KR101971393B1 - Composition for producing a metal nanoparticle comprising ginseng extract and use thereof - Google Patents

Abstract

The present invention relates to a composition for preparing metal nanoparticles comprising ginseng extract as an active ingredient, a method for producing metal nanoparticles using the composition; Metal nanoparticles prepared by the method; It relates to antimicrobial, biofilm degradation and anticoagulant use of the metal nanoparticles. The composition for preparing metal nanoparticles of the present invention and the method for preparing metal nanoparticles using the same using the root or leaf extract of ginseng as a natural extract as an active ingredient, the production time is significantly shorter than before, and without the addition of additional reducing agents or stabilizers. Metal nanoparticles of uniform size can be prepared. In addition, the composition comprising the metal nanoparticles prepared by the method has antimicrobial activity, biofilm degradation activity, anticoagulant activity, anticancer activity and anti-inflammatory activity can be utilized in various industries.

Description

Composition for producing metal nanoparticles comprising ginseng extract and its use {Composition for producing a metal nanoparticle comprising ginseng extract and use}

The present invention comprises a ginseng extract as an active ingredient, a composition for preparing metal nanoparticles; Method for producing a metal nanoparticle using the composition; Metal nanoparticles prepared by the method; It relates to antimicrobial, biofilm degradation, anticoagulant, anticancer and anti-inflammatory use of the metal nanoparticles.

Nanotechnology is rapidly growing as an important field in modern science with potential effects in electrical and biotechnology. Among them, metal nanoparticles are considered important because of their unique physical and biological particle size and shape (Colloids Surf B Biointerfaces, 2013, 107, 227-34), and their unique electronic, magnetic, catalytic and optical properties. Due to its various applications in the fields of biomedical and optics, its importance is gradually increasing.

Among other metals, silver has long been used for the treatment of medical diseases to treat many bacterial infections due to its antimicrobial properties, and silver nanoparticles as nanoparticle forms have reduced cytotoxicity and less antibacterial effects than their ionic forms. It is known to increase, and is known to be applied to many fields for antibacterial, anti-inflammatory or infection prevention. In addition, its unique and harmonious surface plasmon oscillation allows drug delivery, tissue or cancer imaging, photo thermal therap, immune chromatographic identification of pathogens in a sample, biosensor, DNA labeling, or It has been used in various fields such as vapor sensing.

In general, chemical, physical, or biological synthesis methods are used to prepare such metal nanoparticles, but the chemical synthesis method is relatively simple but expensive and has a problem of side effects of reagents. In addition, the physical method has a problem in that it is difficult to control the size of the nanoparticles, and expensive manufacturing equipment is required, which makes it difficult to manufacture effective metal nanoparticles.

For this reason, a lot of research has recently been conducted on the production method of environmentally friendly metal nanoparticles containing a natural extract as an active ingredient or a bacteria or strain. However, conventional methods for synthesizing metal nanoparticles by bacteria or strains require a long time for the synthesis from a short day to a few days, and capping agents such as additional reducing agents or stabilizers for stabilization of the synthesized nanoparticles. The disadvantage was the need for a capping agent. Therefore, there is still a need for development of metal nanoparticles that are environmentally friendly and stable and overcome the limitations of manufacturing time compared with chemical or physical synthesis methods.

On the other hand, Panax ginseng is a plant belonging to the Araliaceae family, which is very slow and perennial compared to other plants, and most of them are found in Korea, northeast of China, or Siberian Far East. Ginseng has been cultivated and used in traditional medicines such as tonics, tonics, and anti-aging agents for a long time. The main active ingredient is ginsenoside, which is known to exist more than 40. Most of them are anti-cancer, anti-diabetic, antioxidant, anti-inflammatory, It is known to have biological and pharmaceutical activities, such as alteration and control of immune function, radiation protection, anti forgetfulness, antiapoptosis or antistress. In addition, other studies have reported that some components of ginseng are effective in aging, disorders of major nervous systems, and neurodegenerative diseases.

Ginseng is mainly used for roots and fruits among many parts. This is because ginsenosides are known to be contained in the roots of ginseng in large quantities. On the other hand, despite the ginseng leaves occupy more areas than the roots, unlike the roots of ginseng, the frequency of manufacture of the medicine using it was significantly lower than that of the roots or fruits. Recently, however, the leaves of ginseng contain many active ingredients such as flavonoids, triterpenoids, polyacetylenic, alcohols, ginsenosides, amino acids, peptides, polysaccharides, volatile oils and fatty acids. These active ingredients produce many important pharmacological effects on the cardiovascular, reproductive and metabolic systems, as well as on the major nervous system, as well as the properties of anti-fatigue, anti-hyperglycemic, antidiabetic, anticancer, antioxidant and anti-aging (Biomaterials, 2009, 30 (10), 1857-69), research using ginseng leaves is increasing. However, metal nanoparticles have not yet been prepared using the leaf extract of ginseng.

Under these backgrounds, the inventors of the present invention, in the production of metal nanoparticles using natural extracts, have significantly shorter manufacturing time and uniform size than conventional metal nanoparticles that can be stably produced without the addition of a separate reducing agent or stabilizer. The present invention was completed by developing a method for preparing particles and confirming that the composition comprising the metal nanoparticles prepared by the method has antimicrobial activity, biofilm degradation activity, anticoagulant, anticancer and anti-inflammatory activity.

One object of the present invention is to provide a composition for preparing metal nanoparticles comprising ginseng extract as an active ingredient.

Another object of the present invention is to provide a method for producing a metal nanoparticle, comprising the step of reacting the metal precursor composition and the metal precursor.

Still another object of the present invention is to provide a metal nanoparticle produced by the above production method.

Still another object of the present invention is to provide an antimicrobial composition comprising the metal nanoparticles.

Still another object of the present invention is to provide a biofilm decomposition composition comprising the metal nanoparticles.

Still another object of the present invention is to provide an anticoagulant composition comprising the metal nanoparticle.

Still another object of the present invention is to provide a pharmaceutical composition for preventing or treating cancer, including the metal nanoparticles.

Another object of the present invention to provide an anti-inflammatory composition comprising the metal nanoparticles.

One aspect of the present invention for achieving the above object provides a composition for preparing metal nanoparticles, comprising a ginseng extract.

The composition for preparing metal nanoparticles of the present invention and the method for preparing metal nanoparticles using the same are significantly shorter than the conventional manufacturing time by using the root or leaf extract of ginseng which is a natural extract, or the red ginseng root extract as an active ingredient, and a separate reducing agent. Alternatively, metal nanoparticles having a uniform size can be produced without the addition of a stabilizer. In addition, the composition comprising the metal nanoparticles prepared by the method has antimicrobial activity, biofilm degradation activity, anticoagulant activity, anticancer activity and anti-inflammatory activity can be utilized in various industries.

In the present invention, the term "ginseng" is a plant belonging to the Araliaceae, and compared with other plants, the growth is very slow and perennial, and most of them are found in Korea, the northeast of China, or the Siberian Far East. Means plants. Specifically, the ginseng may be selected from ginseng (panax ginseng) or red ginseng (red ginseng), but is not limited thereto.

As used herein, the term "ginseng extract" means a component or substance belonging to ginseng obtained by a conventional method using a ginseng solvent.

The ginseng extract may be prepared by a manufacturing method including the following steps, but is not limited thereto:

1) extracting by adding an extraction solvent to ginseng;

2) cooling the extract of step 1) and then filtering; And

3) drying the filtered extract of step 2) under reduced pressure.

In the above method, ginseng of step 1) may be used without limitation, such as grown or commercially available, and specifically, ginseng (panax ginseng) or red ginseng (red ginseng) may be used, but is not limited thereto. In addition, the site of ginseng may include all the organs of natural, hybrid or cultivar training, for example, all ground parts such as roots, branches, stems, leaves, flowers, fruits, and more specifically, roots or leaves Can be used, but is not limited thereto.

In one embodiment of the present invention, the root or leaves of the collected ginseng (panax ginseng) and then washed and dried, cut into pieces and heated in sterile water to extract the components of the ginseng in water or red ginseng (red ginseng) root An extract was prepared.

In the method, the extract may be extracted with water, a solvent of C1 to C4 alcohol or a mixture thereof, and specifically, the alcohol may be ethanol or methanol, but is not limited thereto. More specifically, distilled water may be extracted as a solvent.

