KR102562276B1 - Composition of repellent containing plant oil and related chemical compounds for Controlling Haemaphysalis longicornis - Google Patents

Abstract

The present invention relates to a natural repellent composition for controlling small sturgeon ticks of Saturella montanya essential oil and Amorasia rusticana essential oil, and more specifically, to Saturella montanya essential oil, a It relates to a natural repellent composition comprising Morasia rusticana essential oil and their active ingredient or a derivative compound of the active ingredient as an active ingredient.

Description

Composition of repellent containing plant oil and related chemical compounds for Controlling Haemaphysalis longicornis}

The present invention relates to a composition for controlling small ticks, and more specifically, active ingredients of Satureja montana essential oil and Amoracia rusticana essential oil, which have repellent activity against small ticks Or it relates to a repellent composition comprising a derivative compound of an active ingredient.

Ticks were the first arthropods to be recognized as vectors of pathogens, and are now recognized as major arthropod vectors of zoonotic pathogen-related diseases for humans and livestock, along with mosquitoes. In particular , Haemophysalis longicorni , a member of the family of true mites, is found throughout the country from May to August and is known to parasitize animals living outdoors.

The small sophie tick is a major blood-sucking tick that mediates infection with Sever Fever with Thrombocytopenia Symdrome Virus (SFTSV). In Korea, a total of about 1,000 patients infected with SFTSV occurred over the past five years, and 180 deaths, threatening the lives of the people.

The fatality rate of SFTS patients in Korea is about 18% due to the small sophie tick, but there is no existing preventive vaccine or antiviral drug worldwide, so it poses a threat to human survival. Moreover, it takes more than 10 years, including clinical trials, to develop new drugs such as vaccines or treatments that can treat patients infected with SFTSV. It is imperative to align

DEET ( N,N -diethyl- m -toluamide) is a compound that protects humans and animals from mosquitoes/mites and is known as a repellent with excellent repellent activity and durability. According to the US Centers for Disease Control and Prevention, approximately 30% of the US population uses DEET and DEET-based products as mosquito/tick repellents, and estimates that approximately 23-29% of children are exposed to DEET. DEET is effective as a repellent, but its use is limited due to reports of adverse effects on children, pregnant women, and fetuses. Therefore, it is expected that it will be able to make a great contribution to the fields of public health, agricultural food, and disease science by developing a repellent using plant-derived biomaterials to effectively control small sophid ticks.

Patent Document 1: Korean Patent Publication KR 20090040269 A

The inventors of the present invention, while searching for a method for controlling small scale ticks, confirmed that Saturella montana essential oil and Amorasia rusticana essential oil exhibit high repelling effects against small scale ticks, and completed the present invention.

Accordingly, an object of the present invention is to provide a repellent composition for controlling small sophid ticks comprising active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil, and derivative compounds of these active ingredients.

In order to achieve the above object, the present invention relates to a natural repellent for small sophie tick, 5-isopropyl-2-methylphenol, an active ingredient of Saturella Montaña essential oil, and its derivative 5-isopropyl- Provides a repellent active composition comprising at least one or more compounds selected from 2-methylaniline, phenethylisothiocyanate, which is an active ingredient of Amorasia rusticana essential oil, and benzylisothiocyanate, which is a derivative compound thereof, as an active ingredient .

In addition, the present invention provides a tick repellent prepared using the repellent active composition.

The Saturella montana essential oil, Amorasia rusticana essential oil, and their active ingredients and derivative compounds of the active ingredients according to the present invention are plant-derived natural repellents for small ticks, while having excellent repelling activity against small scale ticks, It is effective as a simple, economical and environmentally friendly formulation.

1 is a diagram showing a climbing test assay performed in the present invention.
Figure 2 shows the compounds carried out in the present invention (1) 5-isopropyl-2-methylphenol, (2) 5-isopropyl-2-methylaniline, (3) benzylisothiocyanate and (4) phenethylisothio It is a graph showing the repelling activity after 60 minutes according to 1: 1 mixing of each compound of cyanate.

