KR102181117B1 - Three-phase hybrid nanoparticles and method for manufacturing the same - Google Patents

Abstract

The present invention relates to a three-phase hybrid nanoparticle containing all of the liquid phase, gas phase, and solid phase.

Description

Three-phase hybrid nanoparticles and a method of manufacturing the same TECHNICAL FIELD [THREE-PHASE HYBRID NANOPARTICLES AND METHOD FOR MANUFACTURING THE SAME}

The present invention relates to a three-phase hybrid nanoparticle and a method for producing the same, and more specifically, a three-phase hybrid nanoparticle having a core-shell structure and including a gaseous core, a liquid shell, and a solid core and an interface between the shell and It relates to a method of manufacturing the same.

It also relates to the development of a new nano-bio material that will dramatically increase the effect of cancer treatment using three-phase hybrid nanoparticles.

Current standards for cancer treatment are based on surgical removal of cancer tissues, anticancer drugs, and radiation therapy. However, in the case of cancer tissues that cannot be operated, anticancer drugs and radiation therapy are not effective, so there are limitations. Vascular embolization surgery is a strategy to pursue necrosis of cancer tissues by blocking the blood supply by blocking blood vessels heading to the cancer tissues with an external input material. The currently developed materials for vascular embolization are polymer materials and inorganic materials, and have limitations that are not suitable for commercialization in terms of intravascular adhesion, possibility of penetrating into microvessels, and biocompatibility.

One object of the present invention is to provide a three-phase hybrid nanoparticle.

Another object of the present invention is to provide a method for preparing a three-phase hybrid nanoparticle.

In more detail, an object of the present invention is to provide a therapeutic agent for vascular embolism and a therapeutic agent for cancer tissues through the functions of selective vascular embolization, drug release, and generation of active oxygen of the three-phase hybrid nanoparticles of the present invention.

The three-phase hybrid nanoparticles for one object of the present invention have a core-shell structure, oxygen bubbles in gaseous form form the core, a shell layer containing liquid metal in liquid form surrounds the core, and the interface in solid state is It is disposed between the core and the shell layer, and the solid interface is formed of phospholipids and metal nanoparticles, and the phospholipids form a micelle structure.

The phospholipid may include a drug-carrying ligand.

The drug-carrying ligand contains any one of cyclodextrin, DSPC (1,2-distearoyl-sn-glycero-e-phosphocholine, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine) and cholesterol. do.

The phospholipid includes a photothermal ligand, and the photothermal ligand increases the temperature of the photothermal ligand when light is irradiated.

The phospholipid is characterized in that oxygen bubbles as the core are disposed on the hydrophobic portion.

An additional ligand may be modified outside the shell layer including the liquid metal in the liquid phase.

The additional ligand is at least one of a drug-carrying ligand and a photothermal ligand.

The metal nanoparticles include gold (Au), silver (Ag), copper (Cu), and copper oxide (Cu ₂ O, CuO).

The metal nanoparticles are characterized by having good affinity with liquid metal and high biocompatibility.

The liquid metal includes a gallium-based liquid metal.

The gallium-based liquid metal is any of gallium (Ga) liquid metal, gallium-indium (Ga-In) alloy liquid metal, gallium-tin (Ga-Sn) alloy liquid metal, and gallium-zinc (Ga-Zn) alloy liquid metal. Includes one.

The shell layer containing the liquid metal is characterized in that when light is irradiated, the temperature is increased by heat, thereby causing a shape change.

In the oxygen bubble, gaseous oxygen bubbles may be released by a liquid metal that changes shape.

The three-phase hybrid nanoparticles are characterized in that the shape of the three-phase hybrid nanoparticles is changed by the liquid metal whose shape is changed by increasing the temperature by heat when light is irradiated.

A method of preparing a three-phase hybrid nanoparticle for another object of the present invention comprises the steps of preparing a phospholipid; Preparing metal nanoparticles; Preparing a dispersion by dispersing the phospholipid and metal nanoparticles in an aqueous solution; Injecting oxygen gas into the dispersion; Ultrasonicating the dispersion liquid injected with the oxygen gas; And blowing the sonicated dispersion in a liquid metal after centrifuging, wherein the oxygen gas in the gaseous phase and the liquid metal in the liquid form form a core-shell structure, and the phospholipids in a solid state and the It is characterized in that the metal nanoparticles form an interface between the core and the shell.

