KR20190057010A - Pest control composition comprising Codonopsis pilosula (Franch.) Nannf. extract as active ingredient - Google Patents

Abstract

The present invention relates to a pest control composition comprising a Codonopsis pilosula (Franch.) Nannf. extract as an active ingredient, to a pest control agent, and to a pest control method using the same. The pest control composition comprising the Codonopsis pilosula (Franch.) Nannf. extract or a fraction thereof as an active ingredient shows a pest control effect minimizing harm to humans and harm to environment by suppressing the activity of receptors of insect-specific small neuropeptides, and thus the pest control agent comprising the pest control composition and the pest control method using the same can be utilized for eco-friendly pest control.

Description

[0001] The present invention relates to a pest control composition comprising Codonopsis pilosula (Franch.) Nannf. extract as active ingredient}

The present invention relates to a composition for controlling insect pests, an insect pest controlling agent and a pest controlling method using the same.

Conventional pest control agents including organic phosphorus have problems such as environmental harm and human harmfulness, and thus use of pesticides including insecticides is limited. As interest in environmentally friendly agricultural products is increased, production of high quality organic agricultural products Farms are increasing. Most pest control technologies that have a significant impact on the production and quality of agricultural products are not eco-friendly or ineffective.

Among the pest control techniques, there is a method of using an insect hormone as a biological control method. Insect hormones are organic compounds that are produced in the body of insects and act effectively in trace amounts. Insect hormones act as chemical messengers that transmit information in and out of the body to specific target cells and regulate the activity of the target cells, allowing insects to maintain their normal life cycle. Insect hormones include neuropeptide hormones synthesized and secreted by neurons in the central nervous system and hormones synthesized and secreted by the endocrine glands.

The common functional groups of neuropeptide hormones in insects are as follows. Neuropeptide hormones bind to receptors in specific target cells with high specificity and affinity to regulate cellular function. In particular, short neuropeptide F, which regulates the various physiological activities of insects, binds to the G protein coupled receptor (GPCR) and activates the adenylate cyclase inside the cell membrane to release cAMP (cyclic AMP) from ATP This cAMP, as a secondary messenger, transmits information to the intracellular enzymes or molecular systems.

Small neuropeptide receptors are known to regulate feeding, reproduction and individual growth by activating various sub-signaling pathways, and are distributed throughout several senses. The present inventors for the first time reported using a Drosophila melanogaster model belonging to the family of Dicloviridae (Drosophila melanogaster ) for the first time that a small neuropeptide, a neuropeptide, modulates the expression of an insulin gene and is involved in metabolic function, individual growth, and lifespan control of Drosophila lee et al., Nature Cell Biol ., 2008), especially PKA-CREB-DYRK path to the latest in sub-signaling pathways in a small neuropeptide reported to induce feeding behavior (Hong et al. PLoS Genetics, 2012).

In particular, insect neuropeptides are very well conserved among insect species, but they differ in amino acid sequence and structure from mammalian neuropeptides with similar functions. Therefore, it is known that compounds or agents that inhibit insect neuropeptide signals do not act on mammalian neuropeptide signals or have very low activity. When insect-specific neuropeptide receptor inhibitors are discovered, But it does not work on mammals, including humans, and can reduce human toxicity.

Codonopsis pilosula (Franch.) Nannf.) Is a perennial plant of the bellflower and its roots are especially called ginseng.

Manchans have been used as treatments for increasing red blood cells, decreasing white blood cells, improving immunity, tonic action, and peptic ulcer due to excessive acid in the private sector. In addition, Manzan is a medicine used as a substitute for ginseng to people who have side effects of ginseng.

Korean Patent Registration No. 10-0681702 discloses a cosmetic composition for preventing skin aging which comprises extract of manchus as an active ingredient and Korean Patent Registration No. 10-1090319 discloses a cosmetic composition for preventing aging of skin, The present invention relates to a pharmaceutical composition for the prevention and treatment of diabetes, which comprises a mixed extract of herbal medicines consisting of oyster oil, yin yang, rhubarb, lobule, licorice, persicaria, taxa,

Thus, there is no invention of a pest control agent containing the extract of Manchurus. Especially, the action of the extract of the ginseng extract against the insect neuron peptide and the endocrine system is not known at all.

Under these circumstances, the present inventors have made intensive efforts to develop an environmentally-friendly pest control agent, and as a result, they have found that the extract of Manx ginseng effectively regulates the activity of neuropeptide receptors, thereby reducing insect feeding and increasing food avoidance, And confirming that there is almost no toxicity to humans, thereby completing the present invention.

