Anticancer composition comprising metal-organic framework having immuno-anticancer agent bonded thereto, and use thereof

Abstract

The present invention relates to an anticancer composition comprising a metal-organic framework having an immuno-anticancer agent bonded thereto, and a use thereof. The metal-organic framework having an immuno-anticancer agent bonded thereto can continuously release the immuno-anticancer agent bonded thereto, can increase the efficiency of delivery to a site where cancer cells, not normal cells, are located, and can exhibit the effect of photodynamic therapy of killing cancer cells by producing singlet oxygen under light radiation. In addition, the present invention provides a method for treating cancer, comprising the steps of administering the anticancer composition to an individual and irradiating with light.

Description

Anticancer composition comprising a metal-organic framework combined with an immuno-oncology agent and use thereof

[0001] The present invention relates to an anticancer composition comprising a metal-organic framework combined with an immuno-oncology agent and a use thereof.

[0002] Cancer is one of the incurable diseases that humanity must overcome, and massive amounts of capital are being invested in the development of a cure for it worldwide. In Korea, it is the number one cause of death from disease, with over 100,000 people diagnosed with the disease and over 60,000 people dying from it annually.

[0003] Common cancer treatments include surgery, chemotherapy, and radiation therapy. Chemotherapy is primarily used for patients who are not amenable to surgery or radiation therapy (approximately 50% of all cancer cases) and those whose cancer has already metastasized. However, due to drug resistance, recurrence, metastasis, and aftereffects, the development of cancer treatment technologies that minimize side effects is crucial.

[0004] Third-generation cancer treatments, known as immunotherapy, work by activating the body's immune system, encouraging immune cells to attack cancer cells. Unlike first-generation treatments like chemotherapy and second-generation targeted therapies, these treatments are expected to reduce side effects and maximize therapeutic efficacy.

[0005] However, even in the case of third-generation immunotherapy, there is a side effect of immune disease symptoms in which immune cells attack not only cancer cells but also normal cells when the immune system is excessively activated due to the immunotherapy.

[0006] There is a need to develop a treatment that can significantly improve patient safety by solving the above problems while minimizing systemic exposure of the immunotherapy agent and exposure to normal cells.

[0007] [Prior Art Literature]

[0008] [Patent Document]

[0009] KR 10-2022-0151333 A1

[0010] The purpose of the present invention is to provide an anticancer composition comprising a metal-organic framework combined with an immuno-oncology agent and its use.

[0011] Another object of the present invention is to provide an anticancer composition comprising a metal-organic framework having an immuno-cancer agent bound thereto, which can continuously release the immuno-cancer agent bound to the metal-organic framework, increase the delivery efficiency to a site where cancer cells are located rather than normal cells, and exhibit a photodynamic therapeutic effect in which the metal-organic framework generates singlet oxygen upon light irradiation to kill cancer cells.

[0012] Another object of the present invention is to provide a method for treating cancer, comprising a step of irradiating light after administering the anticancer composition to a subject.

[0013] To achieve the above purpose, the present invention relates to an anticancer composition comprising a metal-organic framework (MOF) to which an immuno-oncology agent is bound.

[0014] Additionally, the metal-organic framework may include a metal cluster and a ligand compound represented by the following chemical formula 1 that coordinately bonds to the metal cluster:

[0015] [Chemical Formula 1]

[0016]

[0017] Here,

[0018] X1 and X3 are N(R9),

[0019] X2 and X4 are N,

[0020] R1 to R9 are the same or different from each other, and each independently represent hydrogen, deuterium, a cyano group, a nitro group, a halogen group, a hydroxy group, a substituted or unsubstituted alkylthio group having 1 to 4 carbon atoms, a substituted or unsubstituted alkyl group having 1 to 30 carbon atoms, a substituted or unsubstituted cycloalkyl group having 3 to 20 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 24 carbon atoms, a substituted or unsubstituted aralkyl group having 7 to 30 carbon atoms, a substituted or unsubstituted aryl group having 6 to 30 carbon atoms, a substituted or unsubstituted heteroaryl group having 1 to 60 carbon atoms, a substituted or unsubstituted heteroarylalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 60 carbon atoms, a substituted or unsubstituted arylalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl ... It is selected from the group consisting of a 30-carbon alkylamino group, a substituted or unsubstituted arylamino group having 6 to 30 carbon atoms, a substituted or unsubstituted aralkylamino group having 7 to 30 carbon atoms, a substituted or unsubstituted heteroarylamino group having 1 to 24 carbon atoms, a substituted or unsubstituted alkylsilyl group having 1 to 30 carbon atoms, a substituted or unsubstituted arylsilyl group having 6 to 30 carbon atoms, and a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms.

[0021] In addition, the metal-organic framework is a metal selected from the group consisting of Li, Na, Mg, Ca, Sr, Ba, Sc, Y, Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, W, Mn, Re, Fe, Ru, Os, Co, Rh, Ir, Ni, Pd, Pt, Cu, Ag, Au, Zn, Cd, Hg, Al, Ga, In, Tl, Si, Ge, Sn, Pb, As, Sb and Bi or Li + , Na + , Mg 2+ , Ca 2+ , Sr 2+ , Ba 2+ , Sc 3+ , Y 3+ , Ti4+ , Zr 4+ , Hf 4+ , V 4+ , V 3+ , V 2+ , Nb 3+ , Pitch 3+ , Cr 3+ , Mo 3+ , W 3+ , Mn 3+ , Mn 2+ , Re 3+ , Re 2+ , Fe 3+ , Fe 2+ , Ru 3+ , Ru 2+ , Home 3+ , Home 2+ , Co 3+ , Co 2+ , Rh 2+ , Rh + , Ir 3+ , Ir 2+ , Ir + , Knee 2+ , Knee + , Pd 2+ , Pd + , Pt 2+ , Pt + , Cu 2+ , Cu + , Ag + , Au + , Zn 2+ , Cd 2+ , Hg 2+ , Al 3+ , Ga 3+ , His 3+ , Tl 3+ , See 4+ , See 2+ , Ge 4+ , Ge 2+ , Sn 4+ , Sn 2+ , Pb 4+ , Pb 2+ , As 5+ , As 3+ , As + , Sb 5+ , Sb 3+ , Sb + , Be5+ , Bi 3+ and Bi + It may include a metal ion selected from the group consisting of:

[0022] Additionally, the metal-organic framework may be selected from the group consisting of aluminum-based metal-organic frameworks, iron-based metal-organic frameworks, zirconium-based metal-organic frameworks, and mixtures thereof.

[0023] Additionally, the above-mentioned immuno-oncology agent can coordinately bind to the unsaturated metal site of the metal cluster.

[0024] Additionally, the above immunotherapy agent may be an immune-modulating agent.

[0025] Additionally, the immunomodulator may be an agonist of TLR7 (Toll-like receptor 7) or TLR8.

[0026] Additionally, the cancer may be colon cancer, liver cancer, lung cancer, breast cancer, melanoma, stomach cancer, colon cancer, skin cancer, ovarian cancer, cervical cancer, thyroid cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, esophageal cancer, or fibrosarcoma.

