

As a result, Compound 1 was determined as 3-bromo-4,5-dihydroxybenzyl methyl ether, and described in Chem. Pharm. Bull ., 34: 4083-4091, 1986) was identified (formula 1). Compound 2 was determined as 3-bromo-4,5-dihydroxybenzaldehyde, and described in Chin. J. Oceanol. Limnol., 23 (2): 226-229 (2005)]. And Compound 3 was determined as urceolatol and found in J. Chem. Asian Nat. Prod. Res., 8 (4): 379-383 (2006)].

<Formula 1>

<Formula 2>

<Formula 3>

Experimental Example 1: Antiscutica Activity of Morrow Red Thread Extract ( in vitro )

Antiscutica activity of the samples obtained in Examples 1 and 2 was measured.

The cultured Scutica insects were centrifuged at 500 X g for 5 minutes, and DMEM medium (or seawater) was added to the pellet to float the insects, and the concentration of the insects was adjusted to 1-2 × 10 ³ insects / 50 uL. Morrow red thread extracts and fractions were made into 2-fold dilution series and placed in 50 uL in 96-well microtight plate. Here, the filling fluid was adjusted to the set concentration shown in Table 1 by putting 50 uL per well. Table 1 shows the results of microscopic observation after 30 minutes, 1 hour, 2 hours and 20 hours, and after 20 hours.

TABLE 1

Antiscutica Activity of Morrow Red Thread Extract ( in vitro ) ¹

MLC: minimal lethal concentration

The n -hexane fraction showed the highest antiscutica activity at 62.5 ppm under both media and seawater conditions. The MLC of formalin used as a positive control is considered to be a fairly good activity compared to 31.3. In comparison, the 90% methanol fraction was 250 ppm in the medium condition and 125 ppm in the seawater condition, which was 2-4 times lower than the n -hexane fraction.

Experimental Example 2: Antiscutica Activity of Compounds 1 to 3 and Synthetic Structure Analogues thereof

Antisuctica activity of Compounds 1 to 3 and 4 synthetic structural analogs (Compounds 4 to 7) isolated from 90% aqueous methanol fractions was investigated.

<Formula 4>

Compound 4: 3,4-dihydroxybenzaldehyde

<Formula 5>

Compound 5: 2-bromo-4-methoxyphenol

(6)

Compound 6: 5-bromovanillin

<Formula 7>

Compound 7: 3,5-dibromo-4-hydroxybenzaldehyde

As a result, Compound 1 showed the highest activity at 62.5 ppm MLC under seawater conditions. In addition, compound 5 of the synthetic structural analogue showed the same effect as compound 1. This is considered to be a very good effect when compared to the MLC of formalin 31.3 ppm. In addition, considering that the MLC of the 90% aqueous methanol fraction of the mother fraction was 125 ppm under the same conditions, it was determined that Compound 1 was an important material for the activity of the mother fraction. In comparison, the anti-scutica activity of Compound 2 was shown to be relatively low at 500 ppm, the highest set concentration. In addition, the activity of Compound 3 did not appear even at the highest setting concentration of 250ppm. In medium condition, Compound 1 showed 125 ppm of MLC, showing the highest activity, but the effect was lowered overall under medium condition.

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TABLE 2

In vitro antiscutica activity of compounds 1 to 3 and synthetic structural analogs thereof

MLC: minimal lethal concentration

Claims (5)

A composition for preventing, treating or inhibiting Scutika infection, which contains an extract of Polysiphonia morrowii Harvey as an active ingredient. The composition for preventing, treating or inhibiting Skutika infection of fish according to claim 1, wherein the extract of Morrow red thread is a non-polar solvent soluble extract. 3. The composition for preventing, treating or inhibiting Skutika infection of fish according to claim 2, wherein the Morrow red thread nonpolar solvent soluble extract is any one selected from the group consisting of MeOH, n -hexane or CH ₂ Cl ₂ extract. . Squeeze of fish containing 3-bromo-4,5-dihydroxybenzyl methyl ether of the formula (1) or derivatives thereof as an active ingredient Composition for preventing, treating or inhibiting tika infection.
<Formula 1>

The composition for preventing, treating or inhibiting Skutika infection of fish according to claim 4, wherein the derivative of the compound of Formula 1 is 2-bromo-4-methoxyphenol of Formula 5.
&Lt; Formula 5 >

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