Korean Patent Publication No. 10-0681702 Korean Patent Publication No. 10-1090319

It is an object of the present invention to provide a composition for controlling insects, which comprises the extract of L. ganoderma or a fraction thereof as an active ingredient.

Another object of the present invention is to provide a pest controlling agent comprising the composition for pest control.

It is still another object of the present invention to provide a pest control method comprising the step of treating the insect pest controlling composition.

It is still another object of the present invention to provide a quasi-drug composition comprising the composition for controlling insects.

Each description and embodiment disclosed in the present application can be applied to each other description and embodiment. That is, all combinations of the various elements disclosed in this application fall within the scope of the present application. Further, the scope of the present application is not limited by the specific description described below.

As one embodiment of the present invention, the present invention provides a composition for controlling insects, which comprises extract of L. ganoderma or a fraction thereof as an active ingredient.

In the present invention, it has been confirmed that the extract of L. ganoderma or a fraction thereof has an effect of inhibiting the activity of small neuropeptide receptors known to regulate feeding, reproduction and individual growth of insects, and can be used for pest control.

In the present invention, the term " ginseng " is a perennial plant of the bellflower, and its root is particularly called ginseng, and its scientific name is Codonopsis pilosula (Franch.) Nannf. Or Codonopsis tangshen Oliv. to be. In addition, manchans are also called ginseng or perennial herb.

In China, dried roots of C. pilosula (Franch.) Nannf. And C. tangshem Oliv are not called manzan but called ginseng . It is called heaven.

Mancham is native to Gangwon-do and Gyeongsangnam-do in the Republic of Korea.

For the purpose of the present invention, manzan is not limited to its kind, and cultivated or commercially available manzan can be used without limitation in its source.

Furthermore, the specific efficacy of manzan as a pest control agent is not known. Accordingly, the present inventors confirmed that the extract of L. ginseng inhibited insect-specific small neuropeptide receptors, reduced insect feeding and increased insect pest resistance to the food containing L. extract, This is significant in that it is the first time that an environmentally-friendly pest control composition was developed.

The term " extract " in the present invention means an extract obtained by extracting ginseng, a diluted or concentrated solution of the extract, a dried product obtained by drying the extract, a controlled preparation or a purified product of the extracted solution, And extracts of all formulations that can be formed using an extract.

In the present invention, the extract of Manx ginseng comprises the steps of washing, drying and pulverizing ginseng; And extracting the crushed ginseng with a solvent selected from the group consisting of water, C ₁ to C ₆ alcohols and a mixed solvent thereof. Specifically, the extract may be methanol, ethanol, propanol, butanol, pentanol or hexanol, and more specifically ethanol extract.

The extraction method is not particularly limited, and extraction can be carried out at room temperature or with heating under conditions where the active ingredient is not destroyed or minimized. More specifically, the method for obtaining ginseng extract in the present invention is as follows. The dried mannan is dissolved in a polar solvent of a C ₁ to C ₆ alcohol such as water, methanol, ethanol, propanol, butanol, pentanol and hexanol, etc., of about 2 to 20 times, Or a mixed solvent thereof having a mixing ratio of about 1: 0.1 to 1:10 may be used as an elution solvent, but the present invention is not limited thereto. The extraction temperature may be 1 to 100 ° C, specifically 25 ° C, and the extraction period may be about 1 hour to 10 days, specifically, 2 to 50 hours. The extraction method may include shaking extraction, hot water extraction, Reflux cooling extraction, ultrasonic extraction, or a combination thereof. However, the present invention is not limited thereto. In a specific embodiment of the present invention, the ginseng crushed material was immersed in 95% ethanol, sonicated at 45 캜 for 15 minutes, and extracted at a frequency of 10 times per day for 72 hours. The extract was filtered with an incompatible cotton and then lyophilized to obtain ginseng extract.

The ginseng extract can be extracted from natural, hybrid, and variant plants and extracted from plant tissue cultures.

The term " fraction " in the present invention means a product obtained by performing fractionation to separate a specific component or a specific component group from a mixture containing various components.

The fractionation method for obtaining the fraction in the present invention is not particularly limited and may be carried out according to a method commonly used in the art. As a non-limiting example of the above-mentioned fractionation method, a fraction obtained from the extract can be obtained by treating the extract obtained by extracting ginseng with a predetermined solvent.

The kind of the fraction solvent used for obtaining the fraction in the present invention is not particularly limited, and any solvent known in the art can be used. Non-limiting examples of the fraction solvent include polar solvents such as water, alcohol and the like; And non-polar solvents such as hexane, ethylacetate, acetone, and chloroform. These may be used alone or in combination of two or more. In addition, a method of extracting ginseng and passing the ginseng through an ion exchange resin may be used. In the present invention, the ion exchange resin used for fractionating ginseng is more specifically a synthetic adsorbent, which is an aromatic non-substituted resin HP-20 series, SP825 series, AXT204 series, XAD1600T series, MN200 series).

