Drug Delivery System Comprising Protac Incapsulated Metal-Organic Framework And Pharmaceutical Composition Comprising The Same

Abstract

The present invention provides a drug delivery system comprising metal-organic framework particles loaded with Protac, and a pharmaceutical composition comprising the same.

Description

Technology Field

[0001] The present invention relates to a drug delivery system comprising metal-organic framework particles loaded with Protac, and a pharmaceutical composition comprising the same. Background Technology

[0002] PROTACs (Proteolysis-targeting-chimeras) are small molecules designed to selectively degrade disease-causing proteins within cells. By breaking down disease-causing target proteins and effectively eliminating cancer cells or disease proteins, PROTACs are attracting attention as novel drugs applicable to intractable diseases that were difficult to treat with existing medications.

[0003] However, despite these excellent effects, PROTACs have limitations in that their efficacy is limited due to low in vivo drug delivery rates, which are disadvantages such as the large chemical structure of the drug itself, poor pharmacokinetic properties, and limited ability to penetrate cell membranes. Additionally, if proteases are present in the bloodstream, PROTAC molecules may be degraded, which can limit their effectiveness.

[0004] Developing effective delivery methods for PROTACs to overcome these problems is emerging as a critical issue in new drug development utilizing targeted protein degradation.

[0005] Patent Publication No. 2019-0101406 discloses a chimeric molecule targeting EGFR protein degradation and a related method of use. However, no effective delivery method for PROTACs is disclosed at all.

[0006] The inventors focused on the low drug delivery rate of such PROTACs and endeavored to develop a drug delivery system capable of delivering drugs more efficiently. By discovering that loading PROTACs onto a metal-organic framework improves the drug delivery rate, the inventors completed the present invention. Prior art literature

[0007] Patent Publication No. 2019-0101406 The problem to be solved

[0008] One objective of the present invention is to provide a drug delivery system with excellent drug delivery rate comprising metal-organic framework particles loaded with Protac, and a pharmaceutical composition comprising the same. means of solving the problem

[0009] To achieve the above objective, one example of the present invention provides a drug delivery system comprising metal-organic framework particles loaded with a protac.

[0010] Another example of the present invention provides a pharmaceutical composition for the treatment or prevention of cancer comprising metal-organic framework particles loaded with PROTAC as an active ingredient.

[0011] Another example of the present invention provides a pharmaceutical composition for the treatment or prevention of neurodegenerative diseases comprising metal-organic framework particles loaded with PROTACs as an active ingredient.

[0012] Another example of the present invention provides a pharmaceutical composition for the treatment or prevention of autoimmune diseases comprising metal-organic framework particles loaded with PROTACs as an active ingredient. Effects of the invention

[0013] A drug delivery system comprising metal-organic framework particles loaded with PROTACs according to the present invention exhibits excellent drug delivery capabilities. A pharmaceutical composition containing such PROTAC-loaded metal-organic framework particles as an active ingredient has the advantage of being able to overcome the limitations of existing drugs by enabling high accessibility and resolving resistance issues, as it can bind to all sites of a target protein rather than specific sites. Brief explanation of the drawing

[0014] FIG. 1 shows a metal-organic framework used in a drug delivery system in one example of the present invention. Figure 2 shows a PROTAC loaded onto a metal-organic framework in a drug delivery system according to one example of the present invention. Figures 3 to 7 show the scanning electron microscopy (SEM) measurement results, X-ray diffraction (XRD) analysis results, N2 adsorption-desorption isotherm measurement results, Zeta potential measurement results, and FTIR measurement results of PCN-223, respectively. Figures 8 and 9 show the SEM measurement results and FTIR measurement results of PCN-223 equipped with ARV-771, respectively. Figures 10 to 13 show the scanning electron microscope measurement results, X-ray diffraction analysis results, N2 adsorption-desorption isotherm measurement results, and Zeta potential measurement results of PCN-224, respectively. Figures 14 and 15 show the SEM measurement results and FTIR measurement results of the PCN-224 equipped with ARV-771, respectively. Figures 16 to 19 show the scanning electron microscope measurement results, X-ray diffraction analysis results, N2 adsorption-desorption isotherm measurement results, and Zeta potential measurement results of MOF-525, respectively. Figures 20 and 21 show the SEM measurement results and FTIR measurement results of MOF-525 equipped with ARV-771, respectively. Figures 22 to 25 show the scanning electron microscope measurement results, X-ray diffraction analysis results, N2 adsorption-desorption isotherm measurement results, and Zeta potential measurement results of MOF-545, respectively. Figures 26 and 27 show the SEM measurement results and FTIR measurement results of MOF-545 equipped with ARV-771, respectively. Figures 28 to 31 show the scanning electron microscope measurement results, X-ray diffraction analysis results, N2 adsorption-desorption isotherm measurement results, and Zeta potential measurement results of UIO-67, respectively. Figures 32 and 33 show the SEM measurement results and FTIR measurement results of PCN-223 equipped with UIO-67, respectively. Figures 34 to 38 show the results of cytotoxicity tests for PCN-223, PCN-224, MOF-525, MOF-545, and UIO-67, respectively. Specific details for implementing the invention

