KR101894969B1 - Pharmaceutical composition for preventing or treating hearing loss comprising an extract of peanut sprout - Google Patents

Abstract

The present invention relates to a pharmaceutical composition for preventing or treating hearing loss caused by cancer, which comprises a peanut sprout extract as an active ingredient and is capable of preventing or treating cancer-induced hearing loss, .

Description

[0001] The present invention relates to a pharmaceutical composition for preventing or treating cancer-induced hearing loss comprising an extract of peanut sprout as an active ingredient,

The present invention relates to a pharmaceutical composition for preventing or treating hearing loss caused by cancer, which comprises a peanut sprout extract as an active ingredient and is capable of preventing or treating cancer-induced hearing loss, .

In general, cisplatin has been used for solid tumor therapy.

However, such cisplatin has side effects that cause hearing loss, which limits the amount of cisplatin used during treatment.

Studies on antioxidants, N-methyl-D-aspartate (NMDA) antagonists, apoptosis inhibitors, growth factors, and the like have been conducted to find effective substances for prevention and treatment of hearing loss. (Prasher D., Lancet, 352, pp. 1240-1242, 1998).

In addition, there has not been reported a pharmaceutical composition and a food composition for prevention or treatment of hearing loss caused by side effects due to the use of cisplatin until recently.

Accordingly, the present inventors have searched a variety of active substances that can prevent or treat hearing loss caused by the use of cisplatin in natural products having excellent safety, and found that peanut sprout extract has an effect of preventing or treating cancer-induced hearing loss And completed the present invention.

It is an object of the present invention to provide a pharmaceutical composition for preventing or treating hearing loss that can prevent or treat cancer-induced hearing loss by containing peanut sprout extract as an active ingredient.

Another object of the present invention is to provide a food composition for preventing or ameliorating hearing loss that can prevent or ameliorate cancer-induced hearing loss by containing peanut sprout extract as an active ingredient.

The objects of the present invention are not limited to the above-mentioned objects, and other objects not mentioned can be clearly understood by those skilled in the art from the following description.

In order to accomplish the above object, the present invention provides a pharmaceutical composition for preventing or treating cancer-induced hearing loss comprising peanut sprout extract as an active ingredient.

In a preferred embodiment, the anti-cancer agent is cisplatin.

In a preferred embodiment, the peanut bud extract contains 50 μg to 600 μg based on 30 μM of cisplatin.

In a preferred embodiment, the peanut sprout extract increases the expression level of the antioxidant enzyme.

In a preferred embodiment, the antioxidant enzyme is NQO1, HO1, GPx2, Gclc or Catalase.

In a preferred embodiment, the peanut bud extract is extracted with an organic solvent.

In a preferred embodiment, the peanut shoot extract has antioxidant activity through the AKt / Nrf-2 pathway of HEI-OC1 cells, which are neuronal cells.

The present invention further provides a food composition for preventing or ameliorating a cancer-induced hearing loss comprising an extract of peanut sprouts as an active ingredient.

In a preferred embodiment, the anti-cancer agent is cisplatin.

In a preferred embodiment, the peanut bud extract contains 50 μg to 600 μg based on 30 μM of cisplatin.

In a preferred embodiment, the peanut sprout extract increases the expression level of the antioxidant enzyme.

In a preferred embodiment, the antioxidant enzyme is NQO1, HO1, GPx2, Gclc or Catalase.

In a preferred embodiment, the peanut bud extract is extracted with an organic solvent.

In a preferred embodiment, the peanut shoot extract has antioxidant activity through the AKt / Nrf-2 pathway of HEI-OC1 cells, which are neuronal cells.

The present invention has the following excellent effects.

According to the pharmaceutical composition for preventing or treating hamster-induced hearing loss of the present invention and the food composition for preventing or ameliorating hamster-induced hearing loss, the peanut bud extract is contained as an effective ingredient, and when the cisplatin is used for treating hamam cancer, By protecting against toxicity, it has the advantage of preventing or treating side effects of hearing loss.

