Metal nanocomposite for photothermal immunotherapy

Abstract

The present invention relates to a metal nanocomposite targeting cancer cells and immune effector cells and exhibiting a photothermal effect, and a use thereof.

Description

Metal nanocomposites for photothermal immunotherapy

[0001] The present invention relates to a metal nanocomposite that targets cancer cells and immune effector cells and exhibits a photothermal effect, and to uses thereof.

[0002] Macrophages are responsible for both adaptive and innate immunity in the human immune system. Macrophages exist in the blood as monocytes and differentiate into macrophages in response to environmental changes, such as disease onset or exposure to foreign substances, and in response to cell signaling molecules like cytokines. As the immune system is activated in a specific area, macrophages undergo two major polarizations. Among them, M1 macrophages (classically activated macrophages) are involved in pro-inflammatory responses and play a crucial role in suppressing the invasion of foreign substances into the body. In particular, M1 macrophages exhibit the characteristic of attacking and eliminating tumor cells. In contrast, M2 macrophages are involved in anti-inflammatory and immune suppression, playing a key role in wound healing and tissue remodeling. Some of the above M2 macrophages, unlike M1 macrophages, have the property of inducing tumor growth and tissue infiltration, which can affect tumor exacerbation. Since these macrophages have a significant impact on the body's immune activity as well as tumor death and growth, immunotherapies utilizing them are being studied. Among these macrophages, macrophages differentiated under the influence of the tumor microenvironment (hereinafter referred to as TME) are known as tumor-associated macrophages (hereinafter referred to as TAMs). In particular, M2 phenotype TAMs have the property of promoting tumor progression and reducing the effectiveness of drug therapy.In addition, during the migration and invasion stages of tumor cells, TAMs are differentiated into M2 macrophages by interleukin-4 (IL-4) expressed by CD4+ T cells and tumor cells, which plays a role in assisting the migration and metastasis of tumor cells (Cell. 2010;141(1)39-51, Curr Opin Immunol. 2010;22(2)231-7, Nature Reviews Immunol. 2015, 15:7386). Tumor cells induce these properties of macrophages to disrupt the human immune system and evade attacks from immune cells. Among the immune activities of macrophages, phagocytosis is also suppressed by the immune evasion function induced by tumor cell-derived expression factors. The immune evasion signal of tumor cells toward macrophages is called the 'Don't eat me signal', and this signal reduces the anti-tumor effect in the body by preventing the tumor cells from attacking macrophages by interfering with the recognition of cancer cells by macrophages.

[0003] The present invention was conceived to solve the above-described problem, and aims to provide a metal nanocomposite comprising a cancer cell targeting moiety, an immune effector cell targeting moiety, and a metal moiety exhibiting a photothermal effect.

[0004] In addition, the present invention aims to provide a drug delivery vehicle comprising a metal nanocomposite and a pharmaceutically active ingredient.

[0005] In addition, the present invention aims to provide a pharmaceutical composition for preventing or treating cancer.

[0006] In addition, the present invention aims to provide a method for producing a metal nanocomposite.

[0007] Furthermore, the present invention aims to provide uses of metal nanocomposites for use in photothermal therapy and photodynamic therapy.

[0008] In addition, the present invention aims to provide a method for treating cancer.

[0009] The technical problems of the present invention are not limited to the technical problems mentioned above, and other technical problems not mentioned will be clearly understood by those skilled in the art from the description below.

[0010] To solve the above problem, the present invention provides a metal nanocomposite comprising a cancer cell targeting moiety, an immune effector cell targeting moiety, and a metal moiety exhibiting a photothermal effect.

[0011] In addition, the present invention provides a drug delivery vehicle comprising a metal nanocomposite and a pharmaceutically active ingredient.

[0012] In addition, the present invention provides a pharmaceutical composition for preventing or treating cancer, comprising a metal nanocomposite or drug delivery vehicle as an active ingredient.

[0013] The present invention also provides the use of metal nanocomposites for use in photothermal therapy and photodynamic therapy.

[0014] The present invention also provides a method for treating cancer.

[0015] The metal nanocomposite of the present invention has the effect of targeting cancer cells and macrophages.

[0016] In addition, the metal nanocomposite of the present invention has the effect of blocking immune evasion signals generated by the interaction between macrophages and cancer cells, thereby enhancing the phagocytosis of macrophages against cancer cells.

[0017] In addition, the metal nanocomposite of the present invention has a photothermal effect that generates heat in a specific wavelength range, and thus has the effect of targeting and killing cancer cells.

[0018] Figure 1 is a diagram analyzing the manufacturing process (A) and its characteristics (B to G) of Man-AuNR-SNA, a metal nanomaterial that targets cancer cells and macrophages and can be applied to photothermal therapy.

[0019] Figure 2 is a diagram analyzing the cancer cell-specific binding ability of Man-AuNR-SNA:

[0020] A: Schematic diagram of cancer cell-specific binding mechanism of Man-AuNR-SNA;

[0021] B: Results of cancer cell-specific binding analysis of SNA using confocal microscopy;

[0022] C: Results of cancer cell-specific binding analysis of SNA using FACS; and

[0023] D and E: Cancer cell-specific binding assay results of Man-AuNR-SNA.

[0024] Figure 3 is a diagram analyzing the macrophage-specific binding ability of Man-AuNR-SNA:

[0025] A: Results of marker expression analysis of macrophages;

[0026] B: Results of specific binding ability analysis of mannose to M2 macrophages; and

[0027] C: Specific binding analysis results of Man-AuNR-SNA particles to M2 macrophages.

[0028] Figure 4 shows the cytotoxicity of Man-AuNR-SNA on various cells (A) and the cancer cell killing effect of Man-AuNR-SNA by photothermal therapy (B and C).

[0029] Figure 5 is a diagram confirming cancer cell death by combined treatment of Man-AuNR-SNA and photothermal therapy:

[0030] A: Expression analysis of cell death-related proteins; and

[0031] B and C: Annexin V / PI staining analysis results.

[0032] Figure 6 is a diagram confirming the immune evasion inhibition effect of cancer cells by combined treatment with Man-AuNR-SNA and photothermal therapy:

[0033] A: Cancer cell phagocytosis of macrophages by SNA treatment;

[0034] B: Macrophagic activity of the Man-AuNR-SNA particle-only treatment group; and

[0035] C: Macrophage activity of the combined treatment group of Man-AuNR-SNA treatment and photothermal therapy.

[0036] Hereinafter, preferred embodiments of the present invention will be described in detail with reference to the attached drawings. The advantages and features of the present invention, and methods for achieving them, will become clear with reference to the embodiments described in detail below together with the attached drawings. However, the present invention is not limited to the embodiments disclosed below, but can be implemented in various different forms. These embodiments are provided only to ensure that the disclosure of the present invention is complete and to fully inform those skilled in the art of the scope of the invention, and the present invention is defined only by the scope of the claims. Like reference numerals refer to like elements throughout the specification.

[0037] Unless otherwise defined, all terms (including technical and scientific terms) used herein may be used in a sense commonly understood by those of ordinary skill in the art to which the present invention pertains. Furthermore, terms defined in commonly used dictionaries are not to be interpreted ideally or excessively unless explicitly and specifically defined otherwise. The terminology used herein is for the purpose of describing embodiments and is not intended to limit the present invention. In this specification, singular forms also include plural forms, unless specifically stated otherwise.

[0038] The terms "comprises" and / or "comprising" as used in the specification do not exclude the presence or addition of one or more other components, steps, operations and / or elements.