[0015] Hereinafter, a drug delivery system comprising metal-organic framework particles loaded with Protac according to an example of the present invention is described.

[0017] Protax

[0018] PROTAC (Proteolysis-targeting-chimera) is a type of small molecule designed to selectively degrade disease-causing proteins within cells and generally has a molecular structure consisting of (1) a ligand that binds to a target protein, (2) a ligand that binds to an E3 ubiquitin ligase enzyme, and (3) a linker that connects the two ligands.

[0019] PROTAC technology utilizes the ubiquitin-proteasome system, one of the body's protein degradation systems. When the enzyme E3 ligase attaches ubiquitin to a protein that needs to be removed from the body, the proteasome (a protein-degrading enzyme) recognizes and degrades the ubiquitin-bound protein. By binding a PROTAC to a target protein and linking the enzyme E3 ligase to it, the target protein can be degraded using the body's ubiquitin system.

[0021] Metal-Organic Framework

[0022] Metal-Organic Frameworks (MOFs) refer to crystalline organic-inorganic hybrid polymer materials that are three-dimensional porous structures formed by the coordination bonding of metal ions or metal ion clusters with organic ligands. Also known as Porous Coordination Polymers (PCPs), they possess a regular pore structure and a high specific surface area.

[0023] Metal-organic frameworks can have various structures with different sizes, porosity, three-dimensional structures, and surface areas depending on the type and binding method of metal ions and organic ligands. Generally, the pore size can be controlled from a few angstroms to a few nanometers by adjusting the size of the organic ligand, and the surface properties of the pores can be controlled by selecting an appropriate metal ion or introducing functional groups into the organic ligand.

[0024] Metal-organic frameworks possess porous characteristics with voids within their structure, resulting in a very large surface area. Furthermore, their open pore structure allows for the large-scale movement of molecules or solvents compared to other known porous materials. When used as catalysts or gas storage materials, they offer the advantage of maximizing efficiency due to the abundance of active sites. Additionally, metal-organic frameworks do not easily deform even at high temperatures and possess a rigid framework, exhibiting excellent thermal and chemical stability.

[0025] These metal-organic frameworks can effectively improve the delivery of PROTACs by protecting the drug from degradation, facilitating transport across the cell membrane, and enhancing accumulation at the site of action.

[0027] The above metal-organic framework can be formed by the binding of metal ions and organic ligands.

[0028] The above metal ions may include one or more metal ions selected from the group consisting of aluminum (Al), iron (Fe), and zirconium (Zr).

[0029] The above organic ligand may have carboxyl or amine groups at both ends. There are no particular limitations on such organic ligands, but examples include 4,4'-biphenyldicarboxilic acid, benzene-1,4-dicarboxylic acid, 9,10-anthracenedicarboxylic acid, biphenyl-3,3,5,5'-tetracarboxylic acid, biphenyl-3,4',5-tricarboxylic acid, 5-bromoisophthalic acid, and 5-cyano-1,3-benzenedicarboxylic acid. 2,2-diamino-4,4'-stilbenedicarboxylic acid, terephthalic acid, 2-aminoterephthalic acid, 2,5-diaminoterephthalic acid, 1,1,2,2-tetra(4-carboxylphenyl)ethylene, 2,5-dihydroxyterephthalic acid, 2,2-dinitro-4,4-stilbenedicarboxylic acid, 5-ethynyl-1,3-benzenedicarboxylic acid 2-hydroxyterephthalic acid, 2,6-naphthalenedicarboxylic acid, 1,2,4,5-tetrakis(4-carboxyphenyl)benzene(1,2,4,5-tetrakis(4-carboxyphenyl)benzene), 4,4,4½-s-triazine-2,4,6-triyltribenzoic acid, 1,3,5-tricarboxybenzene, 1,4,7,10-tetraazacyclododecane-N,N',N'',N''-tetraacetic acid, 1,3,5-tris(4-carboxy[1,1'-biphenyl]-4-yl)benzene), Examples include 1,3,5-tris(4-carboxyphenyl)benzene, 1,3,5-triscarboxyphenylethynylbenzene, and 2-methylimidazole. The above organic ligand preferably includes a porphyin ligand.