A method of preparing a three-phase hybrid nanoparticle for another object of the present invention comprises: preparing an aqueous solution containing a phospholipid; Injecting oxygen gas into the aqueous solution; Ultrasonicating and centrifuging the aqueous solution injected with the oxygen gas; Adding metal nanoparticles to the aqueous solution, mixing, and vaporizing to obtain oxygen bubbles with metal nanoparticles attached to the surface thereof; Including the step of blowing oxygen bubbles attached to the surface of the metal nanoparticles in a liquid metal, wherein the oxygen gas in the gaseous phase and the liquid metal in the liquid phase form a core-shell structure, and the phospholipids and the metal nanoparticles in a solid state It is characterized in that the particles form an interface between the core and the shell.

The phospholipid includes a drug-carrying ligand.

The drug-carrying ligand contains any one of cyclodextrin, DSPC (1,2-distearoyl-sn-glycero-e-phosphocholine, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine) and cholesterol. Include.

The phospholipid includes a photothermal ligand, and the photothermal ligand is characterized in that the temperature of the ligand increases when light is irradiated.

The phospholipid is characterized in that the micro-shell structure is formed, and oxygen bubbles as the core are disposed on the hydrophobic portion.

After the step of blowing in the liquid metal, the step of modifying an additional ligand outside the shell layer containing the liquid metal may be further included.

The step of modifying the additional ligand is performed by adding a solution containing the ligand to the upper layer of the liquid metal and blowing in the liquid metal.

The metal nanoparticles include gold (Au), silver (Ag), copper (Cu), and copper oxide (Cu ₂ O, CuO).

The metal nanoparticles are characterized by having good affinity with liquid metal and high biocompatibility.

The liquid metal includes a gallium-based liquid metal.

The gallium-based liquid metal is any of gallium (Ga) liquid metal, gallium-indium (Ga-In) alloy liquid metal, gallium-tin (Ga-Sn) alloy liquid metal, and gallium-zinc (Ga-Zn) alloy liquid metal. Includes one.

The shell containing the liquid metal is characterized in that when light is irradiated, the temperature is increased by heat, thereby causing a shape change.

In the oxygen bubble, gaseous oxygen bubbles may be released by a liquid metal that changes shape.

It is characterized in that when light is irradiated, the temperature is increased by heat and the shape is changed by the liquid metal that has changed shape.

The present invention provides a three-phase hybrid nanoparticle and a method of manufacturing the same, and thus it can be used for vascular embolization by utilizing the thermodynamic shape change characteristics of liquid metal, and selective drug delivery and active oxygen according to destruction of the liquid metal shell It can be expected to dramatically improve the effectiveness in cancer treatment through occurrence.

1 is a schematic diagram of a three-phase hydride nanoparticle of the present invention.
2 is a flow chart of a method of manufacturing a three-phase hybrid nanoparticle according to an embodiment of the present invention.
3 is a flowchart of a method of manufacturing a three-phase hybrid nanoparticle according to another embodiment of the present invention.
4 is a diagram illustrating a method of manufacturing a three-phase hydride nanoparticle of the present invention and a method of treating cancer using the three-phase hybrid nanoparticle of the present invention.

Hereinafter, exemplary embodiments of the present invention will be described in detail with reference to the accompanying drawings. In the present invention, various modifications may be made and various forms may be applied, and specific embodiments will be illustrated in the drawings and described in detail in the text. However, this is not intended to limit the present invention to a specific form of disclosure, it is to be understood as including all changes, equivalents, or substitutes included in the spirit and scope of the present invention. In describing each drawing, similar reference numerals have been used for similar elements.

The terms used in the present application are only used to describe specific embodiments and are not intended to limit the present invention. Singular expressions include plural expressions unless the context clearly indicates otherwise. In the present application, terms such as "comprise" or "have" are intended to designate the existence of features, steps, actions, components, parts, or a combination thereof described in the specification, but one or more other features or steps It is to be understood that it does not preclude the possibility of addition or presence of, operations, components, parts, or combinations thereof.

Unless otherwise defined, all terms used herein, including technical or scientific terms, have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention belongs. Terms as defined in a commonly used dictionary should be interpreted as having a meaning consistent with the meaning in the context of the related technology, and should not be interpreted as an ideal or excessively formal meaning unless explicitly defined in this application. Does not.