[0039] The term "liposome" used in the specification refers to a hollow droplet-like structure formed by a phospholipid bilayer when phospholipid, a major component of biological membranes, is added to an aqueous solution.

[0040] The term "nucleic acid" as used herein refers to a 5-carbon sugar, a phosphate group, and a nitrogenous base as the basic backbone. When the sugar portion is ribose, it is referred to as RNA, and when it is deoxyribose, it is defined as DNA. The base portion of DNA is composed of four bases: adenine (A), thymine (T), cytosine (C), and guanine (G), and the base portion of RNA contains uridyl (U) instead of thymine (T) in DNA. The nucleic acid includes a nucleic acid derivative or analogue, and the nucleic acid derivative or analogue may be a nucleic acid that has changed any of the components of the nucleic acid defined above.

[0041] As used herein, the term "antibody" includes polyclonal antibodies and monoclonal antibodies, antigen-binding fragments of such antibodies, including Fab, F(ab')2, Fd, Fv, scFv, bispecific antibodies, and the smallest recognition unit of an antibody, as well as single-chain derivatives of such antibodies and fragments. The type of antibody can be selected from IgG1, IgG2, IgG3, IgG4, IgA, IgM, IgE, and IgD. In addition, the term "antibody" includes naturally occurring antibodies and non-naturally occurring antibodies, including, for example, chimeric, bifunctional, and humanized antibodies and related synthetic isoforms. The term "antibody" can be used interchangeably with "immunoglobulin." The term "antibody fragment" refers to a portion of an intact antibody and refers to the antigen-determining variable region of an intact antibody. Examples of antibody fragments include, but are not limited to, Fab, Fab', F(ab')2 and Fv fragments, linear antibodies, scFv antibodies, and multispecific antibodies formed from antibody fragments.

[0042] The terms "polypeptide," "peptide," "oligopeptide," and "protein," as used herein, are used interchangeably herein and refer to a polymeric form of amino acids of any length. The polymeric form may include encoded and non-encoded amino acids, chemically or biochemically modified or derived amino acids, and polypeptides having modified peptide backbones.

[0043] As used herein, the term "lectin" refers to a protein or peptide that binds to carbohydrate structures. Those skilled in the art will understand that a lectin is a protein or peptide that is highly specific for binding to sugar moieties.

[0044] As used herein, “anti-tumor effect” or “anticancer effect” refers to a biological effect that can be manifested by a decrease in tumor volume, a decrease in tumor cell count, a decrease in the number of metastases, an increase in lifespan, or an alleviation of various physiological symptoms associated with a cancerous condition.

[0045] As used herein, "targeting" refers to the ability to target a specific cell or site within a living organism, and refers to the property of a substance being specifically delivered to that site when distributed within the body. Targeting can be a property that is limited to a specific biological site, meaning that a significantly greater amount of the substance is delivered to that site compared to other sites.

[0046] As used herein, "treatment" refers to any action that improves or beneficially alters the symptoms of a disease by administering the composition of the present invention. Those skilled in the art to which the present invention pertains will be able to accurately determine the criteria for diseases for which the composition of the present invention is effective and determine the degree of improvement, enhancement, and treatment by referencing materials provided by the Korean Medical Association and other sources.

[0047] The term "therapeutically effective amount" used in conjunction with the active ingredient in the specification means an amount effective in preventing or treating the target disease, and the therapeutically effective amount of the composition of the present invention may vary depending on various factors, such as the administration method, the target site, the condition of the patient, etc. Therefore, the dosage for use in humans should be determined as an appropriate amount by taking both safety and efficacy into consideration. It is also possible to estimate the amount used in humans from the effective amount determined through animal testing. Such considerations in determining the effective amount are described, for example, in Hardman and Limbird, eds., Goodman and Gilman's The Pharmacological Basis of Therapeutics, 10th ed.(2001), Pergamon Press; and E.W. Martin ed., Remington's Pharmaceutical Sciences, 18th ed.(1990), Mack Publishing Co.

[0048] The term "pharmaceutically effective amount" as used herein means an amount sufficient to treat a disease at a reasonable benefit / risk ratio applicable to medical treatment and not causing side effects. The effective dosage level may be determined based on factors including the patient's health condition, type of disease, cause of disease, severity, activity of the drug, sensitivity to the drug, method of administration, time of administration, route of administration and excretion rate, duration of treatment, drugs used in combination or simultaneously, and other factors well known in the medical field. The composition of the present invention may be administered as an individual therapeutic agent or in combination with other therapeutic agents, may be administered sequentially or simultaneously with conventional therapeutic agents, and may be administered singly or in multiple doses. Taking all of the above factors into consideration, it is important to administer an amount that can achieve the maximum effect with the minimum amount without side effects, and this can be easily determined by those skilled in the art.

[0049] Throughout the specification, when we say that an element is "on" another element, this includes not only cases where the element is in contact with the other element, but also cases where another element exists between the two elements.

[0050]

[0051] In one aspect, the present invention relates to a metal nanocomposite comprising a cancer cell targeting moiety, an immune effector cell targeting moiety, and a metal moiety exhibiting a photothermal effect.

[0052] In one embodiment, it may be used for photothermal therapy and photodynamic therapy by light irradiation.

[0053] In one embodiment, the light irradiation may be irradiating a near-infrared light source within an emission wavelength range of 700 to 1100 nm.

[0054] In one embodiment, the cancer cell targeting moiety can be a lectin that targets cancer cell glycans, such as a galectin, a siglec, a selectin, a C-type lectin, CD301, L-PHA (Phaseolus vulgaris leukocyte agglutinin), E-PHA (Phaseolus vulgaris erythroagglutinin), a tomato lectin (Lycopersicon esculentum lectin, LEA), a peanut lectin (Arachis hypogaea agglutinin, PNA), a potato lectin (Solanum tuberosum lectin), a pork mitogen (Phytolacca American lectin), a wheat germ agglutinin (Triticum Vulgaris lectin), an Artocarpus Artocarpus polyphemus lectin (Jacalin retin), Vicia villosa agglutinin (VVA), Helix pomatia agglutinin (HPA), Wisteria floribunda agglutinin (WFA), Sambucus nigra agglutinin (SNA), BC2LCM (lectin from the gram-negative bacterium Burkholderia cenocepacia), Maackia amurensis leukoglutinin (MAL), Psathyrella velutina (PVL), Sclerotium rolfsii lectin (SRL), Eucheuma serra agglutinin (ESA), CLEC17A (prolectin), Aleuria aurantia lectin, Sambucus sieboldiana lectin (SSA), Glechoma hederacea lectin (Gleheda), Morus nigra agglutinin (Morniga G),Salvia sclarea lectin, Salvia bogotensis lectin, Salvia horminum lectin, Clerodendrum trichotomum lectin, Moluccella laevis lectin, Griffonia simplicifolia (GsLA4), Psophocarpus tetragonolobus (acid WBAI), Abrus precatorius lectin, Molucella laevis lectin (MLL), Amaranthus caudatus lectin, Amaranthus leucocarpus It can be a lectin of Laelia autumnalis, a lectin of Artocarpus integrifolia, a lectin of Maclura pomifera, a lectin of Artocarpus lakoocha, agglutinin of Dolichos biflorusa, agglutinin of Dolichos biflorus, agglutinin of Glycine max or a lectin of Agaricus bisporus, with Sambucus nigra agglutinin being the most preferred.

[0055] In one embodiment, the immune effector cell may be a T cell, a natural killer (NK) cell, a natural killer T (NKT) cell, a macrophage, a monocyte, a dendritic cell, or a neutrophil, most preferably a macrophage.