[0031] There are no particular limitations on the above metal-organic frameworks, but examples include UIO-66, UIO-66-NH2, UIO-67, UIO-67-NH2, PCN-128, PCN-222, PCN-223, PCN-224, MOF-525, MOF-545, MOF-801, MOF-808, MOF-867, Al-MIL-53, Al-MIL-53-NH2, Al-MIL-88, Al-MIL-88-NH2, Al-MIL-100, Al-MIL-101, Al-MIL-101-NH2, Al-MIL-125, Al-MIL-125-NH2, Fe-MIL-53, Fe-MIL-53-NH2, Fe-MIL-88, Fe-MIL-88-NH2, It may be one or more selected from the group consisting of Fe-MIL-100, Fe-MIL-101, Fe-MIL-101-NH2, Fe-MIL-125, and Fe-MIL-125-NH2. Preferably, it may be PCN-223, PCN-224, MOF-525, MOF-545, or UIO-67.

[0033] For example, PCN-223 is a material having a three-dimensional porous structure composed of zirconium (Zr) and an organic ligand ((4-carboxyphenyl)porphyrin), and can load PROTACs into the pores of PCN-223. In particular, in the metal-organic framework particles loaded with PROTACs according to one example of the present invention, PROTACs are loaded via coordination bonding to unsaturated metal sites of the Zr clusters of PCN-223. Due to this bonding, the release period of PROTACs is long and exposure to normal cells is minimized, thereby maximizing delivery efficiency to the disease site. Other metal-organic frameworks can also have their delivery efficiency maximized by these characteristics.

[0035] One example of the present invention also provides a pharmaceutical composition for the treatment or prevention of cancer comprising a metal-organic framework particle loaded with a PROTAC as an active ingredient. Herein, the cancer refers to colorectal cancer, liver cancer, lung cancer, breast cancer, melanoma, gastric cancer, colon cancer, skin cancer, ovarian cancer, cervical cancer, thyroid cancer, kidney cancer, prostate cancer, bladder cancer, pancreatic cancer, esophageal cancer, or fibrosarcoma.

[0036] The above pharmaceutical composition may include a pharmaceutically acceptable carrier. The term "pharmaceuticalally acceptable carrier" may refer to a carrier or diluent that does not irritate living organisms and does not impair the biological activity and properties of the injected compound. Here, "pharmaceuticalally acceptable" means that it does not inhibit the activity of the active ingredient and does not possess toxicity exceeding the level to which the subject of application (prescription) can tolerate it. Any type of carrier that is commonly used in the relevant technical field and is pharmaceutically acceptable may be used in the above pharmaceutical composition. Non-limiting examples of the above carriers include lactose, dextrose, maltodextrin, sucrose, sorbitol, mannitol, xylitol, erythritol, maltitol, glycerol, ethanol, starch, acacia gum, alginate, gelatin, calcium phosphate, calcium silicate, cellulose, methyl cellulose, microcrystalline cellulose, polyvinylpyrrolidone, water, saline solution, sterile water, Ringer's solution, buffered saline solution, albumin injection solution, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. These may be used alone or in a mixture of two or more. The above pharmaceutical composition may be prepared into an oral or parenteral formulation according to the route of administration by conventional methods known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The above pharmaceutical compositions may each be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, and aerosols, as well as topical preparations, suppositories, or sterile injectable solutions, according to conventional methods. When formulating the above pharmaceutical compositions, they may be prepared using commonly used diluents or excipients such as fillers, extenders, binders, wetting agents, disintegrants, or surfactants, but are not limited thereto.

