KR102372822B1 - Composition for preventing or treating kidney damage on heavy metal comprising extracts from Dendropanax morbifera - Google Patents

Abstract

The present invention relates to a composition for preventing or treating kidney damage caused by heavy metals containing hwangchil extract, hwangchil complex extract or hwangchil complex fermented extract as an active ingredient, LLC- As a result of inducing toxicity by treating PK1 kidney cells with cadmium, it was confirmed that cytotoxicity and intracellular active oxygen generation were reduced compared to untreated kidney cells, so the Hwangchil extract, Hwangchil complex or Hwangchil complex fermented product The composition contained as a component exhibits the possibility of alleviating kidney damage caused by cadmium, and thus can be provided as a useful pharmaceutical composition and health food for preventing and treating kidney damage caused by heavy metals.

Description

Composition for preventing or treating kidney damage caused by heavy metals containing Hwangchil extract as an active ingredient {Composition for preventing or treating kidney damage on heavy metal comprising extracts from Dendropanax morbifera}

The present invention relates to a composition for preventing or treating kidney damage caused by heavy metals containing hwangchil extract, hwangchil complex extract or hwangchil complex fermented extract as an active ingredient.

Cadmium (Cd) is a first-class carcinogen and highly toxic metal. It is very harmful to the human body and causes symptoms such as breathing difficulties and liver dysfunction. It is the cause of 'itai-itai disease', one of the four major pollution diseases. Since it has similar properties to zinc required by the human body, it is ingested into the body and causes kidney failure or osteomalacia by interfering with calcium absorption. In contrast to zinc (Zn), which is in the same group on the periodic table, as an essential trace element that plays an important role in various processes in the body, cadmium is very toxic and accumulates in the body, causing various poisoning symptoms.

Cadmium enters the human body through some foods (eg, fish and shellfish, animal kidneys or liver) or contaminated water, and accumulates in the liver and kidneys (kidneys), causing damage to these organs. In particular, kidney damage causes various diseases by excreting various proteins and sugars essential for human body function. Cadmium poisoning is accompanied by diarrhea, abdominal pain, severe vomiting, and can lead to impaired reproductive function and infertility, liver and kidney disorders, damage to the central nervous system and immune system, mental illness, high blood pressure, and cancer. Long-term ingestion even in small amounts can cause poisoning symptoms.

Hwangchil tree (Dendropanax morbifera Lev.) is a tree of the Araliaceae family, and it is a medicinal plant with excellent pharmacological effects to the extent that it has the scientific name Dendropanax, which means a panacea tree. In the world, it is distributed in the southern coastal area of Korea and Japan's temperate, subtropical and Taiwanese regions. Hwangchil, the sap from the bark of the hwangchil tree, contains benzoin and has been used as a natural paint in the past. It has been found to be effective in promoting hard tissue regeneration, preventing hypertension, stroke, and dementia, but the renal protective effect of Hwangchil extract by heavy metals is not yet known.

Republic of Korea Patent Publication No. 10-2019-0120880 (published on 10.08. 2019)

It is an object of the present invention to provide a composition for protecting kidney damage caused by heavy metals containing hwangchil extract, hwangchil complex extract or hwangchil complex fermented extract as an active ingredient.

The present invention provides a pharmaceutical composition for preventing or treating kidney damage caused by heavy metals containing Hwangchil extract as an active ingredient.

The present invention provides a health food for preventing or improving kidney damage caused by heavy metals containing Hwangchil extract as an active ingredient.

The present invention provides a pharmaceutical composition for the prevention or treatment of kidney damage caused by heavy metals containing, as an active ingredient, two or more complex extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract, and cornus extract.

The present invention provides a health food for preventing or improving kidney damage caused by heavy metals containing, as an active ingredient, two or more complex extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract, and cornus extract.

The present invention provides a pharmaceutical composition for preventing or treating kidney damage caused by heavy metals containing, as an active ingredient, two or more complex fermented extracts selected from the group consisting of Hwangchil extract, Sukjihwang extract, Goji tea extract, and Cornus officinalis extract.

In addition, it provides a health food for preventing or improving kidney damage caused by heavy metals containing, as an active ingredient, two or more complex fermented extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract, and cornus extract.

According to the present invention, as a result of inducing toxicity by treating LLC-PK1 kidney cells pretreated with hwangchil extract, hwangchil complex or hwangchil complex fermented product, kidney cells that are not treated with hwangchil extract, hwangchil complex or hwangchil complex fermented product As it was confirmed that cytotoxicity and intracellular active oxygen generation are reduced compared to that, the composition containing the hwangchil extract, hwangchil complex or fermented hwangchil complex as an active ingredient shows the possibility of alleviating kidney damage caused by cadmium, so that the kidney caused by heavy metals It can be provided as a pharmaceutical composition and health food useful for the prevention and treatment of damage.