[0056] In one embodiment, the immune effector cell targeting moiety may be a sugar, and may be mannose (Man).

[0057] In one embodiment, the metal moiety may be one or more of gold, silver, copper, and iron of the metal series, (1) one or more of SiO2-metal and SiN-metal of the metal-insulator series, (2) one or more of TiO2-metal, ZnO-metal, and SnO2-metal of the metal-semiconductor series, or a composite thereof, and gold is most preferred.

[0058] In one embodiment, the metal nanocomposite may be in the form of a rod, particle or shell.

[0059] In one embodiment, the metal nanocomposite may further include a labeling material, and the labeling material may be at least one selected from the group consisting of a chromogenic enzyme, a radioisotope, a chromophore, a chromogenic material, a luminescent material, a fluorescent material, a superparamagnetic particle, and an ultrasuper paramagnetic particle.

[0060] In one embodiment, the chromogenic enzyme can be a streptavidin-conjugated horseradish peroxidase (HRP), alkaline phosphatase (AP), β-galactosidase, luciferase, acetylcholinesterase, urease, catalase, asparginase, ribonuclease, malate dehydrogenase, staphylococcal nuclease, triose phosphate isomerase, glucose oxidase, cytochrome P450, and peroxidase compound.

[0061] In one embodiment, the fluorescent material may be a fluorescent protein, a photoprotein, luciferase, a fluorescent dye or a time-resolved fluorescence (TRF), and the fluorescent material may be Alexa Fluor 350, 405, 430, 488, 500, 514, 633, 647, 660, 680, 700, cy3, cy5, cy7, Rubpy (tris(2,2-bipyridyl)ruthenium(Ⅱ)), FITC (fluoresein isothiocyanate), rhodamine 6G, rhodamine B, TAMRA (5(6)-Carboxytetramethylrhodamine), Texas Red, It may be at least one selected from the group consisting of DAPI (4,6-diamidino-2-phenylindole) and coumarin.

[0062] In one aspect, the present invention relates to a drug delivery system comprising a metal nanocomposite and a pharmaceutically active ingredient.

[0063] In one embodiment, the pharmaceutically active ingredient may be a nucleic acid, an antibody, an effector, a drug, a prodrug, a toxin, a peptide, or a small molecule compound.

[0064] In one embodiment, the drug delivery system of the present invention can covalently bind a pharmaceutically active ingredient to a metal nanocomposite via a linker or non-covalently bind the pharmaceutically active ingredient without the linker.

[0065] In one embodiment, the drug can be a peptide drug, a protein drug, an anticancer agent, an antibacterial agent, an antiviral agent, an antidementia agent, an immunomodulator, a desensitizing agent, an antigen, a non-steroidal anti-inflammatory agent, an anti-inflammatory drug, an anesthetic, an antioxidant, an anti-infective agent, a chemotherapeutic agent, an anti-nociceptive agent, a DMOAD, an anabolic agent, an anti-catabolic agent, an autophagy modulator, an anti-osteoclast-mediated bone loss agent, a nutraceutical agent, an analgesic, a biologic, or a mixture thereof.

[0066] In one embodiment, the drug delivery system can deliver a drug specifically to cancer cells.

[0067] In one embodiment, the drug may be an immunogenic apoptosis inducer, a pro-apoptotic peptide, an inhibitor of microtubulin structure formation, a meiosis inhibitor, a topoisomerase inhibitor, a DNA intercalator, a toxin, or an anticancer agent.

[0068] In one embodiment, the pro-apoptotic peptide can be selected from the group consisting of KLA, alpha-defensin-1, BMAP-28, Brevenin-2R, Buforin IIb, cecropin A-Magainin 2 (CA-MA-2), Cecropin A, Cecropin B, chrysophsin-1, D-K6L9, Gomesin, Lactoferricin B, LLL27, LTX-315, Magainin 2, Magainin II-bombesin conjugate (MG2B), Pardaxin, and combinations thereof.

[0069] In one embodiment, the immunogenic apoptosis inducer may be selected from the group consisting of anthracycline-based anticancer drugs, taxane-based anticancer drugs, anti-EGFR antibodies, BK channel agonists, bortezomib, cardiac glycosides, cyclophosphamide-based anticancer drugs, GADD34 / PP1 inhibitors, LV-tSMAC, Measles virus, bleomycin, mitoxantrone, oxaliplatin, and combinations thereof.

[0070] In one embodiment, the anticancer agent is SN-38 (7-Ethyl-10-hydroxy-camptothecin), daunorubicin, doxorubicin, epirubicin, idarubicin, pixantrone, sabarubicin, valrubicin, paclitaxel, docetaxel, mechloethamine, chlorambucil, phenylalanine, mustard, cyclophosphamide, ifosfamide, carmustine (BCNU), lomustine (CCNU), Streptozotocin, busulfan, thiotepa, cisplatin, carboplatin, dactinomycin (actinomycin D), plicamycin, mitomycin C, vincristine, vinblastine, teniposide, topotecan, iridotecan, uramustine, melphalan, bendamustine, dacarbazine, temozolomide, altretamine, duocarmycin, nedaplatin, oxaliplatin, satraplatin, triplatin tetranitrate, 5-fluorouracil, 6-mercaptopurine,Capecitabine, cladribine, clofarabine, cystarbine, floxuridine, fludarabine, gemcitabine, hydroxyurea, methotrexate, pemetrexed, pentostatin, thioguanine, etoposide, mitoxantrone, izabepilone, vindesine, vinorelbine, estramustine, maytansine, DM1 (mertansine), DM4, dolastatin, auristatin E, It may be selected from the group consisting of auristatin F, monomethyl auristatin E (MMAE), monomethyl auristatin F, and derivatives thereof, and may be DM1 or MMAE.

[0071] In the present invention, the drug is preferably a cytotoxic anticancer agent. Cytotoxic anticancer agents can be broadly classified into antimetabolites, microtubulin targeting agents (tubulin polymerase inhibitors and tubulin depolymerization agents), alkylating agents, antimitotic agents, DNA cleavage agents, DNA cross-linker agents, DNA intercalator agents, and DNA topoisomerase inhibitors. As for antimetabolites, folic acid derivatives such as methotrexate, purine derivatives such as cladribine, pyrimidine derivatives such as azacitidine, doxifluridine, fluorouracil, etc. are known in the art, and as for microtubule targeting agents, monomethylauristatin Auristatin-based drugs such as E(MMAE), monomethylauristatin F(MMAF), and dolastatin, and maytansines are known in the art, and as alkylating agents, alkyl sulfonate preparations such as busulfan and treosulfan, nitrogen mustard derivatives such as bendamustine, and platinum preparations such as cisplatin and heptaplatin are known in the art.Also known in the art as antimitotic agents are taxanes such as docetaxel and paclitaxel, vinca alkalis such as vinflunine, and podophyllotoxin derivatives such as etoposide. Calicheamicins are known as DNA cleavage agents, and PBD duplexes are known as DNA cross-linker agents. Also known in the art are doxorubicin as DNA intercalator agents, and SN-28 as DNA topoisomerase inhibitors.

[0072] In addition, the drug may be a gene, plasmid DNA, an antisense oligonucleotide, siRNA, a peptide, a ribozyme, a viral particle, an immunomodulator, a protein, a contrast agent, etc. More specifically, the drug may be a gene encoding a tumor suppressor gene or a variant thereof, or a gene encoding apoptin, which induces apoptosis only in tumor cells, an antisense oligonucleotide (sequence: 5'-TCC ATG GTG CTC ACT-3') for HER-2, which is a cancer treatment target, or a diagnostic contrast agent such as Gd-DTPA, which is an MRI contrast agent.

