PHARMACEUTICAL COMPOSITION FOR TREATING INFLAMMATORY DISEASES COMPRISING GERMANIUM TELLURIDE NANOSHEETS COATED WITH POLYVINYLPYRROLIDONE

Abstract

The present invention relates to a composition for treating inflammatory diseases comprising germanium telluride nanosheets coated with polyvinylpyrrolidone, and the nanosheets have excellent anti-inflammatory and thus are excellent in treating inflammatory bowel disease and psoriasis.

Claims

1. A method of treating inflammatory diseases, comprising administering a pharmaceutically effective amount of a pharmaceutical composition to a subject in need thereof, wherein the pharmaceutical composition comprise single layer of germanium telluride nanosheets coated with polyvinylpyrrolidone as an active ingredient.

2. The method of claim 1, wherein the inflammatory disease is one or more diseases selected from the group consisting of inflammatory bowel disease, psoriasis, atopic dermatitis, psoriatic arthritis, rheumatoid arthritis, ankylosing spondylitis and systemic lupus erythematosus.

3. The method of claim 1, wherein the inflammatory bowel disease is Crohn's disease or ulcerative colitis.

4. The method of claim 1, wherein the nanosheet suppresses the expression of pro-inflammatory factors and increases the expression of anti-inflammatory factors.

5. The method of claim 4, wherein the pro-inflammatory factor is selected from the group consisting inducible nitric oxide synthase (iNOS), tumor necrosis factor-α(TNF-α) and interleukin-1β.

6. The method of claim 4, wherein the anti-inflammatory factor is Arginasel (Arg 1) or cluster of differentiation 206 (CD206).

7. The method of claim 1, wherein the pharmaceutical composition is a formulation for oral or parenteral administration.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0041] The above and other objects, features and advantages of the present invention will become more apparent to those of ordinary skill in the art by describing in detail exemplary embodiments thereof with reference to the accompanying drawings, in which:

[0042] FIG. 1 illustrates the process of fabricating germanium telluride nanosheets coated with polyvinylpyrrolidone (hereinafter referred to as GeTe-PVP NS) according to an exemplary embodiment of the present invention;

[0043] FIG. 2A illustrates the results of confirming the size of the fabricated GeTe-PVP NS;

[0044] FIG. 2B illustrates the results of confirming the thickness of the fabricated GeTe-PVP NS;

[0045] FIG. 3A illustrates the results of morphological analysis of the fabricated GeTe-PVP NS;

[0046] FIG. 3B illustrates the results of X-ray diffraction analysis of the fabricated GeTe-PVP NS;

[0047] FIG. 4A illustrates the results of confirming whether the surface of the germanium telluride nanosheet is coated with polyvinylpyrrolidone by energy-dispersive X-ray spectroscopy;

[0048] FIG. 4B illustrates the result of confirming whether the surface of the germanium telluride nanosheet is coated with polyvinylpyrrolidone by X-ray photoelectron spectroscopy;

[0049] FIG. 5A illustrates the results of confirming the absorption spectra of GeTe-PVP NS, gold nanorods (GNRs), and an indocyanine green (ICG) solution in the ultraviolet to near-infrared regions;

[0050] FIG. 5B illustrates the results of confirming the fluorescence spectra of the GeTe-PVP NS solution;

[0051] FIG. 6A illustrates the results of confirming the photoacoustic absorbance of the GeTe-PVP NS solution;

[0052] FIG. 6B illustrates the results of confirming the photoacoustic absorbance of solutions containing GeTe-PVP NS at various concentrations;

[0053] FIG. 7 shows a chicken breast stacked on top of a tube containing a GeTe-PVP NS solution;

[0054] FIGS. 8A-F illustrates the results of confirming the changes in photoacoustic absorbance of GeTe-PVP NS according to the thickness of chicken breast tissue in the regions of λ=800 nm (A and B), λ=950 nm (C and D) and λ=680 to 980 nm (E and F);

[0055] FIG. 9 illustrates the results of recording the temperature change while exposing the aqueous dispersion of GeTe-PVP NS to a laser;

[0056] FIG. 10 illustrates the results of recording the temperature change of gelatin containing GeTe-PVP NS while irradiating the gelatin with a laser;

[0057] FIG. 11 illustrates the results of confirming cell viability according to treatment concentration after treating various types of normal cells with GeTe-PVP NS;

[0058] FIG. 12A illustrates the results of confirming the expression changes of iNOS (inducible nitric oxide synthase) mRNA after treating lipopolysaccharide (LPS)—stimulated macrophage cells with GeTe-PVP NS;

[0059] FIG. 12B illustrates the results of confirming the expression changes of TNF-α (tumor necrosis factor-α) mRNA after treating lipopolysaccharide (LPS)—stimulated macrophage cells with GeTe-PVP NS;

[0060] FIG. 12C illustrates the results of confirming the expression changes of IL-1β(interlukin-1β) mRNA after treating lipopolysaccharide (LPS)—stimulated macrophage cells with GeTe-PVP NS;

[0061] FIG. 12D illustrates the results of confirming the expression changes of Arg 1 (Arginasel) mRNA after treating lipopolysaccharide (LPS)—stimulated macrophage cells with GeTe-PVP NS;

[0062] FIG. 12E illustrates the results of confirming the expression changes of CD 206 (Cluster of Differentiation 206) mRNA after treating lipopolysaccharide (LPS)—stimulated macrophage cells with GeTe-PVP NS;

[0063] FIG. 12F illustrates the results of confirming the expression changes of nitro oxide (NO) mRNA after treating lipopolysaccharide (LPS)—stimulated macrophage cells with GeTe-PVP NS;

[0064] FIG. 13A illustrates the results of confirming cell viability according to treatment concentration and treatment time after treating colorectal cancer cell DLD-1 with GeTe-PVP NS;