[0027] Additionally, the metal-organic complex can generate singlet oxygen upon light irradiation.

[0028] A method for treating cancer according to another embodiment of the present invention may include administering the anticancer composition to a subject in an amount effective for treating cancer and irradiating the subject with light.

[0029] In the present invention, the term "metal-organic framework (MOF)" refers to a porous material in which metal clusters and organic linkers (or organic bridging ligands) are connected by coordination bonds to form a three-dimensional structure, and various MOFs can be created depending on the selection of metal ions and organic ligands. The MOF is characterized by porosity in which empty spaces exist within the structure, and the pore size, porosity, three-dimensional structure, surface area, etc. can be designed in various ways depending on the type and bonding method of the metal ions and organic ligands that make up the MOF. Due to this porosity, the MOF not only has a very large surface area but also has an open pore structure, so it can transport a large amount of molecules or solvents compared to other porous materials known in the art, and when used as a catalyst or gas storage, it has the advantage of having many active sites, which can maximize efficiency. In addition, the MOF is not easily deformed at high temperatures and has a rigid skeleton, so it has excellent chemical and thermal stability.

[0030] In the present invention, “subject” may include mammals, birds, reptiles, farmed fish, etc., including dogs, cats, rats, livestock, humans, etc., without limitation, and the subject may exclude humans.

[0031] In the present invention, “hydrogen” is hydrogen, light hydrogen, deuterium or tritium, unless specifically limited.

[0032] In the present invention, “halogen group” is fluorine, chlorine, bromine or iodine.

[0033] In the present invention, “alkyl” refers to a monovalent substituent derived from a straight or branched saturated hydrocarbon having 1 to 40 carbon atoms. Examples thereof include, but are not limited to, methyl, ethyl, propyl, isobutyl, sec-butyl, pentyl, iso-amyl, and hexyl.

[0034] In the present invention, “alkenyl” refers to a monovalent substituent derived from a straight or branched unsaturated hydrocarbon having 2 to 40 carbon atoms and at least one carbon-carbon double bond. Examples thereof include, but are not limited to, vinyl, allyl, isopropenyl, and 2-butenyl.

[0035] In the present invention, “alkynyl” refers to a monovalent substituent derived from an unsaturated hydrocarbon having 2 to 40 carbon atoms and a straight or branched chain having at least one carbon-carbon triple bond. Examples thereof include, but are not limited to, ethynyl and 2-propynyl.

[0036] In the present invention, “alkylthio” means the above-described alkyl group bonded via a sulfur linkage (-S-).

[0037] In the present invention, “aryl” refers to a monovalent substituent derived from an aromatic hydrocarbon having 6 to 60 carbon atoms, which is a single ring or a combination of two or more rings. In addition, it may also include a form in which two or more rings are simply attached to each other (pendant) or condensed, and specifically, it may be a naphthyl group, anthracenyl group, phenanthryl group, triphenyl group, pyrenyl group, phenalenyl group, perylenyl group, chrysenyl group, fluorenyl group, etc., but is not limited thereto. The fluorenyl group may be substituted, and adjacent groups may be bonded to each other to form a ring.

[0038] In the present invention, “heteroaryl” refers to a monovalent substituent derived from a monoheterocyclic or polyheterocyclic aromatic hydrocarbon having 6 to 30 carbon atoms. At this time, at least one carbon atom in the ring, preferably 1 to 3 carbon atom(s), is substituted with a heteroatom such as N, O, S, or Se. In addition, a form in which two or more rings are simply attached to each other (pendant) or condensed may be included, and a form condensed with an aryl group may also be included. Examples of such heteroaryls include, but are not limited to, 6-membered monocyclic rings such as pyridyl, pyrazinyl, pyrimidinyl, pyridazinyl, and triazinyl; polycyclic rings such as phenoxathienyl, indolizinyl, indolyl, purinyl, quinolyl, benzothiazole, and carbazolyl; and 2-furanyl, N-imidazolyl, 2-isoxazolyl, 2-pyridinyl, and 2-pyrimidinyl.

[0039] In the present invention, “aryloxy” is a monovalent substituent represented by RO-, wherein R represents aryl having 6 to 60 carbon atoms. Examples of such aryloxy include, but are not limited to, phenyloxy, naphthyloxy, and diphenyloxy.

[0040] In the present invention, “alkyloxy” is a monovalent substituent represented by R'O-, wherein R' means alkyl having 1 to 40 carbon atoms, and may include a linear, branched, or cyclic structure. Examples of alkyloxy include, but are not limited to, methoxy, ethoxy, n-propoxy, 1-propoxy, t-butoxy, n-butoxy, and pentoxy.

[0041] In the present invention, “alkoxy” may be straight chain, branched chain, or cyclic chain. The carbon number of the alkoxy is not particularly limited, but is preferably 1 to 20 carbon atoms. Specifically, it may be methoxy, ethoxy, n-propoxy, isopropoxy, i-propyloxy, n-butoxy, isobutoxy, tert-butoxy, sec-butoxy, n-pentyloxy, neopentyloxy, isopentyloxy, n-hexyloxy, 3,3-dimethylbutyloxy, 2-ethylbutyloxy, n-octyloxy, n-nonyloxy, n-decyloxy, benzyloxy, p-methylbenzyloxy, etc., but is not limited thereto.

[0042] As used herein, "aralkyl" refers to an aryl-alkyl group, where aryl and alkyl are as defined above. Preferred aralkyl groups include lower alkyl groups. Non-limiting examples of suitable aralkyl groups include benzyl, 2-phenethyl, and naphthalenylmethyl. Bonding to the parent moiety is via the alkyl group.

[0043] In the present invention, “arylamino group” means an amine substituted with an aryl group having 6 to 30 carbon atoms.

[0044] In the present invention, “alkylamino group” means an amine substituted with an alkyl group having 1 to 30 carbon atoms.

[0045] In the present invention, “aralkylamino group” means an amine substituted with an aryl-alkyl group having 6 to 30 carbon atoms.

[0046] In the present invention, “heteroarylamino group” means an amine group substituted with an aryl group and a heterocyclic group having 6 to 30 carbon atoms.

[0047] In the present invention, “heteroaralkyl group” means an aryl-alkyl group substituted with a heterocyclic group.

[0048] In the present invention, “cycloalkyl” refers to a monovalent substituent derived from a monocyclic or polycyclic non-aromatic hydrocarbon having 3 to 40 carbon atoms. Examples of such cycloalkyl include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, norbornyl, and adamantine.

[0049] In the present invention, “heterocycloalkyl” means a monovalent substituent derived from a non-aromatic hydrocarbon having 3 to 40 carbon atoms, wherein at least one carbon atom in the ring, preferably 1 to 3 carbon atom(s), is substituted with a heteroatom such as N, O, S or Se. Examples of such heterocycloalkyl include, but are not limited to, morpholine and piperazine.

[0050] In the present invention, “alkylsilyl” means silyl substituted with alkyl having 1 to 40 carbon atoms, and “arylsilyl” means silyl substituted with aryl having 6 to 60 carbon atoms.

[0051] In the present invention, “fused ring” means a fused aliphatic ring, a fused aromatic ring, a fused heteroaliphatic ring, a fused heteroaromatic ring, or a combination thereof.

