Nanocomposite for cancer diagnosis and treatment, composition containing same for cancer therapy, and contrast agent for cancer diagnosis

Abstract

The present invention relates to a nanocomposite for cancer diagnosis and treatment, a composition comprising same for cancer treatment, and an ultrasonic contrast agent for cancer diagnosis and, more specifically, to a nanocomposite comprising at least one coordination nanoparticle for cancer diagnosis and treatment, a composition comprising same for cancer treatment, and a contrast agent for cancer diagnosis, wherein the coordination nanoparticle includes: a perfluorocarbon coordinating iron ions; at least one dopamine compound selected from the group consisting of dopamine and dopamine analogs; and a nucleophile.

Description

Nanocomposites for cancer diagnosis and treatment, compositions for cancer treatment comprising the same, and contrast agents for cancer diagnosis

[0001] The present invention relates to a nanocomposite for diagnosing and treating cancer, a composition for treating cancer comprising the same, and a contrast agent for diagnosing cancer, and more particularly, to a fluorescent nanocomposite based on perfluorocarbons (PFCs) capable of simultaneously performing the functions of diagnosing and treating cancer cells, and its use.

[0002] Conventional cancer treatments have primarily involved surgery, radiation therapy, and drug therapy. However, these traditional cancer treatments, including surgery, chemotherapy, and radiation therapy, not only have extremely limited therapeutic effects but also damage normal tissue, resulting in serious side effects. Therefore, newer approaches, such as targeted cancer therapy and photodynamic therapy (PDT), are attracting attention. The fundamental concept of PDT is to exploit the principle that photosensitizers, when exposed to light, generate singlet oxygen and other reactive oxygen species. Various photosensitizers are continuously being developed in countries such as the United States and Japan, and are being applied to treat various types of cancer.

[0003] Most of the photosensitizers currently used in photodynamic cancer therapy have limitations in selective delivery to cancer tissues due to nonspecific damage to normal tissues, decomposition into inactive forms, low solubility when administered in vivo, inappropriate interaction with biomolecules, and mutual entanglement between photosensitizers. They also exhibit low photosensitivity. Therefore, an appropriate delivery system is required to enhance the therapeutic effect.

[0004] Photosensitizers for effective treatment require the following characteristics: stability in vivo, cancer targeting, minimal toxicity when not irradiated with light, efficient light penetration into the diseased area in vivo, and high photosensitivity.

[0005] Currently, the only photosensitizers available domestically are first-generation porphyrins, Photofrin (630 nm, Axcan, Canada) and Photogem (630 nm, Lomonosov Institute of Fine Chemicals, Russia). However, their high supply price (approximately 3 million won per treatment) is a problem. Furthermore, various photosensitizers are continuously being developed overseas, including in the US and Japan, and are being applied to treat various types of cancer. However, clinical application requires considerable time and financial constraints.

[0006] Meanwhile, iron oxide nanoparticles possess magnetization properties, making them highly valuable in biomedical applications. To be used as an in vivo contrast agent, iron oxide nanoparticles must be well-dispersed in water and possess a uniform, small particle size. However, even if these prerequisites are met, most iron oxide nanoparticles injected in vivo are rapidly removed from the blood by the reticuloendothelial system in organs such as the lungs, liver, and spleen, accumulating in these organs and thus failing to function as a contrast agent.

[0007] Therefore, surface modification of iron oxide nanoparticles is also an important factor in reducing the degree of removal of iron oxide nanoparticles by the reticuloendothelial system. Currently, many studies have been reported to increase the blood circulation time in the body by coating iron oxide nanoparticles with water-soluble polymers. For example, many studies have been reported on water-soluble iron oxide nanoparticles coated with polyvinylpyrrolidone (Korean Patent Registration No. 10-0746312), contrast agents using polysuccinimide polymers (Korean Patent Registration No. 10-0635026), polyethylene glycol-coated iron oxide nanoparticles, silicon-coated iron oxide nanoparticles, dextran-coated iron oxide nanoparticles, etc.

[0008] However, most conventional water-soluble iron oxide nanoparticles have the disadvantage of requiring a step of chemically modifying the polymer to provide a ligand introduction site in order to diagnose specific organs or diseases in the body. In particular, even when surface-modified water-soluble iron oxide nanoparticles using such polymers, the iron oxide nanoparticles are still not taken up and delivered to the target site, such as the liver or spleen, or even if they are delivered, they are delivered in small amounts. Furthermore, conventional contrast agents using iron oxide nanoparticles have been limited to diagnostic applications, and it is very difficult to find contrast agents developed using iron oxide nanoparticles that can deliver large enough doses of the agent to target cells for simultaneous treatment. Another problem with surface modification of iron oxide nanoparticles using polymers is that it is difficult to conduct research on the fate and toxicity of the polymers themselves used for surface modification in vivo, making it difficult to evaluate the biocompatibility of polymer-modified iron oxide nanoparticles.