In order to achieve the above object, the present invention relates to a natural repellent for small sophie tick, 5-isopropyl-2-methylphenol, an active ingredient of Saturella Montaña essential oil, and its derivative 5-isopropyl- Provides a repellent active composition comprising at least one or more compounds selected from 2-methylaniline, phenethylisothiocyanate, which is an active ingredient of Amorasia rusticana essential oil, and benzylisothiocyanate, which is a derivative compound thereof, as an active ingredient .

In the present invention, “Saturella Montaña” belongs to the Lamiaceae family and is a perennial semi-shrub native to Europe, Russia, Turkey, and northern Africa. Saturella montana is a fragrant herb that has long been used as a spice, tea, and condiment. “Amorasia Rusticana” belongs to the Brassicaceae family and is a perennial plant that grows widely in southeastern Europe and West Asia. The root of Amorasia rusticana is called mustard radish and is widely used as a spicy spice.

The essential oil of Saturella Montaña in the present invention may be used that is distributed and sold on the market, and the essential oil of Amorasia rusticana obtained by extraction by continuous steam distillation is used.

In the present invention, the '5-isopropyl-2-methylphenol' compound is represented by Formula (1), and '5-isopropyl-2-methylaniline' is a compound represented by Formula 2 below, The compound 'phenethylisothiocyanate' is represented by the following formula (3), and 'benzylisothiocyanate' is a compound represented by the following formula (4).

<Formula 1>

<Formula 2>

<Formula 3>

<Formula 4>

The 5-isopropyl-2-methylphenol compound of the above formula (1) and the 5-isopropyl-2-methylaniline of the above formula (2) may be commercially distributed and sold, and also, the above formula The phenethyl isothiocyanate compounds and benzyl isothiocyanate compounds of (3) and (4) can also be used commercially available products.

The compounds represented by the above formulas (1) to (4) exhibit excellent repellent activity against the small sophid mite, respectively (see Tables 6 and 7), and also the compounds represented by formulas (1) to (4) Mixtures of two or more of them also show excellent repellent activity against the small sophid tick (see Table 8).

In the present invention, the compounds may be included in a concentration of 0.1 to 10.0% with respect to the repellent composition. Preferred concentrations are 0.5 to 10% concentration.

If the concentration of Saturella Montaña essential oil and Amorasia rusticana essential oil compound is less than 0.1% of the repellent composition, the repellent effect may be low, and if it exceeds 10.0% relative to the total weight, the content of other general additives is relatively high. It may be difficult to show the expected repelling effect in terms of the blending ratio.

In the present invention, the repellent composition may be used in the form of one or more formulations selected from the group consisting of aerosols, sprays, liquids, emulsions, coating agents, creams, capsules, ointments and granules, and preferably aerosols. It can be used in the form of formulations of agents, emulsions, sprays or creams.

Repellents containing active ingredient compounds of Saturella montanya essential oil, Amorasia rusticana essential oil, and derivative compounds thereof according to the present invention are directly sprayed on areas with a large amount of small ticks, or conventional repellent treatment methods It can be used for direct spraying on humans and livestock or for household items including clothing.

The repellent may be prepared and applied in various forms, and may be dissolved and dispersed in one or more solvents for spraying or application, or as an extender using a conventional surfactant, such as a liquid solvent, a pressurized liquefied gas, a lotion, and/or a thickening agent. It is preferably used in the form of sprays, aerosols, emulsions, and creams.

In order to prepare such various types of preparations, the active ingredient compounds of Saturella Montaña essential oil and Amorasia rusticana essential oil, and derivative compounds thereof according to the present invention are solvents and additives commonly used in the field of repellents Can be used there is. In addition, the content of the active ingredient in the repellent is used in an amount effective to exhibit a repellent effect against mites, and the specific content at this time may vary depending on the type of preparation and application method.

Hereinafter, the present invention will be described in more detail through examples. However, these examples are intended to illustrate the present invention by way of example, and the scope of the present invention is not limited by the examples.

