KR101835457B1 - Composition for anti-cancer, anti-inflammation or protecting cell comprising an extract of microalgae - Google Patents

Abstract

In this specification, a composition for protecting cells from anticancer, anti-inflammation or toxicity comprising an extract of a cultured microalgae of Nephroselmis sp. , Specifically, the microalgae culture can use exhaust gas or industrial wastewater. / RTI > Also disclosed herein is a process for making the composition.

Description

TECHNICAL FIELD [0001] The present invention relates to an anticancer, antiinflammatory or cytoprotective composition containing a microalgae extract,

The present invention relates to anticancer, anti-inflammatory or cytoprotective compositions comprising a microalgae cultured extract and a process for their preparation.

Research and development of microalgae strains are changing to production of high-value products such as coloring materials, cosmetic raw materials and health function / medicines in fueling such as biodiesel and bioethanol. The development of technology for the production of medicinal materials using microalgae is still in its early stage, but as the recognition of the concept of aging population and well-being is settled, it is expected to expand the scale of the drug market derived from natural products.

There is a growing interest in physiologically active substances under the above recognition, but research on physiologically active substances using microalgae is still insufficient. Accordingly, studies on microalgae containing physiologically active substances have been demanded.

Korean Patent No. 10-1197101 Korea Patent No. 10-0889973

An aspect of the present invention is to provide an anti-cancer, anti-inflammatory or cytoprotective composition comprising an extract of a microalgae cultured product.

In one aspect of the present invention, the microalgae is Nephroselmis sp. KGE 1 to provide an anti-cancer, anti-inflammatory or cytoprotective composition.

According to an aspect of the present invention, there is provided a composition for treating cancer, anti-inflammation or cell protection, which is a cultured product obtained by treating a carbon source with at least one of carbon sources of exhaust gas and industrial wastewater.

In one aspect of the present invention, there is provided an anticancer, anti-inflammatory or cytoprotective composition, wherein the extract is an extract, which is extracted with an extraction solvent of water, C1-C4 lower alcohol, hexane or chloroform as an extraction solvent.

Another aspect of the present invention is to provide a method for producing the above anti-cancer, anti-inflammatory or cytoprotective composition.

An aspect of the present invention provides an anticancer composition comprising an extract of a Nephroselmis sp. Microalgae culture as an active ingredient.

In one aspect of the present invention, the composition provides an anti-cancer composition that is for skin cancer suppression.

One aspect of the present invention provides an anti-inflammatory composition comprising an extract of a Nephroselmis sp. Microalgae culture as an active ingredient.

An aspect of the present invention provides a composition for protecting cells from toxicity comprising an extract of a Nephroselmis sp. Microalgae culture as an active ingredient. Specifically, the cell may be a kidney cell. In addition, the toxicity may be drug toxicity. More specifically, the drug may be an anti-cancer agent.

In one aspect of the present invention, the anticancer agent is a platinum-containing anti-cancer drug.

In one aspect of the present invention, the platinum-containing anticancer agent is a platinum-containing anticancer agent selected from the group consisting of cisplatin, transplatin, carboplatin, ornaplastatin, dicycloplatin, do.

In one aspect of the invention, the microalgae is Nephroselmis sp. KGE 1.

In one aspect of the present invention, the microalgae may have the deposit number KCTC12872BP.

In one aspect of the present invention, the extract is an extract of at least one of water, C1-C4 lower alcohol, hexane and chloroform.

In one aspect of the invention, the composition provides a pharmaceutical, cosmetic or health functional food composition.

Another aspect of the present invention provides a method of producing an anti-cancer, anti-inflammatory or cytoprotective composition comprising culturing Nephroselmis sp. Microalgae.

In another aspect of the present invention, the step of culturing is a step of treating and culturing a carbon source of any one or more of exhaust gas and industrial wastewater as a carbon source.

In another aspect of the present invention, the exhaust gas includes carbon dioxide, and the concentration of carbon dioxide is 5 to 15% by volume based on the exhaust gas.

In another aspect of the present invention, the step of culturing comprises culturing at least one of the exhaust gas and the industrial wastewater by treating the culture medium with a carbon source-free synthetic medium.

The extract of the microbial strain according to one aspect of the present invention or the composition containing the same is excellent in physiological activity.

The extract of the microbial strain cultured according to one aspect of the present invention or the composition containing the same has excellent anticancer effect.

The extract of the microbial strain cultured according to one aspect of the present invention or the composition containing the same is excellent in the skin cancer suppressing effect.