1 is a result of confirming the cytotoxicity of LLC-PK1 cells treated with Goji berry extract, Cornus extract, Sukjihwang extract, Hwangchil extract, Hwangchil complex extract and Hwangchil complex fermented extract at concentrations of 50, 100 and 500 μg/ml.
Figure 2 shows the renal cell protective effect of cadmium-treated LLC-PK1 cells treated with Goji extract, Cornus extract, Sukjihwang extract, Hwangchil extract, Hwangchil complex extract and Hwangchil complex fermented extract at 50, 100 and 500 μg/ml concentrations. This is the confirmed result.
Figure 3 is cadmium treatment for LLC-PK1 cells induced reciprocal damage, Goji berry extract, Cornus officinalis extract, Sukjihwang extract, Hwangchil extract, Hwangchil complex extract and Hwangchil complex fermented extract at concentrations of 100 and 500 μg/ml. This is the result of confirming the oxygen suppression effect.

Hereinafter, the present invention will be described in more detail.

The present invention can provide a pharmaceutical composition for preventing or treating kidney damage caused by heavy metals containing Hwangchil extract as an active ingredient.

The hwangchil extract may be extracted with water, C1 to C4 alcohol or a mixed solvent thereof, and more preferably, it may be extracted using hot water extract or 95% ethanol.

The heavy metal may be cadmium, but is not limited thereto.

The present invention can provide a health food for preventing or improving kidney damage caused by heavy metals containing Hwangchil extract as an active ingredient.

The present invention can provide a pharmaceutical composition for preventing or treating kidney damage caused by heavy metals containing, as an active ingredient, two or more complex extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract, and cornus extract.

More preferably, the complex extract may be a complex extract obtained by mixing Hwangchil extract, Goji berry extract, Cornus officinalis extract and Sukjihwang extract in a weight ratio of 1:1:1:2.

The complex extract may be extracted with water, C1 to C4 alcohol or a mixed solvent thereof, and more preferably, it may be extracted using hot water extract or 95% ethanol.

The heavy metal may be cadmium, but is not limited thereto.

The present invention can provide a health food for preventing or improving kidney damage caused by heavy metals containing, as an active ingredient, two or more complex extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract and cornus extract.

The present invention can provide a pharmaceutical composition for preventing or treating kidney damage caused by heavy metals containing, as an active ingredient, two or more complex fermented extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract, and cornus extract.

More preferably, the complex fermented extract is obtained by mixing hwangchil, goji berry, cornflower oil and Sukjihwang, fermenting at 30 to 50° C. for 10 to 20 hours, and then extracting it with water, C1 to C4 alcohol or a mixed solvent thereof. It may be one containing the extract as an active ingredient. More preferably, the complex fermented extract may be extracted using hot water extract or 95% ethanol.

The complex fermented extract may be a complex fermented extract obtained by mixing and fermenting hwangchil extract, goji berry extract, cornus extract, and Sukhumus extract in a weight ratio of 1:1:1:2.

The heavy metal may be cadmium, but is not limited thereto.

In addition, the present invention can provide a health food for preventing or improving kidney damage caused by heavy metals containing, as an active ingredient, two or more complex fermented extracts selected from the group consisting of hwangchil extract, sukjihwang extract, goji tea extract and cornflower extract.

In one embodiment of the present invention, the pharmaceutical composition is any one selected from the group consisting of injections, granules, powders, tablets, pills, capsules, suppositories, gels, suspensions, emulsions, drops or liquids according to a conventional method of the formulation can be used.

In another embodiment of the present invention, suitable carriers, excipients, disintegrants, sweeteners, coating agents, swelling agents, lubricants, flavoring agents, antioxidants, buffers, bacteriostats, diluents, dispersants, One or more additives selected from the group consisting of surfactants, binders and lubricants may be further included.

Specifically, carriers, excipients and diluents include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, gum acacia, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline Cellulose, polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, and mineral oil can be used, and solid preparations for oral administration include tablets, pills, powders, granules, and capsules. agent, and the like, and the solid preparation may be prepared by mixing at least one excipient, for example, starch, calcium carbonate, sucrose or lactose, gelatin, and the like in the composition. In addition to simple excipients, lubricants such as magnesium stearate and talc can also be used. Liquid preparations for oral use include suspensions, solutions, emulsions, syrups, and the like, and various excipients such as wetting agents, sweeteners, fragrances, preservatives, etc. may be included in addition to commonly used simple diluents such as water and liquid paraffin. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, suppositories, and the like. Non-aqueous solvents and suspending agents include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. As the base material for the suppository, witepsol, macrogol, tween 61, cacao butter, laurin, glycerogelatin, and the like can be used.