[0065] FIG. 13B illustrates the results of confirming cell viability according to treatment concentration and treatment time after treating colorectal cancer cell HT-29 with GeTe-PVP NS;

[0066] FIG. 13C illustrates the results of confirming cell viability according to treatment concentration and treatment time after treating breast cancer cell MCF-7 with GeTe-PVP NS;

[0067] FIG. 14 is a photograph taken in the region of λ=6680 to 980 after administering GeTe-PVP NSs, gold nanorods or indocyanine green to Balb/C nude mice;

[0068] FIG. 15A is a set of photographs of each organ taken at predetermined times after administering GeTe-PVP NSs to Balb/C nude mice;

[0069] FIG. 15B is a set of photographs of each organ taken at predetermined times after administering gold nanorods to Balb/C nude mice;

[0070] FIG. 15C is a set of photographs of each organ taken at predetermined times after administering indocyanine green to Balb/C nude mice;

[0071] FIG. 16A illustrates the results of confirming the amounts remaining in the stomach at predetermined times after administering GeTe-PVP NSs, gold nanorods, or indocyanine green to Balb/C nude mice;

[0072] FIG. 16B illustrates the results of confirming the amounts remaining in the intestine at predetermined times after administering GeTe-PVP NSs, gold nanorods, or indocyanine green to Balb/C nude mice;

[0073] FIG. 17A illustrates the results of capturing the biodistribution of GeTe-PVP NS and an ICG solution by an in vivo imaging system (IVIS) at predetermined times after administering GeTe-PVP NSs or indocyanine green to Balb/C nude mice;

[0074] FIG. 17B illustrates the results of quantifying the biodistribution of GeTe-PVP NSs and an ICG solution by in vivo imaging system (IVIS) at predetermined times after administering GeTe-PVP NSs or indocyanine green to Balb/C nude mice;

[0075] FIG. 18A illustrates the results of confirming the amount of GeTe-PVP NSs remaining in each organ at predetermined times after administering GeTe-PVP NSs to mice;

[0076] FIG. 18B illustrates the results of confirming the amount of gold nanorods remaining in each organ at predetermined times after administering gold nanorods to mice;

[0077] FIG. 19 schematically shows the process of making an animal model of inflammatory bowel disease;

[0078] FIG. 20A illustrates the results of confirming the body weight loss index while administering materials corresponding to each group to mice: Group 1—normal control group (Group1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0079] FIG. 20B illustrates the results of confirming the stool consistency while administering materials corresponding to each group to mice: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0080] FIG. 20C illustrates the results of confirming the rectal bleeding (hemocult) while administering materials corresponding to each group to mice: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0081] FIG. 20D illustrates the results of confirming the disease activity index (DAI) while administering materials corresponding to each group to mice: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0082] FIG. 21 illustrates the results of quantifying the degree of body weight loss while administering materials corresponding to each group to mice: Group 1—normal control (Group1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0083] FIG. 22A is a photograph taken by removing the intestine after administering materials corresponding to each group to mice: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0084] FIG. 22B illustrates the results of quantifying the length of the excised intestine: Group 1—normal control group (Group1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0085] FIG. 23A illustrates the results of confirming the thickness of the intestine in the proximal and distal regions of the excised intestine: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0086] FIG. 23B illustrates the results of confirming the degree of inflammation in the proximal and distal regions of the excised intestine: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0087] FIG. 23C illustrates the results of confirming the degree of tissue deformation in the proximal and distal regions of the excised intestine: Group 1—normal control group (Group 1, n=8); Group 2—inflammatory bowel disease-induced group; Group 3—inflammatory bowel disease induction+sulfasalazine (SAZ) administration; and Group 4—inflammatory bowel disease induction+GeTe-PVP NS administration;

[0088] FIG. 24 illustrates the results of confirming the amounts of GeTe-PVP NSs remaining in the stomach, small intestine and large intestine at predetermined times after administering GeTe-PVP NSs to normal mice and inflammatory bowel disease-induced mice;

[0089] FIG. 25 illustrates the results of tissue staining by isolating each organ at predetermined times after administering a high concentration of GeTe-PVP NS to mice;

[0090] FIG. 26A illustrates the results of analyzing the level of alkaline phosphatase (ALP) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0091] FIG. 26B illustrates the results of analyzing the level of alanine aminotransferase (ALT) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0092] FIG. 26C illustrates the results of analyzing the level of aspartate aminotransferase (AST) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0093] FIG. 26D illustrates the results of analyzing the level of total protein (TP) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0094] FIG. 26E illustrates the results of analyzing the level of white blood cells (WBC) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0095] FIG. 26F illustrates the results of analyzing the level of red blood cells (RBC) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0096] FIG. 26G illustrates the results of analyzing the level of mean corpuscular hemoglobin concentration (MCH) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0097] FIG. 26H illustrates the results of analyzing the level of platelets (PLT) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0098] FIG. 26I illustrates the results of analyzing the level of lymphocytes (LY) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0099] FIG. 27A illustrates the results of analyzing the level of blood urea nitrogen (BUN) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0100] FIG. 27B illustrates the results of analyzing the level of creatinine (CRE) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0101] FIG. 27C illustrates the results of analyzing the level of hemoglobin (Hb) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0102] FIG. 27D illustrates the results of analyzing the level of mean corpuscular volume (MCV) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0103] FIG. 27E illustrates the results of analyzing the level of monocytes (MO) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0104] FIG. 27F illustrates the results of analyzing the level of hematocrit (HCT) in blood after administering a high concentration of GeTe-PVP NS to mice;