FIG. 1 is a graph showing cell viability results of cisplatin-treated acinar cells pretreated with peanut bud extract-treated auricular cells and peanut bud extract; FIG.
FIG. 2 is a graph showing cell survival discrimination test results of cisplatin-treated acinar cells, cisplatin-treated acinar cells pretreated with peanut bud extract-treated acinar cells and peanut sprout extract.
FIG. 3 is a photograph showing apoptotic exchange of cisplatin-treated acinar cells treated with cisplatin, peanut sprout extract-treated acinar cells and peanut sprout extract.
FIG. 4 is a graph showing the apoptosis rate of cisplatin-treated acinar cells, cisplatin-treated acinar cells pretreated with peanut sprout extract treated with peanut bud extract-treated acinar cells.
FIG. 5 is a graph showing the expression levels of apoptosis-related and anti-apoptotic-related markers of cisplatin-treated acinar cells pretreated with cisplatin-treated acinar cells, peanut sprout-treated acinar cells and peanut sprout extracts.
FIG. 6 is a graph showing the results of reactive oxygen species (ROS) analysis of cisplatin-treated acinar cells treated with cisplatin, peanut sprout extract-treated acinar cells and peanut sprout extract.
FIGS. 7 to 11 show the results of comparing the amounts of antioxidant enzymes expressed in cisplatin-treated aural neurons, cisplatin-treated neurons after pretreatment with peanut bud extract and peanut bud extract, FIG. 7 shows NQO1 , FIG. 8 shows HO1, FIG. 9 shows GPx2, FIG. 10 shows Gclc, and FIG. 11 shows mRNA levels of Catalase.

Although the terms used in the present invention have been selected as general terms that are widely used at present, there are some terms selected arbitrarily by the applicant in a specific case. In this case, the meaning described or used in the detailed description part of the invention The meaning must be grasped.

Hereinafter, the technical structure of the present invention will be described in detail with reference to preferred embodiments shown in the accompanying drawings.

However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Like reference numerals designate like elements throughout the specification.

The pharmaceutical composition for the prevention or treatment of induced hearing loss according to one embodiment of the present invention is a composition for preventing or treating hearing loss, which is a side effect caused by the use of a cancer drug, and contains peanut bud extract as an active ingredient.

Here, the anti-cancer agent may be cisplatin.

The peanut sprout extract is prepared by extracting with an organic solvent using dried powder of peanut sprouts, and the organic solvent present in the peanut sprout extract is removed with a rotary vacuum concentrator. The organic solvent may be ethanol, but is not limited thereto.

The peanut sprout extract enhances the expression of antioxidant enzymes in the nerve cells, and the antioxidant enzymes are NQO1, HO1, GPx2, Gclc or Catalase.

It is preferable that the peanut sprout product contains 50 μg to 600 μg based on 30 μM of cisplatin.

The reason for this is that when the peanut sprout extract is less than 50 μg, the protective effect of the auditory neurons is insignificant, and when the peanut sprout extract is more than 600 μg, the protective effect of the auditory neurons is not enhanced It is not.

That is, the pharmaceutical composition for the prevention or treatment of cancer-induced hearing loss according to the present invention maximizes the expression level of antioxidant enzyme through the AKt / Nrf-2 pathway of HEI-OC1 cells, which are neuronal cells, when using cisplatin for chemotherapy Thereby protecting the aural cell from cytotoxicity caused by cisplatin.

Such an effect has the advantage of preventing or treating side effects of hearing loss caused by the use of cisplatin.