[0073] In one embodiment, the drug delivery system of the present invention may further comprise a substance that targets cancer cells in the cancer cell targeting moiety, wherein the substance that targets cancer cells may comprise a small molecule, virus, antibody, antibody fragment, aptamer, hormone, cytokine, chemokine, ligand, peptide that is a portion of a cytokine, or peptide that is a portion of a ligand that specifically binds to cancer cells.

[0074] In one embodiment, the drug delivery system of the present invention can target a cancer cell-specific biomarker, wherein the cancer cell-specific biomarker is an antigen specific for 17-1A antigen, GD3 ganglioside R24, EGFRvⅢ, PSMA, PSCA, HLA-DR, EpCAM, MUC1 core protein, aberrantly glycosylated MUC1, fibronectin isoform containing ED-B domain, HER2 / neu, carcinoembryonic antigen (CEA), gastrin-releasing peptide (GRP) receptor antigen, mucin antigen, epidermal growth factor receptor (EGF-R), HER3, HER4, MAGE antigen, SART antigen, MUC1 antigen, c-erb-2 antigen, TAG 72, carbonic anhydrase IX, alpha-fetoprotein, A3, A33 antibody, Ba 733, BrE3-antigen, CA125, CDl, CD1a, CD3, CD5, CDl5, CD16, CD19, CD20, CD21, CD22, CD23, CD25, CD30, CD33, CD38, CD40, CD45, CD52, CD74, CD79a, CD80, CD138, colon-specific antigen-p (CSAp), CSAp, EGP-1, EGP-2, Ep-CAM, FIt-1, Flt-3, folate receptor, HLA-DR, human chorionic gonadotropin (HCG) and its subunits, hypoxia-inducible factor (HIF-I), Ia, IL-2, IL-6, IL-8, insulin growth factor-1 (IGF-1), KC4-antigen, KS-1-antigen, KSl-4, Le-Y, macrophage inhibition factor (MIF), MAGE, MUCl, MUC2, MUC3, MUC4, NCA66, NCA95, NCA90, antigen specific for PAM-4 antibody, placental growth factor, p53, prostatic acid phosphatase, PSA, RS5, SlOO, TAC, tenascin, TRAIL receptors, Tn antigen,It may be at least one selected from the group consisting of Thomson-Friedenreich antigens, tumor necrosis antigens, VEGF, ED-B fibronectin, angiogenesis marker, oncogene marker or oncogene product, and the oncogene may be an apoptosis-related gene, a transcription factor gene, a metastasis-related gene, angiogenesis-related gene or a tyrosine-kinase gene.

[0075] In one aspect, the present invention relates to a pharmaceutical composition for preventing or treating cancer, comprising the metal nanocomposite or drug delivery vehicle of the present invention as an active ingredient.

[0076] In one embodiment, the pharmaceutical composition for preventing or treating cancer may be a targeted anticancer agent.

[0077] In one embodiment, the cancer may be an immune-evading cancer, more preferably non-small cell lung cancer, triple-negative breast cancer (TNBC), or pancreatic cancer.

[0078] In one embodiment, the cancer may be a solid cancer or a blood cancer, and may include colon cancer, breast cancer, uterine cancer, cervical cancer, ovarian cancer, prostate cancer, brain tumor, head and neck carcinoma, melanoma, myeloma, leukemia, lymphoma, stomach cancer, lung cancer, pancreatic cancer, non-small cell lung cancer, liver cancer, esophageal cancer, small intestine cancer, anal cancer, fallopian tube carcinoma, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, bladder cancer, kidney cancer, ureteral cancer, renal cell carcinoma, renal pelvic carcinoma, bone cancer, skin cancer, head and neck cancer, cutaneous melanoma, intraocular melanoma, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, central nervous system (CNS) tumor, primary CNS lymphoma, spinal cord tumor, glioblastoma multiforme, and pituitary adenoma. It can be any one selected from the group consisting of cancers, more preferably breast cancer, cervical cancer or pancreatic cancer.

[0079] In one embodiment, the composition may further comprise an anticancer agent.

[0080] In one embodiment, the anticancer agent is selected from the group consisting of acibaicin, aclarubicin, acodazole, acronicin, adozelesin, alanosine, aldesleukin, allopurinol sodium, altretamine, aminoglutethimide, amonafide, ampligen, amsacrine, androgens, anguidine, aphidicolin glycinate, asare, asparaginase, 5-azacytidine, azathioprine, Bacillus Calmette-Guerin (BCG), Baker's Antipol, beta-2-deoxythioguanosine, bisanthrene HCl, bleomycin sulfate, bulseophan, buthionine sulfoximine, BWA 773U82, BW 502U83 / HCl, BW 7U85 mesylate, cerasemide, carbetimer, carboplatin, carmustine, chlorambucil, Chloroquinoxaline-sulfonamide, chlorozotocin, chromomycin A3, cisplatin, cladribine, corticosteroids, Corynebacterium parvum, CPT-11, crisnatol, cyclocytidine, cyclophosphamide, cytarabine, Cytembena, Davis maleate, decarbazine, dactinomycin, daunorubicin HCl, diazauridine, dexrazoxane, dianhydrogalactitol, diaziquone, dibromodulcitol, didemine B, diethyldithiocarbamate, diclycoaldehyde, dihydro-5-azacytine, doxorubicin, echinomycin, dedatrexate, edelfosine, efloxacin, Elliot's solution, elsamitrucine, epirubicin, esorubicin, estramustine Phosphate, Estrogen, Etanidazole, Ethiophos, Etoposide, Padrazole, Fazarabine, Fenretinide, Filgrastim, Finasteride, Flavone Acetate, Floxuridine, Fludarabine Phosphate, 5'-Fluorouracil, Fluosol™, Flutamide, Gallium Nitrate, Gemcitabine, Goserelin Acetate, Hefsulfam, Hexamethylene Bisacetamide, Homoharringtonine, Hydrazine Sulfate, 4-Hydroxyandrostenedione, Hydrodiurea, Idarubicin HCl, Ifosfamide, 4-Ipomeanol, Iproplatin, Isotretinoin, Leucovorin Calcium, Leuprolide Acetate, Levamisole, Liposomal Daunorubicin, Liposomal Entrapped Doxorubicin, Lomustine,Lonidamine, Maytansine, Mechlorethamine Hydrochloride, Melphalan, Menogaril, Merbaron, 6-Mercaptopurine, Mesna, Methanol Extract of Bacillus calete-guerins, Methotrexate, N-Methylformamide, Mifepristone, Mitoguazone, Mitomycin-C, Mitotane, Mitoxantrone Hydrochloride, Monocyte / Macrophage Colony-Stimulating Factor, Nabilone, Nafoxidine, Neocarzinostatin, Octreotide Acetate, Ormaplatin, Oxaliplatin, Paclitaxel, Palla, Pentostatin, Piperazinedione, Pipobroman, Pirarubicin, Piritrexim, Piroxantrone Hydrochloride, PIXY-321, Plicamycin, Porfimer Sodium, Prednimustine, Procarbazine, progestins, pyrazole furin, razoxane, sargramostim, semustine, spirogermanium, spiromustine, streptonigrin, streptozocin, sulfenor, suramin sodium, tamoxifen, taxolere, tegafur, teniposide, terephthalamidine, teroxirone, thioguanine, thiotepa, thymidine injection, tiazofurin, topotecan, toremifene, tretinoin, trifluoperazine hydrochloride, trifluridine, trimetrexate, tumor necrosis factor (TNF), uracil mustard, vinblastine sulfate, vincristine sulfate, vindesine, vinorelbine, vinzolidine, Yoshi 864, zorubicin, cytosine arabinoside, etoposide, melphalan, taxol and their It may include a mixture. Preferably, it may be cisplatin, paclitaxel, 5-FU (5-fluorouracil), methotrexate, doxorubicin, daunorubicin, cytosine arabinoside, etoposide, melphalan, chlorambucil, cyclophosphamide, vindesine, mitomycin, bleomycin, tamoxifen and taxol, and more preferably, it may be cisplatin, paclitaxel or 5-FU (5-fluorouracil).