As the extraction method, conventional methods in the art such as filtration, hot water extraction, dipping extraction, reflux cooling extraction, and ultrasonic extraction may be used. The hot water extraction method may be extracted from 1 to 5 times, specifically 3 times may be extracted repeatedly, but is not limited thereto. The extraction solvent may be added 0.1 to 10 times to the dried ginseng, specifically 0.3 to 5 times, but is not limited thereto. In addition, the extraction temperature may be specifically 20 ℃ to 40 ℃ but not limited to this, the extraction time may be specifically 12 to 48 hours, but is not limited thereto.

In the above method, the reduced pressure concentration of step 3) may specifically use a vacuum decompression concentrator or a vacuum rotary evaporator, but is not limited thereto. The drying may be performed under reduced pressure, vacuum drying, boiling drying, spray drying or freeze drying. But it is not limited thereto.

As used herein, the term "metal nanoparticle" is an ultrafine particle having a size of at least 1 to 100 nm, which is made of a metal, and refers to a particle having specific and various properties due to its small size. Metal nanoparticles in the present invention may have a different physiological activity according to the manufacturing method, specifically prepared using ginseng root or leaf extract or red ginseng root extract, the metal nanoparticles of the present invention is a bioactive material of each extract It includes, but may have a very excellent antimicrobial activity, biofilm degradation activity, anticoagulant activity, anticancer activity and anti-inflammatory activity than the metal nanoparticles prepared by other methods, but is not limited thereto.

Specifically, the "metal" may be one or more selected from the group consisting of gold (Au), silver (Ag), copper (Cu), palladium (Pd), platinum (Pt), and rhodium (Rd), and more specifically, As, it may be gold (Au) or silver (Ag), but is not limited thereto.

In the present invention, gold nanoparticles and AuNP (s) are mixed, and silver nanoparticles and AgP (s) are mixed.

In one embodiment of the present invention, the ginseng root or leaf extract or the red ginseng root extract and the extract is observed gold or silver by visually observing the color change of the reaction mixture resulting from reduction reaction with tetrachlorogold (III) acid or silver nitrate solution. Synthesis of nanoparticles was confirmed (Example 3).

Therefore, the composition of the present invention is to prepare metal nanoparticles using ginseng extract as an active ingredient, in particular, gold or silver nanoparticles may be prepared using ginseng leaf or root extract or red ginseng root extract.

Another aspect of the present invention is to provide a method for producing a metal nanoparticle comprising the step of reacting a metal precursor with a composition for producing a metal nanoparticle comprising the ginseng extract.

The method of the present invention manufactures metal nanoparticles using ginseng extract, which is a natural extract, which is environmentally friendly and significantly shortens the manufacturing time compared to chemical synthesis and physical synthesis, and at the same time maintains stabilization without a separate reducing agent or stabilizer. Metal nanoparticles can be produced, which can be used very usefully in various industrial fields.

Hereinafter, the method for producing a metal nanoparticle of the present invention will be described in detail.

In the present invention, the terms "ginseng", "ginseng extract" and "metal nanoparticles" are as described above.

The manufacturing method of the present invention can produce a variety of metal nanoparticles according to the metal precursor to be used, specifically, gold (Au), silver (Ag), copper (Cu), palladium (Pd), platinum (Pt) and It may be one or more selected from the group consisting of rhodium (Rd), but is not limited thereto. More specifically, gold (Au) or silver (Ag) can be selected.

As used herein, the term "metal precursor" means a compound added to prepare metal nanoparticles. Specifically, as the metal precursor, various kinds of compounds may be selected and used according to the metal type of the nanoparticles to be prepared. For example, at least one selected from the group consisting of tetrachlorogold (III) acid (HAuCl ₄ ), NaAuCl ₃ , and AuCl ₃ may be used to prepare gold nanoparticles, and more specifically, tetrachlorogold. Acid (III) (HAuCl ₄ ) may be used, but is not limited thereto.

In addition, at least one selected from the group consisting of AgBF ₄ , AgCF ₃ SO ₃ , AgClO ₄ , AgNO ₃ , AgPF _6, and Ag (CF ₃ COO) may be used to prepare silver nanoparticles, and more specifically, Silver nitrate (AgNO ₃ ) may be used, but is not limited thereto.

In addition, the production of copper nanoparticles are selected from the group consisting of CuCl ₂ , CuF ₂ , CuBr ₂ , CuI ₂ , (CH ₃ COO) ₂ Cu, Cu (ClO ₄ ) ₂ , Cu (NO ₃ ) ₂ and CuSO ₄ . One or more may be used, but is not limited thereto.

In the production method of the present invention, the concentration of the metal precursor used for the reaction may affect the production of metal nanoparticles. The concentration of the metal precursor of the present invention is not limited as long as the metal nanoparticles can be prepared by the composition for preparing metal nanoparticles, but specifically the concentration of the metal precursor is 0.01 to 100 mM, more specifically 0.01 to 50 mM, most specifically May be 0.01 to 10 mM.

In the production method of the present invention, the reaction temperature may affect the production of metal nanoparticles. The reaction temperature of the present invention is not limited as long as the metal nanoparticles can be prepared as a composition for preparing metal nanoparticles, but specifically, the reaction temperature may be 10 to 100 ° C, more specifically 50 to 100 ° C.

In the production method of the present invention, the reaction time may affect the production of metal nanoparticles. The reaction time of the present invention is not limited as long as the metal nanoparticles can be prepared as a composition for preparing metal nanoparticles, but specifically, when the metal is gold, the reaction time may be 1 minute to 1 hour, more specifically 1 minute to 50 minutes. When the metal is silver, the reaction time may be 30 minutes to 5 hours, more specifically 30 minutes to 2 hours.

In a specific embodiment of the present invention, when preparing gold or silver nanoparticles using leaf extract of panax ginseng, the reaction mixture containing tetrachlorogold (III) acid turned dark purple within 3 minutes. (A of FIG. 1), the reaction mixture containing silver nitrate turned completely brown within 45 minutes (b of FIG. 1). In addition, in the case of using the root extract of Panax ginseng, the reaction mixture containing silver nitrate took 2 hours to turn from white to brown (FIG. 1 d), and the reaction mixture containing tetrachlorogold (III) acid. Took 5 minutes (e of FIG. 1). In addition, when prepared using the red ginseng root extract, it was confirmed that the reaction mixture turns brown and ruby red within 1 hour for the production of silver nanoparticles and within 10 minutes for the production of gold nanoparticles, respectively. (F and g in Fig. 1).

Through the above results, the gold or silver nanoparticles prepared using the leaves or roots of the ginseng and the iris root extract are synthesized in a very short time compared to the nanoparticles prepared using other natural extracts or other site extracts of ginseng. In particular, when using the leaf extract of ginseng, it can be seen that a stable metal nanoparticles are synthesized in an economical and environmentally friendly way in a remarkably fast time.

The production method of the present invention may further comprise the step of recovering the metal nanoparticles by centrifuging the reaction solution.

In a specific embodiment of the present invention, the composition for preparing metal nanoparticles containing ginseng extract is added to 1 mM tetrachlorogold (III) acid and silver nitrate solution, respectively, and then maintained at 80 ° C. and centrifuged at 16,000 rpm for 20 minutes. To prepare a metal nanoparticle (Example 3).

Yet another aspect of the present invention is to provide a metal nanoparticle produced by the above production method.

Metal nanoparticles of the present invention is prepared using ginseng root or leaf extract or red ginseng root extract, containing the bioactive material of each extract, antimicrobial activity, biofilm very superior to the metal nanoparticles prepared by other methods It has degradation activity, anticoagulant activity, anticancer activity and anti-inflammatory activity and can be usefully used in various industries.

The "metal nanoparticle" is as described above.

The metal nanoparticles of the present invention can be used for medical, biological analysis, fuel, and electronic component materials. Specifically, the gold nanoparticles are used for organic solar cells, sensor probes, and biomedical medicines due to their inherent light conducting properties. It can be used as drag delivery, conductive material and catalyst, and silver nanoparticles can be used for antibacterial agent, biosensor for quantitative detection, optical analysis method and the like due to optical properties, conductivity and antimicrobial properties. In addition, the metal nanoparticles of the present invention can control the optical and electronic characteristics of the particles by changing the size or shape, the chemical properties or the aggregation state of the surface.

The shape of the metal nanoparticles prepared by the manufacturing method of the present invention may be generally spherical, but is not limited thereto.