In addition, the pharmaceutical composition of the present invention may contain a pharmaceutically acceptable carrier and may be formulated into a liquid formulation, suspension, or the like according to a conventional method. The pharmaceutically acceptable carriers include aqueous diluents or solvents such as phosphate buffered saline, purified water, sterilized water, etc., and include non-aqueous diluents or solvents such as propylene glycol, olive oil and the like . Further, if necessary, it may contain a wetting agent, a fragrance, a preservative, and the like. The peanut sprout extract contained in the pharmaceutical composition may be appropriately selected by a person skilled in the art depending on the condition and body weight of the individual, the degree of disease, the drug form, the administration route and the period.

The present invention also provides a food composition for preventing or ameliorating a cancer-induced hearing loss comprising an extract of peanut buds as an active ingredient.

Meanwhile, the anti-cancer agent and the peanut bud extract are the same as those described above, so duplicate descriptions are omitted.

There is no particular limitation on the type of food contained in the food composition of the present invention. Examples of the food include dairy products including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen and other noodles, gums and ice cream, various soups, drinks, tea, drinks, alcoholic beverages and vitamin complex And includes all health foods in a conventional sense.

[Production Example] Preparation of peanut sprout extract of the present invention

The peanut sprouts were cleaned with water to remove foreign matter, and then steam was added to cool the peanut sprouts.

Then, the peanut sprouts were lyophilized. The lyophilized peanut sprouts were extracted with ethanol as an organic solvent at a temperature of 60 ° C for 90 minutes, and then the ethanol was removed with a rotary vacuum concentrator to obtain a peanut sprout extract.

[Experimental Example 1]

In this experiment, the cells were pretreated with peanut sprout extracts and peanut sprout extracts, and cell differentiation test of cisplatin treated acinar cells was performed.

In this experiment, we used the House Ear Institute-Organ of Corti 1 (HEI-OC1) as an auditory neuron and cultured by conventional method.

Then, the neuron cells were exposed to peanut sprout extract for 24 hours.

The prepared neurons were pretreated with peanut bud extracts of 0, 50, 100, 200, 300, 400, 500, 600 and 700 μg for 2 hours, and the neurons pretreated with peanut bud extract were exposed to cisplatin for 24 hours .

At this time, cisplatin was used at 30 μM.

Thereafter, a normal MTT assay was performed, and the results are shown in Fig.

1, when the peanut bud extract extracts were 50, 100, 200, 300, 400, 500, 600 and 700 μg, the extracts of peanut sprout extracts were 100.0 ± 1.2%, 103.3 ± 0.9% Survival rates were 96.5 ± 0.2%, 109.2 ± 2.5%, 112.2 ± 2.9%, 106.5 ± 0.4%, 109.7 ± 3.4% and 118.8 ± 1.5%, respectively.

In the case of cisplatin-treated acinar cells pretreated with peanut sprout extract, 54.7 ± 1.0% and 59.2 ± 1.2% of peanut shoot extracts were obtained at 50, 100, 200, 300, 400, 500, 600 and 700 μg, respectively %, 60.6 ± 1.1%, 75.1 ± 3.0%, 67.6 ± 0.7%, 71.8 ± 2.6%, 69.1 ± 2.4%, and 69.6 ± 1.4%, respectively.

This means that the peanut sprout extract according to the present invention does not affect cell cytotoxicity.

In addition, the peanut sprout weight indicates that an effective range of 50 μg to 600 μg based on 30 μM of cisplatin is effective.

The reason for this is as follows. When the peanut bud extract is less than 50 μg, the protective effect of the neuron cell is insignificant. When the peanut bud extract is more than 600 μg, the protective effect of the neuron is not enhanced. Because.

[Experimental Example 2]

In this experiment, the cells were pretreated with cisplatin-treated acinar cells, peanut sprout extract-treated acinar cells, and peanut sprout extracts, and cell-lysing tests were performed on cisplatin-treated acinar cells.

In this experiment, we used the House Ear Institute-Organ of Corti 1 (HEI-OC1) as an auditory neuron and cultured by conventional method.