According to an embodiment of the present invention, the pharmaceutical composition is administered in a conventional manner via intravenous, intraarterial, intraperitoneal, intramuscular, intrasternal, transdermal, intranasal, inhalational, topical, rectal, oral, intraocular or intradermal routes. can be administered to the subject.

The preferred dosage of the hwangchil extract, the complex extract or the complex fermented extract may vary depending on the condition and weight of the subject, the type and extent of the disease, the drug form, the administration route and period, and may be appropriately selected by those skilled in the art. According to an embodiment of the present invention, although not limited thereto, the daily dose may be 0.01 to 200 mg/kg, specifically 0.1 to 200 mg/kg, and more specifically 0.1 to 100 mg/kg. Administration may be administered once a day or may be administered in several divided doses, thereby not limiting the scope of the present invention.

In the present invention, the 'subject' may be a mammal including a human, but is not limited to these examples.

In another embodiment of the present invention, the health food is used with other foods or food additives in addition to the hwangchil extract, complex extract or complex fermented extract, and may be appropriately used according to a conventional method. The mixed amount of the active ingredient may be suitably determined according to the purpose of its use, for example, prophylactic, health or therapeutic treatment.

The effective dose of the compound contained in the health food may be used according to the effective dose of the therapeutic agent, but in the case of long-term intake for health and hygiene or health control, it may be less than or equal to the above range, It is clear that the ingredient can be used in an amount beyond the above range because there is no problem in terms of safety.

The type of health food is not particularly limited, and examples include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, alcoholic beverages, and vitamin complexes.

Hereinafter, examples will be described in detail to help the understanding of the present invention. However, the following examples are merely illustrative of the content of the present invention, and the scope of the present invention is not limited to the following examples. The embodiments of the present invention are provided to more completely explain the present invention to those of ordinary skill in the art.

<Example 1> Preparation of extracts and complexes, fermented products

1) Preparation of Hwangchil extract

After quantifying hwangchil, distilled water was added 6 times, and hot water extraction was performed at 80° C. for 6 hours. The extract was filtered to remove solids, concentrated using a concentrator, and then freeze-dried and used as a sample.

2) Manufacture of Hwangchil Complex

Hwangchil, goji berries, cornflower oil, and sukjihwang were mixed in a ratio of 1:1:1:2, distilled water was added 6 times, and hot water was extracted at 105° C. for 6 hours. The extract was filtered to remove solids, concentrated using a concentrator, and then freeze-dried and used as a sample.

3) Manufacture of Hwangchil Complex Fermented Product

Hwangchil, goji berry, cornflower oil, and sorghum yellow were mixed in a 1:1:1:2 ratio, and distilled water was hydrolyzed 6 times. Rhodobacter sphaeroides) and filamentous fungi (Aspergillus oryzae, Mucor hiemalis) mixed with effective microorganisms (EM) were added and fermented at 39°C for 15 hours, followed by hot water extraction and sterilization at 105°C for 6 hours. was filtered to remove solids. After concentration using a concentrator, lyophilization was used as a sample.

4) Manufacture of extracts from Goji berry, Cornus officinalis, and Sukhumi

Goji berry, cornus nutmeg, and Sukhumi were quantified, and 95% ethyl alcohol was hydrolyzed 10 times, followed by extraction at room temperature for 6 hours. The extract was filtered to remove solids, concentrated using a concentrator, and then freeze-dried and used as a sample.

<Example 2> Confirmation of cytotoxicity of extracts and complexes, fermented products against LLC-PK1 cells

MTT assay was performed to measure the cytotoxicity of the sample to LLC-PK1 cells. LLC-PK1 cells were aliquoted to 2 × 10 ⁵ cells/well in a 24-well plate and cultured for 20 to 24 hours in a 5% CO ₂ incubator, and then treated for each concentration of each extract, followed by 20 in a 5% CO ₂ incubator. Incubated for ~24 hours. 50 μl of MTT (5 mg/ml) solution dissolved in PBS was put into each well and reacted for 4 hours in an incubator. Then, the supernatant was removed, and 200 μl of DMSO was added to each well to dissolve the formed formazan crystal, and absorbance was measured with an ELISA reader at a wavelength of 550 nm. The measured formazan production level was compared with the value of normal cells and expressed as a percentage (%).

As a result, as shown in FIG. 1 , it was confirmed that cytotoxicity of goji berries, cornflower oil, Sukjihwang, hwangchil extract and hwangchil complex, and fermented hwangchil complex against LLC-PK1 cells did not appear at any concentration.

<Example 3> Confirmation of renal protective effect of extracts and complexes and fermented products against cadmium in LLC-PK1 cells

MTT assay was performed to measure the cytoprotective ability of the sample against cadmium-treated LLC-PK1 cells. LLC-PK1 cells were aliquoted to 2×10 ⁵ cells/well in a 24-well plate and cultured for 20 to 24 hours in a 5% CO ₂ incubator, and then each sample was treated by concentration for 20 to 20 in a 5% CO ₂ incubator. Pre-incubation for 24 hours.