[0009] Accordingly, if a diagnostic and therapeutic contrast agent that can simultaneously perform imaging and treatment without introducing a polymer to modify the surface of iron oxide nanoparticles is developed, it is expected to be widely applied in related fields.

[0010] One aspect of the present invention relates to a nanocomposite that simultaneously performs diagnostic and therapeutic functions on cancer cells.

[0011] Another aspect of the present invention is to provide a cancer treatment composition that provides excellent anticancer effects by killing cancer cells.

[0012] Another aspect of the present invention is to provide a tumor-targeting contrast agent.

[0013] According to one aspect of the present invention, a nanocomposite for diagnosing and treating cancer is provided, comprising: an iron ion; a perfluorocarbon coordinating the iron ion; at least one dopamine compound selected from the group consisting of dopamine and dopamine analogues; and one or more coordination nanoparticles comprising a nucleophile.

[0014] According to another aspect of the present invention, a composition for treating cancer is provided, comprising the nanocomposite of the present invention.

[0015] According to another aspect of the present invention, a contrast agent for cancer diagnosis is provided, comprising the nanocomposite of the present invention.

[0016] The fluorescent nanocomposite containing perfluorocarbons (PFCs) of the present invention is specifically delivered to tumor cells, and can selectively and precisely distinguish tumor cells through dual imaging of fluorescence and ultrasound for tumor cells. It exhibits high dispersion stability under aqueous conditions and very low cytotoxicity. In addition, the perfluorocarbon-fluorescent nanocomposite (PFC@FPNPs-Fe) of the present invention is an anticancer agent that is delivered to tumor cells and self-replenishes hydrogen peroxide (H2O2) through oxygen (O2) release under acidic conditions of endosomes and lysosomes, and not only has the function of enhanced chemodynamic therapy, but also can provide the function of photothermal therapy through irradiation with near infrared (NIR) light, and is therefore expected to be widely applied to anticancer combination therapy technologies.

[0017] Figure 1 schematically illustrates the manufacturing process of perfluorocarbon-fluorescent nanocomposite (PFC@FPNPs-Fe).

[0018] Figure 2(a) illustrates the cancer-specific imaging diagnostic principle of PFC@FPNPs-Fe, and Figure 2(b) schematically illustrates the tumor cell death principle.

[0019] Figure 3 shows the iron ions (Fe) of PFC@FPNPs-Fe 2+ ) is a graph showing the change in nanoparticle size according to addition.

[0020] Fig. 4(a) shows the FT-IR analysis result of PFC@FPNPs-Fe, Fig. 4(b), Fig. 4(c), and Fig. 4(d) show the XPS analysis result of PFC@FPNPs-Fe, Fig. 4(e) shows the TEM analysis result of PFC@FPNPs-Fe, and Fig. 4(f) shows the TEM-mapping analysis result of PFC@FPNPs-Fe.

[0021] Figure 5 shows the fluorescence change of PFC@FPNPs-Fe according to (a) EDA treatment and (b) addition of iron ions, (c) Stern-Volmer relationship according to fluorescence quenching of PFC@FPNPs-Fe, (d) fluorescence recovery principle of PFC@FPNPs-Fe according to pH change, (e) fluorescence change of PFC@FPNPs-Fe according to time and pH change, and (f) graph of iron ion release ability of PFC@FPNPs-Fe according to pH using inductively coupled plasma mass spectrometer (ICP-MS).

[0022] Figure 6 shows (a) intracellular fluorescence changes of PFC@FPNPs-Fe in acidic and neutral pH environments and (b) quantitative fluorescence intensity, (c) microbubble images of PFC@FPNPs-Fe with and without acoustic pulses, (d) bright signals by microbubbles, and (e) quantitative signal intensity of bright signals.