[0052] In the present invention, “forming a ring by bonding with adjacent groups” means forming a substituted or unsubstituted aliphatic hydrocarbon ring; a substituted or unsubstituted aromatic hydrocarbon ring; a substituted or unsubstituted aliphatic heterocycle; a substituted or unsubstituted aromatic heterocycle; or a condensed ring thereof by bonding with adjacent groups.

[0053] Examples of the “aromatic hydrocarbon ring” in the present invention include, but are not limited to, a phenyl group, a naphthyl group, an anthracenyl group, etc.

[0054] In the present invention, “aliphatic heterocycle” means an aliphatic ring containing at least one heteroatom.

[0055] In the present invention, “aromatic heterocycle” means an aromatic ring containing at least one heteroatom.

[0056] In the present invention, "substitution" means that a hydrogen atom bonded to a carbon atom of a compound is replaced with another substituent, and the position of substitution is not limited as long as it is a position where a hydrogen atom is replaced, i.e., a position where a substituent can be substituted, and when two or more are substituted, the two or more substituents may be the same or different from each other. The above substituent is hydrogen, a cyano group, a nitro group, a halogen group, a hydroxy group, an alkyl group having 1 to 30 carbon atoms, an alkenyl group having 2 to 30 carbon atoms, an alkynyl group having 2 to 24 carbon atoms, a heteroalkyl group having 2 to 30 carbon atoms, an aralkyl group having 6 to 30 carbon atoms, an aryl group having 5 to 30 carbon atoms, a heteroaryl group having 2 to 30 carbon atoms, a heteroarylalkyl group having 3 to 30 carbon atoms, an alkoxy group having 1 to 30 carbon atoms, an alkylamino group having 1 to 30 carbon atoms, an arylamino group having 6 to 30 carbon atoms, an aralkylamino group having 6 to 30 carbon atoms, a heteroarylamino group having 2 to 24 carbon atoms, a substituted or unsubstituted alkylsilyl group having 1 to 30 carbon atoms, a substituted or unsubstituted alkyl group having 6 to It may be substituted with one or more substituents selected from the group consisting of 30 arylsilyl groups and substituted or unsubstituted aryloxy groups having 6 to 30 carbon atoms, but is not limited to the above examples.

[0057] The present invention relates to a metal-organic framework having an immuno-cancer agent bound thereto, which can continuously release the immuno-cancer agent bound to the metal-organic framework, and can increase the delivery efficiency to a site where cancer cells are located rather than normal cells, and the metal-organic framework can exhibit a photodynamic therapeutic effect capable of killing cancer cells by generating singlet oxygen upon light irradiation.

[0058] In addition, the present invention relates to a method for treating cancer, comprising a step of irradiating light after administering the anticancer composition to a subject.

[0059] FIG. 1 is a front view of a metal-organic framework according to one embodiment of the present invention.

[0060] FIG. 2 is a side view of a metal-organic framework according to one embodiment of the present invention.

[0061] Figure 3 shows the results of scanning electron microscopy measurements on a metal-organic framework according to one embodiment of the present invention.

[0062] FIG. 4 is an X-ray diffraction analysis result for a metal-organic framework according to one embodiment of the present invention.

[0063] Figure 5 shows the N2 adsorption / desorption isotherm, specific surface area, and pore measurement results for a metal-organic framework according to one embodiment of the present invention.

[0064] Figure 6 shows the results of zeta potential measurement for a metal-organic framework according to one embodiment of the present invention.

[0065] Figure 7 shows the FT-IR measurement results for a metal-organic framework according to one embodiment of the present invention.

[0066] FIG. 8 is a front view of a metal-organic framework according to one embodiment of the present invention.

[0067] FIG. 9 is a side view of a metal-organic framework according to one embodiment of the present invention.

[0068] Figure 10 shows the results of scanning electron microscopy measurements on a metal-organic framework according to one embodiment of the present invention.

[0069] Figure 11 shows the results of X-ray diffraction analysis for a metal-organic framework according to one embodiment of the present invention.

[0070] Figure 12 shows the N2 adsorption / desorption isotherm, specific surface area, and pore measurement results for a metal-organic framework according to one embodiment of the present invention.

[0071] Figure 13 shows the results of zeta potential measurement for a metal-organic framework according to one embodiment of the present invention.

[0072] Figure 14 shows the FT-IR measurement results for a metal-organic framework according to one embodiment of the present invention.

[0073] FIG. 15 is a front view of a metal-organic framework according to one embodiment of the present invention.

[0074] FIG. 16 is a side view of a metal-organic framework according to one embodiment of the present invention.

[0075] Figure 17 shows the results of scanning electron microscopy measurements on a metal-organic framework according to one embodiment of the present invention.

[0076] Figure 18 is an X-ray diffraction analysis result for a metal-organic framework according to one embodiment of the present invention.

[0077] Figure 19 shows the results of N2 adsorption / desorption isotherm, specific surface area, and pore measurement for a metal-organic framework according to one embodiment of the present invention.

[0078] Figure 20 shows the results of zeta potential measurement for a metal-organic framework according to one embodiment of the present invention.

[0079] Figure 21 shows the FT-IR measurement results for a metal-organic framework according to one embodiment of the present invention.

[0080] Figure 22 shows the results of a cytotoxicity test for a metal-organic framework according to one embodiment of the present invention.

[0081] Figure 23 shows the results of a cytotoxicity test for a metal-organic framework according to one embodiment of the present invention.

[0082] Figure 24 shows the results of a cytotoxicity test for a metal-organic framework according to one embodiment of the present invention.

[0083] Figure 25 shows the results of a 5-minute PDT test according to the power of a light source when a metal-organic framework according to one embodiment of the present invention is irradiated with light.

[0084] Figure 26 shows the results of a cumulative 3-minute PDT test following light irradiation on a metal-organic framework according to one embodiment of the present invention.

[0085] Figure 27 shows the results of a cumulative 10-minute PDT test following light irradiation on a metal-organic framework according to one embodiment of the present invention.

[0086] Figure 28 shows the results of a toxicity test on colon cancer cells for a metal-organic framework according to one embodiment of the present invention.

[0087] Figure 29 shows the results of scanning electron microscopy measurements of a metal-organic framework and an immuno-cancer agent combined therewith according to one embodiment of the present invention.

[0088] Figure 30 shows the results of a cumulative 10-minute PDT test following light irradiation on a metal-organic framework combined with an immuno-oncology agent according to one embodiment of the present invention.

[0089] Figure 31 shows the results of a toxicity test on colon cancer cells for a metal-organic framework combined with an immuno-oncology agent according to one embodiment of the invention.

[0090] Figure 32 shows the results of a cytotoxicity test for a metal-organic framework combined with an immuno-oncology agent according to one embodiment of the present invention.

[0091] Figure 33 shows the results of a cytotoxicity test for a metal-organic framework combined with an immuno-oncology agent according to one embodiment of the present invention.

[0092] Figure 34 shows the results of a cytotoxicity test for a metal-organic framework combined with an immuno-oncology agent according to one embodiment of the present invention.