[0105] FIG. 28A illustrates the results of confirming the psoriasis area severity index (PASI) after applying or administering each test material to the back of mice: NC=negative control group (petrolatum application); IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0106] FIG. 28B illustrates the results of confirming the psoriasis area severity index (PASI) after applying or administering each test material to the ears of mice: NC=negative control group (petrolatum application); IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0107] FIG. 29 illustrates the results of isolating tissues and staining them with Masson trichrome after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0108] FIG. 30 illustrates the results of isolating tissues and staining them with hematoxylin & eosin after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0109] FIG. 31A illustrates the results of isolating blood and confirming the level of IL-6 after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0110] FIG. 31B illustrates the results of isolating blood and confirming the level of nitrite after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0111] FIG. 32A illustrates the results of isolating spleen cells and confirming Th1 cell populations through flow cytometry after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0112] FIG. 32B illustrates the results of isolating spleen cells and confirming the CD4.sup.+T-bet.sup.+T cell populations among Th1 cells after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0113] FIG. 32C illustrates the results of isolating spleen cells and confirming the CD4.sup.+IFN.sub.Y.sup.+T cell populations among Th1 cells after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0114] FIG. 33A illustrates the results of isolating spleen cells and confirming Th17 cell populations through flow cytometry after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0115] FIG. 33B illustrates the results of isolating spleen cells and confirming the CD4.sup.+ROR.sub.Y.sup.+T cell populations among Th1 cells after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0116] FIG. 33C illustrates the results of isolating spleen cells and confirming the CD4.sup.+IL17.sup.+T cell populations among Th1 cells after applying or administering each test material to the back and ears of mice: NC=negative control; IMQ=induction of psoriasis by applying imiquimod on the back and ears; Anti-17A=psoriasis induction and anti-interleukin 17A administration; and GeTe-PVP NS=psoriasis induction and GeTe-PVP NS administration;

[0117] FIG. 34 shows the results of observing cell morphology after treating macrophage cells with LPS and GeTe-PVP NS and irradiating the macrophage cells with laser;

[0118] FIG. 35A illustrates the results of confirming the expression change of nitric oxide synthase (INOS) after treating macrophage cells with LPS and GeTe-PVP NS and irradiating the macrophage cells with laser; and

[0119] FIG. 35B illustrates the results of confirming the expression change of monocyte chemoattractant protein-1 (MCP-1) after treating macrophage cells with LPS and GeTe-PVP NS and irradiating the macrophage cells with laser.

DETAILED DESCRIPTION OF EXEMPLARY EMBODIMENTS

[0120] Exemplary embodiments of the present invention will be described in detail below with reference to the accompanying drawings. While the present invention is shown and described in connection with exemplary embodiments thereof, it will be apparent to those skilled in the art that various modifications can be made without departing from the spirit and scope of the invention.

[0121] Hereinafter, one or more specific exemplary embodiments will be described in more detail through Examples. However, these Examples are provided only for exemplarily explaining the one or more specific exemplary embodiments, and the scope of the present invention is not limited to these Examples.

Preparation Example: Preparation of Germanium Telluride Nanosheets (GeTe-PVP NSs) Coated with Polyvinylpyrrolidone

[0122] 1-1. Preparation of GeTe-PVP NSs

[0123] Bulk germanium telluride (GeTe) powder was purchased from TRUNNANO (Henan, China). Phosphate buffered saline (PBS, pH 7.4), pH 2 and 8 buffers, ethanol and polyvinylpyrrolidone (PVP) with a molecular weight of about 29 K were purchased from Sigma-Aldrich (St. Louis, USA). All other agents used were of the highest commercial quality available. Deionized water was used to prepare all solutions.

[0124] Germanium telluride nanosheets (hereinafter referred to as GeTe-PVP NSs) coated with polyvinylpyrrolidone were prepared using liquid-phase exfoliation.

[0125] First, GeTe powders were sonicated in ethanol for exfoliation. However, GeTe NSs quickly aggregated and precipitated in the solution. To prevent aggregation, GeTe NSs were coated with polyvinylpyrrolidone (PVP) by sonicating.

[0126] Specifically, GeTe powder was dispersed in a 1 mg/ml mixture of ethanol and PVP with an initial concentration of 5 mg/ml. Then, the suspension was sonicated for 10 h in an ice-bath at 500 W and centrifuged at 3,000 rpm for 10 min to remove the clumps of un-exfoliated bulk GeTe. The supernatant was then centrifuged at 10,000 rpm for 30 min and washed with deionized water for three times. The final precipitate was dispersed in PBS and stored at 4° C. for future use.

[0127] The process of preparing GeTe-PVP NSs is schematically illustrated in FIG. 1.

[0128] 1-2. Characterization of GeTe-PVP NSs

[0129] The thicknesses of GeTe-PVP NSs were measured using direct light scattering analysis and atomic-force microscopy, AFM; NX-10, Park Systems, Korea). As a result, it could be seen that bulk GeTe was successfully exfoliated into a single layer by confirming that the GeTe-PVP NS had a thickness of about 1.4 nm with a single layer (FIG. 2B) and an average size of 166 nm (FIG. 2A).

[0130] In addition, as a result of observing the morphologies of the prepared GeTe-PVP NSs with a transmission electron microscope (TEM; JEM-F200, JEOL, Japan), a selected area electron diffraction (SAED) pattern could be observed.

[0131] In the X-ray diffractometry (XRD) spectrum results, a sharp peak was observed at 2 θ=29.8°, and this peak is a characteristic peak of rhombohedral GeTe for both bulk GeTe and GeTe-PVP NSs (FIG. 3B). Therefore, the observation of this peak indicates that the crystallinity is well maintained even after exfoliation of bulk GeTe.

[0132] Next, it was confirmed whether GeTe NSs were coated with PVP.

[0133] Energy-dispersive X-ray spectroscope (EDS) mapping images showed that the surfaces of GeTe NSs were well coated with PVP (FIG. 4A).