[0023] Figure 7 shows (a) the enhanced chemodynamic and photothermal mechanisms of PFC@FPNPs-Fe for cancer treatment, (b) the temperature change of PFC@FPNPs-Fe under NIR irradiation, (c) the live-dead staining results of 4T1 cells treated with PFC@FPNPs-Fe over time after NIR irradiation, (d) the thermal imaging camera images (top) and optical microscope images (bottom) of PFC@FPNPs-Fe under NIR irradiation over time, (e) the oxygen release amount of PFC@FPNPs-Fe under NIR irradiation over time, (f) the quantification of hydroxyl radicals according to the presence or absence of NIR irradiation of PFC@FPNPs and PFC@FPNPs-Fe, (g) the quantification images of intracellular reactive oxygen species of PFC@FPNPs-Fe under NIR irradiation in 4T1 cells, and (h) the anticancer effect of PFC@FPNPs-Fe through FACS analysis.

[0024] Figure 8 shows (a) the tumor-specific bright signal and (b) the quantitative intensity by ultrasound of balb / c nude mice injected with PFC@FPNPs-Fe, (c) the tumor-specific fluorescence signal image of balb / c nude mice injected with PFC@FPNPs-Fe, (d) the fluorescence image and (e) the quantitative intensity of major organs extracted from balb / c nude mice 24 hours after PFC@FPNPs-Fe injection.

[0025] Figure 9 shows (a) a cancer cell treatment method for confirming the anticancer effect of PFC@FPNPs-Fe, (b) a thermal imaging camera image taken after irradiating a near-infrared laser on a mouse injected with PFC@FPNPs-Fe, (c) changes in the weight of the mouse during the 14-day treatment course, (d) changes in the size of the tumor, and (e) comparative photographs of the size of cancer cells obtained through excision after cancer treatment.

[0026] Hereinafter, preferred embodiments of the present invention will be described with reference to the attached drawings. However, the embodiments of the present invention may be modified in various other forms, and the scope of the present invention is not limited to the embodiments described below.

[0027] According to the present invention, a nanocomposite for cancer diagnosis and treatment is provided, which can be used as a tumor-targeting contrast agent capable of selective and precise diagnosis of tumor cells and exhibits an anticancer effect through excellent cancer cell killing.

[0028] The nanocomposite for cancer diagnosis and treatment of the present invention comprises a coordination nanoparticle that coordinates an iron ion, and more specifically, comprises at least one coordination nanoparticle comprising an iron ion; a perfluorocarbon that coordinates the iron ion; at least one dopamine compound selected from the group consisting of dopamine and dopamine analogues; and a nucleophile.

[0029] The coordination nanoparticle of the present invention is in the form of a perfluorocarbon (PFC) core; and a shell including a dopamine compound and a nucleophile; wherein the nanocomposite may be in the form of one to three coordination nanoparticles coordinating iron ions. In this case, it is preferable that the iron ion has an oxidation state of +3.

[0030] The nanocomposite of the present invention comprises a gas-generating core and an outer shell including a fluorescent dopamine compound, and can be used as a tumor-targeting contrast agent, i.e., a cancer diagnostic and cancer therapeutic agent, which is specifically delivered to tumor cells and enables selective and precise diagnosis of tumor cells through dual imaging of fluorescence and ultrasound, and at the same time, provides an excellent anticancer effect through cancer cell death through photothermal therapy and enhanced chemodynamic therapy.

[0031] In the present invention, the perfluorocarbon may be at least one selected from the group consisting of perfluorobutane (PFB), perfluoropentane (PFP), perfluorohexane (PFH), perfluoromethylcyclohexane (PFmH), and perfluorooctane (PFO).

[0032] Meanwhile, the dopamine analogue may be at least one selected from the group consisting of levodopa (LVD), nornephrine (NPP), 6-hydroxydopamine (HDA), dopamine (DA), and epinephrine (EPP).

[0033] In addition, the nucleophile may be a compound containing an amine group and / or a hydroxyl group, and more specifically, at least one selected from the group consisting of ethylenediamine, polyethyleneimine, 1,8-diaminooctane, 1,4-diaminobutane, and isopropanol, for example, ethylenediamine.

[0034] The above-described coordination nanoparticles of the present invention may be in the form of nanoparticles having a particle diameter of 10 to 100 nm, for example, 15 to 50 nm. Meanwhile, the nanoparticle complex may have a particle diameter of 100 to 400 nm, for example, 150 to 300 nm.

[0035] According to another aspect of the present invention, a composition for treating cancer is provided, comprising the nanocomposite of the present invention described above.

[0036] The composition for treating cancer of the present invention may contain the nanocomposite of the present invention in an amount of 10 μg / mL to 1 ng / mL. However, the present invention is not limited thereto, and the appropriate dosage of the composition for treating the present invention varies depending on factors such as the formulation method, administration method, patient's age, weight, sex, pathological condition, food, administration time, administration route, excretion rate, and response sensitivity, and a generally skilled physician can easily determine and prescribe an effective dosage for the desired treatment or prevention.