In addition, the size of the metal nanoparticles prepared by the method of the present invention may be specifically 3 to 80 nm, more specifically 5 to 50 nm, most specifically 10 to 40 nm, but is not limited thereto. Compared with the known gold nanoparticles having a size of 2 to 40 nm, and silver nanoparticles having a size of 100 nm, the size of gold or silver nanoparticles prepared using the ginseng extract of the present invention is conventionally manufactured. It is small compared to metal nanoparticles, and its distribution is uniform. In particular, silver nanoparticles of the present invention are much smaller than conventional sizes.

In a specific embodiment of the present invention, it was confirmed that the metal nanoparticles prepared by using ginseng extract to produce nanoparticles of uniform size in the range of 10 to 40 nm (Example 3). Specifically, in the case of gold nanoparticles, the ginseng) leaf extract was 10 to 30 nm in size, the root extract of ginseng was 10 to 40 nm, and the red ginseng root extract was 10 to 30 nm. The size was very uniform.

In addition, in the case of silver nanoparticles, the size of 10 to 20nm when using the leaf extract of ginseng, the size of 10 to 30nm when using the root extract of ginseng, 10 to 30nm of the root extract of red ginseng It was confirmed that the size of the silver nanoparticles is also in a very uniform range.

Another aspect of the invention provides an antimicrobial composition comprising the metal nanoparticles.

The metal nanoparticles of the present invention are prepared using ginseng root or leaf extract or red ginseng root extract, and contain a bioactive substance of each extract, and thus have very superior antimicrobial activity than metal nanoparticles prepared by other methods.

The "metal nanoparticle" is as described above.

The antimicrobial composition of the present invention may have an antimicrobial activity against bacteria, fungi or yeast, in particular, Vibrio (Vibrio) into the microorganism is Salmonella (Salmonella) microorganism of the genus, stearyl Pyrococcus (Staphylococcus) in the microorganism is Escherichia (Escherichia) in the microorganism Bacillus (Bacillus) in microorganisms, and Candida may have an antimicrobial activity for at least one selected from the group consisting of (Candida) spp, more specifically, Vibrio para H. Molina T kusu (Vibrio parahaemolyticus , Salmonella enterica , Staphylococcus aureus , Escherichia coli , Bacillus anthracis , Bacillus cereus and Candida albicans Candida albicans) as it may have the antimicrobial activity for at least one selected from the group consisting of, whereby Not one.

Specifically, the silver nanoparticles of the present invention or the composition containing the silver nanoparticles are very excellent in antimicrobial activity against pathogenic microorganisms than the general silver nanoparticles, it can be widely used in various fields such as cosmetics, quasi-drugs, pharmaceuticals, foods, etc. have.

In a specific embodiment of the present invention, gold nanoparticles using the ginseng leaf extract was confirmed to have an antimicrobial activity against Vibrio parahemoliticus, Salmonella enterica, Escherichia coli, Stephylococcus aureus, Silver nanoparticles using ginseng root extract showed antimicrobial activity against Escherichia coli, Bacillus cereus, Stephylococcus aureus, Vibrio parahemolyticus and Bacillus anthracis. In addition, the silver nanoparticles using the red ginseng root extract have been confirmed to have antimicrobial activity against Vibrio parahemolyticus, Stephylococcus aurus, Bacillus cereus and Candida albicans, which is the largest against candida albicans, Next, it was confirmed that Vibrio parahemoliticus, Stephylococcus aureus, Bacillus cereus in the order of having antimicrobial activity. Through the above results, it can be seen that the silver nanoparticles of the present invention have antimicrobial activity.

Yet another aspect of the present invention is to provide a biofilm decomposition composition comprising the metal nanoparticles.

The metal nanoparticles of the present invention are prepared by using ginseng root or leaf extract or red ginseng root extract, and contain bioactive substances of each extract, and thus have much better biofilm degradation activity than the metal nanoparticles prepared by other methods. Have

The "metal nanoparticle" is as described above.

In the present invention, the term "biofilm" refers to a group of bacteria that bacteria are thinly adhered like a film. In other words, biofilms are produced by bacteria, which in turn provide a habitat for the bacteria, thereby accelerating bacterial growth.

The biofilm degrading composition of the present invention has a biofilm degrading activity and inhibits biofilm-related infection, thereby reducing the cost and pain of the patient. Specifically, the surface of plastic, metal, etc. is corroded. It can be used in a variety of fields, such as the industry to protect from, dental equipment industry, plumbing industry, but is not limited thereto.

In a specific embodiment of the present invention, as a result of confirming the biofilm degradation activity of the present invention by colorimetric method ( Sphyrococcus aureus ) and Pseudomonas aeruginosa ( Pseudomonas aeruginosa ), ginseng leaf extract Silver nanoparticles prepared using the highest biofilm degrading activity against Stepilococcus aureus and Pseudomonas eruzinosa at 4 g / ml concentration of silver nanoparticles were prepared using red ginseng root extract. In the above concentrations, it was confirmed that there is biodegradation activity against Stepilococcus aureus and Pseudomonas aeruginosa.

Through this, it can be confirmed that the nanoparticles prepared using ginseng extract may have a biofilm-degrading activity on the strain, thereby inhibiting biofilm-related infections, and thus may play a role in reducing cost and patient pain.

Yet another aspect of the present invention is to provide an anticoagulant composition comprising the metal nanoparticles.

The metal nanoparticles of the present invention are prepared using ginseng root or leaf extract or red ginseng root extract, and contain a bioactive substance of each extract, and thus have an excellent anticoagulant activity than the metal nanoparticles prepared by other methods. .

The "metal nanoparticle" is as described above.

In the present invention, the term "anticoagulant" means inhibiting the normal blood coagulation properties of the body, it is also used as anticoagulant. Specifically, there are anticoagulants that inhibit coagulation by adding them to the blood, and antithrombin III and heparin that inhibit the actions of various coagulation factors as blood physiological anticoagulants for the active coagulation factors present in the blood. It is divided into anticoagulant which inhibits the activity of the coagulation factor by oral administration, and anticoagulant which appears in the pathological state and reversibly and irreversibly inhibits the activity of the coagulation factor.

The anticoagulant composition of the present invention includes metal nanoparticles prepared using ginseng extract, specifically, ginseng leaf or root extract or red ginseng root extract, thereby preventing blood coagulation. It may be used in the field of biotechnology, and more specifically, it may be used as a carrier of drugs and genes in direct contact with blood or as a biosensor, an in vivo nanocarrier, but is not limited thereto. It can also be used for experimental purposes in vitro.

In the present invention, the anticoagulant composition may be used as a pharmaceutical, quasi-drug, cosmetics, food and feed composition.

In a specific embodiment of the present invention, the blood of healthy male and female volunteers ranging in age from 25 to 45 years old is collected in two vacuum collectors and labeled as A and B, respectively. Observed. As a result, the gold nanoparticles prepared using the ginseng leaf extract was inhibited coagulation formation when gold nanoparticles are present, it was confirmed that blood is completely coagulated in the control group that does not contain gold nanoparticles. Through these results, it can be seen that the gold nanoparticles biosynthesized by the leaf extract of ginseng has anticoagulant activity.

Another aspect of the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising the metal nanoparticle of the present invention.

The metal nanoparticles of the present invention are prepared using ginseng root or leaf extract or red ginseng root extract, and contain a bioactive substance of each extract, and thus have very superior anticancer activity than metal nanoparticles prepared by other methods.

The "metal nanoparticle" is as described above.

The term "prevention" of the present invention means any action that inhibits or delays cancer disease by administration of the pharmaceutical composition.

In addition, the term "treatment" of the present invention means all the actions that improve or cure the symptoms of cancer disease by administration of the pharmaceutical composition.

The cancer may include lung cancer, non-small cell lung cancer, colon cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, skin or intraocular melanoma, uterine cancer, ovarian cancer, rectal cancer, gastric cancer, anal muscle cancer, colon cancer, breast cancer, fallopian tube carcinoma, Endometrial carcinoma, cervical carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, small intestine cancer, endocrine adenocarcinoma, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, Chronic or acute leukemia, lymphocyte lymphoma, bladder cancer, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system (CNS) tumor, primary central nervous system lymphoma, spinal cord tumor, brain stem glioma, pituitary adenoma, etc. May be, but is not limited thereto.

The pharmaceutical composition may comprise a pharmaceutically acceptable carrier.

As used herein, the term “pharmaceutically acceptable carrier” may refer to a carrier, excipient or diluent that does not stimulate the biological activity and properties of the compound to be injected without stimulating the organism, and in particular, an unnatural carrier (non-naturally occuring carrier). The kind of the carrier usable in the present invention is not particularly limited, and any carrier can be used as long as it is a conventionally used and pharmaceutically acceptable carrier in the art. Non-limiting examples of the carrier include saline, sterile water, Ringer's solution, buffered saline, albumin injection solution, dextrose solution, maltodextrin solution, glycerol, ethanol and the like. These may be used alone or in combination of two or more thereof.