[0038] A pharmaceutical composition according to one example of the present invention may be administered parenterally during clinical administration and may be used in the form of a general pharmaceutical formulation. Parenteral administration may refer to administration via routes other than oral administration, such as rectal, intravenous, peritoneal, intramuscular, arterial, transdermal, nasal, inhalation, ocular, and subcutaneous. When formulating, the composition is prepared using diluents or excipients such as commonly used fillers, volume expanders, binders, wetting agents, disintegrants, and surfactants. Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solvents, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solvents and suspension solvents may include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Witepsol, Macrogol, Tween 61, Tween 80, cacao gelatin, laurin gelatin, glycerogelatin, etc., may be used as bases for suppositories.

[0039] In addition, when the above-described pharmaceutical composition of the present invention is used as a medicine, it may contain one or more additional components exhibiting the same or similar functions in addition to the active ingredient of the present invention. The components exhibiting the same or similar functions may preferably be anticancer agents. The types of the above anticancer drugs are not particularly limited, but include actinomycin D, bleomycin sulfate, daunomycin, daunorubicin, doxorubicin, epirubicin, idarubicin, mitomycin, mitomycin-C, mitramycin, irinotecan, camptothecin, novobiocin, epirubicin, dactinomycin, amsacrine, teniposide, etoposide, cisplatin, and carboplatin. One or more selected from oxaliplatin, paclitaxel, docetaxel, gefitinib, erlotinib, and afatinib may be included.

[0040] In addition, the above pharmaceutical composition may be used in combination with various carriers permitted as pharmaceuticals, such as physiological 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 absorption.

[0042] Another example of the present invention provides a pharmaceutical composition for the treatment or prevention of neurodegenerative diseases comprising metal-organic framework particles loaded with PROTACs as an active ingredient.

[0043] Another example of the present invention provides a pharmaceutical composition for the treatment or prevention of autoimmune diseases comprising metal-organic framework particles loaded with PROTACs as an active ingredient.

[0044] In the above composition, the PROTAC and metal-organic framework may be selected from the aforementioned PROTAC and metal-organic framework.

[0046] The present invention will be explained in more detail through the following examples. However, these examples are intended to illustrate the invention and the scope of the invention is not limited to these examples.

[0048] Examples

[0049] Synthesis of Metal-Organic Frameworks (MOFs) and Preparation of PROTAC-Loaded Metal-Organic Frameworks

[0050] Devices used

[0051] Scanning Electron Microscope (SEM): A Zeiss Ultra Plus was used.

[0052] X-ray diffraction analysis: A Bruker D2 Phaser was used.

[0053] N2 Adsorption-Desorption Isotherm: Microtaac's Belsorp Mini X was used.

[0055] Preparation Example 1: Synthesis of PCN-223

[0056] 196 mg of tetrakis(4-carboxyphenyl)porphyrin (TCPP) was added to 60 ml of N,N-dimethylformamide (DMF) and stirred. 104 mg of zirconyl chloride octahydrate (ZrOCl2*8H2O) was added to this solution and stirred. Then, the mixture was reacted at 105°C for 17 hours. The synthesized particles were washed three times with dimethylformaldehyde (DMF) and three times with ethanol (EtOH), and then dried at 80°C to obtain a powder.

[0057] The results of measuring the obtained PCN-223 with a scanning electron microscope (Zeiss, Ultra Plus) are shown in Figure 3.

[0058] The results of the X-ray diffraction analysis (Bruker, D2 Phaser) of PCN-223 are shown in Figure 4.

[0059] The results of the N2 adsorption-desorption isotherm measurements of PCN-223 are shown in Fig. 5 (Microtaac, Belsorp Mini X).

[0060] The surface area of ​​PCN-223 is 2273.6 m² 2 / g, the pore size (Pore Diameter) was measured to be 1.42 nm.

[0061] The results of the Zeta Potential measurement of PCN-223 are shown in Figure 6.

[0063] Example 1: Manufacture of PCN-223 equipped with ARV-771

[0064] PCN-223 prepared according to Preparation Example 1 was pretreated in a vacuum oven at 150°C for 24 hours. 100 mg of the pretreated powder was weighed and placed in an amber vial. Meanwhile, ARV-771 was dissolved in DMSO at a concentration of 1 mg / ml, and 5 ml of this solution was added to the powder sample in the amber vial. After 48 hours, the powder particles were washed three times with fresh DMSO and three times with ethanol (EtOH). The powder was dried at 80°C to obtain the powder.

[0065] The SEM measurement results of PCN-223 equipped with ARV-771 are shown in Fig. 8.