Example 1. Extraction of essential oil from plants

In the present invention, essential oil sold and distributed on the market was used for Saturella montana essential oil, and essential oil was obtained by extracting the root of Amorasia rusticana by continuous steam distillation extraction.

The roots of Amorasia rusticana in a plant state were washed with distilled water, dried for 48 hours or more in a well-ventilated place, and then ground. 1000 ml of distilled water was injected into 100 g of the pulverized sample, and extraction was performed at 100 to 150 ° C for 3 to 6 hours using a steam distillation extraction (SDE). At this time, hexane 300, a non-polar organic solvent, was used as the extraction solvent. ml was used. The extracted hexane extract was dried over anhydrous magnesium sulfate, concentrated under reduced pressure, sealed and stored in a refrigerator.

Example 2. Repellent activity test for small sophid mite

Example 2-1. Pygmy nymphs used for repellent activity assay

The nymph stage, which has the highest density among larvae, nymphs, and adults, was used, and wild individuals were collected from the grass near the ranch in Jeju Island. The collected mites were used after being stored at a temperature of 26 ° C and a relative humidity of 70 ± 5.

Example 2-2. Repellent activity test method using climbing test

A climbing test was used to test the repellent activity of the Amorasia rusticana essential oil extracted in Example 1 and the commercially available Saturella montana essential oil against small sophid mites (see FIG. 1).

The climbing test is a method that utilizes the behavioral response of ticks climbing vertically.

After designating the upper part of the filter paper with the size of 8.0 cm in length × 1.0 cm in width as the sample treatment area (height 1.0 cm) and the central part as the mite treatment area (height 1.0 cm), 20 μl of diluted samples of various concentrations are uniformly distributed in area A. processed. The treated sample was volatilized at room temperature for 30 and 60 minutes, respectively, and then mounted on a cradle to inoculate 10 small tick nymphs in area B. As a negative control group, ethanol, a dilution solvent of the sample, was used.

After recording the number of ticks that crossed the area A where each sample was treated, the avoidance rate (%) was expressed by the following formula. Here, T is the number of ticks that are not evaded in the sample treatment area (ticks crossing area A), and C is the number of ticks that are not evaded in the negative control group.

Avoidance rate (%) = (C-T)/C × 100

The avoidance rate was calculated using the SPSS statistical program (version 12.0), and the average It is expressed as standard error.

Example 2-3. Evaluation of repellent activity of Saturella montana essential oil and Amorasia rusticana essential oil

As a result of examining the repellent activity of Saturella montana essential oil and Amorasia rusticana essential oil using a climbing test, as shown in Table 1 below, both essential oils had excellent repellent activity of 100% for up to 60 minutes at 5.0% and 3.0% concentrations showed Table 1 below shows the repelling activity results of Saturella montana essential oil and Amorasia rusticana essential oil against small sophid mite nymphs using a climbing test.

sample density(%) Avoidance rate (average standard error, %) Elapsed time (minutes) after sample application 30 60 Saturella Montana Essential Oil 5.0 100.0±0.0 100.0±0.0 3.0 100.0±0.0 100.0±0.0 1.0 86.7±3.3 63.3±3.3 Amorasia Rusticana Essential Oil 5.0 100.0±0.0 100.0±0.0 3.0 100.0±0.0 100.0±0.0 1.0 76.7±3.3 56.7±3.3

Example 3. Component Analysis of Saturella Montana Essential Oil and Amorasia Rusticana Essential Oil and Exploration of Active Ingredients

Example 3-1. Component Analysis of Saturella Montaña Essential Oil and Amorasia Rusticana Essential Oil

Components of Saturella montana essential oil and Amorasia rusticana essential oil were analyzed using a gas chromatography-mass spectrometer (Hewlett-Packard HP 6890, H5973 IV series).

As for the column, HP Innowax capillary column and DB5 column (0.25 mm i.d. × 0.25 μm thickness × 2,990 cm) were used, and helium was used as the carrier gas and the temperature was 51-201℃ (2℃). /min).