[0037] Meanwhile, the cancer to which the present invention can be applied may be selected from the group consisting of gastric cancer, liver cancer, lung cancer, colon cancer, breast cancer, prostate cancer, ovarian cancer, pancreatic cancer, cervical cancer, thyroid cancer, laryngeal cancer, acute myeloid leukemia, brain tumor, neuroblastoma, retinoblastoma, head and neck cancer, salivary gland cancer, and lymphoma, and may be, for example, breast cancer.

[0038] More specifically, the pharmaceutical composition for treating cancer of the present invention can be prepared as an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to an active ingredient.

[0039] Here, the route of administration may be any suitable route including topical route, oral route, intravenous route, intramuscular route, and direct absorption through mucosal tissue, and preferably, the composition is in the form of a parenteral dosage form, for example, it may be a composition for intravenous administration, and two or more routes may be combined. An example of a combination of two or more routes is a case where two or more dosage forms of drugs are combined according to the route of administration, for example, one drug is administered intravenously first and the other drug is administered topically second. For example, the composition for treating cancer of the present invention is a composition for intravenous administration.

[0040] Pharmaceutically acceptable carriers are well known in the art depending on the route of administration or formulation, and specific examples can be found in the pharmacopoeias of each country, including the “Korean Pharmacopoeia.”

[0041] When the pharmaceutical composition of the present invention is prepared as a parenteral dosage form, it can be formulated in the form of injections, transdermal administration, nasal inhalants, and suppositories using a suitable carrier according to a method known in the art. When formulated as an injection, an aqueous isotonic solution or suspension can be used as a suitable carrier, and specifically, an isotonic solution such as PBS (phosphate buffered saline) containing triethanolamine, sterile water for injection, or 5% dextrose can be used. When formulated as a transdermal dosage form, it can be formulated in the form of an ointment, cream, lotion, gel, external solution, paste, liniment, aerosol, etc. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, or carbon dioxide, and in the case of formulating it as a suppository, the carrier can be witepsol, tween 61, polyethylene glycol, cacao butter, laurin butter, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearate, sorbitan fatty acid ester, or the like.

[0042] Specific formulations of pharmaceutical compositions are known in the art.

[0043] Meanwhile, the cancer treatment composition of the present invention may be used for photothermal cancer treatment in which the composition is administered to a subject and then irradiated with near-infrared light. In this case, the near-infrared light may have a wavelength of 0.75 to 3 μm.

[0044] According to another aspect of the present invention, a contrast agent for cancer diagnosis is provided, which comprises the nanocomposite of the present invention described above.

[0045] The nanocomposite used in the present invention exhibits excellent local cancer targeting properties and can be used as an image-guided surgical contrast agent that accurately indicates the resection margin during cancer imaging diagnosis and cancer surgery using a near-infrared fluorescence signal.

[0046] Furthermore, due to its excellent local cancer targeting and singlet oxygen generation performance, it can be used as a phototherapy agent to non-surgically cure cancer or reduce the size of cancer tissue to minimize the surgical resection area. Furthermore, because it exhibits near-infrared fluorescence along with excellent local and singlet oxygen generation performance, it can be used simultaneously for bioimaging diagnostics, such as optical imaging, and cancer-targeted photodynamic therapy.

[0047] The contrast agent composition of the present invention may further include the pharmaceutically acceptable carrier described above, and may be administered parenterally or orally, as the case may be. In the case of parenteral administration, it may be administered by intravenous injection, intramuscular injection, intra-articular injection, intrasynovial injection, intramembranous injection, intrahepatic injection, intralesional injection, or intracranial injection.

[0048] The fluorescent nanocomposite containing perfluorocarbons (PFCs) of the present invention is specifically delivered to tumor cells, and can selectively and precisely distinguish tumor cells through dual imaging of fluorescence and ultrasound for tumor cells, and exhibits high dispersion stability under aqueous conditions and very low cytotoxicity.

[0049] In addition, the nanocomposite of the present invention is an anticancer agent that is delivered to tumor cells and self-replenishes hydrogen peroxide (H2O2) through oxygen (O2) release under acidic conditions of endosomes and lysosomes, for example, at pH 4.5 to pH 5.5, and has the function of enhanced chemodynamic therapy, and can also provide the function of photothermal therapy through irradiation with near infrared (NIR) light, so it is expected to be widely applied to anticancer combination treatment technology using the same.

