Drug delivery system comprising metal-organic framework particles loaded with protac and pharmaceutical composition comprising same

Abstract

The present invention relates to a drug delivery system comprising metal-organic framework particles loaded with a Protac, and a pharmaceutical composition comprising same. The drug delivery system can exhibit excellent drug delivery ability. With an ability to bind to all sites rather than specific sites of a target protein, the pharmaceutical composition is highly accessible and can solve a resistance problem, thereby overcoming the limitations of existing drugs.

Description

Drug delivery system comprising metal-organic framework particles loaded with PROTAC and pharmaceutical composition comprising the same

[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.

[0002] Proteolysis-targeting-chimera (PROTAC) is a small molecule designed to selectively degrade disease-causing proteins within cells. PROTAC is attracting attention as a new drug that can be applied to incurable diseases previously difficult to treat with existing drugs, as it degrades disease-causing target proteins, thereby eliminating cancer cells and disease-causing proteins.

[0003] However, despite these promising effects, Protac suffers from limitations such as its large chemical structure, poor pharmacokinetic properties, and limited ability to penetrate cell membranes. This results in low drug delivery rates in vivo, limiting its effectiveness. Furthermore, the presence of proteases in the bloodstream can degrade Protac molecules, limiting their effectiveness.

[0004] Developing an effective delivery method for proteases that can overcome these problems is emerging as a key issue in the development of new drugs utilizing targeted protein degradation.

[0005] Patent Publication No. 2019-0101406 discloses a chimeric molecule targeting EGFR protein degradation and related methods of use. However, it does not disclose any effective delivery method for Protac.

[0006] The inventor of the present invention, noting the low drug delivery rate of such protac, endeavored to develop a drug delivery system capable of delivering drugs more efficiently, and discovered that the drug delivery rate is improved by loading the protac onto a metal-organic framework, thereby completing the present invention.

[0007] [Prior Art Literature]

[0008] [Patent Document]

[0009] Patent Publication No. 2019-0101406

[0010] One object of the present invention is to provide a drug delivery system having an excellent drug delivery rate comprising metal-organic framework particles loaded with Protac, and a pharmaceutical composition comprising the same.

[0011] To achieve the above object, one example of the present invention provides a drug delivery system comprising metal-organic framework particles loaded with Protac.

[0012] Another example of the present invention provides a pharmaceutical composition for treating or preventing cancer, comprising metal-organic framework particles loaded with protac as an active ingredient.

[0013] Another example of the present invention provides a pharmaceutical composition for treating or preventing a neurodegenerative disease, comprising a metal-organic framework particle loaded with a protac as an active ingredient.

[0014] Another example of the present invention provides a pharmaceutical composition for treating or preventing an autoimmune disease, comprising a metal-organic framework particle loaded with a protac as an active ingredient.

[0015] The drug delivery system comprising the metal-organic framework particles loaded with Protac according to the present invention exhibits excellent drug delivery capacity. Pharmaceutical compositions containing these Protac-loaded metal-organic framework particles as active ingredients have the advantage of being able to bind to all sites of a target protein, rather than just specific ones, thereby increasing accessibility and addressing resistance issues, thereby overcoming the limitations of existing drugs.

[0016] Figure 1 illustrates a metal-organic framework used in a drug delivery system according to one example of the present invention.

[0017] Figure 2 shows an example of a drug delivery system according to the present invention in which a protac is loaded onto a metal-organic framework.

[0018] Figures 3 to 7 show the results of scanning electron microscopy (SEM) measurement, X-ray diffraction (XRD) analysis, N2 adsorption / desorption isotherm measurement, Zeta Potential measurement, and FTIR measurement, respectively, of PCN-223.

[0019] Figures 8 and 9 show the SEM and FTIR measurement results of PCN-223 loaded with ARV-771, respectively.

[0020] Figures 10 to 13 show the results of scanning electron microscope measurement, X-ray diffraction analysis, N2 adsorption / desorption isotherm measurement, and Zeta Potential measurement of PCN-224, respectively.

[0021] Figures 14 and 15 show the SEM and FTIR measurement results of PCN-224 loaded with ARV-771, respectively.

[0022] Figures 16 to 19 show the results of scanning electron microscope measurement, X-ray diffraction analysis, N2 adsorption / desorption isotherm measurement, and Zeta Potential measurement of MOF-525, respectively.

[0023] Figures 20 and 21 show the SEM and FTIR measurement results of MOF-525 loaded with ARV-771, respectively.

[0024] Figures 22 to 25 show the results of scanning electron microscope measurement, X-ray diffraction analysis, N2 adsorption / desorption isotherm measurement, and Zeta Potential measurement of MOF-545, respectively.

[0025] Figures 26 and 27 show the SEM and FTIR measurement results of MOF-545 loaded with ARV-771, respectively.

[0026] Figures 28 to 31 show the results of scanning electron microscope measurement, X-ray diffraction analysis, N2 adsorption / desorption isotherm measurement, and Zeta Potential measurement, respectively, of UIO-67.

[0027] Figures 32 and 33 show the SEM and FTIR measurement results of PCN-223 equipped with UIO-67, respectively.

[0028] Figures 34 to 38 show the cytotoxicity test results of PCN-223, PCN-224, MOF-525, MOF-545, and UIO-67, respectively.