The pharmaceutical composition comprising a pharmaceutically acceptable carrier may be in various oral or parenteral formulations. When formulated, diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrating agents, and surfactants are usually used.

Specifically, solid preparations for oral administration include tablets, pills, powders, granules, capsules and the like, and such solid preparations include at least one excipient such as starch, calcium carbonate, sucrose, lactose in the compound. , Gelatin and the like can be mixed. In addition to simple excipients, lubricants such as magnesium stearate, talc can also be used. Oral liquid preparations include suspensions, solvents, emulsions, and syrups.In addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, fragrances, and preservatives may be included. have. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations and suppositories. As the non-aqueous solvent and suspending agent, propylene glycol, polyethylene glycol, vegetable oils such as olive oil, injectable esters such as ethyl oleate and the like can be used. As the base of the suppository, utopsol, macrogol, tween 61, cacao butter, laurin butter, glycerogelatin and the like can be used.

The pharmaceutical composition may be administered in a pharmaceutically effective amount.

The "pharmaceutically effective amount" means an amount sufficient to treat the disease at a reasonable benefit / risk ratio applicable to medical treatment, and the effective dose level is the type of subject and its severity, age, sex, type of virus infected, drug Activity, sensitivity to drug, time of administration, route of administration and rate of release, duration of treatment, factors including concurrent use of drugs, and other factors well known in the medical arts. For example, the pharmaceutical composition may be administered such that the metal nanoparticles of the present invention are each administered at 0.0001 to 1000 mg / kg, specifically, at 0.001 to 100 mg / kg.

The administration means introducing the composition of the present invention to the patient in any suitable manner, and the route of administration of the composition can be administered via any general route as long as it can reach the target tissue. Intraperitoneal administration, intravenous administration, intramuscular administration, subcutaneous administration, intradermal administration, oral administration, topical administration, intranasal administration, but is not limited thereto.

The pharmaceutical composition of the present invention may be administered daily or intermittently, and the number of administrations per day may be administered once or divided into two to three times. The frequency of administration in the case where the two active ingredients are single drugs may be the same or different times. In addition, the compositions of the present invention can be used alone or in combination with other drug treatments for the prevention or treatment of cancer diseases. Taking all of the above factors into consideration, it is important to administer an amount that can obtain the maximum effect in a minimum amount without side effects, and can be easily determined by those skilled in the art.

The subject means all animals, such as rats, mice, and livestock, including humans who may develop or may develop cancer. Specifically, it may be a mammal including a human.

In a specific embodiment of the present invention, it was confirmed that the silver and gold nanoparticles showed cytotoxicity at concentrations of 100 μg / mL and 2 μg / mL, respectively, against human lung cancer cells (A549) (FIG. 25). This suggests that silver and gold nanoparticles biosynthesized by the leaf extract of ginseng have anticancer activity.

Another aspect of the present invention provides a food composition for preventing or improving cancer comprising the metal nanoparticles of the present invention.

The "metal nanoparticle" is as described above.

The term "food" of the present invention is a dairy product including meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, chewing gum, ice cream, various soups, beverages, tea, drinks, alcoholic beverages, Vitamin complexes, nutraceuticals and health foods and the like, and include all foods in the conventional sense.

The functional food (functional food) is the same term as a food for special health use (Food for special health use, FoSHU), in addition to the nutritional supply, the processed food, so that the bioregulatory function appears efficiently, high medical effect Means food. Here, the term "functionality" means obtaining useful effects for health purposes such as nutrient control or physiological action on the structure and function of the human body. The food of the present invention can be prepared by a method commonly used in the art, and the preparation can be prepared by adding the raw materials and ingredients commonly added in the art. In addition, the formulation of the food may also be prepared without limitation as long as the formulation is recognized as a food. Food composition of the present invention can be prepared in various forms of formulation, unlike the general medicine has the advantage that there is no side effect that can occur when taking a long-term use of the drug as a raw material, and excellent portability, the present invention Can be taken as a supplement to enhance the effect of preventing or ameliorating inflammatory diseases.

The health food refers to foods having active health maintenance or promotion effect as compared to general foods, and the health supplement food refers to foods for health supplementation purposes. In some cases, the terms nutraceutical, health food, dietary supplement are used.

Specifically, the health functional food is a food prepared by adding the compound of the present invention to food materials such as beverages, teas, spices, gums, confections, or the like, encapsulated, powdered, suspensions, etc. Means to bring effect, but unlike the general medicine has the advantage that there is no side effect that can occur when taking long-term use of the drug as a raw material.

Since the food composition of the present invention can be consumed on a daily basis, since a high effect can be expected for the prevention or improvement of cancer diseases, it can be very usefully used.

The food composition may further include a physiologically acceptable carrier, and the type of carrier is not particularly limited and may be any carrier that is commonly used in the art.

In addition, the food composition may include additional ingredients that are commonly used in food compositions to improve the smell, taste, time and the like. For example, it may include vitamins A, C, D, E, B1, B2, B6, B12, niacin, biotin, folate, panthotenic acid, and the like. In addition, minerals such as zinc (Zn), iron (Fe), calcium (Ca), chromium (Cr), magnesium (Mg), manganese (Mn), copper (Cu) and chromium (Cr) may be included. It may also contain amino acids such as lysine, tryptophan, cysteine, valine and the like.

In addition, the food composition is a preservative (potassium sorbate, sodium benzoate, salicylic acid, sodium dehydroacetic acid, etc.), fungicides (bleaching powder and highly bleaching powder, sodium hypochlorite, etc.), antioxidants (butylhydroxyanisol (BHA), butylhydride) Oxytoluene (BHT), etc.), colorant (such as tar pigment), colorant (sodium nitrite, sodium nitrite, etc.), bleach (sodium sulfite), seasoning (MSG glutamate, etc.), sweetener (ducin, cyclate, saccharin Foods such as sodium, etc.), fragrances (vanillin, lactones, etc.), swelling agents (alum, potassium D-tartrate, etc.), reinforcing agents, emulsifiers, thickeners (foils), coatings, gum herbicides, foam inhibitors, solvents, modifiers, etc. It may include food additives. The additive may be selected according to the type of food and used in an appropriate amount.

The compound of the present invention may be added as it is or used with other food or food ingredients, and may be appropriately used according to conventional methods. The mixed amount of the active ingredient can be suitably determined according to the purpose of use (prevention, health or therapeutic treatment). In general, the food composition of the present invention may be added in an amount of 50 parts by weight or less, specifically 20 parts by weight or less based on the food or beverage in the manufacture of food or beverage. However, when ingested for a long time for the purpose of health and hygiene, it may include the content below the above range, and since there is no problem in terms of safety, the active ingredient may be used in an amount above the above range.

An example of the food composition of the present invention may be used as a health beverage composition, in which case it may contain various flavors or natural carbohydrates and the like as additional ingredients, such as a general beverage. The above-mentioned natural carbohydrates include monosaccharides such as glucose and fructose; Disaccharides such as maltose and sucrose; Polysaccharides such as dextrin, cyclodextrin; Sugar alcohols such as xylitol, sorbitol, and erythritol. Sweeteners include natural sweeteners such as taumartin, stevia extract; Synthetic sweeteners such as saccharin and aspartame; The ratio of the natural carbohydrate may be generally about 0.01 to 0.04 g, specifically about 0.02 to 0.03 g per 100 mL of the health beverage composition of the present invention.

In addition to the above, the health beverage composition includes various nutrients, vitamins, electrolytes, flavors, colorants, pectic acid, salts of pectic acid, alginic acid, salts of alginic acid, organic acids, protective colloid thickeners, pH regulators, stabilizers, preservatives, glycerin, Alcohol or carbonation agent and the like. Others may contain fruit flesh for the production of natural fruit juices, fruit juice drinks, or vegetable drinks. These components can be used independently or in combination. Although the ratio of such an additive is not critical, it is generally selected from the range of 0.01 to 0.1 parts by weight per 100 parts by weight of the health beverage composition of the present invention.

The food composition of the present invention may include various weight percents as long as it can exhibit a prophylactic or ameliorating effect of cancer disease, but specifically, the metal nanoparticles of the present invention may be 0.00001 to 100% by weight or 0.01 to 80 to the total weight of the food composition. It may include by weight, but is not limited thereto.