[0093] The present invention relates to an anticancer composition comprising a metal-organic framework (MOF) to which an immuno-oncology agent is bound.

[0094] Hereinafter, embodiments of the present invention will be described in detail so that those skilled in the art can easily implement them. However, the present invention may be implemented in various different forms and is not limited to the embodiments described herein.

[0095] Recently, light therapy has been attracting attention as a new cancer treatment technology that is noninvasive and has minimal side effects. This treatment, known as phototherapy, can be divided into photothermal therapy and photodynamic therapy.

[0096] The above photothermal therapy is a technology that treats cancer by generating heat when light of a specific wavelength is irradiated on a light-absorbing material, and the above photodynamic therapy is a technology that kills cancer through a secondary chemical reaction when light is absorbed.

[0097] More specifically, photodynamic therapy involves a photosensitizer that is activated by light of a specific wavelength and binds to oxygen in the tissue, producing singlet oxygen (singlet oxygen). 1 O2) is generated, and their strong chemical reaction is carried out by utilizing the principle of necrosis of surrounding cell molecules. Reactive oxygen species such as singlet oxygen are essential for various physiological functions such as muscle contraction, fat metabolism, stem cell differentiation, and cell maintenance when present in small amounts, but when overexpressed, they can cause cell death, carcinogenesis, and tissue aging.

[0098] In particular, photodynamic therapy has the great advantage of being non-invasive and allowing for repeated treatments, making it easy to increase its effectiveness, and being able to be used in conjunction with other treatments such as chemotherapy, making it possible to use it as an adjuvant treatment.

[0099] Accordingly, the present invention relates to an anticancer composition, characterized in that it includes a metal-organic framework capable of photodynamic therapy and includes an immuno-cancer agent.

[0100] That is, as described above, photodynamic therapy can be used as an adjuvant therapy that can be used in conjunction with other anticancer treatments, and thus, the present invention relates to a composition that can perform photodynamic therapy using a metal-organic framework as well as anticancer treatment by an immunotherapy agent.

[0101] The anticancer composition of the present invention may include a metal-organic framework (MOF) to which an immuno-cancer agent is bound. Specifically, the metal-organic framework is characterized in that an immuno-cancer agent is bound to the MOF.

[0102] As described above, immunotherapy drugs work by activating the body's immune system to cause immune cells to attack cancer cells, which is expected to reduce the side effects of anticancer drugs and maximize the therapeutic effect. However, if the immune system is excessively activated due to immunotherapy drugs, there is a side effect of immune disease symptoms in which immune cells attack not only cancer cells but also normal cells.

[0103] In order to solve the above problems, in the present invention, by binding an immuno-cancer agent to a metal-organic framework, the immuno-cancer agent can be continuously released by a single administration, and an excessive amount of the immuno-cancer agent is not exposed to the entire body, and can be released at the site where cancer cells are located by the metal-organic framework, thereby solving the problems of conventional immuno-cancer agents.

[0104] The above-mentioned immuno-oncology agent is an immune-modulating agent, and the immune-modulating agent may be an agonist of TLR7 (Toll-like receptor 7) or TLR8.

[0105] The above-mentioned Toll-like receptors (TLRs) are mainly expressed in immune cells and play a crucial role in the innate immune response of mammals. TLRs detect pathogen-associated molecular patterns (PAMPs) and stimulate immune cells through the MyD88-dependent interleukin 1 receptor (IL-1R)-TLR signaling pathway, resulting in the activation of the transcription factor NF-κB2. Ten functional TLR families (TLR1 to TLR10) have been identified in humans (Akira S. et al., Nature Immunol., (2001) 2:675-680). TLR targets currently undergoing clinical trials for application in anticancer immunotherapy include TLR3, 4, 7, 8, and 9 (SE Lee and JH Rhee, J. Bacteriol. Virol. (2012), 42(3):255-262).

[0106] The above immunotherapy agent may be an imidazoquinoline compound, more specifically, Resiquimod (R-848), but is not limited to the above compound.

[0107] Resiquimod (R-848) is an immune response modulator with antiviral and antitumor effects. It is used as a topical gel for the treatment of skin lesions caused by herpes simplex virus and cutaneous T-cell lymphoma, and can be used as an adjuvant to enhance the efficacy of vaccines. In animal disease models, systemic administration of nanoparticles loaded with resiquimod has been shown to enhance the response rate to checkpoint inhibitor cancer immunotherapy through stimulation of tumor-associated macrophages. This may be related to multiple mechanisms of action, including agonism of Toll-like receptor 7 (TLR7) and TLR8, as well as upregulation of opioid growth factor receptors.

[0108] The above immuno-oncology agent can coordinately bind to the unsaturated metal site of the metal cluster.

[0109] The metal-organic framework of the present invention may include a metal cluster and a ligand compound represented by the following chemical formula 1 that coordinately bonds to the metal cluster:

[0110] [Chemical Formula 1]

[0111]

[0112] Here,

[0113] X1 and X3 are N(R9),

[0114] X2 and X4 are N,

[0115] R1 to R9 are the same or different from each other, and each independently represent hydrogen, deuterium, a cyano group, a nitro group, a halogen group, a hydroxy group, a substituted or unsubstituted alkylthio group having 1 to 4 carbon atoms, a substituted or unsubstituted alkyl group having 1 to 30 carbon atoms, a substituted or unsubstituted cycloalkyl group having 3 to 20 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 24 carbon atoms, a substituted or unsubstituted aralkyl group having 7 to 30 carbon atoms, a substituted or unsubstituted aryl group having 6 to 30 carbon atoms, a substituted or unsubstituted heteroaryl group having 1 to 60 carbon atoms, a substituted or unsubstituted heteroarylalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 60 carbon atoms, a substituted or unsubstituted arylalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl ... It is selected from the group consisting of a 30-carbon alkylamino group, a substituted or unsubstituted arylamino group having 6 to 30 carbon atoms, a substituted or unsubstituted aralkylamino group having 7 to 30 carbon atoms, a substituted or unsubstituted heteroarylamino group having 1 to 24 carbon atoms, a substituted or unsubstituted alkylsilyl group having 1 to 30 carbon atoms, a substituted or unsubstituted arylsilyl group having 6 to 30 carbon atoms, and a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms.

[0116] The metal cluster included in the above metal-organic framework contains an unsaturated metal site capable of forming a coordination bond. The above-described immuno-cancer agent can be bound to the unsaturated metal site by a coordination bond. The metal-organic framework to which the immuno-cancer agent is bound can be actively decomposed by irradiating light at a site where the release of the immuno-cancer agent is required, thereby controlling the release of the loaded immuno-cancer agent. This enables release at a target (specific region) and, at the same time, can be controlled to enable sustained release by controlling the amount of light irradiation.