[0050] Hereinafter, the present invention will be described in more detail through specific examples. The following examples are merely illustrative examples to aid understanding of the present invention and are not intended to limit the scope of the present invention.

[0051] Example

[0052] 1. Preparation and working principle of the perfluorocarbon fluorescent nanocomposite of the present invention

[0053] The perfluorocarbon fluorescent nanocomposite (PFC@FPNPs-Fe) of the present invention was prepared using perfluorocarbon (PFC) and epinephrine (EPP). Specifically, a solution of 120 μL of perfluorocarbon and 2.8 mL of tertiary distilled water was sonicated for about 2 minutes to form a perfluorocarbon emulsion. Then, a solution of ethanol and ammonia water (NH4OH) in a volume ratio of about 1:10 was mixed with the perfluorocarbon emulsion solution, and then about 15 mg of epinephrine and 1 mL of ethylenediamine (EDA) were added and reacted for 24 hours. Thereafter, 1 mg / mL of iron(II) chloride (FeCl2) was added to the reacted nanocomposite to form iron ions (Fe 2+) prepared a perfluorocarbon-fluorescent nanocomposite combined with a catechol group on the surface of the nanocomposite (Fig. 1).

[0054] The fluorescent nanocomposite containing the perfluorocarbon (PFC) of the present invention manufactured in this way is injected into the body, and then is selectively adsorbed to tumor cells by the enhanced permeability and retention (EPR) effect, and then catechol-iron ions (Fe) on the surface of the fluorescent nanocomposite are released in the environment of endosomes and lysosomes. 2+ ) releases iron ions from the bond, and imaging of tumor cells is possible simultaneously with the fluorescence signal switching from the "OFF" to the "ON" state. In addition, in terms of the effect when used as an ultrasound contrast agent, the core gas-generating perfluorocarbon of the perfluorocarbon-fluorescent nanocomposite of the present invention vibrates in response to an acoustic pulse at an in vivo temperature (> 37°C) and provides bright signals by generating microbubbles, thereby providing ultrasound images of tumor cells (Fig. 2(a)).

[0055] In addition, in terms of anticancer effect, the perfluorocarbon-fluorescent nanocomposite of the present invention simultaneously has the functions of chemodynamic therapy and photothermal therapy, and thus provides effective tumor cell growth inhibition and death effects. Specifically, iron ions released from the perfluorocarbon-fluorescent nanocomposite in the lysosomal environment of tumor cells decompose hydrogen peroxide (H2O2) inside cancer cells into hydroxyl radicals (·OH) through the Fenton reaction, thereby inducing apoptosis (cell death) of tumor cells. In addition, the perfluorocarbon-fluorescent nanocomposite exhibits excellent photothermal conversion properties that absorb light in the near-infrared wavelength range to generate heat, thereby generating high heat inside tumor cells and inducing necrosis of tumor cells. In particular, the temperature increase due to near-infrared light irradiation promotes the release of oxygen from perfluorocarbon within the perfluorocarbon-fluorescent nanocomposite, and the hydrogen peroxide generated within tumor cells through the redox reaction of the released oxygen and iron ions enhances the generation of hydroxyl radicals through the Fenton reaction, thereby inducing a more effective anticancer effect (Fig. 2(b)).

[0056] 2. Structural analysis of the perfluorocarbon-fluorescent nanocomposite (PFC@FPNPs-Fe) of the present invention

[0057] A. Analysis using dynamic light scattering (DLS)

[0058] The size of the PFCs@FPNPs-Fe nanoparticles of the present invention was confirmed through DLS analysis, and iron ions (Fe 2+ ) It was confirmed that the perfluorocarbon-fluorescent nanocomposite before bonding had a spherical shape with a size of approximately 50 nm.

[0059] Meanwhile, in the case of the PFCs@FPNPs-Fe nanocomposite of the present invention, which was measured after binding iron ions to the surface of the perfluorocarbon-fluorescent nanocomposite, an aggregation phenomenon occurred between the perfluorocarbon-fluorescent nanocomposite, forming a nanocomposite of about 200 nm in size. It was confirmed that this was due to chelation between the iron ions and the catechol group on the surface of the perfluorocarbon-fluorescent nanocomposite (Fig. 3).