1 shows a schematic diagram of the three-phase hydride nanoparticles of the present invention.

According to FIG. 1, the three-phase hybrid nanoparticles have a core-shell structure, gaseous oxygen bubbles form the core 10, and a shell layer 30 containing a liquid metal, which is a liquid phase, surrounds the core, A solid interface 20 is disposed between the core and the shell layer, and the solid interface is formed of phospholipids and metal nanoparticles, and the phospholipids form a micelle structure.

The phospholipid includes a drug-carrying ligand. The drug-carrying ligand is formed of any one of cyclodextrin, DSPC (1,2-distearoyl-sn-glycero-e-phosphocholine, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine) and cholesterol. The drug-carrying ligand material includes a functional group of at least one of a thiol group, a carboxiyl group, an amine group, and a hydroxyl group. The cancer treatment substance used may include at least one of doxorubicin, 5-fluorouracil, paclitaxel, docetaxel, and gemcitabine.

The phospholipid includes a photothermal ligand, and the photothermal ligand increases the temperature of the ligand when light is irradiated. The photothermal ligand may include carbon nanotubes (CNT) and graphene. Depending on the type of the photothermal ligand, the absorption wavelength indicating photothermal characteristics is different and the photothermal efficiency is also different. In addition, light-heating efficiency may vary according to the light irradiation time and the location of the target.

The phospholipid includes at least one of a drug-carrying ligand and a photothermal ligand.

The phospholipids can be classified into a hydrophobic portion and a hydrophilic portion, and are characterized in that they form a microshell structure. An oxygen bubble, which is the core 10, is disposed on the hydrophobic portion of the phospholipid. The hydrophilic portion of the phospholipid may form a solid interface 20 between the core and the shell together with the metal nanoparticles. The solid interface 20 is disposed between the core and the shell layer containing the liquid metal. The solid interface 20 is formed of phospholipids and metal nanoparticles. The metal nanoparticles contain any one of gold (Au), silver (Ag), copper (Cu), and copper oxide (Cu ₂ O, CuO), and are characterized by having good affinity with liquid metal and high biocompatibility. .

A shell layer 30 including a liquid metal in a liquid state is disposed to surround the core 10. The liquid metal includes a gallium-based liquid metal. The gallium-based liquid metal is any of gallium (Ga) liquid metal, gallium-indium (Ga-In) alloy liquid metal, gallium-tin (Ga-Sn) alloy liquid metal, and gallium-zinc (Ga-Zn) alloy liquid metal. Includes one. According to the phase equilibrium, the gallium-indium (Ga-In) alloy may have an indium content of 0 wt% to 50 wt% in a liquid at a human body temperature (~37°C). Depending on the composition of gallium and indium, it can have a great effect on the surface density and cytotoxicity of the ligand.

An additional ligand may be modified outside of the shell layer 30 including the liquid metal in the liquid phase, and the additional ligand includes at least one of a drug-carrying ligand and a photothermal ligand.

The shell layer 30 including the liquid metal is characterized in that when light is irradiated, the temperature is increased by heat, thereby causing a shape change. In the oxygen bubble, gaseous oxygen bubbles may be released from the core 10 by the liquid metal that changes shape.

When the three-phase hybrid nanoparticles are irradiated with light, the temperature of the liquid metal rises, and a part of the temperature of the liquid metal moves to the photothermal ligand and the metal nanoparticle forming the boundary between the core and the shell, and the photothermal ligand becomes photothermal. Due to the characteristics, the temperature of the three-phase hybrid nanoparticles is increased locally, and the metal nanoparticles further increase the temperature of the three-phase hybrid nanoparticles by the reflective properties of the metal. As the temperature rises, the liquid metal particles of the shell layer containing the liquid metal change shape from a one-dimensional circular shape to a rod shape with a large aspect ratio. Due to the large surface area compared to the unit volume, the liquid metal that has been transformed can effectively block the passage of blood to local areas, and thus can be effectively used for vascular embolization. In addition, the boundary between the core and the shell is released due to the change in the shape of the liquid metal, and as the boundary is released, the active oxygen forming the core is released to the outside of the three-phase hybrid nanoparticles. The active oxygen helps to increase the concentration of active oxygen in the body and is effective in necrosis of cancer tissues. In addition, as the boundary between the core and the shell is released, the drug-carrying ligand is released to the outside of the three-phase hybrid nanoparticles, which may help to necrosis the cancer tissue.