[0117] The metal-organic framework is a metal selected from the group consisting of Li, Na, Mg, Ca, Sr, Ba, Sc, Y, Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, W, Mn, Re, Fe, Ru, Os, Co, Rh, Ir, Ni, Pd, Pt, Cu, Ag, Au, Zn, Cd, Hg, Al, Ga, In, Tl, Si, Ge, Sn, Pb, As, Sb and Bi, or Li. + , Na + , Mg 2+ , Ca 2+ , Sr 2+ , Ba 2+ , Sc 3+ , Y 3+ , Ti 4+ , Zr 4+ , Hf 4+ , V 4+ , V 3+ , V 2+ , Nb 3+ , Ta 3+ , Cr 3+ , Mo 3+ , W 3+ , Mn 3+ , Mn 2+ , Re 3+ , Re 2+ , Fe3+ , Fe 2+ , Ru 3+ , Ru 2+ , Os 3+ , Os 2+ , Co 3+ , Co 2+ , Rh 2+ , Rh + , Ir 3+ , Ir 2+ , Ir + , Ni 2+ , Ni + , Pd 2+ , Pd + , Pt 2+ , Pt + , Cu 2+ , Cu + , Ag + , Au + , Zn 2+ , Cd 2+ , Hg 2+ , Al 3+ , Ga 3+ , In 3+ , Tl 3+ , Si 4+ , Si 2+ , Ge 4+ , Ge 2+ , Sn 4+ , Sn 2+ , Pb 4+ , Pb 2+ , As 5+ , As 3+ , As + , Sb 5+ , Sb 3+ , Sb + , Bi 5+ , Bi 3+ and Bi + It may include a metal ion selected from the group consisting of:

[0118] The above metal-organic framework can be selected from the group consisting of aluminum-based metal-organic frameworks, iron-based metal-organic frameworks, zirconium-based metal-organic frameworks, and mixtures thereof.

[0119] However, preferably, it may be a metal-organic framework formed by the bonding of a ligand represented by the above-described chemical formula 1 and a metal ion, and more specifically, it may be PCN-221, PCN-222, PCN-223, PCN-224, MOF-545 or MOF-525, but is not limited to the above examples.

[0120] The above ligand may be a compound in which all of R1 to R9 are hydrogen. Specifically, it may be a compound represented by the following chemical formula 2:

[0121] [Chemical Formula 2]

[0122]

[0123] -COO of the ligand compound represented by the above chemical formula 2 - The metal can form coordination bonds with metal clusters to form metal-organic frameworks.

[0124] As described above, the metal-organic framework of the present invention can exhibit the effect of actively decomposing and releasing a bound immuno-anticancer agent by irradiating light at a targeted location, and can also exhibit a photodynamic therapeutic effect in which the metal-organic framework generates singlet oxygen by irradiating light, thereby killing cancer cells.

[0125] The photodynamic therapy described above is a three-way non-toxic process that requires the simultaneous presence of a photosensitizer (PS), light, and molecular oxygen. Although each component is individually harmless, when the photosensitizer is activated by light, energy is transferred to molecular oxygen, generating highly cytotoxic reactive oxygen species (ROS). Among these ROS, singlet oxygen ( 1 O2) can damage tumor cells. Photosensitizers are an important component in effective photodynamic therapy.

[0126] The ligand compound represented by the above chemical formula 1 is a very effective photosensitizer.

[0127] In addition, the metal-organic framework having a porous structure as in the present invention can integrate the photosensitizer into a periodic array exhibiting a high photosensitizer loading, 1 Easy to improve O2 production efficiency 1 It is an excellent photosensitizer because it can provide an O2 diffusion path.

[0128] The metal-organic framework of the present invention is highly efficient as a photosensitizer, as it includes high stability, a large surface area, and pores of an appropriate size for rapid oxygen diffusion.

[0129] The anticancer composition of the present invention is characterized by including a metal-organic framework to which an immuno-cancer agent is bound, as described above.

[0130] The above metal-organic framework can generate singlet oxygen upon light irradiation, thereby exhibiting a photodynamic therapeutic effect. Furthermore, it can be actively decomposed upon light irradiation, thereby exhibiting a release effect of the bound immuno-oncology agent at a specific site.

[0131] Due to these characteristics, conventional immuno-oncology agents have been problematic due to side effects from systemic exposure, but as in the present invention, by releasing the immuno-oncology agent by light irradiation only at the site where cancer cells are located, not through systemic exposure but through local exposure, the cancer cell killing effect by the immuno-oncology agent can be enhanced, and the anticancer effect can be further enhanced through photodynamic therapy using a metal-organic framework.

[0132] The cancer may be colon cancer, liver cancer, lung cancer, breast cancer, melanoma, stomach cancer, colon cancer, skin cancer, ovarian cancer, cervical cancer, thyroid cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, esophageal cancer, or fibrosarcoma.

[0133] The anticancer composition of the present invention can be provided as an anticancer pharmaceutical composition.

[0134] The pharmaceutical composition of the present invention can be administered parenterally during clinical administration and can be used in the form of a general pharmaceutical preparation. Parenteral administration can mean administration via routes other than oral administration, such as rectal, intravenous, peritoneal, intramuscular, intraarterial, transdermal, nasal, inhalation, ocular, and subcutaneous. When formulated, it is prepared using diluents or excipients such as commonly used fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants. Preparations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspending solvents can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases that can be used include witepsol, macrogol, Tween 61, cocoa butter, laurin butter, and glycerogelatin.

[0135] In addition, when the pharmaceutical composition of the present invention is used as a medicine, it may contain one or more components having the same or similar function in addition to the effective ingredient of the present invention. The component exhibiting the same or similar function may preferably be an anticancer agent, and the anticancer agent is actinomycin D, bleomycin sulfate, daunomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitomycin, mitomycin-C, mithramycin, irinotecan, camptothecin, novobiocin, epirubicin, dactinomycin, amsacrine, teniposide, etoposide, cisplatin, It may include, but is not limited to, one or more selected from carboplatin, oxaliplatin, paclitaxel, docetaxel, gefitinib, erlotinib, and afatinib, or pharmaceutically acceptable salts thereof, and mixtures thereof.

[0136] In addition, the pharmaceutical composition may be used by mixing with various carriers acceptable as pharmaceuticals, such as saline or organic solvents, and carbohydrates such as glucose, sucrose or dextran, antioxidants such as ascorbic acid or glutathione, chelating agents, low-molecular-weight proteins or other stabilizers may be used as pharmaceuticals to increase stability or absorbability.

[0137] According to another embodiment of the present invention, a method for treating cancer may include administering to a subject the composition of the present invention in an amount effective for treating cancer, and irradiating the subject with light.

[0138] In the method for treating cancer according to the present invention, the composition comprises an immuno-oncology agent combined with a metal-organic framework capable of generating singlet oxygen as an active ingredient, thereby simultaneously exhibiting anticancer immunotherapy and photodynamic therapy effects, and thus can be used for the treatment of cancer. The immuno-oncology agent and the metal-organic framework are as described above.

[0139] Manufacturing Example 1

[0140] Synthesis of PCN-223

[0141] A TCPP mixed solution was prepared by adding 200 mL of N,N-Dimethylformamide (DMF) and 96 mg of Tetrakis(4-carboxyphenyl)porphyrin (TCPP) and stirring. 54 mg of zirconyl chloride octahydrate (ZrOCl2*8H2O) was added to the TCPP mixed solution and stirred.