The term " insect pests " in the present invention refers to all insects which directly or indirectly harm human life. In the present invention, the insect pests may be insects harmful to animals or plants, and may also be human disease-mediated insects, farm insects, horticultural insects and horticultural insects and wooden insects. And inhibiting the activity of the neuropeptide receptor.

Specifically, the pests are selected from the group consisting of Egyptian forest mosquito, white mosquito mosquito, school mosquito, Chinese spotted mosquito, tropical house mosquito, mite, thripworm, Aphids, pea aphids, tobacco may be Louis or potato beetles, but are not limited thereto.

The term " pest control " in the present invention is a series of actions for reducing harm caused by a pest. The pest control method specifically includes physical (mechanical) control, poison control, chemical control, biological control, legal control, But are not limited to, solar heating, hot water impregnation, steam heating, pyrogen heating, infrared cooling, cooling, chopping, ultrasound, embedding, immersion and eco-control.

"Control" is a series of actions to reduce the harm caused by a pest, which can be distinguished from an "insect" that necessarily involves killing the pest. Therefore, using the composition for pest control of the present invention may have little or no effect of disturbing the ecosystem. In addition, since the above-mentioned " control " may include " avoidance ", the composition for controlling insects of the present invention may also be used as a composition for avoiding insects.

In the present invention, the extract of L. ganoderma or a fraction thereof is used in an amount of 0.5 to 5 wt% (w / w), specifically 0.5 to 4 wt%, 0.5 to 3 wt%, 0.5 to 2 wt%, 0.5 to 5 wt% 1 to 5 wt.%, 2 to 5 wt.%, 3 to 5 wt.% Or 4 to 5 wt.%.

If the extract of lanx or extract thereof contains less than 0.5% by weight (w / w) of the total weight of the composition, insect control effect is lowered due to insufficient inhibition of small neuropeptide receptors of insect pests, There is a problem in that the production cost of the homogeneous solution preparation and the controlling agent is increased.

The composition for controlling insect pests may further comprise extracts other than the ginseng extract. As an example, it may further include, but is not limited to, passenger extract.

The composition for insect control of the present invention can inhibit the activity of an insect-specific neuropeptide receptor, and the neuropeptide may be but is not limited to a short neuropeptide F (sNPF).

The term " short neuropeptide F (sNPF) " in the present invention is a peptide hormone of insects. The receptors of small neuropeptides activate various sub-signaling pathways, regulating insect feeding, reproduction and individual growth It is known and distributed evenly in several senses. The composition for controlling pests may exhibit a controlling effect specifically for insects having activity against small neuropeptides, but the present invention is not limited thereto.

In one embodiment of the present invention, it was confirmed that the extract of L. ganoderma suppresses the activity of insect-specific small neuropeptide receptors, resulting in a decrease in feeding of Drosophila and an increase in the avoidance of Drosophila against the food containing L. extract .

The above results confirm that the extract of L. ganoderma can be used as an insect control agent.

In another aspect of the present invention, the present invention provides an insect control agent comprising the composition for controlling insects. Pest and pest control are as described above.

In another aspect of the present invention, the present invention provides a pest control method comprising the step of treating the insect pest control composition. Pest and pest control are as described above.

The method may be a method of directly or indirectly treating the insect pest controlling composition of the present invention. However, the insect pest controlling composition may be placed, for example, in the vicinity of humans, animals, Lt; / RTI >

According to another aspect of the present invention, there is provided a quasi-drug composition comprising the above composition for controlling insects.

The quasi-drug is a preparation used for sterilization, insecticide and similar uses for the prevention of the infectious diseases described in the Article 2 (7) of the Pharmaceutical Affairs Act. It is used as an insect repellent for the human or animal health such as flies, mosquitoes, May mean an antiseptic or an attractant insecticide.

The quasi-drug may also include external preparations for skin and personal hygiene products. But is not limited to, a disinfectant cleaner, a shower foam, a guarine, a wet tissue, a detergent soap, a hand wash, or an ointment.

When the composition for controlling insect pests according to the present invention is used as a quasi-drug additive, the composition may be added as it is, or may be used together with other quasi-drugs or quasi-drugs, and may be suitably used according to a conventional method. The amount of the active ingredient to be mixed can be appropriately determined depending on the purpose of use.