The temperature of the injector was set at 211° C., and the ionization potential was performed at 70 eV.

The results of component analysis of Saturella Montaña essential oil and Amorasia rusticana essential oil are shown in Table 2 and Table 3, respectively.

Table 2 below shows the chemical composition of Saturella Montaña essential oil.

Retention Time (min) Library Search Composition ratio (%) 4.935 alpha-pinene 1.37 5.222 Kampen 0.13 5.735 beta-pinene 0.27 5.910 beta-myrcene 1.06 6.445 alpha-terpinene 0.65 6.590 ortho-cymene 11.59 6.675 limonene 0.56 6.735 eucalyptol 0.48 7.230 gamma-terpinene 1.37 7.930 Linanol 1.62 9.215 bornell 0.14 9.600 alpha-terpineol 0.32 10.585 para-manth-6-n-2-one 0.15 11.170 thymol 0.53 11.370 5-isopropyl-2-methylphenol 76.40 13.265 Beta-Caryophyllene 2.45 13.740 alpha-caryophyllene 0.30 15.500 Caryophyllene Oxide 0.41 26.810 13-Dococenamide 0.20

As shown in Table 2, as a result of analyzing the compounds contained in Saturella Montana essential oil, 5-isopropyl-2-methylphenol (76.40%), ortho-cymene (11.59%), beta-caryophyllene (2.45%) %), linanol (1.62%), alpha-pinene (1.37%) and gamma-terpinene (1.37%).

Table 3 below shows the chemical composition of Amorasia rusticana essential oil.

Retention Time (min) Library Search Composition ratio (%) 4.841 Butyl isothiocyanate 1.43 6.035 Decane 20.54 6.218 1,1-dimethoxycyclohexane 3.77 9.631 dodecane 2.13 13.805 phenethylisothiocyanate 65.86 24.956 1,2-benzenedicarboxylic acid 6.28

As shown in Table 3, as a result of analyzing the compounds contained in Amorasia rusticana essential oil, phenethylisothiocyanate (65.86%), decane (20.54%), 1,2-benzenedicarbosyl acid (6.28%) ), 1,1-dimethoxycyclohexane (3.77%), dodecane (2.13%) and butylisothiocyanate (1.43%) were identified.

Example 3-2. Evaluation of repellent activity of essential oil components of Saturella Montaña

The repellent activity of the components of the essential oil of Saturella montana analyzed in Example 3-1 against the small sturgeon mite was tested using a climbing test. The results are shown in Table 4 below.

In order to evaluate the repellent activity of the components of the Saturella Montaña essential oil, 5-isopropyl-2-methylphenol, ortho-cymene, beta-caryophyllene, linanol, beta-myrcene, alpha-pinene and gamma - Terpinene was purchased from Sigma-Aldrich and used. Table 4 below shows the results of the repellent activity of the components of the essential oil of Saturella Montaña against the small sophid mite nymphs using the climbing test.

sample density(%) Avoidance rate (average standard error, %) Elapsed time (minutes) after sample processing 30 60 5-isopropyl-2-methylphenol 1.0 100.0±0.0 83.3±3.3 ortho-cymene 1.0 6.7±3.3 0.0±0.0 Beta-Caryophyllene 1.0 26.7±3.3 10.0±10.0 Linanol 1.0 0.0±0.0 0.0±0.0 beta-myrcene 1.0 10.0±10.0 0.0±0.0 alpha-pinene 1.0 0.0±0.0 0.0±0.0 gamma-terpinene 1.0 0.0±0.0 0.0±0.0

As shown in Table 4, 5-isopropyl-2-methylphenol at a concentration of 1.0% showed excellent repelling activity of 83.3% up to 60 minutes. On the other hand, ortho-cymene, beta-caryophyllene, linol, beta-myrcene, alpha-pinene and gamma-terpinene showed low or no repellent activity at a concentration of 1.0%. Therefore, it was confirmed that the active ingredient showing the repelling activity of Saturella montana essential oil was 5-isopropyl-2-methylphenol.