[0142] Afterwards, the reaction was performed in an oven at 105℃ for 17 hours. After the reaction, the reactant was obtained, centrifuged at 15,000 rpm for 10 minutes, and the supernatant was removed. 30 ml of DMF was added to the settled lower layer fraction and resuspended to wash the obtained product. Afterwards, the washing process was performed three times in the same manner, and 30 ml of ethanol was treated three times. After discarding the ethanol in the upper layer in the last centrifugation process, the PCN-223 particles in the lower layer were obtained by drying in an oven at 80℃ for 8 hours. Powder X-ray diffraction (PXRD) was measured to confirm whether the particles were well formed.

[0143] Synthesis of PCN-224

[0144] Particles were obtained in the same manner as PCN-223, except that the reaction was carried out at 120°C for 24 hours in an oven.

[0145] Synthesis of MOF-525

[0146] A TCPP mixed solution was prepared by adding 80 mL of N,N-Dimethylformamide (DMF), 75 mg of Tetrakis(4-carboxyphenyl)porphyrin (TCPP), and 1.75 g of benzoic acid and stirring. 2220 mg of ZrOCl was added to the TCPP mixed solution and stirred.

[0147] Afterwards, the reaction was performed in a 100℃ oven for 24 hours. After the reaction, the reactant was obtained, centrifuged at 15,000 rpm for 10 minutes, and the supernatant was removed. 30 ml of DMF was added to the lower layer fraction and resuspended to wash the obtained product. Afterwards, the washing process was performed three times using the same method, and 30 ml of ethanol was treated three times. After discarding the ethanol in the upper layer in the last centrifugation process, the MOF-525 particles in the lower layer were obtained by drying in an 80℃ oven for 8 hours. Powder X-ray diffraction (PXRD) was measured to confirm whether the particles were well formed.

[0148] Experimental example

[0149] Analysis of synthetic results

[0150] For scanning electron microscope (SEM) measurements, powder samples were placed in 2 mg to 3 mg Eppen tubes, 100 μl of ethanol was added, and bath sonication was performed. 10 μl was then spotted onto a silicon wafer and dried in an oven at 80°C. The dried silicon wafer was then attached to an SEM mount using carbon tape and moved to an SEM device for image capture (Company name: Zeiss / Model name: ULTRA PLUS).

[0151] Next, for X-ray diffraction (XRD) measurement, the powder sample was carefully placed on the XRD sample holder, and the sample was compressed by pressing it with an appropriate force using a glass slide glass. After that, all powder scattered around the sample holder was carefully removed, and the sample holder was moved to the equipment, set up, and measurements were performed in the range of 2℃ to 30℃ (Company name: Bruker / Model name: D2 phaser).

[0152] Next, for N2 adsorption isotherm and BET specific surface area-pore measurement, about 40 mg of completely dried sample was prepared and carefully placed in a glass sampler using a glass funnel, and all powder adhering around the glass tube except for the lower sample container was removed. After that, the upper filter cap was attached, and degas was performed by vacuum and heat treatment at 120℃ for 12 hours in a pretreatment device. The weight of the pretreated sample was measured using a microbalance, and the weight of the empty cell that was weighed in advance was subtracted to calculate the weight of the intact sample. The glass sampler was attached to the N2 adsorption-desorption device and the measurement was performed (Company name: MICROTAAC / Model name: BELSORP MINI X).

[0153] The structural images for PCN-223 particles are shown in Figs. 1 and 2.

[0154] Figure 1 is a structure for PCN-223 from the b side (front), and Figure 2 is an image of the side structure.

[0155] The scanning electron microscope observation results for PCN-223 are as shown in Fig. 3. In addition, the X-ray diffraction analysis results are as shown in Fig. 4, which confirmed that the manufactured particles were synthesized with the crystal structure of PCN-223. In addition, the BET (Brunauer-Emmett-Teller) analysis results through N2 adsorption showed that the specific surface area of ​​the PCN-223 particles was 2273.6 m 2 / g, and the pore diameter is 1.42 nm (Fig. 5).

[0156] The zeta potential of PCN-223 is 4.26 mV (Fig. 6), and the FT-IR measurement results are as shown in Fig. 7.

[0157] Structural images of PCN-224 particles are shown in Figs. 8 and 9. Fig. 8 is a structure of PCN-224 from the b-side (front), and Fig. 9 is an image of the side structure.

[0158] The scanning electron microscope observation results for PCN-224 are as shown in Fig. 10. In addition, the X-ray diffraction analysis results are as shown in Fig. 11, which confirmed that the manufactured particles were synthesized with the crystal structure of PCN-224. In addition, the BET (Brunauer-Emmett-Teller) analysis results through N2 adsorption showed that the specific surface area of ​​the PCN-224 particles was 1894 m 2 / g, and the pore diameter is 1.61 nm (Fig. 12).

[0159] The zeta potential of PCN-224 is 10.43 mV (Fig. 13), and the FT-IR measurement results are as shown in Fig. 14. The structural images for MOF-525 particles are as shown in Figs. 15 and 16. Fig. 15 is the structure of MOF-525 from the b-side (front), and Fig. 16 is an image of the side structure.

[0160] The scanning electron microscope observation results for MOF-525 are as shown in Figure 17. In addition, the X-ray diffraction analysis results are as shown in Figure 18, which confirmed that the manufactured particles were synthesized with the crystal structure of MOF-525. In addition, the BET (Brunauer-Emmett-Teller) analysis results through N2 adsorption showed that the specific surface area of ​​the MOF-525 particles was 1356.3 m 2 / g, and the pore diameter is 1.68 nm (Fig. 19).

[0161] The zeta potential of MOF-525 is 14.51 mV (Fig. 20), and the FT-IR measurement results are as shown in Fig. 21.

[0162] Cytotoxicity assessment

[0163] L929 (fibroblasts) were seeded in a 96-well plate at 9,000 cells per well and incubated at 37°C for 24 hours. MOF suspensions were prepared at concentrations of 50 ug / mL, 25 ug / mL, 10 ug / mL, and 1 ug / mL using cell culture medium. The existing cell culture medium in the 96-well plate was removed, and 100 μl of the diluted suspension was added to each well. Incubated at 37°C for 24 hours. The suspension in the well was removed and washed with PBS. 100 μL of culture medium containing 10% Ez-Cytox was added to each well. After incubation for about 1 hour, the absorbance was measured at 450 nm using a microplate reader (reference wavelength 600 nm to 650 nm).

[0164] The results of evaluating cytotoxicity are as shown in Figures 22 to 24.

[0165] Figure 22 shows the cytotoxicity evaluation for PCN-223, confirming that there was no cytotoxicity up to a MOF suspension at a concentration of 50 ug / mL.

[0166] Figure 23 shows the cytotoxicity evaluation for PCN-224, confirming that there was no cytotoxicity up to a MOF suspension at a concentration of 50 ug / mL.

[0167] Figure 24 shows a cytotoxicity evaluation for MOF-525, confirming that there was no cytotoxicity up to a MOF suspension at a concentration of 50 ug / mL.