[0066] The FTIR measurement results of PCN-223 equipped with ARV-771 are shown in Fig. 9.

[0068] Preparation Example 2: Synthesis of PCN-224

[0069] 100 mg of TCPP was added to 80 ml of DMF and stirred. 300 mg of zirconyl chloride octahydrate (ZrOCl2*8H2O) was added to this solution and stirred. Then, the reaction was carried out at 125°C for 24 hours. The particles obtained from the reaction were washed three times with DMF and three times with ethanol, and then dried at 80°C to obtain a powder.

[0070] The results of measuring the obtained PCN-224 with a scanning electron microscope are shown in Figure 10.

[0071] The results of the X-ray diffraction analysis (Bruker, D2 Phaser) of PCN-224 are shown in Fig. 11.

[0072] The results of the N2 adsorption-desorption isotherm measurements of PCN-224 are shown in Fig. 12 (Microtaac, Belsorp Mini X).

[0073] The surface area of ​​PCN=224 is 1894 m² 2 / g, the pore size (Pore Diameter) was measured to be 1.61 nm.

[0074] The results of the Zeta Potential measurement of PCN-224 are shown in Figure 13.

[0076] Example 2: Manufacture of PCN-224 equipped with ARV-771

[0077] PCN-224 synthesized according to Preparation Example 2 was pretreated in a vacuum oven at 150°C for 24 hours. 100 mg of the pretreated powder was weighed and placed in an amber vial. Meanwhile, a drug solution was prepared by dissolving ARV-771 in DMSO at a concentration of 1 mg / ml, and 5 ml of this solution was added to the powder sample weighed in the amber vial. After 48 hours, the powder particles were washed three times with fresh DMSO and three times with ethanol (EtOH). The powder was dried at 80°C to obtain the powder.

[0078] The SEM measurement results of PCN-224 equipped with ARV-771 are shown in Fig. 14.

[0079] The FTIR measurement results of the PCN-224 equipped with ARV-771 are shown in Fig. 15.

[0081] Preparation Example 3: Synthesis of MOF-525

[0082] First, 235 mg of TCPP and 6.75 g of benzoic acid were added to 80 ml of DMF and stirred. 520 mg of zirconyl chloride octahydrate (ZrOCl2*8H2O) was added to this solution and stirred. Then, the mixture was reacted at 100°C for 24 hours. After washing three times with DMF and three times with ethanol, the mixture was dried at 80°C to obtain a powder.

[0083] The results of measuring the obtained MOF-525 with a scanning electron microscope are shown in Fig. 16.

[0084] The results of the X-ray diffraction analysis of MOF-525 are shown in Fig. 17.

[0085] The results of the N2 adsorption-desorption isotherm measurement of MOF-525 are shown in Fig. 18 (Microtaac, Belsorp Mini X).

[0086] The specific surface area of ​​MOF-525 is 1,356.3 m² 2 / g, the pore diameter was measured to be 1.68 nm.

[0087] The results of measuring the Zeta Potential of MOF-525 are shown in Fig. 19.

[0089] Example 3: Manufacture of MOF-525 equipped with ARV-771

[0090] MOF-525 synthesized according to Preparation Example 3 was pretreated in a vacuum oven at 150°C for 24 hours. 100 mg of the pretreated powder was weighed and placed in an amber vial. Meanwhile, a drug solution was prepared by dissolving ARV-771 in DMSO at a concentration of 1 mg / ml, and 5 ml of this solution was added to the powder sample weighed in the amber vial. After 48 hours, the powder particles were washed three times with fresh DMSO and three times with ethanol (EtOH). The powder was dried in an oven at 80°C to obtain the powder.

[0091] The SEM measurement results of MOF-525 equipped with ARV-771 are shown in Fig. 20.

[0092] The FTIR measurement results of MOF-525 equipped with ARV-771 are shown in Fig. 21.

[0094] Preparation Example 4: Synthesis of MOF-545

[0095] 32.5 mg of TCPP and 187.5 mg of zirconyl chloride octahydrate (ZrOCl2*8H2O) were added to 80 ml of DMF and stirred. 1.25 ml of dichloroacetic acid was added. Then, the mixture was reacted at 130°C for 18 hours. After washing three times with DMF and three times with ethanol, the mixture was dried at 80°C to obtain a powder.

[0096] The results of measuring the obtained MOF-545 with a scanning electron microscope are shown in Fig. 22.