The extract of the microbial strain culture according to one aspect of the present invention or the composition containing the same is excellent in anti-inflammatory effect.

The extract of the microbial strain according to one aspect of the present invention or the composition containing the same is excellent in cytoprotective effect against toxicity.

The extract of the microbial strain cultured according to one aspect of the present invention or the composition containing the same is excellent in the effect of protecting the kidney cells against drug toxicity. Specifically, the drug toxicity may be an anticancer agent, preferably a platinum-containing anticancer agent.

The method according to another aspect of the present invention can produce a composition for anticancer, anti-inflammation or cell protection by cultivating microalgae by treating carbonaceous gas of a thermal power plant with carbon source as a carbon source to microalgae and reducing the source of greenhouse gas .

Fig. 1 is a result of confirming the growth amount of each microalgae according to the culture method according to the present invention.
FIG. 2 is a table showing the results and the results of the skin cancer activity of the culture extract cultured by the culture method according to the present invention.
FIG. 3 shows the results of the anti-inflammatory activity of the culture extract cultured by the culture method of the present invention and the numerical values thereof.
FIG. 4 is a graph showing cytotoxicity of a culture extract cultured by the culture method according to the present invention on brain cancer cells.
FIG. 5 shows the items and numerical values of the protective effect of the kidney cells of the culture extract cultured by the culture method according to the present invention.

An aspect of the present invention provides an anticancer composition comprising an extract of a Nephroselmis sp. Microalgae culture as an active ingredient.

In one aspect of the present invention, the composition provides an anti-cancer composition that is for skin cancer suppression.

One aspect of the present invention provides an anti-inflammatory composition comprising an extract of a Nephroselmis sp. Microalgae culture as an active ingredient.

An aspect of the present invention provides a composition for protecting cells from toxicity comprising an extract of a Nephroselmis sp. Microalgae culture as an active ingredient. Specifically, the cell may be a kidney cell. In addition, the toxicity may be drug toxicity. More specifically, the drug may be an anti-cancer agent.

In one aspect of the present invention, the anticancer agent is a platinum-containing anti-cancer drug.

In one aspect of the present invention, the platinum-containing anticancer agent is a platinum-containing anticancer agent selected from the group consisting of cisplatin, transplatin, carboplatin, ornaplastatin, dicycloplatin, do.

In one aspect of the invention, the microalgae is Nephroselmis sp. KGE 1.

In one aspect of the present invention, the microalgae may have the deposit number KCTC12872BP.

In one aspect of the present invention, the extract is an extract of at least one of water, C1-C4 lower alcohol, hexane and chloroform.

In one aspect of the invention, the composition provides a pharmaceutical, cosmetic or health functional food composition.

Another aspect of the present invention provides a method of producing an anti-cancer, anti-inflammatory or cytoprotective composition comprising culturing Nephroselmis sp. Microalgae.

In another aspect of the present invention, the step of culturing is a step of treating and culturing a carbon source of any one or more of exhaust gas and industrial wastewater as a carbon source.

In another aspect of the present invention, the exhaust gas includes carbon dioxide, and the concentration of carbon dioxide is 5 to 15% by volume based on the exhaust gas.

In another aspect of the present invention, the step of culturing comprises culturing at least one of the exhaust gas and the industrial wastewater by treating the culture medium with a carbon source-free synthetic medium.

As used herein, the term " cultivation "means that microalgae grow while accepting factors necessary for growth among any one or more components of the synthetic medium, exhaust gas and industrial wastewater. Specifically, the cultivation of the present specification can take place in an incubator shaker at a temperature of 27 ° C, an initial pH of 5.4-5.7, an illuminance of 100 μmol / m 2 -sec and a 150 rpm, but is not limited thereto.

As used herein, the term "exhaust gas" may include all gaseous materials that are generated when a specific substance is combusted, synthesized, or decomposed. Specifically, the exhaust gas of the present specification may include gas discharged from a thermal power plant, and such exhaust gas may include, but is not limited to, carbon dioxide.

As used herein, "treatment" includes "contacting" microalgae with the carbon source. That is, the treatment means that one or more cross sections of the carbon source (preferably, exhaust gas) and the microalgae are in contact with each other, whereby the substances necessary for growth of the microalgae in the carbon source can move to the microalgae.