Another aspect of the present invention provides a feed composition for cancer prevention or improvement comprising the metal nanoparticles of the present invention.

The term "feed" of the present invention is any natural or artificial diet, meal, or the like for the animal to eat, ingest and digest or as appropriate, comprising the compound of the present invention as an active ingredient The feed may be prepared in various forms of feed known in the art, and may specifically include rich feed, forage and / or special feed.

The metal nanoparticles of the present invention included in the feed composition of the present invention may vary depending on the purpose of use and conditions of use of the feed. For example, 0.01 to 100% by weight, more specifically 1 to 1, based on the total weight of the livestock feed composition. 80 wt% may be included, but is not limited thereto.

Another aspect of the invention provides an anti-inflammatory composition comprising the metal nanoparticles.

The metal nanoparticles of the present invention are prepared using ginseng root or leaf extract or red ginseng root extract, and contain a bioactive substance of each extract, and thus have very superior anti-inflammatory activity than metal nanoparticles prepared by other methods. .

The "metal nanoparticle" is as described above.

As used herein, the term “anti-inflammatory” refers to an action of inhibiting or reducing inflammation, wherein the term “inflammation” refers to a protective response occurring in the body when a living tissue is damaged, causing an inflammatory disease. to be.

In the present invention, the anti-inflammatory composition can be used for the prevention, treatment or amelioration of inflammatory diseases by showing the activity of inhibiting or reducing inflammation.

The inflammatory disease is not particularly limited as long as the symptoms can be alleviated, alleviated, improved or treated by the anti-inflammatory composition of the present invention. Specific examples include Crohn's disease, erythema, atopy, rheumatoid arthritis, Hashimoto's thyroiditis, and malignancy. Anemia, Edison's disease, type 1 diabetes, lupus, chronic fatigue syndrome, fibromyalgia, hypothyroidism and hypertension, scleroderma, Behcet's disease, inflammatory bowel disease, multiple sclerosis, myasthenia gravis, Meniere's syndrome, Gillian Guilian-Barre syndrome, Sjogren's syndrome, vitiligo, endometriosis, psoriasis, vitiligo, systemic scleroderma or ulcerative colitis.

The anti-inflammatory composition in the present invention can be used as a pharmaceutical, quasi-drug, cosmetics, food and feed composition.

In a specific embodiment of the present invention, silver nanoparticles were found to have no cytotoxicity to RAW264.7 cells up to 1000 μM concentration and inhibit NO production in a concentration-dependent manner (FIG. 26). In addition, it was confirmed that gold nanoparticles have no cytotoxicity to RAW264.7 cells up to a concentration of 100 μM and inhibit NO production in a concentration-dependent manner (FIG. 26). Through this, it was confirmed that silver and gold nanoparticles biosynthesized by the leaf extract of ginseng have anti-inflammatory activity without cytotoxicity.

The composition for preparing metal nanoparticles of the present invention and the method for preparing metal nanoparticles using the same using the root or leaf extract of ginseng as a natural extract as an active ingredient, the production time is significantly shorter than before, and without the addition of additional reducing agents or stabilizers. Metal nanoparticles of uniform size can be prepared. In addition, the composition comprising the metal nanoparticles prepared by the method has antimicrobial activity, biofilm degradation activity, anticoagulant activity, anticancer activity and anti-inflammatory activity can be utilized in various industries.

1 is a reaction of ginseng leaf extract (a to c), ginseng root extract (d and e) or red ginseng (f and g) extract and tetrachlorogold (III) acid (HAuCl ₄ · 3H ₂ O) or silver nitrate solution The following color change is shown (control: control, AgNPs: silver nanoparticles, AuNPs: gold nanoparticles).
Figure 2 is a view confirming the production of gold or silver nanoparticles prepared using ginseng leaf extract through UV-vis.
3 is a view confirming the production of gold or silver nanoparticles prepared using the ginseng root extract through UV-vis.
Figure 4 is a view confirming the production of gold or silver nanoparticles prepared using the red ginseng root extract through UV-vis.
5 is a view showing the structural characteristics of gold or silver nanoparticles prepared using ginseng leaf extract through FE-TEM.
Figure 6 is a view showing the structural characteristics of gold or silver nanoparticles prepared using ginseng root extract through FE-TEM.
Figure 7 shows a simple dispersion of the structural features of the gold or silver nanoparticles prepared using ginseng root extract through FE-TEM, fringe spacing of nanoparticles (fringe spacing).
8 is a view showing the structural characteristics of gold or silver nanoparticles prepared using the red ginseng root extract through FE-TEM.
Figure 9 shows the purity of the gold or silver nanoparticles prepared using ginseng leaf extract through EDAX analysis.
10 shows the purity of gold or silver nanoparticles prepared using ginseng root extract through EDAX analysis.
Figure 11 shows the purity of gold or silver nanoparticles prepared using red ginseng leaf extract through EDAX analysis.
12 is an analysis of element mapping results of gold or silver nanoparticles prepared using ginseng leaf extract through TEM.
FIG. 13 is an analysis of element mapping results of gold or silver nanoparticles prepared using ginseng root extract through TEM.
14 is an analysis of element mapping results of gold or silver nanoparticles prepared using red ginseng root extract through TEM.
15 shows a diffraction pattern of gold or silver nanoparticles prepared using ginseng leaf extract through X-ray diffraction.
Figure 16 shows the diffraction pattern of gold or silver nanoparticles prepared using ginseng root extract, through X-ray diffraction.
FIG. 17 is a diagram illustrating a size distribution profile result based on volume, intensity, and number of gold or silver nanoparticles prepared using ginseng leaf extract through dynamic light scattering analysis.
FIG. 18 is a diagram showing a size distribution profile result based on the volume, intensity, and number of gold or silver nanoparticles prepared using red ginseng root extract through dynamic light scattering analysis.
Figure 19 shows the antimicrobial activity of gold or silver nanoparticles prepared using the leaf extract of ginseng.
20 shows the antimicrobial activity of silver nanoparticles prepared using the root extract of ginseng.
Figure 21 shows the antimicrobial activity of the silver nanoparticles prepared using the root extract of red ginseng.
22 shows biofilm degradation activity of silver nanoparticles prepared using the leaf extract of ginseng.
Figure 23 shows the biofilm degradation activity of silver nanoparticles prepared using the root extract of red ginseng.
Figure 24 shows the anticoagulant activity of gold or silver nanoparticles prepared using the leaf extract of ginseng.
FIG. 25 shows cytotoxicity against Keratinocyte cells and lung cancer cells (A549 cells) of gold or silver nanoparticles prepared using leaf extracts of ginseng.
Figure 26 shows the anti-inflammatory activity of gold or silver nanoparticles prepared using the leaf extract of ginseng.

Hereinafter, the present invention will be described in more detail with reference to Examples. These examples are intended to illustrate the present invention more specifically, but the scope of the present invention is not limited to these examples.

Example 1. Materials

Tetrachlorogold (III) acid (HAuCl ₄ .3H ₂ O) and silver nitrate (AgNO ₃ ) were purchased from Sigma-Aldrich Chemicals, USA, media were purchased and purchased from Difco (MB cell, Seoul, Republic of Korea). Also, the roots and leaves of ginseng were collected from 4 year old ginseng in Gochang, Korea.

Standard antibiotic discs (discs) are commercially available vancomycin (VA30) 30 μg / disc, rifampicin (RD5) 5 μg / disk, oleandomycin (OL15) 15 μg / disk, penicillin G (P10) 10 Μg / disk, 30 μg / disk of novobiocin (NV30), 15 μg / disk of lincomycin (MY15) and 30 μg / disc of tetracycline (TE30) were purchased (Oxoid Ltd., England). .

Pathogenic bacterial strains include Vibrio parahaemolyticus [ATCC 33844], Salmonella enteric [ATCC 13076], Escherichia coli [ATCC 10798], Stepirococcus aureus Staphylococcus aureus [ATCC 6538], Bacillus anthracis [NCTC 10340], Bacillus cereus [ATCC 14579], Candida albicans [KACC 30062], and Pseudomonas aeruginosa ( Pseudomonas aeruginosa ) [ATCC 27853] was used in the group consisting of. The bacterial strains were cultured in agar nutrient medium at 37 ° C. and stored at −0 ° C. in glycerol stock vials.

Other human blood was collected from healthy male and female volunteers and stored in a vacuum collector tube (Greiner Bio-One).