The three-phase hybrid nanoparticles are formed by a change in the shape of the liquid metal caused by a local increase in temperature due to the light-irradiation, the reflective properties of the metal nanoparticles forming the boundary between the core and the shell, and the photothermal ligand. It is characterized in that the change.

The time for irradiating light to the three-phase hybrid nanoparticles may be 1 minute to 24 hours.

2 shows a flow chart of a method of manufacturing a three-phase hybrid nanoparticle according to an embodiment of the present invention.

According to FIG. 2, a method of preparing a three-phase hybrid nanoparticle according to an embodiment of the present invention includes preparing a phospholipid (S210); Preparing metal nanoparticles (S220); Dispersing the phospholipid and metal nanoparticles in an aqueous solution to prepare a dispersion (S230); Injecting an oxygen gas into the dispersion (S240); Ultrasonicating the dispersion liquid injected with the oxygen gas (S250); And blowing the sonicated dispersion in a liquid metal after centrifuging (S260), wherein the oxygen gas in the gaseous form and the liquid metal in the liquid form form a core-shell structure, and the phospholipids in a solid state and the It is characterized in that the metal nanoparticles form an interface between the core and the shell.

In step S210, phospholipids are prepared. The phospholipid includes a drug-carrying ligand. The drug-carrying ligand is formed of any one of cyclodextrin, DSPC (1,2-distearoyl-sn-glycero-e-phosphocholine, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine) and cholesterol. The drug-carrying ligand material includes a functional group of at least one of a thiol group, a carboxiyl group, an amine group, and a hydroxyl group. The cancer treatment substance used may include at least one of doxorubicin, 5-fluorouracil, paclitaxel, docetaxel, and gemcitabine. The phospholipid is a photothermal ligand. The photothermal ligand is characterized in that the temperature of the ligand is increased when light is irradiated, and the photothermal ligand may include carbon nanotubes (CNT) and graphene. The absorption wavelength indicating the photothermal characteristic is different and the photothermal efficiency may be different depending on the type of the phospholipid, and the photothermal efficiency may be different depending on the light irradiation time and the location of the target, and the phospholipid includes at least one of a drug-carrying ligand and a photothermal ligand. The phospholipid can be divided into a hydrophobic portion and a hydrophilic portion, and has a microshell structure, characterized in that the core oxygen bubble is disposed on the hydrophobic portion of the phospholipid. The part side can form a solid interface between the core and the shell with the metal nanoparticles.

In step S 220, metal nanoparticles are prepared. The metal nanoparticles contain any one of gold (Au), silver (Ag), copper (Cu), and copper oxide (Cu ₂ O, CuO), and are characterized by having good affinity with liquid metal and high biocompatibility. .

In step S 230, a dispersion is prepared by dispersing phospholipids and metal nanoparticles in an aqueous solution.

In step S 240, oxygen gas is injected into the dispersion in which the phospholipid and metal nanoparticles are dispersed. The oxygen gas is blown into the dispersion and injected.

In step S250, the dispersion liquid blown with the oxygen gas is subjected to ultrasonic treatment. Depending on the ultrasonic irradiation time, the radius distribution of the nanoparticles may change.