[0081] The pharmaceutical composition of the present invention is administered in a pharmaceutically effective amount, and the pharmaceutical composition of the present invention may include a carrier, diluent, excipient, or a combination of two or more thereof commonly used in biological preparations.

[0082] In one embodiment, the pharmaceutical composition may be in one or more dosage forms selected from the group consisting of oral dosage forms, topical preparations, suppositories, sterile injectable solutions and sprays, with oral or injectable dosage forms being more preferred.

[0083] The pharmaceutical composition of the present invention can be administered parenterally (for example, intravenously, subcutaneously, intraperitoneally, or locally in the form of an injection) or orally, depending on the intended method. The dosage range varies depending on the patient's weight, age, sex, health condition, diet, administration time, administration method, excretion rate, and disease severity. Liquid preparations for oral administration of the composition of the present invention include suspensions, oral solutions, emulsions, syrups, etc., and in addition to commonly used simple diluents such as water and liquid paraffin, various excipients such as wetting agents, sweeteners, flavoring agents, and preservatives can be included. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, suppositories, etc. The pharmaceutical composition of the present invention can be administered by any device that allows the active ingredient to move to target cells. Preferred administration methods and preparations include intravenous injections, subcutaneous injections, intradermal injections, intramuscular injections, drip injections, etc. Injectables can be manufactured using aqueous solvents such as saline solution and Ringer's solution, non-aqueous solvents such as vegetable oils, higher fatty acid esters (e.g., ethyl oleate, etc.), alcohols (e.g., ethanol, benzyl alcohol, propylene glycol, glycerin, etc.), and pharmaceutical carriers such as stabilizers to prevent deterioration (e.g., ascorbic acid, sodium bisulfite, sodium pyrosulfite, BHA, tocopherol, EDTA, etc.), emulsifiers, buffers to adjust pH, and preservatives to inhibit microbial growth (e.g., phenylmercuric nitrate, thimerosal, benzalkonium chloride, phenol, cresol, benzyl alcohol, etc.).

[0084] The pharmaceutical composition of the present invention may further include a pharmaceutically acceptable additive. At this time, the pharmaceutically acceptable additive may include starch, gelatinized starch, microcrystalline cellulose, lactose, povidone, colloidal silicon dioxide, calcium hydrogen phosphate, lactose, mannitol, maltose, gum arabic, pregelatinized starch, corn starch, powdered cellulose, hydroxypropyl cellulose, Opadry, sodium starch glycolate, carnauba wax, synthetic aluminum silicate, stearic acid, magnesium stearate, aluminum stearate, calcium stearate, sucrose, dextrose, sorbitol, and talc. The pharmaceutically acceptable additive according to the present invention is preferably included in the composition in an amount of 0.1 to 90 parts by weight, but is not limited thereto.

[0085] In one aspect, the present invention relates to a cosmetic composition comprising the metal nanocomposite of the present invention.

[0086] In one embodiment, the cosmetic composition of the present invention may be for skin improvement, and the skin improvement may be at least one selected from the group consisting of skin wrinkle improvement, skin moisturizing improvement, skin whitening, skin aging prevention, skin immunity or defense enhancement, pore reduction, skin barrier improvement, pigmentation inhibition, sebum inhibition or acne improvement, atopy improvement, or skin soothing.

[0087] In one embodiment, the cosmetic composition may be for removing sebum.

[0088] The cosmetic composition of the present invention can be manufactured by including a cosmetically effective amount of the above-described active ingredients of the present invention and a cosmetically acceptable carrier.

[0089] The cosmetic composition of the present invention can be manufactured by including a cosmetically effective amount of the above-described active ingredients of the present invention and a cosmetically acceptable carrier.

[0090] The cosmetic composition may be in any form suitable for topical application, including solutions, gels, solids, pastes, anhydrous products, emulsions obtained by dispersing an oil phase in an aqueous phase, suspensions, microemulsions, microcapsules, microgranules, or vesicular dispersions of ionic (liposomes) and nonionic types, or in the form of creams, toners, lotions, powders, ointments, sprays, or concealer sticks. These compositions may be prepared according to methods conventional in the art. The composition according to the present invention may also be used in the form of a foam or an aerosol composition further containing a compressed propellant.

[0091] The cosmetic composition according to one embodiment of the present invention is not particularly limited in its formulation, and may be formulated as cosmetics such as, for example, a softening toner, an astringent toner, a nourishing toner, a nourishing cream, a massage cream, an essence, an eye cream, an eye essence, a cleansing cream, a cleansing foam, a cleansing water, a pack, a powder, a body lotion, a body cream, a body oil, and a body essence.

[0092] When the formulation of the cosmetic composition of the present invention is a paste, cream or gel, animal fiber, plant fiber, wax, paraffin, starch, tragacanth, cellulose derivative, polyethylene glycol, silicone, bentonite, silica, talc or zinc oxide may be used as a carrier component.

[0093] When the cosmetic composition of the present invention is in the form of a powder or spray, lactose, talc, silica, aluminum hydroxide, calcium silicate or polyamide powder may be used as a carrier component, and particularly in the case of a spray, a propellant such as chlorofluorohydrocarbon, propane / butane or dimethyl ether may be additionally included.

[0094] When the formulation of the cosmetic composition of the present invention is a solution or emulsion, a solvent, solvating agent or emulsifying agent is used as a carrier component, and examples thereof include water, ethanol, isopropanol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyl glycol oil, glycerol aliphatic ester, polyethylene glycol or fatty acid ester of sorbitan.

[0095] When the formulation of the cosmetic composition of the present invention is a suspension, a liquid diluent such as water, ethanol or propylene glycol, a suspending agent such as ethoxylated isostearyl alcohol, polyoxyethylene sorbitol ester and polyoxyethylene sorbitan ester, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar or tragacanth, etc. can be used as a carrier component.

[0096] When the formulation of the cosmetic composition of the present invention is a surfactant-containing cleansing, aliphatic alcohol sulfate, aliphatic alcohol ether sulfate, sulfosuccinic acid monoester, isethionate, imidazolinium derivative, methyl taurate, sarcosinate, fatty acid amide ether sulfate, alkylamidobetaine, fatty alcohol, fatty acid glyceride, fatty acid diethanolamide, vegetable oil, linolenic derivative, or ethoxylated glycerol fatty acid ester may be used as a carrier component.

[0097] The cosmetic composition of the present invention can be applied to cosmetics such as skin, lotion, cream, essence, pack, foundation, color cosmetics, sunscreen, two-way cake, face powder, compact, makeup base, skin cover, eye shadow, lipstick, lip gloss, lip fix, eyebrow pencil, toner, and cleansing agents such as shampoo and soap.

[0098] A cosmetic composition according to one embodiment of the present invention may further include, in addition to the above-described effective ingredient, a functional additive and an ingredient included in a general cosmetic composition. The functional additive may include an ingredient selected from the group consisting of water-soluble vitamins, oil-soluble vitamins, high molecular weight peptides, high molecular weight polysaccharides, sphingolipids, and seaweed extract.