Example 3-3. Evaluation of repellent activity of essential oil components of Amorasia rusticana

The repellent activity of the components of the essential oil of Amorasia rusticana analyzed in Example 3-1 against the small sophid mite was assayed using a climbing test.

In order to evaluate the repelling activity of Amorasia rusticana essential oil components, butyl isothiocyanate, decane, dodecane and phenethyl isothiocyanate were purchased from Sigma-Aldrich and used.

Table 5 below shows the repellent activity results of the components of the Amorasia rusticana essential oil against the small sophid mite nymphs using the climbing test.

sample density(%) Avoidance rate (average standard error, %) Elapsed time (minutes) after sample processing 30 60 Butyl isothiocyanate 1.0 13.3±3.3 0.0±0.0 Decane 1.0 0.0±0.0 0.0±0.0 dodecane 1.0 6.7±3.3 0.0±0.0 phenethylisothiocyanate 1.0 100.0±0.0 93.3±3.3

As shown in Table 5, 5-isopropyl-2-methylphenol at a concentration of 1.0% showed excellent repelling activity of 93.3% up to 60 minutes. On the other hand, butyl isothiocyanate, decane and dodecane showed low or no repellent activity at a concentration of 1.0%. Therefore, it was confirmed that the active ingredient showing the repelling activity of Amorasia rusticana essential oil was phenethylisothiocyanate.

Example 5. Evaluation of repellent activity of active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil, and their derivatives

Example 5-1. Comparison of repelling activity of 5-isopropyl-2-methylphenol and its derivatives

'5-isopropyl-2-methylphenol compound', an active ingredient of the essential oil of Saturella Montaña identified in Example 3-2, and '5-isopropyl-2-methylaniline', a derivative compound thereof, The repellent activity of each concentration (0.5%, 1.0%, 2.0% and 3.0%) against mites was compared using a climbing test.

Table 6 below shows the results of repelling activity of 5-isopropyl-2-methylphenol and its derivatives against nymphs.

sample density(%) Avoidance rate (average standard error, %) Elapsed time (minutes) after sample processing 30 60 5-isopropyl-2-methylphenol 3.0 100.0±0.0 100.0±0.0 2.0 100.0±0.0 100.0±0.0 1.0 100.0±0.0 83.3±3.3 0.5 63.3±5.8 26.7±5.8 5-isopropyl-2-methylaniline 3.0 100.0±0.0 100.0±0.0 2.0 86.7±8.8 63.3±6.7 1.0 76.7±8.8 40.0±5.8 0.5 56.7±5.8 23.3±5.8

As shown in Table 6, 5-isopropyl-2-methylphenol showed 100% repelling activity up to 60 minutes at concentrations of 3.0% and 2.0%, and 83.3% at 1.0% and 0.5% concentrations up to 60 minutes. showed repellent activity. In addition, 5-isopropyl-2-methylaniline exhibited repellent activity of 100%, 63.3%, and 40%, respectively, at concentrations of 1.0%, 2.0%, and 3.0% up to 60 minutes.

Example 5-2. Comparison of repellent activity of phenethylisothiocyanate and its derivatives

Concentrations of 'phenethylisothiocyanate', an active ingredient of the essential oil of Amorasia rusticana identified in Example 3-3, and 'benzylisothiocyanate', a derivative compound thereof, against small sophid ticks (0.5%, 1.0%, 2.0% and 3%) were compared using a climbing test.

Table 7 shows the results of the repelling activity of phenethylisothiocyanate and its derivatives against small scorpion mite nymphs.

sample density(%) Avoidance rate (average standard error, %) Elapsed time (minutes) after sample processing 30 60 Benzylisothiocyanate 3.0 100.0±0.0 100.0±0.0 2.0 100.0±0.0 86.7±6.7 1.0 96.7±3.3 73.3±3.3 0.5 63.3±5.8 46.7±5.8 phenethylisothiocyanate 3.0 100.0±0.0 100.0±0.0 2.0 100.0±0.0 100.0±0.0 1.0 100.0±0.0 93.3±3.3 0.5 73.3±5.8 53.3±5.8

As shown in Table 7, phenethylisothiocyanate exhibited 100% repelling activity for up to 60 minutes at concentrations of 3.0% and 2.0%, and 93.3% repelling activity for up to 60 minutes at 1.0% concentration. . Benzyl isothiocyanate exhibited 100% repelling activity up to 60 minutes at a concentration of 3.0%, and 86.7% and 73.3% repelling activity up to 60 minutes at 2.0% and 1.0% concentrations, respectively.