[0168] Evaluation of photodynamic therapy

[0169] A 50 ug / ml PCN-223 suspension was prepared using cell medium. 100 μl of the PCN-223 suspension was injected into a 96-well black plate. 100 μM singlet oxygen sensor green (SOSG) reagent was added to the cell medium to prepare a 10 μM SOSG suspension for ROS activity detection. 100 μl of the 10 μM SOSG suspension was injected into the well containing the PCN-223 suspension, and pipetted to ensure that the two suspensions were well mixed. For the control well, only 100 μl of the cell medium was injected instead of the PCN-223 suspension, and 100 μl of the 10 μM SOSG suspension was injected. Each well was irradiated with a laser at 100 mW, 200 mW, 300 mW, 400 mW, and 500 mW for 5 minutes. The amount of ROS released was evaluated by measuring the intensity of the SOSG reagent using a microplate reader.

[0170] The test results are as shown in Fig. 25. It was confirmed that as the laser power increased, the amount of ROS emission increased.

[0171] As an additional test, the test was conducted in the same manner as the previous evaluation test for photodynamic therapy, with the laser intensity fixed at 500 mW, and the irradiation was performed for 30 seconds, 30 seconds (cumulative 1 minute), 1 minute (cumulative 2 minutes), and 1 minute (cumulative 3 minutes), and the intensity of the SOSG reagent was measured using a microplate reader at each time point to evaluate the amount of ROS released.

[0172] The test results are as shown in Fig. 26. It can be confirmed that as the light irradiation time increases, the amount of ROS released increases.

[0173] As an additional test, the test was conducted in the same manner as the previous evaluation test for photodynamic therapy, with the laser intensity fixed at 500 mW, and the irradiation was performed for 1 minute, 2 minutes (cumulative 3 minutes), 2 minutes (cumulative 5 minutes), and 5 minutes (cumulative 10 minutes), and the intensity of the SOSG reagent was measured using a microplate reader at each time point to evaluate the amount of ROS released.

[0174] The test results are as shown in Fig. 27. It can be confirmed that ROS activity increases with increasing time, and in particular, in the case of PCN-224, ROS activity increases rapidly.

[0175] Cytotoxicity test on colon cancer cells

[0176] MC38 (colon cancer cells) were seeded in a 96-well plate at 9,000 cells per well and incubated at 37°C for 24 hours. PCN-223, PCN-224, and MOF-525 suspensions at 50 ug / ml each were prepared using cell medium. The existing cell medium in the 96-well plate was removed, and 100 μl of the 50 ug / ml PCN-223, PCN-224, and MOF-525 suspensions were added to each well. However, for the two controls, only 100 μl of cell medium was injected instead of MOF particles. Each well was irradiated with a 500 mW laser for the first 5 minutes, followed by a rest period and then irradiated once more for a second 5 minutes (total of 10 minutes). The control group was not irradiated with the laser. Incubation was performed at 37°C for 24 hours. The suspension in the well was removed and washed with PBS. 100 uL of medium containing 10% Ez-Cytox was added to each well. After incubation for 1 hour, the absorbance was measured at 450 nm and 600 nm using a microplate reader, and the cell viability was calculated based on the conversion formula (reference wavelength: 600 nm).

[0177] The test results are as shown in Fig. 28.

[0178] Compared to the control group and the group irradiated with only laser, the groups treated with PCN-223, PCN-224, and MOF-525 showed superior killing effects against colon cancer cell lines.

[0179] Manufacturing Example 2

[0180] Preparation of metal-organic frameworks conjugated with immuno-oncology agents

[0181] PCN-223 particles were pretreated in a vacuum oven for 12 hours to secure unsaturated metal sites. 5 mg of Resiquimod was dissolved in 5 ml of ethanol, 50 mg of PCN-223 was added, and the mixture was stirred for 24 hours. After 24 hours, the particles were precipitated by centrifugation at 17,000 rpm for 10 minutes, and the supernatant was removed. 10 ml of new ethanol was added, and the washing process was repeated three times. After the final centrifugation, the sample was collected and dried in an 80°C oven for 1 hour to manufacture PCN-223 loaded with Resiquimod (hereinafter referred to as PCN-223 / Resiquimod).

[0182] PCN-224 / Resiquimod and MOF-525 / Resiquimod were manufactured using the same method as the manufacturing method of PCN-223 / Resiquimod above, using PCN-224 and MOF-525, respectively.

[0183] Experimental Example 2

[0184] Resiquimod payload evaluation

[0185] 5 mg of PCN-223 / Resiquimod, PCN-224 / Resiquimod, or MOF-525 / Resiquimod was weighed and placed in 20 ml of deionized water, then suspended using a sonicator, 1 ml of 37% hydrochloric acid was added, and the particles were disrupted by placing the mixture in an oven at 37°C for 24 hours to obtain 1 ml of a sample. The amount of Resiquimod in the sample was measured using high performance liquid chromatography (HPLC), and this was converted to determine the Resiquimod loading of the PCN-223 / Resiquimod, PCN-224 / Resiquimod, or MOF-525 / Resiquimod samples.

[0186] The shape of PCN-223 / Resiquimod was observed using a scanning electron microscope, and no changes in particle size or shape were observed due to drug loading (Fig. 29). In addition, the amount of Resiquimod loaded in PCN-223 / Resiquimod was analyzed by HPLC, and the loading amounts of Resiquimod in PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod were confirmed to be 12.8 ug / mg, 9.3 ug / mg, and 13.1 ug / mg, respectively.

[0187] Evaluation of photodynamic therapy

[0188] A 50 ug / ml PCN-223 suspension was prepared using cell medium. 100 μl of the PCN-223 suspension was injected into a 96-well black plate. A 10 μM SOSG suspension was prepared using cell medium to contain 100 μM SOSG reagent for ROS activity. 100 μl of the 10 μM SOSG suspension was injected into the well containing the PCN-223 suspension, and pipetted to ensure that the two suspensions were well mixed. For the control well, only 100 μl of the cell medium was injected instead of the PCN-223 suspension, and 100 μl of the 10 μM SOSG suspension was injected. Each well was irradiated with a laser at 500 mW for 1 min, 2 min (cumulative 3 min), 2 min (cumulative 5 min), and 5 min (cumulative 10 min). The amount of ROS released was evaluated by measuring the intensity of the SOSG reagent using a microplate reader.

[0189] The test results are as shown in Fig. 30. It was confirmed that as the laser irradiation time increased, the amount of ROS released increased.

[0190] Cytotoxicity test on colon cancer cells

[0191] CT26 (colon cancer cells) were seeded in a 96-well plate at 9,000 cells per well and incubated at 37°C for 24 hours. PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod suspensions were prepared at 50 ug / ml each using cell medium. The existing cell medium in the 96-well plate was removed, and 100 μl of 50 ug / ml PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod suspensions were added to each well. However, for the two controls, only 100 μl of cell medium was injected instead of MOF particles. Each well was irradiated with a 500 mW laser for the first 5 minutes, followed by a rest period and then irradiated once more for a second 5 minutes (cumulative 10 minutes). One control group was not irradiated with the laser. Incubated at 37°C for 24 hours. The suspension in the well was removed and washed with PBS. 100 uL of medium containing 10% Ez-Cytox was added to each well. After incubation for 1 hour, the absorbance was measured at 450 nm using a microplate reader, and the cell viability was calculated based on the conversion formula (reference wavelength 600 nm).