[0099] The cosmetic composition of the present invention may also contain, as necessary, ingredients included in general cosmetic compositions in addition to the above-described effective ingredients. Examples of other ingredients to be contained include fat components, moisturizers, emollients, surfactants, organic and inorganic pigments, organic powders, ultraviolet absorbers, preservatives, sterilizers, antioxidants, plant extracts, pH adjusters, alcohols, pigments, fragrances, blood circulation promoters, cooling agents, antiperspirants, purified water, and the like.

[0100] In one aspect, the present invention relates to a method for preparing a metal nanocomposite, comprising the steps of: synthesizing a metal nanomaterial; conjugating a cancer cell targeting moiety; and conjugating an immune effector cell targeting moiety.

[0101] In one embodiment, a metal nanomaterial can be carboxylated and then conjugated with a cancer cell targeting moiety.

[0102] In one aspect, the present invention relates to the use of the metal nanocomposite of the present invention for photothermal therapy and photodynamic therapy.

[0103] In one aspect, the present invention relates to a method of treating cancer comprising administering to a subject suffering from cancer a metal nanocomposite or drug delivery vehicle of the present invention.

[0104] Below, specific embodiments of the present invention will be described.

[0105]

[0106] Example 1: Preparation of metal nanocomposites

[0107] A metal nanomaterial, Man-AuNR-SNA, containing Sambucus nigra agglutinin (SNA) lectin as a targeting factor for cancer cell glycans, mannose (Man) as a targeting factor for the surface of macrophages, and gold nanorods (AuNR) as a targeting factor for photothermal therapy was prepared. First, gold nanorods (AuNR) were synthesized using the 'seed-mediated growth method'. Specifically, CTAB (5 mL, 0.2 M) and HAuCl4 (2.5 mL, 1 mM) were mixed with stirring, and then NaBH4 (600 mL, 0.01 M) was added. A seed solution was formed at 30 °C and stirred until the color changed from yellow to dark brown. To prepare the growth solution, 9 g of CTAB, 1.23 g of NaOL, and 24 mL of AgNO34 M were mixed in 250 mL of triple-distilled water, HAuCl4 (250 mL, 1 mM) was added, and the mixture was kept without stirring for 15 min and then stirred for 90 min. 3 mL of 37 wt% HCl was injected and stirred for 15 min, and ascorbic acid (1.25 mL, 64 mM) was added and stirred until the solution became transparent. 800 mL of the seed solution prepared above was injected into the growth solution, stirred for 30 s, and maintained at 30 °C for 12 h. The finally synthesized AuNRs were concentrated by centrifugation to remove CTAB and stored in a refrigerator. To conjugate SNA lectin to the prepared AuNR, SH-PEG-COOH (heterobifunctional polyethylene glycol) was added to the prepared AuNR and stirred. DW containing 0.02% SDS was added and stirred overnight. To remove unbound SH-PEG-COOH, the AuNR was centrifuged twice and adjusted to pH 5.AuNRs were functionalized with carboxyl groups by redispersing in MES buffer at pH 5. To conjugate SNA lectin protein to carboxylated AuNRs, EDC / NHS cross-linker was added, stirred for 20 min, centrifuged to remove unbound cross-linker, and redispersed in MES buffer at pH 7. SNA was then injected, stirred overnight, and unbound SNA was removed by centrifugation and redispersed in DW (AuNR-SNA). SH-PEG-Mannose was added and stirred overnight to conjugate mannose to AuNR-SNA, preparing Man-AuNR-SNA conjugates (Fig. 1A).

[0108]

[0109] Example 2: Characterization of metal nanocomposites

[0110] The optical properties of the Man-AuNR-SNA nanocomposites prepared above were confirmed by analyzing the absorption spectrum from 300 to 999 nm using a microplate reader (BioTek / Agilent, USA). The particle size and surface charge were measured using a Zetasizer Pro (Malvern, UK). In addition, the particle concentration was measured using a NanoSight LM10 nanoparticle tracking analysis (Malvern, UK), and the particle number was calculated using this. The shape and morphology of AuNR and Man-AuNR-SNA were confirmed by TEM analysis using a JEM-F200 Transmission Electron Microscope (JEOL Ltd, Japan). In addition, for the qualitative analysis of the conjugates, AuNR, Man-AuNR-SNA, and SH-PEG-Mannose were freeze-dried, and FTIR (Fourier transform infrared spectrum) analysis was performed using an FTIR Spectrophotometer (ALPHA II, Germany). For the qualitative analysis,1 H Nuclear Magnetic Resonance ( 1 H NMR) 600 MHz (VNS, Varian) was used to measure the spectra. In addition, the concentration of SNA in the Man-AuNR-SNA conjugate was measured using a BCA protein assay kit, and the concentration of mannose was measured by the thiol group quantification method of Ellman's assay using HPLC 1260 Infinity II (Agilent, CA, USA) on a Poreshell 120 EC-C18 column (Agilent, CA, USA). In addition, to confirm whether Man-AuNR-SNA has the photothermal effect, which is an inherent characteristic of gold-based materials, Man-AuNR-SNA was exposed to an 850 nm NIR laser within the 700-1100 nm NIR wavelength range, which is optimal for tissue penetration through photothermal therapy, and its photothermal effect was analyzed. In addition, the photothermal stability of Man-AuNR-SNA was evaluated through three heating and cooling cycles under the laser exposure.

[0111] As a result, it was observed that the surface of AuNR changed due to the binding of SNA and mannose, and it was confirmed that the rod shape and nano size were maintained even after binding (Fig. 1B). As a result of surface charge analysis, the surface charge value decreased (-20.29 mV) due to the introduction of a carboxyl group, and the surface charge value of AuNR-SNA increased due to the amine group present in the lectin protein by the binding of SNA (-6.77 mV), and the surface charge value decreased again due to the binding of mannose (-15.99 mV), and finally, it was confirmed that the nonspecific binding to cells would be less due to the characteristic of the Man-AuNR-SNA particles having a negative surface charge (Fig. 1C). 1H NMR spectral analysis revealed that the characteristic peak of mannose was not observed in AuNR, whereas a peak was observed at 3.55 ppm in AuNR-Mannose (Fig. 1D). FTIR spectra of SH-PEG-Mannose, AuNR, and Man-AuNR-SNA also revealed that the characteristic peak of PEG was not observed in AuNR, whereas C=O (1709 cm -1 ) and -CH2(2942 cm -1 ) and SH-PEG-Mannose, all characteristic peaks were consistent (Fig. 1E). As a result of quantitative analysis of all substances contained in the manufactured Man-AuNR-SNA by NTA, the particle concentration was approximately 4x10 11 particles / mL, and the SNA concentration was measured by BCA analysis, and approximately 1.25 mg / mL was bound, and the mannose concentration was analyzed by Ellman's assay, and approximately 1 mg / mL was bound. In addition, in the case of DW and cell culture medium, the temperature slightly increased to approximately 32 °C upon exposure to 850 nm NIR laser, whereas Man-AuNR-SNA increased to approximately 55 °C (Figs. 1F to G), and it was confirmed that it had excellent photothermal stability (Fig. 1H). Therefore, it was confirmed that Man-AuNR-SNA is suitable as a material for inducing photothermal therapy by 850 nm NIR irradiation because it absorbs maximum light at approximately 840 nm.