Example 2. Preparation of Ginseng Extract

2-1. Preparation of Leaf Extract of Ginseng

15 g of the ginseng leaves collected in Example 1 was cleaned, and then washed several times in distilled water to remove dust and suspended substances, and the shade was dried at room temperature for 6-8 hours. The leaves were then cut into small pieces and ground with mortar and pestle. Then, the pieces were heated in 100 ml of sterile water for 20 minutes, and the extract obtained by leaching the components of ginseng into water was filtered through Whatman filter paper, followed by centrifugation at 10,000 rpm for 15 minutes. The filtered liquid was placed in 100 ml of sterile water and stored at 4 ° C.

2-2. Root Extract Preparation of Ginseng

25 g of the ginseng roots collected in Example 1 was cleaned, and then washed with distilled water several times to remove dust or suspended substances. The roots were then cut into small pieces and ground with mortar and pestle. Then, the pieces were heated in 100 ml of sterile water for 30 minutes, and the extract obtained by leaching the components of ginseng into water was filtered with Whatman filter paper, and then centrifuged at 10,000 rpm for 10 minutes. The filtered liquid was placed in 100 ml of sterile water and stored at 4 ° C.

2-3. Root Extract Preparation of Red Ginseng

After red ginseng root was ground to obtain red ginseng root powder, 10 g of root powder was mixed with 100 ml of sterile water, heated for 30 minutes, and the liquid obtained by leaching the red ginseng component in water was filtered through Whatman filter paper and centrifuged at 10,000 rpm for 10 minutes. The filtered liquid was placed in 100 ml of sterile water and stored at 4 ° C.

Example 3. Synthesis of Gold or Silver Nanoparticles

3-1. Synthesis method of gold or silver nanoparticles using ginseng leaf extract

In 100 ml of ginseng leaf filtrate prepared in Example 2-1, 5 ml of the filtrate was mixed with 25 ml of sterile water, and a final concentration of 1 mM tetrachlorogold (III) acid and silver nitrate solution were added to the reaction mixture, respectively. Added. Thereafter, the reaction mixture was maintained at 80 ° C. such that Au ³⁺ was reduced to Au º, and Ag ⁺ was reduced to Ag º atoms, and the color change indicating the formation of nanoparticles was continuously observed. The reaction mixture was centrifuged at 2,000 rpm for 10 minutes to remove unwanted components, centrifuged at 16,000 rpm for 15 minutes to pellet, washed several times with sterile water, and then dried in air overnight. Here, the color change of the reaction mixture was visually observed to confirm the synthesis of gold or silver nanoparticles (AuNP, AgNP).

As can be seen in Figures 1 a to c, the reaction mixture with tetrachlorogold (III) acid turned dark purple within 3 minutes (a in FIG. 1), and the reaction mixture with silver nitrate was completely removed within 45 minutes. Turned brown (b in FIG. 1). On the other hand, the color change was not observed in the control group, which is present in the same ratio as the ginseng leaf extract and sterile water and similar to the reaction mixture solution (FIG. 1 c). The color change is caused by surface plasmon vibration and is caused by the formation of gold or silver nanoparticles in the reaction mixture.

That is, through the above results, it can be seen that gold or silver nanoparticles prepared using the leaf extract of ginseng are synthesized in a very short time compared to nanoparticles prepared using other natural extracts or other site extracts of ginseng. Using the leaf extract of ginseng, it can be seen that it is possible to synthesize a stable metal nanoparticles in an economical and environmentally friendly way in a very fast time, through the phenolic acid, flavonoids, ginsenosides present in the ginseng leaves It can be indirectly confirmed that, and polysaccharides affect the formation of metal nanoparticles.

3-2. Synthesis method of gold and silver nanoparticles using ginseng root extract

In 100 ml of ginseng root filtrate prepared in Example 2-2, 5 ml of the filtrate was mixed with 25 ml of sterile water, and a final concentration of 1 mM tetrachlorogold (III) acid and silver nitrate were added to the reaction mixture, respectively. The temperature was maintained at 80 ° C. The reaction mixture was centrifuged at 2,000 rpm for 10 minutes to remove unwanted components, centrifuged at 16,000 rpm for 20 minutes to pellet, washed several times with sterile water, and then dried in air overnight. Here, the color change of the reaction mixture was visually observed to confirm the synthesis of gold and silver nanoparticles.

As can be seen in Figure 1 d and e, the reaction mixture with silver nitrate took 2 hours to turn from white to brown, the reaction mixture with tetrachloro gold (III) acid is completely changed to pink after 5 minutes It was. On the other hand, no color change was observed in the control group. In addition, the color change is caused by the surface plasmon (surfacde plasmon) vibration, which is caused by the formation of gold or silver nanoparticles in the reaction mixture, through which the synthesis time is slower than the ginseng leaf extract, but other natural extracts As compared with the synthesis time of the gold or silver nanoparticles prepared by using it was confirmed that the synthesis in a short time.

3-3. Synthesis of Gold and Silver Nanoparticles Using Red Ginseng Root Extract

In 100 ml of red ginseng root filtrate prepared in Example 2-3, 5 ml of the filtrate was mixed with 25 ml of sterile water, and a final concentration of 1 mM tetrachlorogold (III) acid and silver nitrate were added to the reaction mixture, respectively. It was maintained at 80 ℃ so that the reduction reaction can occur. Then, centrifugation at 2,000rpm for 10 minutes to remove the unwanted components, centrifugation at 16,000rpm for 15 minutes to pellet, washed several times with sterile water, and then dried in air overnight. Here, the color change of the reaction mixture was visually observed to confirm the synthesis of gold and silver nanoparticles.

As a result, as can be seen in the f and g of Figure 1, the reaction mixture turned brown and ruby red within 1 hour and 10 minutes, respectively. Through this, gold or silver nanoparticles prepared using the red ginseng root extract are synthesized more slowly than using the ginseng leaf extract, but are synthesized in a significantly faster time than the synthesis time of the metal nanoparticles prepared using other natural extracts. It could be confirmed.

Example 4. Characterization of Metal Nanoparticles Prepared Using Ginseng Extract

4-1. Analysis method

Nanoparticles prepared using the ginseng extract are ultraviolet / visible spectroscopy (UV-vis), field-emission transmission electron microscopy (FE-TEM), energy dispersive X Nanoparticle size, shape, composition and characterization were analyzed using energy dispersive X-ray spectrometer (EDX spectra), elemental mapping, dynamic light scattering (DLS) and stability analysis.

Specifically, the reduction of gold and silver ions was confirmed by scanning each solution with absorbance over a wavelength range of 300-800 nm in a UV-vis spectrophotometer (Ultrospec 2100 Pro, Amersham, Biosciences). In addition, field emission transmission electron microscopy (FE-TEM), energy dispersive X-ray spectroscopy (EDAX), and JEM-2100F (JEOL) devices operating at 200 kV The size, shape, morphology and distribution of the prepared nanoparticles were analyzed using elemental mapping.

In addition, for FM-TEM, EDAX and elemental mapping analysis, a small amount of purified nanoparticles was added dropwise onto a carbon coated copper grid, and dried in an oven at 60 ° C. to prepare a sample. The prepared samples were analyzed for structural properties of nanoparticles through FE-TEM, and the distribution of elements through EDX and element mapping.

In addition, the size analysis profile of the nanoparticles was analyzed by using dynamic light scattering (DLS), Particles size analyzer, Photal, Otsuka Electronics, Japan. Specifically, hydrodynamic diameters and polydispersity index (PDI) were analyzed at 25 ° C., and pure water having a refractive index of 1.3328, viscosity 0.8878 and dielectric constant 78.3 was used as a reference dispersion medium. It was.

X-ray diffraction (XRD) analysis of the prepared nanoparticles was carried out with a CuKα radiation of 40 kv and 40 mA on an X-ray diffractometer (DX Advance, Bruker, Germany). A scan rate of 6 ° / min, a step site of 0.02, and a 2θ range of 20 to 80 ° were performed.

In addition, the stability analysis of the nanoparticles was observed by dividing before and after adding sodium hydroxide to the nanoparticles. The effect of changing the pH on the stability of the nanoparticles was observed in the pH range 3 to 12.