In another aspect of the present invention, the exhaust gas may have a concentration of carbon dioxide with respect to the exhaust gas of 5 to 15% by volume. In one embodiment, the concentration of carbon dioxide relative to the exhaust gas is greater than 5 vol%, greater than 6 vol%, greater than 8 vol%, greater than 10 vol%, greater than 12 vol%, greater than 13 vol%, greater than 13.5 vol% Or more or 14.5% by volume or more. Also, in one embodiment, the concentration of carbon dioxide relative to the exhaust gas is less than or equal to 15 vol%, less than or equal to 14.5 vol%, less than or equal to 14.4 vol%, less than or equal to 14.3 vol%, less than or equal to 14.2 vol%, less than or equal to 14.1 vol%, less than or equal to 14 vol% 13 vol% or less, 12 vol% or less, 11 vol% or less, 10 vol% or 9 vol% or less. When the concentration of carbon dioxide is within the above range, the microalgae culture is excellent.

In one aspect of the present invention, the above-mentioned cultured organism is cultured by treating the carbon source with a synthetic medium containing no carbon source, thereby providing an anticancer, anti-inflammatory or cytoprotective composition.

In one aspect of the present invention, the synthetic medium is one of KH ₂ PO ₄ , CaCl ₂ 2H ₂ O, MgSO ₄ 7H ₂ O, NaNO ₃ , K ₂ HPO ₄ , NaCl, H ₃ BO ₃ and Na ₂ EDTA And may include one or more. The synthetic medium may be but is not limited to BBM medium. The BBM medium is a medium containing KH ₂ PO ₄ , CaCl ₂ 2H ₂ O, MgSO ₄ 7H ₂ O, NaNO ₃ , K ₂ HPO ₄ , NaCl, H ₃ BO ₃ , Na ₂ EDTA and trace elements, Are ZnSO ₄ 7H ₂ O, MnCl ₂ 4H ₂ O, MoO ₃ , CuSO ₄ 5H ₂ O, and Co (NO ₃ ) ₂ 6H ₂ O.

In one aspect of the present invention, the harvesting method of the microalgae culturing is preferably carried out by centrifugation, but not limited thereto, and general cell separation methods can be used.

In one or another aspect of the present invention, the extract provides an anti-cancer, anti-inflammatory or cytoprotective composition which is an extracted extract of at least one of water, C1-C4 lower alcohol, hexane and chloroform.

According to one or more aspects of the present invention, the extraction method may be such that the cultured microalgae is crushed and the extraction solvent is added in an amount of 10 to 30 times the amount of the microalgae culture, and the extraction temperature is most preferably room temperature But is not limited thereto. The extraction time is preferably 12 to 48 hours, and the extraction time is most preferably 1 to 3 times, but not always limited thereto. It is preferable to use a vacuum decompressing concentrator or a vacuum rotary evaporator, but not always limited thereto.

In one aspect of the invention, the pharmaceutical composition may further comprise, in one aspect, suitable carriers, excipients and diluents conventionally used in the manufacture of pharmaceutical compositions. In one aspect, the carrier, excipient and diluent which may be included in the composition include lactose, dextrose, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, starch, acacia rubber, alginate, gelatin, calcium phosphate, calcium silicate, Cellulose, methylcellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate and mineral oil.

The pharmaceutical composition may be formulated in the form of oral, granule, tablet, capsule, suspension, emulsion, syrup, aerosol or the like, oral preparation, suppository and sterilized injection solution according to a conventional method .

In the case of formulation, a diluent such as a filler, an extender, a binder, a wetting agent, a disintegrant, a surfactant, or an excipient is usually used. Solid formulations for oral administration may include tablets, pills, powders, granules, capsules, and the like. Such solid preparations may contain at least one excipient such as starch, calcium carbonate, sucrose or lactose, and gelatin in addition to the active ingredient. In addition to simple excipients, lubricants such as magnesium stearate talc may also be included. Examples of the liquid preparation for oral use include suspensions, solutions, emulsions, syrups and the like. In addition to water and liquid paraffin which are commonly used simple diluents, various excipients such as wetting agents, sweeteners, fragrances, preservatives and the like may be included. Formulations for parenteral administration may include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, freeze-dried preparations, and suppositories. Examples of the suspending agent include propylene glycol, polyethylene glycol, vegetable oil such as olive oil, injectable ester such as ethyl oleate, and the like. Examples of suppositories include witepsol, macrogol, tween 61, laurin, cacao butter, glycerogelatin and the like.

Dosages of the extracts disclosed herein may vary depending on the condition and the weight of the patient, the degree of disease, the type of drug, the route of administration, and the period of time, and may be selected within a range generally used in the art. In one aspect, the daily dose of the active ingredient may be from 0.0001 to 1000 g / kg on a dry weight basis, and in one aspect from 0.001 to 1 g / kg.