[0134] X-ray photoelectron spectroscopy (XPS) spectra displayed Ge 2p.sub.3, Te 3d.sub.3 and Te3 d.sub.5 peaks at 1218, 583 and 573 eV, respectively, for both GeTe and GeTe-PVP NSs. In contrast, O 1s, N 1s, and C 1s peaks originating from PVP were observed at 531, 398, and 285 eV, respectively, only in GeTe-PVP NSs (FIG. 4B). This result clearly indicates that GeTe NSs were well coated with PVP.

Comparative Example: Preparation of Gold Nanorods

[0135] Step 1 (gold seed solution): 5 ml of HAuCl.sub.4 (0.01 M) was added to 5 ml of cetrimonium bromide (CTAB) (0.2 M). Thereafter, 60 μl of cold NaBH.sub.4 was added thereto and stirred for 2 minutes to prepare a gold seed solution. The prepared gold seed solution was stored at room temperature for 3 hours or more, and then used.

[0136] Step 2 (growth of gold nanoparticles): 30 μl of AgNO.sub.3 (0.01 M) and 5 ml of HAuCl.sub.4 (0.01 M) were added to 5 ml of CTAB (0.2 M), and then the resulting mixture was stirred. When 55 μl of ascorbic acid (0.1M) was added to the reaction solution, a yellow reaction solution turned colorless. The reaction solution was reacted at 30° C. for 1 hour while being stirred at 150 rpm (shaking incubation), and 12 ml of the gold seed solution of Step 1 was added thereto. Gold nanorods (GNR) were grown by reacting the mixture at 30° C. with stirring at 150 rpm for 24 hours. The color of the solution changed from colorless to reddish-brown during this process. Thereafter, the reaction time was adjusted (900 ml of CTAB solution: 30° C., 18 hours) to standardize the length of the gold nanoparticles.

Example 1: Optical Spectral Analysis of GeTe-PVP NS

[0137] Optical spectra for GeTe-PVP NS (Preparation Example), gold nanorods (Comparative Example 1), and Indocyanine green (ICG; Sigma-Aldrich, St. Louis, USA) (Comparative Example 2) were confirmed by the Cary5000 UV-Vis-NIR spectrophotometer (Agilent, USA) and FL and PL fluorescence spectrophotometers (F-2500 fluorescence spectrophotometer; Hitachi, Japan).

[0138] As a result, absorption spectra and fluorescence properties in the ultraviolet (UV) to near-infrared (NIR) regions were confirmed for the solution. GeTe-PVP NSs exhibited absorbance similar to gold nanorods (GNRs) and the ICG solution at the same concentration in the NIR I region (750 to 900 nm), but exhibited much higher absorbance in the NIR II region (950 to 1350 nm) (FIG. 5A).

[0139] Further, as a result of measuring fluorescence (FL) spectra for GeTe-PVP NSs (25 μg/ml) at different wavelengths, an FL peak could be observed at approximately 830 nm, and its intensity systematically increased at longer excitation wavelengths (FIG. 5B). These results suggest that GeTe-PVP NSs can be used as contrast agents for FL and multispectral optoacoustic tomography (MSOT) imaging.

[0140] In addition, the changed optical absorption properties and fluorescence properties of GeTe-PVP NSs were confirmed through PA spectroscopy results using MSOT. As a result, it could be seen that GeTe-PVP NSs can be used for imaging by confirming the photoacoustic effect of GeTe-PVP NSs. The GeTe-PVP NS solution showed higher PA absorbance than GNRs and ICG at wavelengths equal to or higher than 900 μm, which is the NIR II region (FIG. 6A). Furthermore, it was shown that the photoacoustic absorbance further increased at 800 nm and 980 nm as the concentrations of GeTe-PVP NSs increased (FIG. 6B).

Example 2: Penetration Depth Measurement of GeTe-PVP NS s

[0141] Chicken breast tissues were stacked by varying the thicknesses from 1 cm to 5 cm (FIG. 7). A 3 mm diameter tube containing 0.1 ml of 0.5 mg/ml GeTe-PVP NS or gold nanorod solution was placed in the middle of the chicken breasts stacked layer by layer, and the tube was imaged with MSOT inVision 256-TF. The data was reconstructed and conducted using multispectral processing with ViewMSOT software (iThera Medical, Germany)

[0142] It could be confirmed that gold nanorods exhibited slightly higher PA intensities at λ=800 nm than GeTe-PVP NSs at a thickness of up to 5 cm (FIGS. 8A and 8B), but GeTe-PVP NSs exhibited remarkably higher PA intensities than gold nanorods at λ=950 nm at all thicknesses, and images can be acquired up to a depth of 5 cm (FIGS. 8C and 8D). When comparing the images taken in the region of =680 to 980 nm, clearer images and PA intensities could be confirmed at all thicknesses when GeTe-PVP NSs were used rather than gold nanorods (FIGS. 8E and 8F).

Example 3: Photothermal Effect of GeTe-PVP NSs

[0143] 3-1. Photothermal effect of GeTe-PVP NS solution

[0144] GeTe-PVP NS aqueous dispersions at a concentration of 0.25 mg/ml were exposed to an 808-nm NIR laser at power densities of 0.5, 1, 1.5, and 2 W cm.sup.−2 for 100 min, respectively. The laser was turned on for 10 min and then turned off for 10 min, and this was repeated five times. The temperature change of the membranes was recorded every 1 min using an IR thermal camera (FLIR T335 Thermal Imaging Camera, FLIR, USA).

[0145] As a result, it could be confirmed that the temperature of the dispersion increased to 18.4±0.4, 35.2±0.3, 47.8±0.4, and 61.9±1.3° C., respectively, and that the temperature of the dispersion immediately dropped to room temperature when the laser was turned off. Furthermore, it could be seen that the temperature rises and drops in the same way even though the experiment was repeated five times (FIG. 9).

[0146] Since the elevated temperature of the dispersion is the temperature that kills cancer cells, it is expected that GeTe-PVP NSs can be used for phototherapy, which is one method of cancer treatment.