4-2. Characterization Results of Gold or Silver Nanoparticles Prepared Using Ginseng Extract

4-2-1. Ultraviolet / Visible Spectroscopy (UV-vis) Analysis

As a result of confirming the production of gold or silver nanoparticles prepared using the ginseng leaves, root extract and red ginseng root extract prepared in Examples 3-1 to 3-3 through UV-vis analysis, using ginseng leaf extract In the case of gold nanoparticles at 578nm, silver nanoparticles at the main absorption peak was confirmed, it was confirmed that the gold and silver nanoparticles were prepared using the ginseng leaf extract (Fig. 2 a and b). In addition, when the ginseng root extract was used, the main absorption peak was confirmed at 412 nm and the silver nanoparticles at 412 nm, and it was confirmed that gold and silver nanoparticles were prepared even when the ginseng root extract was used. A and b) of 3).

Finally, when the red ginseng root extract was used, the main absorption peak was confirmed at 410 nm for silver nanoparticles and at 560 nm for gold nanoparticles. Thus, gold and silver nanoparticles using red ginseng root extract were prepared. (A and b in FIG. 4).

These main absorption peaks are specific peaks of gold or silver nanoparticles, corresponding to the surface plasmon resonance of the nanoparticles. The gold or silver nanoparticles synthesized using the ginseng leaf, root or red ginseng root extract did not change the UV-vis spectrum for several months even when stored at 20 ° C. It indicates that it is stable.

4-2-2. Field-emission transmission electron microscopy (FE-TEM) analysis

Through FE-TEM, structural features of gold or silver nanoparticles prepared using the leaves, roots or root extracts of red ginseng were prepared in Examples 3-1 to 3-3.

As a result, the gold nanoparticles prepared from the leaf extract of ginseng had a size of 10 to 30 nm and had a spherical shape (a and b in FIG. 5), and the interference fringe spacing was 0.23 nm (FIG. 5, c). In addition, in FIG. 5D, the selected electron diffraction pattern of the purified spherical gold nanoparticles having the corresponding rings on [111], [200], [220], [311], and [222]. diffraction pattern, SEAD) could be confirmed (FIG. 5 d). The silver nanoparticles have a spherical shape of 10 to 20 nm (e and f in Fig. 5), and the interference fringe spacing is 0.24 nm (g in Fig. 5), [111], [200], [220], [ 311], and SEAD images of silver nanoparticles having a ring corresponding to [222] was confirmed (h of Figure 5).

Silver nanoparticles prepared with the root extract of ginseng are 10 to 30nm in size and spherical (a and b in Fig. 6), gold nanoparticles are 10 to 40nm in size, it was confirmed that the sphere (Fig. 6c, d ). In addition, the silver and gold nanoparticles were uniform and simple dispersion type (a of FIG. 7), it was confirmed that the nanoparticle interference fringe spacing (fringe spacing) of 0.23nm (b, c of FIG. 7). Ring-like diffraction patterns showing that the particles were crystals identified SEAD images of silver and gold nanoparticles corresponding to [111], [200], [220], and [311] (FIG. 7). D to f).

Gold and silver nanoparticles prepared using the red ginseng root extract was confirmed to have a size of 10 to 30nm, a spherical shape, and a uniform simple dispersion (FIGS. 8A and 8B).

Through these results, it was confirmed again that the gold and silver nanoparticles prepared using the leaves, root extract of ginseng or root extract of red ginseng. In addition, it was confirmed that the size of the gold and silver nanoparticles produced is uniform and small in size compared to the metal nanoparticles prepared in the past, and in particular, silver nanoparticles are significantly larger than the conventional size of 100nm. It was confirmed that the reduction.

4-2-3. Energy Dispersive X-Ray Analysis (EDAX) Analysis

Through EDAX analysis, the purity of the gold and silver nanoparticles prepared in Example 3 was measured, respectively.

As a result, in the case of gold or silver nanoparticles prepared using the leaf extract of ginseng, as shown in a and b of FIG. 9, gold nanoparticles are optically at 2.15 keV and silver nanoparticles at 3 keV. Absorption was observed (a and b in Fig. 9), and when the root extract of ginseng was used, optical absorption was observed at 3 keV for silver nanoparticles and 2.2 keV for gold nanoparticles (a and b in Fig. 10). ). In addition, in the case of using the root extract of red ginseng, optical absorption was observed at 3 keV for silver nanoparticles and 2.3 keV for gold nanoparticles (a and b in FIG. 11). And as a property of the silver nanocrystals, it can be confirmed that the gold and silver nanoparticles are produced through this.

4-2-4. Transmission electron microscope (TEM) analysis

Through TEM, the distribution of gold and silver nanoparticles prepared in Example 3 was analyzed, and the element mapping result confirmed the synthesized nanoparticle pellet solution through an electron microscope.

As a result, it was confirmed that the distribution of gold nanoparticles prepared using ginseng leaf extract was 54.73% (a and b in FIG. 12) and 47.70% was silver nanoparticles (c and d in FIG. 12). Distribution of silver nanoparticles prepared using the extract was found to be 54.73% (c and d in FIG. 13), and gold nanoparticles were 52.16% (c and d in FIG. 13). In addition, the element mapping results when synthesized using the red ginseng root extract was confirmed through a to d of FIG. The results obtained from the elemental mapping confirmed that the gold and silver nanoparticles of the present invention are excellent particles in each material.

4-2-5. X-ray diffraction (XRD) analysis

Through XRD analysis, the diffraction patterns of the gold and silver nanoparticles prepared in Example 3 were analyzed.

As a result, the gold nanoparticles prepared using the ginseng leaf extract showed intense peaks in the 2θ value full spectrum ranging from 20 to 80 corresponding to [111], [200], [220], [311] and [222]. The silver nanoparticles were able to identify peaks at 2θ values corresponding to [111], [200], [331], [241], and [311] in the XRD spectrum (FIGS. 15A and 15B). When the ginseng root extract was used, intense peaks were observed in the 2θ value full spectrum corresponding to [111], [200], [331], [241] and [311] (FIGS. 16A and 16B). This shows similar values to Bragges's reflections of gold and silver nanocrystals, indicating that gold and silver nanoparticles were prepared from these results.

4-2-6. Dynamic Light Scattering (DLS) Analysis

Through dynamic light scattering analysis, the size distribution of the particles was determined based on the volume, intensity, and number of the gold and silver nanoparticles prepared in Example 3.

As a result, the gold nanoparticles prepared using the ginseng leaf extract ranged from 50 to 150 nm with a polydispersity index (PDI) of 0.191 (a to c in FIG. 17), and the silver nanoparticles to It was confirmed that the dispersion index (polydispersity index) is in the range of 70 to 140 nm, 0.13 (Fig. 17 d to f), the average particle size of the gold and silver nanoparticles, that is, the hydrodynamic diameter of the nanoparticles, respectively It was confirmed that it is 97nm and 80nm. In addition, the silver nanoparticles using the red ginseng root extract has a polydispersity index (PDI) of 0.190, an average diameter of 83 nm (a to c in FIG. 18), and the gold nanoparticles have a polydispersity index of polydispersity. index) was 0.159 and the average diameter was 183 nm (d to f in FIG. 18).

4-2-7. Stability analysis

Finally, for the stability analysis of the gold and silver nanoparticles prepared in Example 3, 0.1M sodium hydroxide was added to the reaction mixture for the synthesis of nanoparticles having pH of 5.6 and 6, respectively.

As a result, gold or silver nanoparticles prepared using the leaf or root extract of ginseng, even if sodium hydroxide was added, it was confirmed that the main shift (shift) does not appear stable, especially, pH 3 to 12 There was no major change in the absorption of the range. In addition, after three weeks after synthesis, half of the mixture was checked for UV-vis spectra, and no change was observed. In addition, even when synthesized using the red ginseng root extract, it was confirmed that the main shift (shift) does not appear in the pH range is stable, and after three weeks after synthesis, half of the result of checking the UV-vis spectrum of the mixed solution Also, no change was observed.

This means that the gold and silver nanoparticles synthesized using the ginseng leaf or root extract or the red ginseng extract exhibit stability without the addition of additional reducing agents or stabilizers. In particular, for leaf extracts, the above results indicate that phenolic acids and flavonoids exhibit antioxidant activity present in ginseng leaves, and ginsenosides such as Rb1, Rb2, Rc, Rd, Re, Rg1, F1, F2, and F4. It is suggested that components such as side play an important role in the stability of the synthesized gold and silver nanoparticles, and accordingly, the ginseng leaf extract acts as a reducing agent and stabilizer by itself without any other reducing agent or stabilizer, Gold and silver nanoparticles, prepared using ginseng leaf extracts, suggest the potential for application to drug carriers over a wide range of pH.