In one aspect of the invention, the composition may be a cosmetic composition. The cosmetic composition of the present invention may be prepared in any formulation conventionally produced in the art and may be in the form of a solution, suspension, emulsion, paste, gel, cream, lotion, powder, soap, surfactant- , Oil, powder foundation, emulsion foundation, wax foundation and spray, but is not limited thereto.

In one aspect of the present invention, the composition may be a food composition, for example, various foods, beverages, gums, tea, vitamin complexes, health supplements, etc., and may be in the form of powder, granule, tablet, capsule or beverage . The food composition of each formulation can be blended with the ingredients commonly used in the field in addition to the active ingredient without difficulty by those skilled in the art depending on the purpose of formulation or use, and synergistic effect can be obtained when the composition is applied simultaneously with other ingredients.

The amount of the active ingredient contained in the food or beverage may be in the range of 1 to 5% by weight of the total food in the case of a health food composition, 0.02 to 10 g in terms of 100 ml and 0.3 To 1 g.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are only for illustrating the present invention and that the scope of the present invention is not construed as being limited by these embodiments.

[Example 1] Analysis of composition of exhaust gas

After exhaust gas was collected from Yeongdong Thermal Power Plant, the components in the exhaust gas were analyzed using KANE-455 combustion efficiency meter (UK). As a result, the NO _X was confirmed to be present in 50 ppm, 200 ppm and SO _X is CO ₂ is 140,000 ppm.

[Example 2] Culture of microalgae

Synthesis of Bold? S Basal medium (BBM, Bischoff, HW, Bold, HC, 1963. Phycological Studies IV. Some soil algae from enchanted rock and related algal species, University of Texas Publication 6318: 1-95) The purging of the exhaust gas was performed after filling 300 mL of the medium. After the microalgae of Table 1 were injected at the same concentration, the initial OD ₆₈₀ concentration in the bottle was adjusted to 0.015-0.016. Each of the bottles was incubated for 6 days at a temperature of 27 DEG C, pH 5.5, 120 mu mol / m ² -sec.

Algal species Strain ID Accession number Nephroselmis sp. KGE1 HE861879 Nephroselmis sp. KGE8 HE861886 Scenedesmus sp. KGE7 HE861882 Chlorella variabilis KGE26 HE974914

 [ Experimental Example  1] Measurement of growth of microalgae

Suspended Solid (SS) method was used to measure the growth rate of microalgae in Example 2 by dry weight. Specifically, GF-C (Whatman) filter paper was dried in a 100 ° C. dry oven for 2 hours and then stored in a desiccator for 30 minutes to measure its weight. 10 mL of the microalgae solution was passed through a pressure- The post-filter paper was dried in a 105 ° C dry oven for 4 hours and weighed.

Calculation of dry weight = (weight of filter dried after filtration - weight of filter dried before filtration) × 100

As a result of the experiment, as shown in FIG. 1, the growth rate of KGE1 strain and KGE8 strain of Nephroselmis spp. Were rapidly increased as in the case of Scenedesmus and Chlorella microalgae according to one aspect of the present invention.

[Example 3] Extraction of anticancer, anti-inflammatory or renal protective substance

The cultured microalgae cultured in Example 2 was separated by centrifugation.

Methanol was used as an extraction solvent for microalgae cultures. For extraction, ultrasonic disruption was performed for about 30 minutes, 30 ml of the extraction solvent was added to the microalgae culture, and the mixture was extracted twice at room temperature for 24 hours. Further, a vacuum decompression concentrator (vacuum rotary evaporator) was used for the concentration.

[Experimental Example 2] Measurement of physiological activity of skin cancer

The physiological activity of SK-MEL-2 (Human Melanoma Cells) of skin cancer cells (ATCC, SK-MEL-2) was investigated by extracting the microalgae of Example 3 and a positive control group of ethoposide (Sigma Aldrich) . As a negative control, skin cancer cells not treated with the drug were used. The experimental results are shown in Fig.

As a result, the positive control group etoposide showed 50% of the skin cancer cell death as compared with the negative control group, and the microalgae cultured extract showed excellent efficacy in KGE1. In addition, it did not show efficacy in KGE8, KGE7 and KGE26 extracts.