In step S260, the sonicated dispersion is centrifuged, and the resulting core is blown in a liquid metal to include a gaseous core, a liquid shell, and a solid boundary surface forming the interface between the core and the shell. Prepare nanoparticles. The liquid metal includes a gallium-based liquid metal, and the gallium-based liquid metal is a gallium (Ga) liquid metal, a gallium-indium (Ga-In) alloy liquid metal, a gallium-tin (Ga-Sn) alloy liquid metal and a gallium -Zinc (Ga-Zn) alloy may contain any one of the liquid metal, and most preferably gallium-indium (Ga-In) alloy liquid metal may be used. According to the phase equilibrium, the gallium-indium (Ga-In) alloy may exist as a liquid in an indium content of 0 wt% to 50 wt% under human body temperature (~37°C). Depending on the composition, it has a great influence on the surface density and cytotoxicity of the ligand. The size of the three-phase hybrid nanoparticles may be 100 nm to 1000 nm. The three-phase hybrid nanoparticles satisfying the above size may be introduced into blood vessels around cancer cells regardless of the size of the blood vessel. An additional ligand may be modified outside of the shell layer containing the liquid metal. The additional ligand is at least one of a drug-carrying ligand and a photothermal ligand. The description of the drug-carrying ligand and the broad-spectrum ligand is the same as described above, and thus will be omitted. A solution including an additional ligand may be filled on the liquid metal, and oxygen bubbles attached to the surface of the metal nanoparticles may be blown. After the blowing, the liquid metal forms a shell along the metal nanoparticle layer attached to the surface of the oxygen bubble to form a three-phase hybrid nanoparticle, and the prepared three-phase hybrid nanoparticle rises past the solution layer containing an additional ligand. An additional ligand is modified on the surface of the shell containing the liquid metal. The three-phase hybrid nanoparticles modified with the additional ligand may help stabilize the dispersion of the three-phase hydride nanoparticles.

3 shows a flowchart of a method of manufacturing a three-phase hybrid nanoparticle according to another embodiment of the present invention.

According to FIG. 3, a method of preparing a three-phase hybrid nanoparticle according to an embodiment of the present invention includes preparing an aqueous solution containing a phospholipid (S310); Injecting oxygen gas into the aqueous solution (S320); Ultrasonicating and centrifuging the aqueous solution injected with the oxygen gas (S330); Adding and mixing metal nanoparticles to the aqueous solution, followed by vaporization to obtain oxygen bubbles with metal nanoparticles attached to the surface (S340); Including the step of blowing oxygen bubbles attached to the surface of the metal nanoparticles in a liquid metal (S350), wherein the oxygen gas in a gaseous form and the liquid metal in a liquid form form a core-shell structure, and the phospholipids in a solid state and It characterized in that the metal nanoparticles form an interface between the core and the shell.

In step S 310, an aqueous phospholipid solution is prepared. Details of the phospholipids are the same as those of the phospholipids used in FIG. 2, so detailed information is omitted.

In step S320, oxygen gas is injected into the aqueous phospholipid solution. Through this step, a core composed of oxygen bubbles in which oxygen gas is disposed in the hydrophobic portion of the phospholipid is formed.

In step S330, the aqueous phospholipid solution injected with the oxygen gas is subjected to ultrasonic treatment and centrifugation. Depending on the ultrasonic irradiation time, the radius distribution of the nanoparticles may change.

In step S340, metal nanoparticles are added to the aqueous solution subjected to the ultrasonic wave and centrifugation and then vaporized. After vaporization, an oxygen burr with metal nanoparticles attached to the surface can be obtained. Details of the metal nanoparticles are the same as those of the metal nanoparticles used in FIG. 2, so detailed information is omitted.

In step S 350, the oxygen bubble to which the metal nanoparticles are attached is blown in the liquid metal. A liquid metal shell is formed along the metal nanoparticle layer attached to the surface of the oxygen bubble, thereby preparing a three-phase hybrid nanoparticle. Details of the liquid metal are the same as those of the liquid metal used in FIG. 2, and thus detailed information will be omitted. The size of the three-phase hybrid nanoparticles may be 100 nm to 1000 nm. The three-phase hybrid nanoparticles satisfying the above size may be introduced into blood vessels around cancer cells regardless of the size of the blood vessel. An additional ligand may be modified outside of the shell layer containing the liquid metal. The additional ligand is at least one of a drug-carrying ligand and a photothermal ligand. The description of the drug-carrying ligand and the broad-spectrum ligand is the same as described above, and thus will be omitted. A solution including an additional ligand may be filled on the liquid metal, and oxygen bubbles attached to the surface of the metal nanoparticles may be blown. After the blowing, the liquid metal forms a shell along the metal nanoparticle layer attached to the surface of the oxygen bubble to form a three-phase hybrid nanoparticle, and the prepared three-phase hybrid nanoparticle rises past the solution layer containing an additional ligand. An additional ligand is modified on the surface of the shell containing the liquid metal. The three-phase hybrid nanoparticles to which the additional ligand has been received may help stabilize the dispersion of the three-phase hydride nanoparticles.