The quasi-drug composition of the present invention may be, for example, one prepared in the form of a general emulsified formulation and a solubilized formulation. For example, they may have formulations such as emulsions such as lotions, creams, ointments, sprays, oil gels, gels, oils, aerosols, and softeners, but they can be used without limitation as long as they exhibit the pest control effect of the present invention .

In addition, the quasi-drug composition may be formulated into each formulation as appropriate, if necessary, in an oil, water, surfactant, humectant, lower alcohol having 1 to 4 carbon atoms, a thickener, a chelating agent, a pigment, Can be used.

The composition for insect control comprising the extract of L. ginseng or the fraction thereof as an active ingredient shows insect pest control effect that minimizes human harm and environmental hazard through inhibition of receptor activity of insect specific small neuropeptides, A pest control agent containing a composition for a pest control agent and a pest control method using the same can be used for environmental pest control.

1 shows a method for measuring the activity of a small neuropeptide and its receptor using a cell line injected with a small neuropeptide receptor expression vector (pAc5.1-sNPFR) and a luciferase expression vector (pGL3-CRE) Fig.
FIG. 2 is a graph showing the effect of inhibiting the activity of the small-scale neuropeptide receptor of the extract of the ginseng extract.
FIG. 3 is a graph showing the ingestion inhibitory ability of the extract of L. ganoderma.
Fig. 4 is a diagram showing the effect of avoiding insects by the extract of ginseng.
FIG. 5 is a diagram showing the insecticidal efficacy of the extract of L. ganoderma.
FIG. 6 is a graph showing the non-toxic effect of the extract of L. ginseng on human neurons.

Hereinafter, the present invention will be described in more detail with reference to Examples and Experimental Examples. However, these examples and experimental examples are intended to illustrate the present invention, and the scope of the present invention is not limited to these examples and experimental examples.

Example 1: Preparation of ginseng extract

The slurry was immersed in 95% ethanol. Were sonicated at 45 ° C for 15 min and extracted at a frequency of 10 times per day for 72 h. The extracts were filtered and then lyophilized using a cotton pad. This construct was dissolved in DMSO at 20 mg / ml and then added to the cell culture medium or insect medium at 1 wt% (w / w) and used in the following examples.

Example 2: Measurement of inhibitory activity of small-scale neuropeptide receptor activity of lanthanum extract

Example 2-1: Production of Insect Cell Lines Overexpressing Small Neuropeptide Receptors

The effect of ginseng extract inhibiting small neuropeptide receptor-specific activity using a luminescent assay technique based on the analysis of the target gene promoter of small neuropeptides to analyze the activity of small neuropeptides and their receptors that regulate feeding behavior .

First, a luciferase expression vector (pGL3-CRE) containing an insect expression vector (pAc5.1-sNPFR) and a CREB response element (CRE) cloned in the fruitfly's small neuropeptide receptor (sNPFR) To produce a cell line capable of measuring the activity of a small neuropeptide receptor.

Treatment of small neuropeptides with the pAc5.1-sNPFR and pGL3-CRE vectors induced PKA-CREB signaling by the over-expressed receptor (sNPFR) and CREB transcription factor binding to pGL3-CRE Luciferase is expressed. When luciferin was treated with luciferin in the cells, luciferin was emitted by luciferase, and the activity of the small neuropeptide and its receptor was measured by measuring the degree of luminescence using a luminometer One).

Example 2-2: Measurement of activity of small neuropeptide receptor

The extract of L. ginseng and the cell line were used to inhibit the activity of the extract of L. ginseng on the small neuropeptide receptors.

Specifically, the degree of luminescence of pAc5.1-sNPFR and pGL3-CRE vector-treated cell line (control group) treated with small neuropeptide alone and cell line (experimental group) treated with small neuropeptide and lanx extract were compared.

In order to compare the effects of the extract of Manchurus japonica with the effects of biological pesticides (including insecticidal effects against moth larvae, and extracts of Manchuria japonica, Seiho extract, Maquina extract and reptile extract) Or the degree of luminescence of the cell line treated with each of its components was also compared.

As a result, compared to the control group treated with small neuropeptide alone, the experimental group treated with the small neuropeptide and the lanthanum extract decreased the luciferin luminescence by 20% and inhibited the activity of the small neuropeptide receptor in the extract of lanthanum (Fig. 2).

In addition, the inhibitory effect of the extract of L. ginseng on the small neuropeptide receptor activity was confirmed to be superior to that of the bio-pesticide and its constituents (FIG. 2).

Example 3: Measurement of inhibition of ingestion of the extract of manchus

Example 3-1: Drosophila preparation and breeding

Drosophila was kept at 25 ℃ and used for experiment after rearing in normal medium for 3-5 days after hatching.