[0147] 3-2. Photothermal Effect of Phantom Containing GeTe-PVP NSs

[0148] To measure the photothermal effect of GeTe-PVP NSs in vivo, gelatin containing GeTe-PVP NSs at a concentration of 0.25 mg/ml was prepared. The gelatin was irradiated with an 808 nm laser in the NIR I region and a 980 nm laser in the NIR II region at an intensity of 1 W/cm.sup.2 for 10 minutes. As a result of measuring the temperature change of gelatin, it could be confirmed that the temperature of gelatin increased to 19.8±1.0 and 20.1±0.3° C. (FIG. 10). Since this is a temperature that sufficiently kills cancer cells, GeTe-PVP NSs can be used for the purpose of photothermal therapy in cancer therapy.

Example 4: Cytotoxicity and Anti-Inflammatory Assays of GeTe-PVP NSs

[0149] 4-1. Cytotoxicity and GeTe-PVP NSs

[0150] Two types of primary cells, human mesenchymal stem cells (hMSCs) and human fibroblast-like synoviocytes (FLSs) and two cell lines of murine macrophage cells (RAW264.7) and African green monkey kidney epithelial cells (Vero CCL-81) were seeded in 96-well plates at a density of 8,000 cells per well. Cells were cultured overnight in an incubator maintained at 37° C. in 5% CO.sub.2 and a humidified atmosphere. The primary cells were cultured in a low glucose DMEM medium (Corning, USA) supplemented with 10% (v/v) FBS and the cell lines were cultured in a high glucose DMEM medium supplemented with 10% (v/v) FBS. The next day, the culture medium was replaced with a fresh culture medium containing different concentrations of GeTe-PVP NSs (0 to 50 μg/ml) and incubated for 24 hours. Cell viability was then determined by CCK-8 Assay (96992, Sigma). 10 μL of CCK-8 reagent was added to each well, and the plate was incubated for 2 hours. Cell viability was assessed by measuring the optical density at 450 nm.

[0151] As a result, it could be confirmed that GeTe-PVP NSs have no cytotoxicity (FIG. 11).

[0152] 4-2. Measurement of In Vitro Anti-Inflammatory Activity of GeTe-PVP NSs

[0153] RAW264.7 macrophage cells were purchased from Korea Cell Line Bank. Cells were seeded at a density of 8×10.sup.5 cells per well in 6-well plates and cultured overnight in DMEM medium (Corning, USA) supplemented with 10% (v/v) FBS. The next day, cells were treated and stimulated with 1 μg/ml of LPS and treated with various concentrations of GeTe-PVP NSs (0, 6.25, 12.5, 25, and 50 μg/ml) for 24 hours. Then, the cells were collected by scraping, and RNA was extracted using the GeneJET RNA Purification Kit (Thermo Scientifics). cDNA was synthesized using the Maxime RT premix kit (iNtRON Biotechnology, Korea). Real-time PCR was performed with the Step One Plus real-time PCR machine (Applied Biosystems, USA) using PCRBIO SYBR Green (PCRBIOSYSTEM, UK). Relative mRNA expression levels for each sample were calculated using the 2.sup.−ΔΔCT method with GAPDH expression. The confirmed genes are pro-inflammatory factors inducible nitric oxide synthase (iNOS), tumor necrosis factor-α(TNF-α) and interleukin-1β, and anti-inflammatory factors Arginasel (Arg 1) and CD 206.

[0154] As a result of confirmation, it could be seen that the expression of pro-inflammatory factors decreased (FIGS. 12A, 12B and 12C) and the expression of anti-inflammatory factors increased depending on the treatment concentration of GeTe-PVP NSs (FIGS. 12D, 12E and 12F). This result means that GeTe-PVP NSs have excellent anti-inflammatory activity.

Example 5: In Vitro Anticancer Activity Assay of GeTe-PVP NSs

[0155] Two types of colorectal cancer cells, DLD-1 and HT-29, and breast cancer cells, MCF-7, were treated with GeTe-PVP NSs at a concentration of 0 to 100 μg/ml and cultured for 24 or 48 hours, and cell activity was confirmed by CCK-8. As a result, unlike the results of Example 4 where GeTe-PVP NSs did not affect the viabilities of the four types of normal cells, it could be confirmed that GeTe-PVP NSs kill cancer cells (FIGS. 13A, 13B and 13C).

[0156] This is presumed to be due to the selective anti-cancer activity of Te caused by the generation of reactive oxygen species (ROS) in cancer cells. In the case of breast cancer treatment, when GeTe-PVP NSs are used as a combined treatment together with phototherapy using optical heat of Example 3, a more effective cancer treatment effect will be obtained.

Example 6: MSOT Imaging of Stomach and Intestine Using GeTe-PVP NSs

[0157] After GeTe-PVP NS (5 mg/kg, 100 μl), gold nanorods (5 mg/kg, 200 μl) and an ICG solution (5 mg/kg, 200 μl) to Balb/C nude mice, images were captured in a region of λ=680 to 980 nm and compared. As a result, it could be confirmed that not only clearer images of GeTe-PVP NSs could be obtained from experimental groups orally administered GeTe-PVP NSs compared to those of experimental groups orally administered gold nanorods or the ICG solution, but also GeTe-PVP NSs remained in the intestine for a longer period of time (FIG. 14).

[0158] As a result of confirming images of each organ with MSOT, it was possible to confirm in vivo images of GeTe-PVP NSs, gold nanorods, and the ICG solution remaining in the stomach and intestine over time (FIG. 15). In addition, changes in the GeTe-PVP NSs, GNR, and ICG solution over time were confirmed using PA intensity. As a result, it could be confirmed that a larger amount of GeTe-PVP NSs remained in the gastrointestinal tract up to 2 hours after administration compared to gold nanorods and the ICG solution, and a larger amount remained in the intestines up to 24 hours after administration (FIGS. 16A and 16B).