Example 6. Evaluation of synergistic effect of repellent activity according to 1: 1 mixing of active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil and derivative compounds thereof

Repelling active compounds identified in Examples 5-1 and 5-2 (1) 5-isopropyl-2-methylphenol, (2) 5-isopropyl-2-methylaniline, (3) benzylisothiocyanate and (4) phenethylisothiocyanate. In order to confirm the synergistic effect of repellent activity according to 1: 1 mixing of each compound, the repellent activity was tested using a climbing test, and the significance of the synergistic effect of repellent activity was verified by Duncan's multiple A range test was used ( P <0.05).

Table 8 shows the results of comparison of repellent activity according to 1: 1 mixing of active ingredients of Saturella montana essential oil and Amorasia rusticana essential oil and derivative compounds thereof.

sample density(%) Avoidance rate (average standard error, %) Elapsed time (minutes) after sample processing 60 5-isopropyl-2-methylphenol (1) 0.5 26.7±5.8ef 5-isopropyl-2-methylaniline (2) 0.5 23.3±5.8f Benzylisothiocyanate (3) 0.5 46.7±5.8cd Phenethylisothiocyanate (4) 0.5 53.3±5.8bc (1)+(2) 0.5 40.0±10.0d (1)+(3) 0.5 63.3±5.8ab (1)+(4) 0.5 66.7±5.8a (2)+(3) 0.5 36.7±5.8de (2)+(4) 0.5 43.3±5.8cd (3)+(4) 0.5 70.0±10.0a

As shown in Table 8, the repelling activity after 60 minutes of single treatment of compounds (1), (2), (3) and (4) and (1) + (2), (1) + (3), As a result of comparing the repelling activity after 60 minutes of (1) + (4), (2) + (3) and (3) + (4) mixtures (mixing in a 1: 1 weight ratio), compounds (1) + ( 2), (1) + (3), (1) + (4) and (3) + (4) mixtures showed a significant increase in repellent activity compared to each single treatment (see Fig. 2).

Claims (6)

a mixture of 5-isopropyl-2-methylphenol (1) and 5-isopropyl-2-methylaniline (2); a mixture of 5-isopropyl-2-methylphenol (1) and phenethylisothiocyanate (3); a mixture of 5-isopropyl-2-methylaniline (2) and phenethylisothiocyanate (3); a mixture of 5-isopropyl-2-methylaniline (2) and benzylisothiocyanate (4); And a mixture of phenethyl isothiocyanate (3) and benzyl isothiocyanate (4); repellent active composition for controlling small sophie ticks, characterized in that it comprises one of as an active ingredient. According to claim 1,
The active ingredient is a mixture of 5-isopropyl-2-methylphenol (1) and 5-isopropyl-2-methylaniline (2); a mixture of 5-isopropyl-2-methylaniline (2) and phenethylisothiocyanate (3); and a mixture of 5-isopropyl-2-methylaniline (2) and benzylisothiocyanate (4); A repellent active composition for controlling small sophie ticks, characterized in that any one of them. delete According to claim 1,
The mixture is included in a concentration of 0.1 to 10.0 wt% of the composition,
The ratio of the components of each mixture is 1: 1 weight ratio, characterized in that the repellent active composition for controlling small sophie ticks. A mite repellent prepared using the repellent active composition for controlling small scorpion mites according to any one of claims 1, 2, and 4. According to claim 5,
The repellent is characterized in that it is used in the form of one or more formulations selected from the group consisting of aerosols, emulsions, sprays and creams.