[0060] B. Physical and chemical structural analysis and evaluation of the perfluorocarbon-fluorescent nanocomposite of the present invention

[0061] The physical and chemical structure analysis of the PFC@FPNPs-Fe of the present invention was evaluated using Fourier transform infrared (FT-IR), X-ray photoelectron spectroscopy (XPS), and transmission electron microscope (TEM). Specifically, PFC@FPNPs-Fe formed through chelation between catechol groups on the surface of the perfluorocarbon-fluorescent nanocomposite and iron ions was confirmed to have iron-oxygen (Fe-O) chemical bonds on the surface through FT-IR analysis (Fig. 4(a)), and the presence of external iron ions and peaks of internal perfluorocarbon were confirmed through XPS analysis (Figs. 4(b), 4(c) and 4(d)). In particular, changes in the physical structure and chemical element composition of the PFC@FPNPs-Fe according to the addition of iron ions were confirmed through TEM analysis of the perfluorocarbon-fluorescent nanocomposite of the present invention (Figs. 4(e) and 4(f)).

[0062] 3. Evaluation of optical properties of the perfluorocarbon-fluorescent nanocomposite of the present invention

[0063] The perfluorocarbon-fluorescent nanocomposite of the present invention has a structural mechanism in which chemical degradation occurs structurally from a nucleophile such as ethylenediamine (EDA) during polymerization in an aqueous basic solution, thereby generating fluorescence. For this reason, the optical properties of the perfluorocarbon-fluorescent nanocomposite of the present invention may vary depending on the concentration of EDA during preparation (Fig. 5(a)).

[0064] In addition, the perfluorocarbon-fluorescent nanocomposite of the present invention exhibits aggregation when combined with iron ions, resulting in fluorescence quenching and an intensity of about 1.91 x 10 -2 It has a Stern-Volmer fluorescence quenching constant of mM (Figs. 5(a), 5(b) and 5(c)).

[0065] In contrast, the perfluorocarbon-fluorescent nanocomposite of the present invention exhibits a property in which the chelate bond between the iron ion and the catechol group is broken under acidic conditions of pH 5.4 or lower, and iron ion is simultaneously released, and through this process, the fluorescent property is restored (Figs. 5(d), 5(e) and 5(f)).

[0066] 4. In vitro fluorescence and ultrasound imaging of the perfluorocarbon-fluorescent nanocomposite of the present invention

[0067] The perfluorocarbon-fluorescent nanocomposite of the present invention is absorbed into 4T1 tumor cells, which are human-derived breast cancer cells, and releases iron ions in the intracellular endosomal and lysosomal environments, enabling observation of fluorescent signals.

[0068] To evaluate this, 1 x 10 4 After treating 4T1 cells cultured in each 96-well plate with PFC@FPNPs-Fe at a concentration of 0.1 mg / mL for about 12 hours, the fluorescence images of the cells were observed.

[0069] As a result, the PFC@FPNPs-Fe of the present invention rapidly released iron ions under acidic conditions (pH 5.4) compared to neutral conditions (pH 7.4), and fluorescence signals were observed in 4T1 tumor cells within 6 hours (Figs. 6(a) and 6(b)).

[0070] In addition, in the ultrasound contrast agent, it was confirmed that the perfluorocarbon of the inner core of the PFC@FPNPs-Fe of the present invention vibrates in response to an acoustic pulse at a body temperature and provides bright signals by generating microbubbles, thereby providing ultrasound images of tumor cells (Figs. 6(c), 6(d) and 6(e)).

[0071] 5. In vitro cancer therapeutic effect of the perfluorocarbon-fluorescent nanocomposite of the present invention

[0072] The PFC@FPNPs-Fe of the present invention not only has the function of chemodynamic therapy by Fenton reaction by releasing iron ions under acidic conditions, for example, pH 4.5 to pH 5.5, but also enables photothermal therapy through irradiation with near-infrared light.

[0073] In particular, the release of oxygen (O2) by near-infrared light irradiation can self-replenish hydrogen peroxide (H2O2) through a redox reaction with iron ions, thereby improving the performance of existing chemotherapy (Fig. 7(a)).

[0074] To confirm the in vitro temperature change due to near-infrared irradiation of PFC@FPNPs-Fe, a near-infrared laser with a wavelength of 808 nm was irradiated on the PFC@FPNPs-Fe solution at an intensity of 1 W for 15 min, and the temperature was confirmed to increase up to 55°C (Fig. 7(b)). Based on these results, 1 x 10 4After treating 4T1 cells cultured in a 96-well plate with PFC@FPNPs-Fe at a concentration of 0.1 mg / mL for approximately 6 hours, the death of cancer cells was confirmed through near-infrared irradiation.