4 shows a diagram of a method for preparing a three-phase hydride nanoparticle of the present invention and a method for treating cancer through the three-phase hybrid nanoparticle of the present invention.

According to FIG. 4, the three-phase hybrid nanoparticles prepared by the above method are prepared as an injection agent or a delivery agent through a catheter and injected into a blood vessel around a target cancer tissue. By using a photon through a micro light source, it induces shape change of the injected three-phase hybrid nanoparticles and simultaneously induces blood vessel occlusion, drug release, and generation of active oxygen.

Example 1

Preparation of three-phase hybrid nanoparticles

Phospholipids and surfactants including DSPC, DSPE-PEG-2000-Amine, DSPE-PEG-2000-Biotin, which are harmless to the human body, are 50:50:0, 80:20:0, 85:8:7, After mixing in a ratio of 90:5:5, sucrose and metal nanoparticles are added to the aqueous solution to attach the metal nanoparticles to the oxygen bubble surface. Add 1.6 times ascorbic acid and mix. The concentration of the metal nanoparticles used at this time is 4×10 ^-6 M ~ 5×10 ^-6 M. The aqueous solution in which the phospholipid and metal nanoparticles are mixed is treated with a vacuum pump for at least 2 hours. After rehydration at 65° C., 99.9% oxygen gas is injected, ultrasonic treatment and centrifugation are performed to prepare oxygen bubbles with metal nanoparticles attached to the surface, and then blown in liquid metal to prepare three-phase hybrid nanoparticles. In order to modify the additional ligand on the surface of the shell containing the liquid metal, a solution containing the additional ligand is filled on the liquid metal, and oxygen bubbles attached to the surface of the metal nanoparticles are blown. In this case, in order to modify the additional ligand on the surface of the shell containing the liquid metal by another method, the oxygen bubble may be blown after forming nanoparticles by filling a solution containing the additional ligand on the liquid metal.

Example 2

Preparation of three-phase hybrid nanoparticles

Phospholipids and surfactants including DSPC, DSPE-PEG-2000-Amine, DSPE-PEG-2000-Biotin, which are harmless to the human body, are 50:50:0, 80:20:0, 85:8:7, After mixing in a ratio of 90:5:5, it is dried at 80℃. After drying, an aqueous solution is added to rehydrate, and phospholipids and surfactants are ultrasonically treated to disperse, and 99.9% oxygen gas is injected to form oxygen bubbles through ultrasonic treatment and sequential centrifugation. To form a liquid metal shell, metal nanoparticles with good affinity with liquid metal and excellent biocompatibility are attached to the surface. After mixing the prepared aqueous solution and the aqueous metal nanoparticle solution, both are vaporized. At this time, it is treated for 2 hours or more with a vacuum pump to remove all solvents. Then, it is rehydrated at 65°C. The oxygen bubbles attached to the surface of the metal nanoparticles are blown in the liquid metal of a gallium-indium (Ga-In) alloy to form a liquid metal shell on the oxygen bubbles attached to the surface of the metal nanoparticles to form a three-phase hybrid nanoparticle. To manufacture. In order to modify the additional ligand on the surface of the shell containing the liquid metal, a solution containing the additional ligand is filled on the liquid metal, and oxygen bubbles attached to the surface of the metal nanoparticles are blown. In this case, in order to modify the additional ligand on the surface of the shell containing the liquid metal by another method, the oxygen bubble may be blown after forming nanoparticles by filling a solution containing the additional ligand on the liquid metal.

Drug loading experiment

Comparative Sample 1

Cytotoxicity and disinfectant toxicity tests for the three-phase hybrid nanoparticles prior to drug loading were performed. The cytotoxicity test for the drug-free three-phase hybrid nanoparticles was performed on human dermal fibroblasts. Three-phase hybrid nanoparticles containing no drug prepared in the two-dimensional cultured fibroblast culture medium were administered at different concentrations (0~500μg/mL), and cytotoxicity within 24 hours was calcein-AM and ethidium homodimer-1 fluorescence. Simultaneous staining was performed to confirm the loss of intercellular esterase activity and plasma membrane integrity. In addition, the growth of fibroblasts during the culture period within 7 days under the same conditions was quantitatively performed using a commercially available measurement kit of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Measure. Additionally, small animal toxicity tests for phase 3 nanoparticles are conducted in rats. 100 μL of saline solution containing 10 mg or less of three-phase hybrid nanoparticles is administered to the animal via intravenous injection. For a period of 2 weeks or less, the amount of three-phase hybrid nanoparticles remaining in the liver, spleen, and kidney is quantitatively analyzed.