Example 7: IVIS Fluorescence Imaging of Stomach and Intestine Using GeTe-PVP NSs

[0159] After GeTe-PVP NSs (5 mg/kg, 100 μl) or an ICG solution (5 mg/kg, 200 μl) were orally administered, the mice were subjected to laparotomy at 1, 6, and 24 hours, and the biodistribution of GeTe-PVP NSs and the ICG solution was photographed with an in vivo imaging system (IVIS). Furthermore, the stomach, intestine, liver, lungs, pancreas, heart, and kidneys were removed at each time to confirm their biodistribution by fluorescence imaging with IVIS and quantify the amounts of GeTe-PVP NSs and ICG accumulated in each organ by FL intensity.

[0160] As a result, it was confirmed that a large amount of GeTe-PVP NSs was accumulated in the intestine until 6 hours after administration, and a large amount of the ICG solution remained in the intestine until 1 hour after administration, but large amounts of GeTe-PVP NSs and the ICG solution were released from the intestine after 6 hours (FIGS. 17A and 17B).

Example 8: Confirmation of Biodistribution of GeTe-PVP NS s

[0161] GeTe-PVP NSs (5 mg/kg, 100 μl) or gold nanorods (5 mg/kg, 200 μl) were orally administered to mice. Thereafter, in order to investigate a biodistribution, the stomach, intestine, liver, lungs, pancreas, heart, and kidneys were excised at 1 hour, 6 hours, and 24 hours after administration, and the biodistribution was confirmed by ICP-MS. As a result of confirmation, it could be seen that a large amount of GeTe-PVP NSs remained in the intestine 6 hours after oral administration, and it could be confirmed that GeTe-PVP NSs were present in a larger amount than the gold nanorods. 24 hours after oral administration, it could be confirmed that large amounts of GeTe-PVP NSs and gold nanorods were released from the body (FIGS. 18A and 18B; ID=injected dose).

Example 9: Evaluation of IBD Therapeutic Effect Using GeTe-PVP NSs

[0162] Since the in vitro anti-inflammatory effect of GeTe-PVP NSs was confirmed in Example 4, the therapeutic effect of GeTe-PVP NSs on inflammatory bowel disease was confirmed in an inflammatory bowel disease model.

[0163] 9-1. Establishment of Inflammatory Bowel Disease Model

[0164] 7- to 8-week-old male C57BL/6 mice (Orient Bio Inc.; Seongnam, Korea) were used for the experiments. Colitis was induced by allowing an acute inflammatory bowel disease model to ingest drinking water to which 2% dextran sodium sulfate (DSS; MP Biochemical, Irvine, Ca, USA) was added for 7 days. A normal control group was allowed to ingest the same amount of water for the same time (FIG. 19).

[0165] The experimental groups were classified into the following four groups (Table 1): a normal control group (Group 1, n=8), a group in which colitis is induced by DSS (Group 2, n=8), a group in which sulfasalazine (SAZ), which is a common therapeutic agent, was administered to a DSS-induced colitis model (Group 3, n=8, SAZ=200 mg/kg/day), and a group in which GeTe-PVP NSs were administered to a DSS-induced colitis model (Group 4, n=8, GeTe-PVP NS=5 mg/kg/day).

TABLE-US-00001 TABLE 1 Group (n = 8) Sample Dosage Group 1 PBS Group 2 dextran sodium sulfate (DSS) 2 wt % * 8 days Group 3 dextran sodium sulfate (DSS) + 2 wt % * 8 days + Sulfasalazine (SAZ) 200 mg/kg * 6 days Group 4 dextran sodium sulfate (DSS) + 2 wt % * 8 days + GeTe PVP NS 5 mg/kg * 6 days

[0166] 9-2. Evaluation of Macroscopic Therapeutic Effect Including Disease Activity Measure

[0167] Body weight changes and the disease activity index (DAI) of mice were confirmed every day for 7 days. Each score of weight loss index (0 to 4; FIG. 20A), rectal bleeding observed with the naked eye in the stool or anus (0 to 3; FIG. 20C), and stool consistency (0 to 3; FIG. 20B) is summed to evaluate the disease activity index on a 10-point scale (FIG. 20D). [Stevceva L, Pavli P, Husband A, Ramsay A, Doe WF. Dextran sulphate sodium-induced colitis is ameliorated in interleukin 4 deficient mice. Genes Immun 2001; 2:309-316.]

[0168] As a result of evaluation, Group 2 had severe intestinal bleeding and Group 4 had relatively minor bleeding (FIG. 20C). When the DAI was compared, Group 2 showed a continuous increase in DAI during the administration period, but Groups 3 and 4 showed a tendency to have a lower DAI compared to Group 2. The DAI was significantly lower in Group 4 compared to Group 3 on day 7 of administration (FIG. 20D).

[0169] Further, in Group 1, a sustained body weight gain was confirmed for 7 days, whereas in Group 2, a body weight loss was confirmed from day 4 of administration. A body weight loss was confirmed in Groups 2 and 4 after day 5 of administration, but it could be confirmed that the body weight was slowly decreased compared to Group 2 (FIG. 21).

[0170] After the corresponding sample administration was completed, the mice were sacrificed by cervical dislocation and the intestine from the cecum to the anus was excised to measure the length of the intestine. As a result, the average colorectal length in Group 1 was 72.2 mm, while the average colorectal length in Group 2 was 44.8 mm, which was significantly shorter compared to Group 1. In contrast, Group 4 had an average colorectal length of 50.9 mm, which was longer compared to Group 2 (FIGS. 22A and 22B).

[0171] 9-3. Histological Evaluation of Therapeutic Effect

[0172] Tissue specimens were obtained from the proximal and distal regions of the excised intestine, fixed in formalin, and stained with hematoxylin-eosin and periodic acid-schiff (PAS).