After treatment so that the final concentration of CdCl ₂ is 10 μM, the reaction was carried out for 24 hours, and 50 μl of MTT (5 mg/ml) solution dissolved in PBS was put into each well and reacted for 4 hours in an incubator. After that, the supernatant was removed, and 200 μl of DMSO was added to each well to dissolve the formed formazan crystal, and absorbance was measured with an ELISA reader at a wavelength of 550 nm. The confirmed formazan production level was compared with the value of normal cells and expressed as a percentage (%).

As a result, the LLC-PK1 cells treated with cadmium as shown in FIG. 2 showed a cell viability of 27.38% compared to the untreated LLC-PK1 cells, whereas the Goji berry extract did not show a significant cytoprotective effect. However, the cornus oil extract showed a weak cytoprotective effect from the treatment concentration of 100 μg/ml. In the case of Sukjihwang extract, the cytoprotective effect was shown in a concentration-dependent manner (cell viability of 60.02% at a treatment concentration of 100 μg/ml), and in the case of Hwangchil extract, 47.51% at a concentration of 100 μg/ml and 61.63 at a concentration of 500 μg/ml % of the cell viability appeared, confirming the protective effect. In the case of Hwangchil complex, the cell viability of 57.74% at 100 μg/ml and 64.63% at 500 μg/ml was confirmed. In addition, in Hwangchil complex fermented product, the cell viability was 74.24% at 100 μg/ml concentration and 84.86% at 500 μg/ml, confirming that the cytoprotective effect on cadmium was increased in a concentration-dependent manner.

<Example 4> Confirmation of active oxygen inhibition of extracts and complexes and fermented products for cadmium in LLC-PK1 cells

In LLC-PK1 cells treated with cadmium to induce oxidative damage, the intracellular active oxygen inhibitory effect of Goji berry, Cornus officinalis, Sukjihwang, Hwangchil extract, Hwangchil complex, and Hwangchil complex fermented product was confirmed.

MTT assay was performed to measure the cytoprotective ability of the sample against cadmium-treated LLC-PK1 cells. LLC-PK1 cells were aliquoted to 2×10 ⁵ cells/well in a 24-well plate and cultured for 20 to 24 hours in a 5% CO ₂ incubator, then each sample was treated by concentration and 20 in a 5% CO ₂ incubator. Pre-incubation for ~24 hours. After treatment so that the final concentration of CdCl ₂ was 10 μM, the reaction was carried out for 24 hours, washed with PBS, and then stained with DCF-DA (5 μM) and DAPI (10 μM). After 20 minutes of reaction, intracellular reactive oxygen species were observed under a fluorescence microscope.

As a result, active oxygen was not observed in the cells not treated with cadmium as shown in FIG. 3, but overproduction of active oxygen was confirmed when cadmium was treated. Oxygen was not suppressed.

On the other hand, in the cells treated with Sukjihwang and Hwangchil extract, Hwangchil complex extract or Hwangchil complex fermented product, it was confirmed that the intracellular active oxygen was effectively inhibited from the concentration of 100 μg/ml.

From the above results, it was found that Sukjihwang and Hwangchil extracts exhibited a renal protective effect against oxidative damage caused by cadmium. As confirmed, a synergistic effect by fermentation was demonstrated.

As described above in detail a specific part of the present invention, for those of ordinary skill in the art, it is clear that this specific description is only a preferred embodiment, and the scope of the present invention is not limited thereby. something to do. Accordingly, the substantial scope of the present invention will be defined by the appended claims and their equivalents.

Claims (14)

delete delete delete delete delete delete delete delete delete A pharmaceutical composition for the prevention or treatment of kidney damage caused by heavy metals, which contains a complex fermented extract mixed with Hwangchil extract, Sukjihwang extract, Goji tea extract, and Cornus officinalis extract in a weight ratio of 1:1:1:2 as an active ingredient. The method according to claim 10, wherein the complex fermented extract is a complex extracted with water, C1 to C4 alcohol, or a mixed solvent thereof, after mixing hwangchil, goji berry, cornflower oil and sukjihwang, fermented at 30 to 50° C. for 10 to 20 hours. A pharmaceutical composition for preventing or treating kidney damage caused by heavy metals, characterized in that it contains a fermented extract as an active ingredient. delete The pharmaceutical composition for preventing or treating kidney damage caused by a heavy metal according to claim 10, wherein the heavy metal is cadmium. A health food for preventing or improving kidney damage caused by heavy metals, containing as an active ingredient a complex fermented extract mixed with Hwangchil extract, Sukjihwang extract, Goji tea extract, and Cornus officinalis extract in a weight ratio of 1:1:1:2.

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