Example 3-2: Measurement of feeding amount of fruit fly

The fruit flies were grown in a general insect rearing medium or in a rearing medium containing 1% by weight of the extract prepared from Example 1 for 4 days, starved for 2 hours, and then the number of food consumed including food coloring was measured, The intake was quantified by analysis.

In addition, the same experiment as described above was carried out by using the flies cultured in a rearing medium containing each of the biocidal pesticide of Example 2-2 or its components.

As a result, compared to the control group grown in the insect rearing medium, the fruit fly grown in the rearing medium containing the extract of Lentil was reduced by 60% and the total ingestion by 72%, indicating that the extract of L. ganoderma (Fig. 3).

In addition, it was confirmed that the suppressive effect of the extract of L. ganoderma lucidum was superior to the effect of the biocides and the constituents thereof (Fig. 3).

Example 4: Measurement of avoidance of insects against the extract of lantern ginseng

The following experiment was carried out to measure the avoidance degree of the food containing the extract of Manchus.

Specifically, the fruit fly was raised in the same manner as in Example 3, red or blue edible pigment was added to the rearing medium, and the selection of the medium was added to the blue edible pigment medium of the experimental group only by adding the extract of Example 1 The degree of avoidance after quantification by the color change of the fruit fly abdomen was measured using the following equation.

Avoidance = (Blue-Red) / (Blue + Red + Purple)

As a result, it was confirmed that the avoidance degree of the food containing the extract of L. ginseng was increased 4 times or more as compared with the control group (Fig. 4).

Example 5: Measurement of Insect Killer Effect of Magnesium Extract

The following experiment was carried out to confirm the lethal effects of insecticides against insects.

Specifically, the lethal effect of the extract of L. ganoderma lucidum was determined by inserting 100 adult Drosophila adults raised in the same manner as in Example 3 on the rearing medium supplemented with the extract prepared as in Example 1, .

As a result, it was confirmed that 15% of the fruit flies of all the fruit flies were dead after 5 days of rearing in the culture of the flies raised in the culture medium supplemented with the extract of L. ginseng, thereby confirming the lethal effect of the L. ginseng extract on the insects 5).

Example 6: Measurement of toxicity of mammalian extract of lanthanum extract

The following experiment was carried out to confirm the cytotoxicity of the extract of Manchus to mammals.

Specifically, the cell extract of SK-N-SH, which is a human neural cell line, was treated for 1 day and cell survival rate was measured by WST-1 assay. And compared with the control group.

As a result, there was no significant difference in the survival rate of the experimental group treated with the human ginseng extract and the survival rate of the control group not treated with the ginseng extract, and thus, the ginseng extract was useful as an environmentally friendly insect pest control agent because it has no toxicity to mammals (Fig. 6).

From the above description, it will be understood by those skilled in the art that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. In this regard, it should be understood that the embodiments described above are illustrative in all aspects and not restrictive. The scope of the present invention should be construed as being included in the scope of the present invention without departing from the scope of the present invention as defined by the appended claims.

Claims (11)

A composition for controlling insects, comprising an extract of Codonopsis pilosula (Franch.) Nannf. Or a fraction thereof as an active ingredient.
The composition according to claim 1, wherein the composition inhibits the receptor activity of an insect-specific neuropeptide.
The insect controlling composition according to claim 1, wherein the extract is extracted with a solvent selected from the group consisting of water, an alcohol having 1 to 6 carbon atoms, and a mixed solvent thereof.
4. The composition for controlling insects according to claim 3, wherein the alcohol is ethanol or methanol.
2. The pest controlling composition according to claim 1, wherein the extract or the fraction thereof is contained in an amount of 0.5 to 5 wt% (w / w) based on the total weight of the composition.
The insect controlling composition according to claim 1, wherein the pest is an insect harmful to an animal or a plant.
7. The pest control composition according to claim 6, wherein the pest is any one selected from the group consisting of human disease-mediated insect pests, homestead insect pests, horticultural insect pests and wood insect pests.
The method of claim 7, wherein the pest is selected from the group consisting of Egyptian forest mosquito, white mosquito mosquito, mosquito mite, mite, thrips, brownflies, fruit flies, house flies, fruit flies, grasshoppers, red fire ants and potato beetles By weight of the composition.
9. A pest controlling agent comprising the pest controlling composition according to any one of claims 1 to 8.
9. A pest control method comprising the step of treating a pest control composition according to any one of claims 1 to 8 to a pest.
A quasi-drug composition comprising the pest control composition of any one of claims 1 to 8.

Images