[0075] As a result, as can be seen in Fig. 7(b), when 4T1 cells treated with PFC@FPNPs-Fe were irradiated with near-infrared light for about 15 minutes, an increase in red fluorescence was confirmed, indicating the death of 4T1 cancer cells due to the photothermal effect (Fig. 7(c)).

[0076] Next, in order to confirm the temperature increase effect and the oxygen release ability according to the near-infrared irradiation of the PFC@FPNPs-Fe of the present invention, a near-infrared laser with a wavelength of 808 nm was irradiated, and the temperature change of PFC@FPNPs-Fe and the presence of gas generation were confirmed using a near-infrared thermal imaging camera and an optical microscope. As a result, as shown in Fig. 7(d), when the near-infrared laser was irradiated for 10 minutes, the temperature of PFC@FPNPs-Fe increased to a maximum of 47.1℃, and the generation of microbubbles according to the temperature increase was observed through an optical microscope.

[0077] The generation of microbubbles due to the temperature-dependent phase transition of PFC@FPNPs-Fe resulted in the temperature-dependent oxygen release of PFC@FPNPs-Fe, and the oxygen release ability according to the temperature increase was verified using a dissolved oxygen meter. As shown in Fig. 7(e), when irradiated with a near-infrared laser for 15 minutes, the oxygen release ability of PFC@FPNPs-Fe was confirmed to be about 10 times greater than that of the group that was not irradiated with the laser (Fig. 7(e)).

[0078] Furthermore, in order to confirm the anticancer therapeutic effect according to the chemodynamic therapy of the above PFC@FPNPs-Fe as an anticancer agent, 1 x 10 4After treating 4T1 cells cultured in each 96-well plate with PFC@FPNPs (negative control) and PFC@FPNPs-Fe at a concentration of 0.1 mg / mL for about 6 hours, the concentrations of hydroxyl radicals (·OH) and reactive oxygen species (ROS) in cancer cells were quantitatively measured by near-infrared light irradiation.

[0079] As a result, as can be seen in Figures 7(f) and 7(g), in 4T1 cells irradiated with near-infrared light to PFC@FPNPs-Fe, the concentrations of both hydroxyl radicals and reactive oxygen species rapidly increased, which means that near-infrared light irradiation of PFC@FPNPs-Fe generates hydrogen peroxide (H2O2) within cancer cells and promotes the Fenton reaction, thereby enhancing the therapeutic effect of chemotherapy.

[0080] To specifically evaluate the cancer cell killing effect of PFC@FPNPs-Fe through chemodynamic and photothermal therapy, the surface of 4T1 cells irradiated with near-infrared light after PFC@FPNPs-Fe treatment was analyzed using fluorescence-activated single cell sorting (FACS). As a result, the surface of 4T1 cells irradiated with near-infrared light after PFC@FPNPs-Fe treatment showed prominent features of apoptosis and necrosis compared to 4T1 cells treated with PBS or PFC@FPNPs, which can be said to demonstrate the effective anticancer effect of PFC@FPNPs-Fe. In particular, when PFC@FPNPs-Fe was irradiated with near-infrared light, the feature of necrosis increased compared to the non-irradiated control group, which directly demonstrated the result of photothermal and enhanced chemodynamic therapy (Fig. 7(h)).

[0081] 6. In vivo tumor cell imaging of the perfluorocarbon-fluorescent nanocomposite of the present invention using fluorescence and ultrasound.

[0082] In order to evaluate the cancer-specific tumor cell imaging of the PFC@FPNPs-Fe of the present invention, 0.1 mL of PFC@FPNPs-Fe was intravenously injected at a concentration of 0.1 mg / mL into balb / c nude mice (6w, female) injected with 4T1 cancer cells, and cancer-specific fluorescence and ultrasound images were confirmed.

[0083] As a result, as can be confirmed in the ultrasound images of Figs. 8(a) and 8(b) and the fluorescence image of Fig. 8(c), cancer-specific fluorescence signals and ultrasound-induced bright signals were observed in the cancer tissues of mice injected with PFC@FPNPs-Fe within 4 hours after injection using ultrasound equipment (Voluson Expert 22, GE Healthcare, USA). In particular, when the organs of balb / c nude mice injected with PFC@FPNPs-Fe were removed 24 hours later and the fluorescence signals were analyzed, fluorescence signals were clearly observed in tumor cells, indicating that PFC@FPNPs-Fe can be specifically delivered to cancer cells when injected into the body (Figs. 8(d) and 8(e)).