[0112]

[0113] Example 3: Analysis of cancer cell-specific binding ability of metal nanocomposites

[0114] In general, cancer cells overexpress sialic acid on their surface, whereas normal cells do not (Fig. 2A). Therefore, the binding ability of Man-AuNR-SNA particles to normal H6c7 cells and three cancer cells, MIA PaCa-2, PANC-1, and AsPC-1, was analyzed using SNA, a sialic acid-binding lectin. Specifically, H6c7, MIA PaCa-2, PANC-1, and AsPC-1 cells were seeded in a 96-well plate at a density of 1 × 10 with medium, respectively. 4 After seeding at a density of 10 cells / well, the cells were cultured for 24 hours. The culture medium was removed, fixed with 2% paraformaldehyde, and Man-AuNR-SNA particles were added to each well at a concentration of 5 μg / mL based on the SNA concentration, and reacted at room temperature for 2 hours. Unbound particles were removed by washing three times with 1XPBS, and the fluorescence of particles bound to cells was measured using a microreader (BioTek / Agilent, USA) at λ Em 490 nm, λ ExIt was confirmed at 520 nm. In addition, a control AuNR-BSA particle treatment condition (BSA concentration standard 5 μg / mL) was also added to MIA PaCa-2 pancreatic cancer cells for comparative analysis. In addition, to confirm whether the immune evasion signal generated by the interaction between macrophages and cancer cells is blocked by SNA treatment, resulting in phagocytosis of cancer cells by macrophages, M0, M1, and M2 phenotype macrophages were prepared using a macrophage polarization method, stained with calcein red-AM, and cancer cells reacted with SNA were stained with calcein-AM and fluorescently labeled, and then macrophages and cancer cells were co-cultured and observed under a confocal microscope. As a result, SNA specifically bound only to cancer cells compared to normal cells (Fig. 2B). To confirm whether this binding is mediated by sialic acid, sialidase (i.e., neuraminidase) was treated, and the degree of SNA binding was analyzed by FACS. As a result, the population of normal cells bound to SNA under the non-sialidase treatment condition was 28.9%, whereas the three cancer cell types showed high binding capacity of 68.5%, 82.5%, and 98.3%, respectively. Unlike the normal cells, which showed no change in binding capacity when treated with sialidase, the binding capacity decreased to 29.4%, 45.4%, and 44.8%, respectively (Fig. 2C), confirming that SNA can specifically bind to pancreatic cancer cells and that this is mediated by sialic acid. To confirm whether such cancer cell-specific binding effect of SNA also occurs by Man-AuNR-SNA, the binding capacity to cells was evaluated using fluorescently labeled Man-AuNR-SNA particles. As a result, a higher fluorescence signal was detected in the three cancer cell types than in the normal cells, and the result showed a particle concentration-dependent result (Fig. 2D and E). In contrast, when particles conjugated with BSA rather than SNA were used as a control, a very low signal was detected in pancreatic cancer cells MIA PaCa-2.

[0115] Therefore, it can be seen that Man-AuNR-SNA particles can specifically bind to cancer cells via SNA.

[0116]

[0117] Example 4: Analysis of macrophage-specific binding capacity of metal nanocomposites

[0118] Since Siglec-10, which forms an immune checkpoint by interacting with sialic acid of cancer cells, is most highly expressed in the M2 macrophage population (M0 34.9%, M1 46.4%, and M2 58.6%), the expression of CD11b, a macrophage-wide marker, CD80, an M1-specific marker, and CD206, an M2-specific marker, was analyzed by FACS in M0, M1, and M2 phenotype macrophages. As a result, CD11b was found to be over 99% of the population in all phenotypes, CD80 was found to be 19.2% in M0, 89.2% in M1, and 23.5% in M2, and CD206 was found to be 27.9% in M0, 29.9% in M1, and 46.0% in M2 (Fig. 3A). Since CD206, which is an M2 marker and mannose receptor, is overexpressed only in the M2 phenotype, it was confirmed that mannose can target M2 phenotype macrophages. Accordingly, to confirm the specific binding ability of mannose to M2 macrophages, confocal microscopy analysis was performed using SH-PEG-Mannose and Maleimide-FITC. As a result, mannose was bound to the surface of M2 macrophages, and FITC was labeled by the binding between the terminal thiol (-SH) and maleimide, and fluorescence was detected (Fig. 3B). To determine whether the specific binding of mannose to M2 macrophages was also observed in Man-AuNR-SNA particles, fluorescently labeled Man-AuNR-SNA particles were treated with M0, M1, and M2 phenotype macrophages, respectively, and the fluorescence of the particles bound to the cells was measured using a microplate reader (BioTek / Agilent, USA) at λ Em 490 nm, λ ExIt was confirmed at 520 nm. In addition, the control group treated with AuNR-SNA particles without mannose conjugation to M2 phenotype macrophages was also analyzed. As a result, a significantly higher fluorescence signal was detected in M2 macrophages compared to M0 and M1 macrophages, and this was dependent on the concentration of the treated Man-AuNR-SNA particles (Fig. 3C). In contrast, in the control group treated with particles without mannose conjugation, a significantly lower signal was detected in M2 macrophages, confirming that Man-AuNR-SNA particles can specifically bind to M2 macrophages via mannose.

[0119]

[0120] Example 5: Cytotoxicity analysis of metal nanocomposites

[0121] Human skin fibroblasts HDF, pancreatic ductal epithelial cells H6c7, M0, M1 and M2 macrophages, and pancreatic cancer cells MIA PaCa-2, PANC-1 and AsPC-1 were treated with Man-AuNR-SNA at concentrations of 2-10 μg / mL for 24 h, and then cytotoxicity was evaluated using the CCK-8 (cell counting kit-8) assay.

[0122] As a result, Man-AuNR-SNA showed a cell viability of more than 80% at all concentrations (Fig. 4A), confirming that it was safe with almost no cytotoxicity in all cells, and that the particle alone had no cell killing ability.

[0123]

[0124] Example 6: Analysis of the cancer cell death-inducing effect of metal nanocomposites through photothermal therapy.

[0125] To confirm the cancer cell death induction effect of Man-AuNR-SNA by photothermal therapy, Man-AuNR-SNA particles were added at low concentrations (SNA: 5 μg / mL, NP: 4x10) as in Example 5. 9 particles / mL) and high concentration (SNA: 10 μg / mL, NP: 8x109 particles / mL) were treated for 2 h in each cancer cell line, and the cells were exposed to NIR laser (850 nm) for 0 to 10 min to perform photothermal therapy, followed by CCK-8 assay. In addition, 5 μg / mL of Man-AuNR-SNA particles (NP: 4x10 9 particles / mL) for 2 h, and the cells were exposed to NIR laser (850 nm) for 0–10 min. The expression of apoptosis-related markers was analyzed by RT-qPCR. Cells were stained using Annexin V-FITC Apoptosis kit (Invitrogen, V13242) and analyzed by flow cytometry using FACS Aria II (BD Biosciences, CA, USA) to analyze apoptosis and necrosis.

[0126] CCK-8 analysis results showed that in the case of normal H6c7 cells, both low- and high-concentration treatment conditions of Man-AuNR-SNA showed similar results to the untreated condition, and cell viability was over 80% even when irradiated with NIR for 10 minutes. On the other hand, in the three cancer cell types, cell viability decreased as the NIR irradiation time increased when treated with low concentrations of Man-AuNR-SNA, and decreased to less than about 60% after 10 minutes of treatment, and under the high-concentration condition, it reached less than 60% even after 2 minutes of NIR irradiation, and decreased to less than 40% when it reached 10 minutes (Fig. 4B and C). In addition, RT-qPCR analysis results showed that, compared to the untreated control group, the expression of survival-related protein Bcl-2 decreased under the photothermal therapy application condition, and the expression of apoptosis-related proteins p53, Bax, and Caspase-3 significantly increased (Fig. 5A). Therefore, when this strategy was applied, it was confirmed that apoptosis was effectively induced in cancer cells MIA PaCa-2. In addition, Annexin V / PI staining analysis results showed that while cell death was hardly induced when Man-AuNR-SNA treatment and NIR irradiation were used alone, cell death was significantly increased when combined (Fig. 5B and C).