Sample 1

Cytotoxicity and disinfectant toxicity tests for drug-supported three-phase hybrid nanoparticles were performed. Cytotoxicity tests for drug-carrying three-phase hybrid nanoparticles were conducted on human dermal fibroblasts. Three-phase hybrid nanoparticles carrying the drug prepared in the two-dimensional cultured fibroblast culture medium were administered at different concentrations (0~500μg/mL), and cytotoxicity within 24 hours was stained with calcein-AM and ethidium homodimer-1. Simultaneously perform cell-to-cell esterase activity and check for loss of plasma membrane integrity. In addition, the growth of fibroblasts during the culture period within 7 days under the same conditions was quantitatively performed using a commercially available measurement kit of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Measure. Additionally, small animal toxicity tests on three-phase hybrid nanoparticles are conducted in rats. 100 μL of saline solution containing 10 mg or less of 3-phase nanoparticles is administered to the animal via intravenous injection. For a period of 2 weeks or less, the amount of three-phase hybrid nanoparticles remaining in the liver, spleen, and kidney is quantitatively analyzed.

Sample 2

By carrying a drug and irradiating light on the core-shell nanoparticles modified with the photothermal ligand, the shape change characteristics of the shell structure were confirmed. The effects of the following variables on the morphology change efficiency are analyzed, and drug release, free radical generation, and vascular occlusion effects are optimized according to the morphology change conditions.

Cancer cell removal effect

The effect on cancer cell elimination through drug release targets at least three human cancer cell lines. In order to exclude the direct effect of the three-phase nanoparticles and analyze cancer cell growth inhibition by the released drug, first, the drug-loaded nanoparticles were prepared in a 100 μL saline solution in an amount of 10 mg or less. The solution is stored at a temperature of 37°C for 24 hours to induce dissolution of the drug. Then, the supernatant containing the eluted drug is separated through centrifugation. The supernatant was administered to the culture medium of a two-dimensional cultured human cancer cell line, and the growth of cancer cells within 24 hours was commercialized with the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) series. Quantitative measurements are made by using the prepared measurement kit. The effect of inhibiting the growth of cancer tissues through drug release targets rats with cancerous tissues. In the case of a rat model with cancer tissue, the animal is grown for at least 30 days after cancer cell injection. The drug-loaded nanoparticles are prepared in an amount of 10 mg or less in 100 μL of saline solution, and then administered to the animal via intravenous injection. For a time of 4 weeks or less, the animals are euthanized, cancer tissues are collected, and the mass change of the cancer tissues is quantitatively measured.

Although the above has been described with reference to preferred embodiments of the present invention, those skilled in the art will be able to variously modify and change the present invention without departing from the spirit and scope of the present invention described in the following claims. You will understand that you can.

Claims (19)