[Experimental Example 3] Measurement of anti-inflammatory activity

In order to confirm the anti-inflammatory effect of the microalgae extract of Example 3, the production of nitric oxide was induced by lipopolysaccharide (LPS) in glioblastoma (ATCC T98G, human glioblastoma cells) producing NO Next, the extract was treated and the degree of production of NO induced by LPS in the cells was measured and compared, and it is shown in FIG. In order to confirm the cytotoxicity, cytotoxicity was confirmed through analysis of methylthiazol-2-yl-2,5-diphenyl tetrazolium bromide (MTT), which is shown in FIG.

As a result, the anti-inflammatory activity showed better anti-inflammatory activity than that of KGE8, which is the same KGE1, and KGE7 and KGE26, as the anti-inflammatory effect exhibits a lower level of NO produced. As a result of the cytotoxicity test, it was confirmed that the microalgae cultured extract had little cytotoxicity.

[Experimental Example 4] Measurement of protective effect on kidney cells

In order to measure the kidney protective efficacy of the microalgae extract of Example 3, the protective effect on kidney toxicity was evaluated as follows using kidney cell LLC-PK1 (CL-101 ^TM of ATCC) as follows. LLC-PK1 cells were treated with 100 μg / ml of 10% fetal bovine serum (Gibco BRL Life Technologies), 100 units / ml penicillin G / ml (Gibco BRL Life Technologies) and streptomycin (Gibco BRL Life Technologies) And cultured in a 95% air / 5% carbon dioxide incubator maintained at 37 ° C using DMEM (Cellgro, Manassas, VA, USA) medium. Including the cultured LLC-PK1 cells, 100 μL of a 96-well culture plate were to be introduced 10 ⁴ per each well, and the cell is stable for 22 hours. Subsequently, the microalgae extract was added to the designated concentration, and after 2 hours, 0.1 ml of 250 μM cisplatin dissolved in a radical generating reagent medium was added and cultured for 24 hours. Then, 10 μL of CCK-8 (CCK -8, Dojindo Laboratories, Japan) reagent was added to each well, followed by incubation at 37 ° C. After 1 hour, the viability of the cells was measured by measuring the absorbance using a 450 nm detection wavelength in a BIO-TEK (Winooski, VT, USA) microplate reader. The experimental results are shown in Fig.

As a result, KGE1 exhibited better efficacy than KGE8 and KGE7, and showed similar efficacy to KGE26, since kidney cell survival rate was higher with better kidney protective efficacy.

Korea Biotechnology Research Institute KCTC12872BP 20150717

Claims (18)

An anticancer pharmaceutical composition comprising an extract of Nephroselmis sp. KGE 1 microalgae culture as an active ingredient. The method according to claim 1,
Wherein said cancer is skin cancer. An anticancer health food composition comprising an extract of a microalgae cultivar of Nephroselmis sp. KGE 1 as an active ingredient. The method of claim 3,
Wherein said cancer is skin cancer. An anti-inflammatory composition comprising an extract of a microalgae culture of Nephroselmis sp. KGE 1 as an active ingredient. A pharmaceutical composition for protecting kidney cells from drug toxicity,
The composition comprises an extract of a microalgae culture of Nephroselmis sp. As an active ingredient,
Wherein said medicament comprises cisplatin. A health food composition for protecting kidney cells from drug toxicity,
The composition comprises an extract of a microalgae culture of Nephroselmis sp. As an active ingredient,
Wherein the medicament comprises cisplatin. The method according to claim 6,
Wherein the drug is an anti-cancer agent. 8. The method of claim 7,
Wherein the drug is an anti-cancer agent. 10. The method according to any one of claims 1 to 9,
Wherein the microalgae has the accession number KCTC12872BP. 10. The method according to any one of claims 1 to 9,
Wherein the extract is an extract of at least one of water, C1-C4 lower alcohol, hexane and chloroform. 6. The method of claim 5,
Wherein the composition is a pharmaceutical composition. 6. The method of claim 5,
Wherein the composition is a cosmetic composition. 6. The method of claim 5,
Wherein the composition is a health functional food composition. 10. A method for producing a composition according to any one of claims 1 to 9, comprising the step of culturing Nephroselmis sp. Microalgae. 16. The method of claim 15,
Wherein the culturing is a step of treating and culturing a carbon source of any one or more of exhaust gas and industrial wastewater as a carbon source. 17. The method of claim 16,
Wherein the exhaust gas comprises carbon dioxide and the concentration of carbon dioxide is 5 to 15 vol% relative to the exhaust gas. 17. The method of claim 16,
Wherein the culturing is performed by treating at least one of the exhaust gas and the industrial wastewater with a synthetic medium containing no carbon source.

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