[0127] Through this, it was confirmed that the combined use of Man-AuNR-SNA particle treatment and NIR irradiation had an excellent anticancer effect.

[0128]

[0129] Example 7: Analysis of the immune evasion inhibition effect of cancer cells by a combination of photothermal therapy and metal nanocomposites.

[0130] Pancreatic cancer cells were treated with SNA, and cancer cells labeled with different fluorescence and macrophages of each phenotype were co-cultured for 6 hours. The cancer cell phagocytosis was evaluated by observing the cancer cell phagocytosis according to the presence or absence of SNA treatment using a confocal microscope. In addition, in order to analyze the phagocytosis and the resulting anticancer effect by the combined treatment of Man-AuNR-SNA (5 μg / mL concentration based on SNA) and NIR irradiation (10 minutes), Man-AuNR-SNA particle-bound and fluorescently labeled cancer cells and fluorescently labeled macrophages were co-cultured as in Example 3, and the cell behavior was photographed at 1-minute intervals from 0 hour to 6 hours later using a STELLARIS 5 confocal microscope (LEICA, Germany) and an Incubator T Series (LCI, Korea).

[0131] SNA treatment alone detected high fluorescence in the co-localized region of cancer cells and macrophages, and 3D z-stack image analysis confirmed that the region was completely occupied by cancer cells within macrophages (Fig. 6A), confirming that SNA blocked the sialic acid of cancer cells and enhanced the phagocytic activity of macrophages. Imaging analysis showed that under the condition of Man-AuNR-SNA particle treatment alone, phagocytosis occurred at approximately 4 hours after imaging (Fig. 6B), confirming that it was phagocytosis induced by SNA. In addition, under the condition of combined treatment with Man-AuNR-SNA and NIR irradiation, phagocytosis occurred at approximately 1 hour after imaging, and relatively more cancer cells were killed (Fig. 6C). This suggests that the combined use of Man-AuNR-SNA treatment and photothermal therapy induces immune activation while blocking a specific immune checkpoint.

Claims

1. A metal nanocomposite comprising a cancer cell targeting moiety, an immune effector cell targeting moiety, and a metal moiety exhibiting a photothermal effect.

2. In paragraph 1, Cancer cell targeting moieties include galectins, siglecs, selectins, C-type lectins, CD301, L-PHA (Phaseolus vulgaris leukocyte agglutinin), E-PHA (Phaseolus vulgaris erythroagglutinin), tomato lectin (Lycopersicon esculentum lectin, LEA), peanut lectin (Arachis hypogaea agglutinin, PNA), potato lectin (Solanum tuberosum lectin), pork mitogen (Phytolacca American lectin), wheat germ agglutinin (Triticum Vulgaris lectin), and Artocarpus polyphemus. Lectins (Jacalin lectin), Vicia villosa agglutinin (VVA), Helix pomatia agglutinin (HPA), Wisteria floribunda agglutinin (WFA), Sambucus nigra agglutinin (SNA), BC2LCM (lectin from the gram-negative bacterium Burkholderia cenocepacia), Maackia amurensis leukoglutinin (MAL), Psathyrella velutina (PVL), Sclerotium rolfsii lectin (SRL), Eucheuma serra agglutinin (ESA), CLEC17A (prolectin), Aleuryia Aleuria aurantia lectin, Sambucus sieboldiana lectin (SSA), Glechoma hederacea lectin (Gleheda), Morus nigra agglutinin (Morniga G), Salvia sclarea lectin,Salvia bogotensis lectin, Salvia horminum lectin, Clerodendrum trichotomum lectin, Moluccella laevis lectin, Griffonia simplicifolia (GsLA4), Psophocarpus tetragonolobus (acid WBAI), Abrus precatorius lectin, Molucella laevis lectin (MLL), Amaranthus caudatus lectin, Amaranthus leucocarpus lectin, Laelia otumralis autumnalis lectin, Artocarpus integrifolia lectin, Maclura pomifera lectin, Artocarpus lakoocha lectin, Dolichos biflorusagglutinin, Dolichos biflorus lectin, glycine max lectin or Agaricus bisporus lectin, metal nanocomposites.

3. In paragraph 1, Immune effector cells include T cells, natural killer (NK) cells, natural killer T (NKT) cells, macrophages, monocytes, dendritic cells or neutrophils, metal nanocomposites.

4. In paragraph 1, A metal nanocomposite, wherein the metal moiety is gold, silver, copper, iron, SiO2-metal, SiN-metal, TiO2-metal, ZnO-metal, SnO2-metal or a complex thereof.

5. In paragraph 1, Metal nanocomposites for photothermal and photodynamic therapy by light irradiation.

6. A drug delivery system comprising the metal nanocomposite of paragraph 1 and a pharmaceutically active ingredient.

7. In paragraph 6, The pharmaceutically active ingredient is a drug carrier, which is a nucleic acid, antibody, effector, drug, prodrug, toxin, peptide or small molecule compound.

8. In paragraph 6, A drug delivery vehicle that delivers drugs specifically to cancer cells.

9. In paragraph 8, The drug is an immunogenic apoptosis inducer, a pro-apoptotic peptide, an inhibitor of microtubulin structure formation, a meiosis inhibitor, a topoisomerase inhibitor, a DNA intercalator, a toxin or an anticancer agent, a drug delivery vehicle.

10. A pharmaceutical composition for preventing or treating cancer, comprising the metal nanocomposite of paragraph 1 or the drug delivery system of paragraph 6 as an active ingredient.

11. In paragraph 10, A pharmaceutical composition for the prevention or treatment of cancer, which is an immune-evading cancer.

12. In paragraph 10, A pharmaceutical composition for the prevention or treatment of cancer, wherein the cancer is glioblastoma, brain tumor, melanoma, myeloma, non-small cell lung cancer, oral cancer, liver cancer, stomach cancer, colon cancer, breast cancer, lung cancer, bone cancer, pancreatic cancer, skin cancer, head or neck cancer, cervical cancer, ovarian cancer, colon cancer, small intestine cancer, rectal cancer, fallopian tube carcinoma, anal cancer, endometrial carcinoma, vaginal carcinoma, vulvar carcinoma, Hodgkin's disease, esophageal cancer, lymphoma, bladder cancer, gallbladder cancer, endocrine cancer, thyroid cancer, parathyroid cancer, adrenal cancer, soft tissue sarcoma, urethral cancer, penile cancer, prostate cancer, chronic or acute leukemia, lymphocytic lymphoma, kidney or ureter cancer, renal cell carcinoma, renal pelvic carcinoma, central nervous system tumor, primary central nervous system lymphoma, spinal cord tumor, brainstem glioma, or pituitary adenoma. 13.1) Step of synthesizing metal nanomaterials; 2) a step of conjugating a cancer cell targeting moiety; and 3) A method for producing a metal nanocomposite, comprising the step of conjugating an immune effector cell targeting moiety.

14. Use of metal nanocomposites for photothermal therapy and photodynamic therapy.

15. A method for treating cancer, comprising administering to a subject suffering from cancer the metal nanocomposite of paragraph 1 or the drug delivery system of paragraph 6.

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