Then, the prepared neuron cells were treated with cisplatin-treated cisplatin-treated cisplatin-treated cisplatin cells and the neuronal cells were treated with 300 μg of peanut sprout extract, and treated with 300 μg of peanut sprout extract, Of cisplatin-treated peanut bud extract and cisplatin-treated acinar cells, respectively.

Thereafter, a normal MTT assay was performed, and the results are shown in FIG.

2, peanut sprout extract and cisplatin-treated acinar cells showed significantly higher cell survival rates than cisplatin-treated acinar cells.

[Experimental Example 3]

In this experiment, the cells were pretreated with cisplatin-treated acinar cells, peanut sprout extract-treated acinar cells, and peanut sprout extracts, and cell-lysing tests were performed on cisplatin-treated acinar cells.

For this, in this experiment, neuron cells were prepared under the same conditions as in Experimental Example 2. The prepared neurons were treated with cisplatin-treated cisplatin cells treated with 30 μM of cisplatin, and the neurons were treated with 300 μg of peanut- Cells and auditory neurons were pretreated with 300 μg of peanut sprout extract, followed by 30 μM of cisplatin-treated peanut sprout extract and cisplatin-treated auditory neuronal cells, respectively.

Thereafter, double staining of Hoechst 33258 and propidium iodide (PI) was performed in a usual manner, and the results are shown in Fig.

3, the peanut bud extract and cisplatin-treated acinar cells showed significantly reduced dead cells compared to cisplatin-treated acinar cells.

[Experimental Example 4]

In this experiment, the apoptosis rate of cisplatin-treated auricular cells treated with cisplatin, peanut sprout extract-treated auricular cells and peanut sprout extract was analyzed.

For this, in this experiment, neuron cells were prepared under the same conditions as in Experimental Example 2. The prepared neurons were treated with cisplatin-treated cisplatin cells treated with 30 μM of cisplatin, and the neurons were treated with 300 μg of peanut- Cells and auditory neurons were pretreated with 300 μg of peanut sprout extract, followed by 30 μM of cisplatin-treated peanut sprout extract and cisplatin-treated auditory neuronal cells, respectively.

Thereafter, cell death rate analysis was performed by a conventional method, and the results are shown in FIG.

4, peanut sprout extract and cisplatin-treated acinar cells showed significantly lower cell death rate than cisplatin-treated acinar cells.

[Experimental Example 5]

In this experiment, we analyzed the expression levels of apoptotic and anti - apoptotic markers of cisplatin - treated acinar cells after pretreatment with cisplatin - treated acinar cells, peanut sprout - treated acinar cells and peanut sprout extracts.

For this, in this experiment, neuron cells were prepared under the same conditions as in Experimental Example 2. The prepared neurons were treated with cisplatin-treated cisplatin cells treated with 30 μM of cisplatin, and the neurons were treated with 300 μg of peanut- Cells and auditory neurons were pretreated with 300 μg of peanut sprout extract, followed by 30 μM of cisplatin-treated peanut sprout extract and cisplatin-treated auditory neuronal cells, respectively.

Thereafter, western blot analysis was performed in a usual manner, and the results are shown in Fig.

5, peanut sprout extracts and cisplatin-treated neurons exhibited decreased expression of cleaved caspase-3 (cleaved PARP) and anti-apoptosis-related markers (Bcl-2) The expression level of the gene was increased.

[Experimental Example 6]

In this experiment, reactive oxygen species (ROS) analysis of cisplatin treated neurons, pretreatment with peanut sprout extract treated neurons and peanut sprout extract, and cisplatin treated neurons were performed.

For this, in this experiment, neuron cells were prepared under the same conditions as in Experimental Example 2. The prepared neurons were treated with cisplatin-treated cisplatin cells treated with 30 μM of cisplatin, and the neurons were treated with 300 μg of peanut- Cells and auditory neurons were pretreated with 300 μg of peanut sprout extract, followed by 30 μM of cisplatin-treated peanut sprout extract and cisplatin-treated auditory neuronal cells, respectively.