[0192] The test results are as shown in Fig. 31. In the media treated with PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod of the present invention, it can be confirmed that the cell viability for CT26 is drastically reduced by laser irradiation. This is due to the photodynamic therapeutic effect of PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod and the effect of the release of Resiquimod by light irradiation.

[0193] Cytotoxicity test of PCN-223 / Resiquimod

[0194] L929 cells (murine fibroblast cells) were cultured in a cell incubator at 37°C with 5% CO2 and seeded at 1×10 in a 96-well plate. 4 After seeding with cells / well and culturing for 24 hours, the medium in each well was removed, and PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod were prepared in fresh medium at concentrations of 1 ug / ml, 10 ug / ml, 25 ug / ml, and 50 ug / ml, respectively, and added at a volume of 100 μl per well. After culturing at 37°C in 5% CO2 for 24 hours, the particle suspension in the plate was removed, and each well was washed three times with DPBS. Finally, 90 μl of fresh medium and 10 μl of EZ-cytox reagent were added to each well, and the plate was incubated in a 37°C cell incubator for 1 hour. The cell viability was confirmed by measuring the absorbance at 450 nm and 600 nm using a microplate reader.

[0195] The results of evaluating the cytotoxicity of PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod are shown in Figures 32, 33, and 34.

[0196] PCN-223 / Resiquimod, PCN-224 / Resiquimod, and MOF-525 / Resiquimod showed no cytotoxicity, with survival rates exceeding 100% in the test concentration range of 1 ug / ml and 10 ug / ml. However, cytotoxicity was observed when treated at concentrations of 25 ug / ml or higher, which was attributed to Resiquimod bound to PCN-223, PCN-224, and MOF-525.

[0197] Although the preferred embodiments of the present invention have been described in detail above, the scope of the present invention is not limited thereto, and various modifications and improvements made by those skilled in the art using the basic concept of the present invention defined in the following claims also fall within the scope of the present invention.

[0198] The present invention relates to an anticancer composition comprising a metal-organic framework combined with an immuno-oncology agent and a use thereof.

Claims

1. Containing a metal-organic framework (MOF) combined with an immuno-oncology agent. Anticancer composition.

2. In paragraph 1, The above metal-organic framework comprises metal clusters and A ligand compound represented by the following chemical formula 1 that coordinates to the above metal cluster. Anticancer composition: [Chemical Formula 1] Here, X1 and X3 are N(R9), X2 and X4 are N, R1 to R9 are the same or different, and each independently represent hydrogen, deuterium, a cyano group, a nitro group, a halogen group, a hydroxy group, a substituted or unsubstituted alkylthio group having 1 to 4 carbon atoms, a substituted or unsubstituted alkyl group having 1 to 30 carbon atoms, a substituted or unsubstituted cycloalkyl group having 3 to 20 carbon atoms, a substituted or unsubstituted alkenyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkynyl group having 2 to 24 carbon atoms, a substituted or unsubstituted aralkyl group having 7 to 30 carbon atoms, a substituted or unsubstituted aryl group having 6 to 30 carbon atoms, a substituted or unsubstituted heteroaryl group having 1 to 60 carbon atoms, a substituted or unsubstituted heteroarylalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 60 carbon atoms, a substituted or unsubstituted aryl group having 2 to 30 carbon atoms, a substituted or unsubstituted aryl group having 2 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl group having 1 to 60 carbon atoms, a substituted or unsubstituted arylalkyl group having 2 to 30 carbon atoms, a substituted or unsubstituted alkoxy group having 1 to 30 carbon atoms, a substituted or unsubstituted aryl ... It is selected from the group consisting of a 30-carbon alkylamino group, a substituted or unsubstituted arylamino group having 6 to 30 carbon atoms, a substituted or unsubstituted aralkylamino group having 7 to 30 carbon atoms, a substituted or unsubstituted heteroarylamino group having 1 to 24 carbon atoms, a substituted or unsubstituted alkylsilyl group having 1 to 30 carbon atoms, a substituted or unsubstituted arylsilyl group having 6 to 30 carbon atoms, and a substituted or unsubstituted aryloxy group having 6 to 30 carbon atoms.

3. In paragraph 1, The above metal-organic framework is a metal selected from the group consisting of Li, Na, Mg, Ca, Sr, Ba, Sc, Y, Ti, Zr, Hf, V, Nb, Ta, Cr, Mo, W, Mn, Re, Fe, Ru, Os, Co, Rh, Ir, Ni, Pd, Pt, Cu, Ag, Au, Zn, Cd, Hg, Al, Ga, In, Tl, Si, Ge, Sn, Pb, As, Sb and Bi, or Li. + , Na + , Mg 2+ , Ca 2+ , Sr 2+ , Ba 2+ , Sc 3+ , Y 3+ , Ti 4+ , Zr 4+ , Hf 4+ , V 4+ , V 3+ , V 2+ , Nb 3+ , Ta 3+ , Cr 3+ , Mo 3+ , W 3+ , Mn 3+ , Mn 2+ , Re 3+ , Re 2+ , Fe 3+ , Fe 2+ , Ru 3+ , Ru 2+ , Os 3+ , Os 2+ , Co 3+ , Co 2+ , Rh 2+ , Rh + , Ir 3+ , Ir 2+ , Ir + , Ni 2+ , Ni + , Pd 2+ , Pd + , Pt 2+ , Pt + , Cu 2+ , Cu + , Ag + , Au +, Zn 2+ , Cd 2+ , Hg 2+ , Al 3+ , Ga 3+ , In 3+ , Tl 3+ , Si 4+ , Si 2+ , Ge 4+ , Ge 2+ , Sn 4+ , Sn 2+ , Pb 4+ , Pb 2+ , As 5+ , As 3+ , As + , Sb 5+ , Sb 3+ , Sb + , Bi 5+ , Bi 3+ and Bi + Containing a metal ion selected from the group consisting of Anticancer composition.

4. In paragraph 3, The above metal-organic framework is selected from the group consisting of aluminum-based metal-organic frameworks, iron-based metal-organic frameworks, zirconium-based metal-organic frameworks, and mixtures thereof. Anticancer composition.

5. In paragraph 2, The above immunotherapy agent coordinates to the unsaturated metal site of the metal cluster. Anticancer composition.

6. In paragraph 1, The above immuno-oncology agents are immune-modulating agents. Anticancer composition.

7. In paragraph 6, The above immunomodulator is an agonist of TLR7 (Toll-like receptor 7) or TLR8. Anticancer composition.

8. In paragraph 1, The above cancers are colon cancer, liver cancer, lung cancer, breast cancer, melanoma, stomach cancer, colon cancer, skin cancer, ovarian cancer, cervical cancer, thyroid cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, esophageal cancer or fibrosarcoma. Anticancer composition.

9. In paragraph 1, The above metal-organic complex generates singlet oxygen upon light irradiation. Anticancer composition.

10. A method for treating cancer, comprising administering to a subject an anticancer composition according to paragraph 1 in an amount effective for treating cancer and irradiating the subject with light.

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