Example 5. Evaluation of Antimicrobial Activity of Nanoparticles

5-1. How to measure

In order to measure the antimicrobial activity of the nanoparticles, it was measured on Muller-Hinton agar (MHA) plate using the disc diffusion method (disc diffusion method). Pathogenic microorganisms used in the evaluation of antimicrobial activity include Vibrio parahaemolyticus , Salmonella enterica , Staphylococcus aureus , Escherichia coli , and Bacillus antra Bacillus anthracis , Bacillus cereus and Candida albicans were selected and used.

Apply each strain to 100 μl of MHA medium plate evenly and incubate for one day, and apply 30 μl (100 mg / L) or 50 μl (100 mg / L) of the prepared silver and gold nanoparticle reaction mixture. Incubated at 37 ° C for 24 hours. After incubation, inhibition zones around the silver and gold nanoparticles were measured and compared with the inhibition zones of each antibiotic disc. As a control, one or more standard antibiotics are selected from the group consisting of vancomycin, novobiocin, tetracycline, rifampicin, oleandomycin, penicillin, and lincomycin. It was used by.

5-2. Antibacterial activity measurement result

Gold nanoparticles prepared using the ginseng leaf extract prepared in Example 3-1 is Vibrio parahemoliticus, Salmonella enterica, Escherichia coli and Stepilococcus ulus, as can be seen in FIG. It was confirmed that it has an antimicrobial activity, and silver nanoparticles were confirmed to have an antimicrobial activity against Vibrio parahemoliticus, Salmonella enterica, and Escherichia coli. The results are shown in Table 1 below.

In addition, as a result of measuring the antimicrobial activity of gold and silver nanoparticles prepared using the ginseng root extract prepared in Example 3-2, as shown in Figure 20, silver nanoparticles are Escherichia coli, Bacillus Antimicrobial activity was shown against Cereus, Stephylococcus aureus, Vibrio parahemolyticus and Bacillus anthracis, but gold nanoparticles did not show any activity at the same concentration.

Finally, the silver nanoparticles prepared using the red ginseng root extract prepared in Example 3-3 are Vibrio parahemolyticus, Stephylococcus aureus, Bacillus cereus and Candida as can be seen in FIG. It was confirmed to have antimicrobial activity against albicans. Among them, in particular, the antibacterial activity against Candida albicans was the greatest, followed by Vibrio parahemoliticus, Stephylococcus aurus, and Bacillus cereus. The results are shown in Table 2 below. Through the above results, it can be seen that the gold or silver nanoparticles of the present invention have antimicrobial activity against microorganisms.

Example 6. Evaluation of Biofilm Degradation Activity of Nanoparticles

6-1. How to measure

Biofilm degradation activity of silver nanoparticles was confirmed by colorimetric method for Staphylococcus aureus and Pseudomonas aeruginosa . Specifically, the wells of a 96-well micro-titer plate were filled with 100 μl of S. aureus strains and Pseudomonas aeruginosa strains and incubated for 24 hours in a log phase. Thereafter, silver nanoparticles were added in the range of 1 to 4 μg, the cell culture plate was incubated at 37 ° C. for 4 hours, the medium was removed, and the wells were washed three times with 200 μl of sterile water. . The micro-titer plate was then air dried for 45 minutes, 200 μl of a solution of 0.1% crystal violet added to water was added to each well for 4 minutes, followed by 3 times with 300 μl of sterile water. Excess strain was removed by washing with water. The stain integrated by adjacent cells through this was lysed together in 200 μl of 95% ethanol. Absorption of each well was measured at 595 nm using a micro-titer ELISA reader, and then the percentage of biofilm activity inhibition was calculated using the following formula: [1- (A595 of test / A595 of control)] × 100 (Gurunathan et al., 2014). The experiment was performed repeatedly and the results analyzed under mean ± SD.

6-2. Biofilm Degradation Activity Measurement Results

The silver nanoparticles of Example 3-1, prepared using ginseng leaf extract, were identified as Stepirococcus Ourus and Pseudomonas eruzinosa at a concentration of 4 μg / ml of silver nanoparticles, as shown in FIG. 22. As shown in FIG. 23, silver nanoparticles prepared using the red ginseng root extract prepared in Example 3-3, 4 ㎍ / ml concentration of silver nanoparticles It was confirmed that there is biodegradation activity against Stepilococcus aureus and Pseudomonas eruzinosa.

Through this, it can be confirmed that the nanoparticles prepared using ginseng extract may have a biofilm-degrading activity on the strain, thereby inhibiting biofilm-related infections, and thus may play a role in reducing cost and patient pain.

Example 7 Evaluation of Anticoagulant Activity of Nanoparticles

7-1. How to measure

Human blood was collected from healthy male and female volunteers (age range 25-45 years) and placed in two vacuum collectors, labeled A and B, respectively. After a period of time, the anticoagulant effect of the synthesized gold nanoparticles was observed.

7-2. Measurement result of anticoagulant activity

Gold nanoparticles of Example 3-1 prepared using ginseng leaf extract, as can be seen in Figure 24, it was confirmed that the presence of gold nanoparticles inhibit the formation of coagulation (Fig. 24a) . In comparison, the control without anticoagulant completely coagulates blood (FIG. 24B). In addition, even after 24 hours, no specific change to the stability of the gold nanoparticles was observed.

Through these results, it is possible to know the anticoagulant activity of the gold nanoparticles biosynthesized by the leaf extract of ginseng, and also the result is a carrier or drug carrier of drugs and genes that directly contact the gold nanoparticles with blood This suggests that the sensor can be used as a nanocarrier in vivo.

Example 8. Evaluation of Cytotoxicity of Nanoparticles to Mother Cells and Cancer Cells

In order to measure the anticancer activity of the nanoparticles of Example 3-1, prepared using ginseng leaf extract, cytotoxicity of gold nanoparticles and silver nanoparticles in human lung cancer cells (A549) and hair cells (HaCaT Skin) was examined. Measurement was made by MTT assay. As a result, it was found that the silver nanoparticles had no cytotoxicity even at a concentration of 1000 µg / mL with respect to hair cells, and had biocompatibility (FIG. 25). However, silver nanoparticles showed cytotoxicity against A549 lung cancer cells at a concentration of 100 μg / mL. In addition, in the case of gold nanoparticles, it was found that there was no cytotoxicity even at a concentration of 20 µg / mL with respect to hair cells, and had biocompatibility (FIG. 25). However, gold nanoparticles showed cytotoxicity against A549 lung cancer cells at a concentration of 2 μg / mL. Therefore, it was found that silver nanoparticles have biocompatibility for drug delivery, and gold nanoparticles can be used as anticancer agents.

Example 9. Assessment of Anti-inflammatory Activity of Nanoparticles

After treating the silver or gold nanoparticles of Example 3-1, prepared using ginseng leaf extract, the viability of RAW264.7 cells was measured, and LPS (1 μg / ml) with silver or gold nanoparticles was RAW264. After treatment with .7 cells, the amount of NO produced was measured. At this time, L-NMMA (monomethylarginine) was used as a positive control. As a result, it was confirmed that silver nanoparticles had no cytotoxicity to RAW264.7 cells up to 1000 μM concentration and inhibited NO production in a concentration-dependent manner (FIG. 26). In addition, it was confirmed that gold nanoparticles have no cytotoxicity to RAW264.7 cells up to a concentration of 100 μM and inhibit NO production in a concentration-dependent manner (FIG. 26). Through this, it was confirmed that silver and gold nanoparticles biosynthesized by the leaf extract of ginseng have anti-inflammatory activity without cytotoxicity.

From the above description, those skilled in the art will appreciate that the present invention can be implemented in other specific forms without changing the technical spirit or essential features. In this regard, the embodiments described above are to be understood in all respects as illustrative and not restrictive. The scope of the present invention should be construed that all changes or modifications derived from the meaning and scope of the following claims and equivalent concepts rather than the detailed description are included in the scope of the present invention.

Claims (19)

A gold nanoparticles prepared by reacting a ginseng leaf extract with a gold (Au) precursor, wherein the anti-inflammatory composition comprises gold nanoparticles containing a bioactive substance of the ginseng leaf extract as an active ingredient.
delete delete The composition of claim 1, wherein the extract is extracted with a solvent selected from the group consisting of water, C1 to C4 alcohols, and mixed solvents thereof.
delete delete The composition of claim 1, wherein the concentration of the gold precursor used in the reaction is 0.01 to 100 mM.
The composition of claim 1, wherein the reaction is carried out at 10 to 100 ° C.
The composition of claim 1, wherein the reaction time of the reaction is 1 minute to 1 hour.
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