Thereafter, reactive oxygen species (ROS) analysis was performed in a conventional manner, and the results are shown in FIG.

Referring to FIG. 6, peanut bud extract and cisplatin-treated acinar cells showed a decrease in active oxygen compared with cisplatin-treated acinar cells.

[Experimental Example 7]

In this experiment, antioxidant enzyme expression levels of cisplatin-treated neurons, pretreatment with peanut sprout-treated cisternae and peanut sprout extracts were analyzed.

For this, in this experiment, neuron cells were prepared under the same conditions as in Experimental Example 2. The prepared neurons were treated with cisplatin-treated cisplatin cells treated with 30 μM of cisplatin, and the neurons were treated with 300 μg of peanut- Cells and auditory neurons were pretreated with 300 μg of peanut sprout extract, followed by 30 μM of cisplatin-treated peanut sprout extract and cisplatin-treated auditory neuronal cells, respectively.

Thereafter, RT-PCR analysis at the mRNA level was performed in a conventional manner, and the results are shown in FIGS. 7 to 11.

In FIG. 7, the target antioxidant enzyme is NQO1, FIG. 8 is HO1, FIG. 9 is GPx2, FIG. 10 is Gclc, and FIG.

7 to 11, it was confirmed that the peanut bud extract and cisplatin-treated acinar cells significantly increased the expression levels of all antioxidant enzymes compared with cisplatin-treated acinar cells.

As described above, the pharmaceutical composition for preventing or treating hamster-induced hearing loss according to the present invention and the food composition for preventing or ameliorating hamster-induced hearing loss include HEI-OC1 cells The AKT / Nrf-2 pathway maximizes the activity of the antioxidant enzyme, thereby protecting the aural cell from the cytotoxicity caused by cisplatin, thereby preventing, ameliorating or treating hearing loss.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it is clearly understood that the same is by way of illustration and example only and is not to be taken by way of limitation, Various changes and modifications will be possible.

Claims (14)

A pharmaceutical composition for preventing or treating cancer-induced hearing loss comprising peanut sprout extract as an active ingredient.
The method according to claim 1,
The anti-
A pharmaceutical composition for preventing or treating cancer-induced hearing loss characterized by being cisplatin.
3. The method of claim 2,
The peanut sprout extract
Wherein the composition contains 50 μg to 600 μg based on 30 μM of cisplatin.
The method of claim 3,
The peanut sprout extract
Wherein the expression level of the antioxidant enzyme is increased.
5. The method of claim 4,
The antioxidant enzyme
NQO1, HO1, GPx2, Gclc or Catalase.
6. The method of claim 5,
The peanut sprout extract
Wherein the composition is extracted with an organic solvent.
The method according to claim 6,
Wherein the peanut sprout extract has an antioxidative activity through the AKt / Nrf-2 pathway of HEI-OC1 cells, an auditory neuron cell.
A food composition for preventing or ameliorating cancer-induced hearing loss comprising peanut sprout extract as an active ingredient.
9. The method of claim 8,
The anti-
Wherein the cisplatin is cisplatin.
10. The method of claim 9,
The peanut sprout extract
Wherein the composition contains 50 μg to 600 μg based on 30 μM of cisplatin.
11. The method of claim 10,
The peanut sprout extract
Wherein the expression level of the antioxidant enzyme is increased.
12. The method of claim 11,
The antioxidant enzyme
NQO1, HO1, GPx2, Gclc, or Catalase.
13. The method of claim 12,
The peanut sprout extract
The composition for preventing or ameliorating a cancer-induced hearing loss characterized by being extracted with an organic solvent.
14. The method of claim 13,
Wherein the peanut sprout extract has an antioxidative activity through AKt / Nrf-2 pathway of HEI-OC1 cells, which is an auditory neuron cell, for preventing or ameliorating hearing loss caused by cancer.

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