[0173] As a result of staining, the thickness of the intestine became thicker in Group 2 compared to Group 1, and inflammatory findings of more than moderate severity and deformation of intestinal tissue were also observed. However, Groups 3 and 4 had similar intestinal tissue morphology to Group 1, and many inflammatory findings were not found (FIGS. 23A, 23B and 23C).

[0174] 9-4. Evaluation of Targeting and Retention of GeTe-PVP NSs in Inflammatory Bowel Disease Model and General Model

[0175] GeTe-PVP NSs were orally administered to inflammatory bowel disease model mice and normal mice. The targeting and retention of the inflammatory site of GeTe-PVP NSs were evaluated by ICP-MS in the stomach, small intestine, and large intestine at 6 and 24 hours after administration. 6 hours after oral administration, more GeTe-PVP NSs were found in the small intestine of inflammatory bowel disease mice (IBD) compared to normal mice. 24 hours after oral administration, GeTe-PVP NSs were scarcely found in the stomach, small intestine, and large intestine of normal mice. However, it was confirmed that 9.6% of the administered GeTe-PVP NSs remained in the large intestine in inflammatory bowel disease model mice (FIG. 24). This result means that GeTe-PVP NSs can target the inflammatory site to stay in the corresponding site for a long time in the inflammatory bowel disease model.

Example 10: Evaluation of GeTe-PVP NS Biocompatibility

[0176] After a 1 mg/ml GeTe-PVP NS solution was orally administered to Balb/C mice (OrientBio Inc.; Seongnam, Korea), each organ and blood were isolated and analyzed over time.

[0177] Blood analysis items were as follows: alkaline phosphatase (ALP), alanine aminotransferase (ALT), aspartate aminotransferase (AST), total protein (TP) count, white blood cells (WBC), red blood cells (RBC), mean corpuscular hemoglobin concentration (MCH), platelets (PLT), lymphocytes (LY), blood urea nitrogen (BUN), creatinine (CRE), hemoglobin (Hb), mean corpuscular volume (MCV), monocytes (MO) and hematocrit (HCT).

[0178] As a result of histological analysis, it could be confirmed that GeTe-PVP NSs are not toxic while being biocompatible by confirming that there was no abnormality in all the isolated tissues (FIG. 25). It could be seen that all blood analysis results were also within normal ranges (FIGS. 26A-26I and 27A-27F).

Example 11: Confirmation of Psoriasis Therapeutic Effect of GeTe-PVP NSs

[0179] 11-1. Establishment of Psoriasis Animal Model

[0180] C57BL/6 mice (7 weeks old, female) were purchased from Orient Bio Inc. (Korea). Mice were sufficiently fed solid food (no antibiotics added) and water until the day of the experiment, and were allowed to adapt to an environment with a temperature of 23±2° C., a humidity of 55±10%, and a light-dark cycle of 12-12 hours for one week, and then used for experiments.

[0181] As a psoriasis animal model, an imiquimod-induced psoriasis-like inflammation model commonly used in psoriasis research was used. Specifically, aldara cream (5% imiquimod (IMQ)), which is a TLR7 agonist, was applied to the depilated back and ears of mice for 7 days to induce psoriatic inflammatory responses such as scaling, redness, and skin thickening.

[0182] 11-2. Evaluation of Therapeutic Effect (with Naked Eye)

[0183] In order to evaluate the therapeutic effect of GeTe-PVP NSs on psoriasis, the psoriasis animal models of 11-1 above were classified into four groups as shown in the following Table 2. Group 1 is the negative control group, and the same amount of Vaseline as imiquimod was topically applied on the back and ears. Anti-interleukin 17A known to be effective for treating psoriasis was intraperitoneally injected into Group 3. GeTe-PVP NS s were intradermally injected into Group 4.

TABLE-US-00002 TABLE 2 Test Administration Administration Denoted material Dose date Method Group 1 NC Vaseline Back 60 mg/mouse 1 to 7 Skin application Ears 40 mg/mouse Group 2 IMQ Imiquimod Back 60 mg/mouse 1 to 7 Skin application Ears 40 mg/mouse Group 3 Anti-17A Anti-IL-17A   3 mg/kg 1, 3, 5 Intraperitoneal injection Group 4 GeTe-PVP NS GeTe-PVP NS 2.5 mg/kg 1, 3, 5 Intradermal injection

[0184] The efficacy of GeTe-PVP NSs for treating psoriasis was visually evaluated using a psoriasis area severity index (PASI) score, which is the most commonly used evaluation method for clinical evaluation of psoriasis. In the animal model, skin thickness, redness and the degree of scaling were visually evaluated on a scale of 4 points each. [0185] Thickness: the middle part of the skin on the back and the middle part of the right ear were measured using a caliper/the pattern of change was quantified as 0 to 3 points by setting the thickness of the brown skin on the first day of the experiment (Day 1) for each mouse as 100% and expressing the change in thickness as % (0 point: 1 to 5%, 1 point: 5 to 50%, 2 points: 50 to 100%, and 3 points: >100% change) [0186] Redness (erythema): evaluated by visual observation [0187] Scaling: evaluated by visual observation

[0188] Erythema and keratin were quantified on a scale of 0 to 3 points (0 point: no symptoms, 1 point: mild, 2 point: moderate, and 3 point: severe).

[0189] As a result of the evaluation, it was confirmed that Group 2 (IMQ) had severe inflammatory responses such as scaling, redness, and skin thickening on both the back and ears, and thus psoriasis was well induced. In contrast, it could be seen that Group 3 (anti-17A) administered anti-IL-17A known to treat psoriasis significantly alleviated scaling and redness induced by imiquimod. It could be confirmed that imiquimod-induced redness and skin thickening were also significantly alleviated in Group 4 (GeTe-PVP NSs) (FIGS. 28A and 28B).