[0097] The results of the X-ray diffraction analysis of MOF-545 are shown in Fig. 23.

[0098] The results of the N2 adsorption-desorption isotherm measurement of MOF-545 are shown in Figure 24.

[0099] The specific surface area of ​​MOF-545 is 1082.2 m² 2 / g, the pore diameter was measured to be 3.09 nm.

[0100] The results of measuring the Zeta Potential of MOF-545 are shown in Figure 25.

[0102] Example 4: Manufacture of MOF-545 equipped with ARV-771

[0103] MOF-545 synthesized according to Example 4 was pretreated in a vacuum oven at 150°C for 24 hours. 100 mg of the pretreated powder was weighed and placed in an amber vial. Meanwhile, a drug solution was prepared by dissolving ARV-771 in DMSO at a concentration of 1 mg / ml, and 5 ml of this solution was added to the powder sample weighed in the amber vial. After 48 hours, the particles were washed three times with fresh DMSO and three times with ethanol (EtOH). The powder was obtained by drying in an oven at 80°C.

[0104] The SEM measurement results of MOF-545 equipped with ARV-771 are shown in Fig. 26.

[0105] The FTIR measurement results of MOF-545 equipped with ARV-771 are shown in Fig. 27.

[0107] Preparation Example 5: Synthesis of UIO-67

[0108] 240 mg of biphenyl-4,4-dicarboxylic acid (BPDC), 230 mg of ZrCl4, and 25 µl of H2O were weighed and mixed to dissolve in 160 ml of DMF. The mixture was then reacted in an oven at 95°C for 17 hours. After washing three times with DMF and three times with ethanol, the mixture was dried at 80°C for 6 hours to obtain a powder.

[0109] The results of measuring the obtained UIO-67 with a scanning electron microscope are shown in Fig. 28.

[0110] The results of the X-ray diffraction analysis of UIO-67 are shown in Fig. 29.

[0111] The results of the N2 adsorption-desorption isotherm measurement of UIO-67 are shown in Fig. 30.

[0112] The surface area of ​​UIO-67 is 2192.8 m² 2 / g, the pore diameter was measured to be 1.88 nm.

[0113] The results of the Zeta Potential measurement of UIO-67 are shown in Fig. 31.

[0115] Example 5: Manufacture of UIO-67 equipped with ARV-771

[0116] UIO-67 prepared according to Preparation Example 5 was pretreated in a vacuum oven at 150°C for 24 hours. 100 mg of the pretreated powder was weighed and placed in an amber vial. Meanwhile, a drug solution was prepared by dissolving ARV-771 in DMSO at a concentration of 1 mg / ml, and 5 ml of this solution was added to the powder sample weighed in the amber vial. After 48 hours, the particles were washed three times with fresh DMSO and three times with ethanol (EtOH). The powder was obtained by drying in an oven at 80°C.

[0117] The SEM measurement results of the UIO-67 equipped with ARV-771 are shown in Fig. 32.

[0118] The FTIR measurement results of the UIO-67 equipped with ARV-771 are shown in Fig. 33.

[0120] Experimental Example 1: Cytotoxicity Test Method

[0121] (1) Mouse fibroblasts (L929) were placed in a 96-well plate at a density of 11,000 per well and cultured for 24 hours in a 5% CO2 incubator at 37 degrees Celsius.

[0122] (2) MOF suspensions at concentrations of 100, 50, 25, 10, and 1 µg / ml were prepared using cell culture medium.

[0123] (3) Remove the existing medium from each well plate cell, and prepare suspensions of the metal-organic framework particles to be tested by diluting them in the medium at concentrations of 100, 50, 25, 10, and 1 µg / ml, and add 100 µl to each well.

[0124] (4) It was cultured for 24 hours in a 37°C, 5% CO2 incubator.

[0125] (5) Remove the suspension from the well and wash the suspension with PBS.

[0126] (6) 100 µl of 10% Ez-Cytox medium was added to each well.

[0127] (7) After reacting in an incubator for about 1 hour, the absorbance at 450 nm was measured using a microplate reader. (Reference wavelength 600 nm)

[0128] The results of the cytotoxicity tests for PCN-223, PCN-224, MOF-525, MOF-545, and UIO-67 are shown in Figures 34 to 38, respectively.