[0084] 7. In vivo anticancer effect of the perfluorocarbon-fluorescent nanocomposite of the present invention

[0085] In order to confirm the direct anticancer effect of the PFC@FPNPs-Fe of the present invention, 0.1 mL of PFC@FPNPs-Fe at a concentration of 0.1 mg / mL was intravenously injected twice into balb / c nude mice (6w, Female) injected with 4T1 cancer cells, and the anticancer effect was confirmed after irradiation with near-infrared light.

[0086] Specifically, as in the exemplary cancer cell treatment method of the present invention shown in Fig. 9(a), 4T1 cancer cells were administered at 5 x 10 6 Injected subcutaneously into balb / c nude mice (5w, Female) at a concentration of approximately 80 mm in size of cancer cells 3Upon reaching this, PFC@FPNPs-Fe was intravenously injected twice at two-day intervals, and then irradiated with near-infrared light to observe changes in the size and body weight of cancer cells for 14 days.

[0087] In addition, to confirm the in vivo photothermal conversion effect, a near-infrared thermal imaging camera was used to irradiate mice injected with PFC@FPNPs-Fe with near-infrared light for about 10 minutes, and it was confirmed that the temperature in the in vivo cancer cells increased to 52.8 degrees Celsius (Fig. 9(b)). As a result, no significant change was observed in the body weight of mice treated with PBS, PFC@FPNPs, PFC@FPNPs-Fe, and PFC@FPNPs-Fe + near-infrared irradiation for 14 days, whereas the cancer cells in mice treated with PFC@FPNPs-Fe + near-infrared irradiation showed the greatest reduction, which can be interpreted as a result of PFC@FPNPs-Fe + near-infrared irradiation being very effective in killing cancer cells in vivo through chemodynamic therapy and photothermal therapy (Figs. 9(c), 9(d), and 9(e)).

[0088] Although the embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and it will be apparent to those skilled in the art that various modifications and variations are possible within a scope that does not depart from the technical spirit of the present invention described in the claims.

Claims

1. Iron ions; and Coordinating the above iron ion, A nanocomposite for diagnosing and treating cancer, comprising at least one coordination nanoparticle comprising a perfluorocarbon; at least one dopamine compound selected from the group consisting of dopamine and dopamine analogues; and a nucleophile.

2. A nanocomposite for cancer diagnosis and treatment, wherein the perfluorocarbon in the first paragraph is at least one selected from the group consisting of perfluorobutane (PFB), perfluoropentane (PFP), perfluorohexane (PFH), perfluoromethylcyclohexane (PFmH), and perfluorooctane (PFO).

3. A nanocomposite for diagnosing and treating cancer, wherein the dopamine analogue in paragraph 1 is at least one selected from the group consisting of levodopa (LVD), nornephrine (NPP), 6-hydroxydopamine (HDA), dopamine (DA), and epinephrine (EPP).

4. A nanocomposite for cancer diagnosis and treatment, wherein the nucleophile in the first paragraph is at least one selected from the group consisting of ethylenediamine, polyethyleneimine, 1,8-diaminooctane, 1,4-diaminobutane, and isopropanol.

5. In the first paragraph, the coordination nanoparticles are Perfluorocarbon (PFC) core; and A shell containing epinephrine (EPP) and a nucleophile; A nanocomposite for cancer diagnosis and treatment, comprising:

6. A nanocomposite for cancer diagnosis and treatment in the first paragraph, wherein the nanocomposite is in the form of one to three coordination nanoparticles coordinating iron ions.

7. A nanocomposite for cancer diagnosis and treatment, wherein the coordination nanoparticles in the first paragraph are in the form of nanoparticles having a particle diameter of 10 to 100 nm.

8. A nanocomposite for cancer diagnosis and treatment, wherein the nanoparticle complex has a particle diameter of 100 to 400 nm in the first paragraph.

9. A composition for treating cancer, comprising a nanocomposite of any one of claims 1 to 8.

10. A composition for treating cancer, comprising the nanocomposite in an amount of 10 μg / mL to 1 ng / mL in accordance with claim 9.

11. A composition for treating cancer, wherein the cancer comprises breast cancer.

12. A composition for treating cancer, in claim 9, wherein the composition for treating cancer is a composition for intravenous administration.

13. In the 9th paragraph, the cancer treatment composition is a photothermal cancer treatment composition in which the composition is administered to a subject and then irradiated with near-infrared light.

14. A composition for treating cancer, wherein the near-infrared rays have a wavelength of 0.75 to 3 μm in the 13th paragraph.

15. A contrast agent for cancer diagnosis, comprising a nanocomposite according to any one of claims 1 to 8.

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