[0190] 11-3. Evaluation of Hematological and Histological Therapeutic Effect

[0191] Mice were sacrificed after visual observation was completed on the final day of the experiment. Blood was obtained through cardiac blood collection, and then organs (back, stomach, and spleen) were excised.

[0192] Paraffin sections were prepared by fixing back skin and ears in 4% formalin, and then stained with Masson trichrome (MT) and hematoxylin & eosin (H&E). Thereafter, histological evaluation was performed by taking photographs under a microscope. Next, the epidermis thickness was measured by analyzing photographs of stained tissues with the Image J program.

[0193] In MT staining, the red-stained area is the epidermis, and the blue-stained area is the dermis. It could be confirmed that the epidermis was thicker in Group 2 (IMQ) compared to Group 1 (NC), but the thickness of the epidermis was thinner in Group 3 (Anti-17A) and Group 4 (GeTe-PVP NSs) compared to the imiquimod-applied group (FIG. 29).

[0194] The same result could be confirmed with H&E staining. In H&E staining, it could be seen that the dark purple-stained area at the top of the tissue is the epidermis, but the epidermis was thicker in Group 2 (IMQ) compared to Group 1 (NC), and inflammatory cells appearing as dots infiltrated the epidermis area to a large extent. In addition, in group 2 (IMQ), many symptoms occurring in psoriasis diseases such as parakeratosis, hyperkeratosis, and microabscesses were observed, indicating that psoriasis was well induced. In contrast, it could be confirmed that in Group 3 (Anti-17A) and Group 4 (GeTe-PVP NSs), the thickness of the epidermis was thinner compared to Group 2 (IMQ), the infiltration of inflammatory cells in the dermis is also reduced, and the pattern of psoriasis diseases appearing in the stratum corneum of the skin, such as parakeratosis, was alleviated (FIG. 30).

[0195] The blood was centrifuged at 13,200 rpm for 5 minutes to isolate only plasma (serum). The levels of interleukin-6 (IL-6) and nitric oxide (NO), which are inflammatory cytokines in plasma, were analyzed with ELISA assay.

[0196] As a result of analysis, levels of all inflammatory factors in Group 2 (IMQ) were increased compared to Group 1 (NC). In contrast, levels of inflammatory factors were decreased in Group 3 (Anti-17A) and Group 4 (GeTe-PVP NSs) compared to Group 2 (IMQ). In particular, it could be confirmed that the NO (measured at the level of nitrite) level was significantly reduced in Group 4 (GeTe-PVP NSs) (FIGS. 31A and 31B).

[0197] After the excised spleen was ground, splenocytes were obtained through a red blood cell lysis process. After incubation of splenocytes with antibodies specific for each cell, Th1, Th17, and Treg cell populations were analyzed through flow cytometry (Sony). Th1 cells include inflammatory cytokines CD4.sup.+, T-bet and IFN-γ positive T cells, and Th17 cells include inflammatory cytokines CD4.sup.+, RAR (retinoic acid receptor)—related orphan receptor gamma (RORγ) and IL-17A positive T cells.

[0198] As a result of the analysis, the number of Th1 cells was remarkably increased in Group 2 (IMQ) compared to Group 1 (NC), and slightly decreased in Group 3 (Anti-17A) compared to Group 2 (IMQ). In contrast, it could be seen that in Group 4 (GeTe-PVP NSs), the number of Th1 cells was remarkably reduced (FIGS. 32A, 32B and 32C). Analysis of Th17 cells also showed a similar tendency (FIGS. 33A, 33B and 33C).

Example 12: Evaluation of Inflammatory Therapeutic Effect Using GeTe-PVP

[0199] NSs and Near-Infrared Rays

[0200] 12-1. Induction of Inflammation, GeTe-PVP NS and Near-Infrared Treatment

[0201] RAW264.7 cells were cultured in an incubator at 5% CO.sub.2, 95% humidity and 37° C. using DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. Cells were sub-cultured when they showed a confluence of 70 to 80%.

[0202] RAW264.7 cells were seeded in 12-well plates at a density of 3×10.sup.5 cells/well and cultured for 24 hours as in the existing experiment. Thereafter, the cells were simultaneously treated with 0.5 μg/ml lipopolysaccharide (LPS) and 12.5 μg/ml GeTe-PVPNS and additionally cultured for 24 hours. The next day, the plate was removed, irradiated with a laser at 42° C. for 10 minutes, and incubated in an incubator. After 12 hours, RNA was isolated by harvesting media and cells, and cDNA was synthesized for further experiments.

[0203] As a result of observing the cell morphology before harvesting the cells, it could be confirmed that there was no change (FIG. 34).

[0204] 12-2. Evaluation of Inflammatory Therapeutic Effect

[0205] RT-PCR was performed on the synthesized cDNA to confirm changes in expression of nitric oxide synthase (INOS) and monocyte chemoattractant protein-1 (MCP-1), which are inflammation-promoting materials. As a result, it could be seen that the levels of INOS and MCP-1, which were increased by LPS treatment, were decreased by GeTe-PVPNS treatment, and were further remarkably decreased by near-infrared irradiation (FIGS. 35A and 35B).

[0206] It was confirmed through macroscopic, blood, and histological evaluations that GeTe-PVP NSs according to the present invention had a therapeutic effect not only on inflammatory bowel disease but also on psoriasis, which is a systemic inflammatory disease through disease animal models. In addition, through in vitro cell experiments, it could be confirmed that when concurrent administration of GeTe-PVP NS and near-infrared irradiation, the production of inflammation promoting materials was further suppressed. This indicates that when both GeTe-PVP NSs and near-infrared irradiation are applied to treat inflammatory diseases, better therapeutic effects can be obtained.

[0207] The germanium telluride nanosheets coated with polyvinylpyrrolidone of the present invention have excellent anti-inflammatory and anticancer activity and absorb light in the ultraviolet to near-infrared region, and thus can be used as a composition for treating inflammatory diseases and cancer and a contrast agent composition.