[0130] Experimental Example 2: Measurement of drug loading

[0131] (1) 10 mg of ARV-771 loaded MOF was weighed and placed in an amber vial.

[0132] (2) 20 ml of PBS (pH 7.4) was added to the measured sample.

[0133] (3) Incubated for 72 hours in a shaking incubator (RPM 100) at 37.5 degrees Celsius.

[0134] (4) After 72 hours, the vial was removed and treated with an ultrasonic device for 6 hours to completely break it down.

[0135] (5) The sample that was completely decomposed and released the drug was centrifuged at 14,000 RPM for 10 minutes and the supernatant was collected.

[0136] (6) The total amount of the drug was quantified using HPLC from the obtained supernatant.

[0137] The results of the analysis of the payload capacity of the ARV-771 are shown in Table 1 below.

[0139] Sample name ARV-771 payload (ug / mg) ARV-771 loaded PCN-223 23.61 ARV-771 loaded PCN-224 29.14 ARV-771 loaded MOF-525 32.53 ARV-771 loaded MOF-545 21.44 ARV-771 loaded UIO-67 28.37

[0141] Experimental Example 3: Evaluation of Cancer Cell Killing Ability of ARV-771-Launched MOF

[0142] To evaluate the enhanced drug delivery efficacy of MOFs loaded with PROTAC drugs, cancer cell killing ability was assessed using MC38, a representative cancer cell line.

[0143] Specifically, as examples of metal-organic frameworks equipped with ARV-771, five types of metal-organic frameworks prepared / synthesized in the above examples were applied, and a control group with only ARV-771 added was also applied for testing.

[0144] 100 µl of DMEM medium and 10,000 MC38 cells / well were dispensed into each well of a 96-well cell culture plate and cultured for 24 hours. After 24 hours, a control group containing only ARV-771 and five experimental MOF groups loaded with ARV-771 were each administered at the same drug concentration of 2 µg / ml, resulting in a total concentration of 200 ng / well per well based on a 96-well plate, with N=8 applied to all groups. Culture was carried out in a CO2 incubator for 24 hours. After 24 hours, absorbance analysis at 450 nm and 600 nm was performed using a microplate reader with EZ-Cytox reagent to evaluate the growth rate of cancer cells compared to the untreated group.

[0145] As a result of the above evaluation of cancer cell killing ability, the cancer cell survival rate was 79.2% in the group treated with ARV-771 alone, which was used as a comparison group for each ARV-771-loaded metal-organic framework, whereas it was 23.6% in ARV-771-loaded PCN-223, 32.53% in ARV-771-loaded PCN-224, 21.44% in ARV-771-loaded MOF-545, and 28.37% in ARV-771-loaded UIO-67, confirming that the cancer cell survival rate was significantly lower compared to the use of ARV-771 alone.

[0146] Table 2 below shows the results of the killing ability of ARV-771 loaded MOF MC38 colorectal cancer cell lines.

[0147] Sample name MC38 survival rate (%) compared to untreated group ARV-771 (2 ug / ml) 86.5 ARV-771 loaded PCN-223 47.1 ARV-771 loaded PCN-224 58.3 ARV-771 loaded MOF-525 39.6 ARV-771 loaded MOF-545 45.1 ARV-771 loaded UIO-67 34.5

Claims

Claim 1 A drug delivery system comprising a metal-organic framework particle loaded with a protac, wherein the metal-organic framework comprises a zirconium (Zr) metal cluster and an organic aromatic ligand having a polycarboxyl group, wherein the protac is loaded by a coordination bond to an unsaturated metal site of the zirconium (Zr) metal cluster, and wherein the metal-organic framework is one or more metal-organic frameworks selected from the group consisting of UIO-66, UIO-66-NH2, UIO-67, UIO-67-NH2, PCN-128, PCN-222, PCN-223, PCN-224, MOF-525, MOF-545, MOF-801, MOF-808, and MOF-867. Claim 2 In claim 1, the metal-organic framework is a drug delivery system formed by the binding of an organic ligand and a metal ion. Claim 3 delete Claim 4 In paragraph 2, the above organic ligand is a drug delivery system comprising a porphyrin ligand. Claim 5 delete Claim 6 delete Claim 7 A pharmaceutical composition for the treatment or prevention of cancer comprising the drug delivery system of claim 1. Claim 8 A pharmaceutical composition according to claim 7, wherein the cancer is colorectal 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. Claim 9 delete Claim 10 delete

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