It has a core-shell structure,
Oxygen bubbles in gaseous form form the core,
A shell layer containing a liquid metal in a liquid phase surrounds the core at a temperature above the temperature present in the liquid phase,
A solid interface is disposed between the core and the shell layer,
The solid interface is formed of phospholipids and metal nanoparticles,
Characterized in that the phospholipids form a micelle structure,
Three-phase hybrid nanoparticles.
The method of claim 1,
The phospholipid contains a drug-carrying ligand,
The drug-carrying ligand is
Containing any one of cyclodextrin, DSPC (1,2-distearoyl-sn-glycero-e-phosphocholine, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine) and cholesterol,
Three-phase hybrid nanoparticles.
The method of claim 1,
The phospholipid includes a photothermal ligand,
The photothermal ligand is characterized in that the temperature of the photothermal ligand increases when light is irradiated,
Three-phase hybrid nanoparticles.
The method of claim 1,
The phospholipid is
Characterized in that the oxygen bubble as the core is disposed on the side of the hydrophobic portion,
Three-phase hybrid nanoparticles.
The method of claim 1,
An additional ligand is modified outside of the shell layer containing the liquid metal in the liquid phase,
Three-phase hybrid nanoparticles.
The method of claim 5,
The additional ligand is,
Any one or more of a drug-carrying ligand and a photothermal ligand,
Three-phase hybrid nanoparticles.
The method of claim 1,
The metal nanoparticles include gold (Au), silver (Ag), copper (Cu), and copper oxide (Cu ₂ O, CuO),
The metal nanoparticles,
Characterized in that it has good affinity with liquid metal and has high biocompatibility,
Three-phase hybrid nanoparticles.
The method of claim 1,
The liquid metal includes a gallium-based liquid metal,
The gallium-based liquid metal,
Including any one of gallium (Ga) liquid metal, gallium-indium (Ga-In) alloy liquid metal, gallium-tin (Ga-Sn) alloy liquid metal, and gallium-zinc (Ga-Zn) alloy liquid metal,
Three-phase hybrid nanoparticles.
The method of claim 1,
The shell layer containing the liquid metal,
When light is irradiated, the temperature rises by heat, causing a change in shape,
The shape of the liquid metal is changed by the shape change,
Characterized in that the oxygen bubble is released by the change in the shape of the liquid metal,
Three-phase hybrid nanoparticles.
Preparing phospholipids;
Preparing metal nanoparticles;
Preparing a dispersion by dispersing the phospholipid and metal nanoparticles in an aqueous solution;
Injecting oxygen gas into the dispersion;
Ultrasonicating the dispersion liquid injected with the oxygen gas; And
Centrifuging the sonicated dispersion and then blowing in the liquid metal to obtain oxygen bubbles in which phospholipids and metal nanoparticles are attached to the surface,
The gaseous oxygen gas and the liquid metal as a liquid form a core-shell structure, and the phospholipid and the metal nanoparticles as a solid form an interface between the core and the shell,
A method of preparing a three-phase hybrid nanoparticle.
Preparing an aqueous solution containing phospholipids;
Injecting oxygen gas into the aqueous solution;
Ultrasonicating and centrifuging the aqueous solution injected with the oxygen gas;
Adding metal nanoparticles to the aqueous solution, mixing, and vaporizing to obtain oxygen bubbles with metal nanoparticles attached to the surface thereof;
Including the step of blowing oxygen bubbles attached to the surface of the metal nanoparticles in a liquid metal,
The gaseous oxygen gas and the liquid metal as a liquid form a core-shell structure, and the phospholipid and the metal nanoparticles as a solid form an interface between the core and the shell,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 10 or 11,
The phospholipid contains a drug-carrying ligand,
The drug-carrying ligand is
Containing any one of cyclodextrin, DSPC (1,2-distearoyl-sn-glycero-e-phosphocholine, DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine) and cholesterol,
Method for preparing three-phase hybrid nanoparticles,
The method of claim 10 or 11,
The phospholipid includes a photothermal ligand,
The photothermal ligand is
Characterized in that the temperature of the ligand increases when irradiated with light,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 10 or 11,
The phospholipids form a myshell structure,
Characterized in that the oxygen bubble as the core is disposed on the side of the hydrophobic portion,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 10 or 11,
After the step of blowing in the liquid metal, further comprising modifying an additional ligand outside the shell layer containing the liquid metal,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 15,
The step of modifying the additional ligand,
Comprising by adding a solution containing a ligand to the upper layer of the liquid metal and blowing in the liquid metal,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 10 or 11,
The metal nanoparticles include gold (Au), silver (Ag), copper (Cu), and copper oxide (Cu ₂ O, CuO),
The metal nanoparticles,
Characterized in that it has good affinity with liquid metal and has high biocompatibility,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 10 or 11,
The liquid metal includes a gallium-based liquid metal,
The gallium-based liquid metal is
Including any one of gallium (Ga) liquid metal, gallium-indium (Ga-In) alloy liquid metal, gallium-tin (Ga-Sn) alloy liquid metal, and gallium-zinc (Ga-Zn) alloy liquid metal,
A method of preparing a three-phase hybrid nanoparticle.
The method of claim 10 or 11,
The shell containing the liquid metal,
When light is irradiated, the temperature rises by heat, causing a change in shape,
The shape of the liquid metal is changed by the shape change,
Characterized in that the oxygen bubble is released by the change in the shape of the liquid metal,
A method of preparing a three-phase hybrid nanoparticle.

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