[0029] The present invention relates to a drug delivery system comprising metal-organic framework particles loaded with Protac.

[0030] Hereinafter, a drug delivery system comprising metal-organic framework particles loaded with Protac according to an example of the present invention is described.

[0031]

[0032] Protak

[0033] PROTAC (Proteolysis-targeting-chimera) is a type of small molecule designed to selectively degrade disease-causing proteins within cells. It generally has a molecular structure consisting of (1) a ligand that binds to the target protein, (2) a ligand that binds to an E3 ubiquitin ligase enzyme, and (3) a linker that connects the two ligands.

[0034] 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 to be removed from the body, the proteasome (a protein-degrading enzyme) recognizes and degrades the ubiquitin-bound protein. When Protac binds to a target protein and links the enzyme E3 ligase to it, the body's ubiquitin system can be used to degrade the target protein.

[0035]

[0036] Metal-organic frameworks

[0037] Metal-organic frameworks (MOFs) are three-dimensional porous structures formed by coordination bonds between metal ions or metal ion clusters and organic ligands. They are crystalline organic-inorganic hybrid polymer materials. Also called porous coordination polymers (PCPs), they possess a regular pore structure and high specific surface area.

[0038] Metal-organic frameworks (MOFs) can possess diverse structures, varying in size, porosity, three-dimensional structure, surface area, and other characteristics, depending on the type and bonding method of the metal ions and organic ligands. Generally, pore size can be controlled from several angstroms to several nanometers by adjusting the size of the organic ligand. The surface properties of the pores can be controlled by selecting appropriate metal ions or introducing functional groups into the organic ligands.

[0039] Metal-organic frameworks (MOFs) possess porous structures with voids within their structures, resulting in extremely large surface areas. Their open pore structure allows for the transport of large quantities of molecules or solvents compared to other known porous materials. Furthermore, MOFs offer numerous active sites, maximizing efficiency when used as catalysts or gas storage systems. Furthermore, MOFs are resistant to deformation even at high temperatures, and their rigid frameworks offer excellent thermal and chemical stability.

[0040] These metal-organic frameworks can effectively improve the delivery of proteases by protecting the drug from degradation, facilitating its transport across cell membranes, and enhancing its accumulation at the site of action.

[0041]

[0042] The above metal-organic framework can be formed by the bonding of a metal ion and an organic ligand.

[0043] The above metal ion may include one or more metal ions selected from the group consisting of aluminum (Al), iron (Fe), and zirconium (Zr).

[0044] The organic ligand may have a carboxyl or amine group at both terminals. There is no particular limitation on such organic ligands, but examples thereof include 4,4'-biphenyldicarboxylic 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, 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)benzene5-tetrakis(4-carboxyphenyl)benzene), 4,4,4½-s-triazine-2,4,6-triyl-tribenzoic 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 thereof include 1,3,5-tris(4-carboxyphenyl)benzene, 1,3,5-triscarboxyphenylethynylbenzene, and 2-methylimidazole. The organic ligand preferably includes a porphyrin ligand.

[0045]

[0046] There is no particular limitation on the above metal-organic frameworks, but for example, 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, It may be at least one selected from the group consisting of Fe-MIL-88-NH2, 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, UIO-67.

[0047]

[0048] 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 Protac can be loaded into the pores of PCN-223. In particular, the metal-organic framework particle loaded with Protac according to an example of the present invention has the advantage that Protac is loaded by coordination bonding on the unsaturated metal site of the Zr cluster of PCN-223, and due to this bond, the release period of Protac is long and exposure to normal cells is minimized, thereby maximizing the delivery efficiency to the diseased site. Other metal-organic frameworks can also maximize the delivery efficiency due to these characteristics.

[0049]

[0050] Another example of the present invention provides a pharmaceutical composition for treating or preventing cancer, comprising metal-organic framework particles loaded with protac as an active ingredient. Here, the cancer refers to 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.

[0051] The pharmaceutical composition may include a pharmaceutically acceptable carrier. The term "pharmaceutically acceptable carrier" may refer to a carrier or diluent that does not stimulate a living organism and does not inhibit the biological activity or properties of the injected compound. Here, "pharmaceutically acceptable" means that the carrier does not inhibit the activity of the active ingredient and does not exhibit toxicity exceeding a level to which the subject of application (prescription) can adapt. Any type of carrier commonly used in the art and pharmaceutically acceptable may be used in the 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, polyvinyl pyrrolidone, water, saline solution, sterile water, Ringer's solution, buffered saline, albumin injection solution, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, or mineral oil. These may be used alone or in combination of two or more. The pharmaceutical composition may be prepared as an oral formulation or a parenteral formulation according to the route of administration by a conventional method known in the art, including a pharmaceutically acceptable carrier in addition to the active ingredient. The above pharmaceutical composition may be formulated and used in the form of oral dosage forms such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, etc., external preparations, suppositories, or sterile injectable solutions, respectively, according to conventional methods. When formulating the above pharmaceutical composition, it may be prepared using diluents or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, or surfactants, but may not be limited thereto.

[0052]

[0053] A pharmaceutical composition according to an example of the present invention can be administered parenterally during clinical administration and can be used in the form of a general pharmaceutical formulation. 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. Formulations 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, tween 80, cocoa butter, laurin butter, and glycerogelatin.

[0054] In addition, when the pharmaceutical composition of the present invention is used as a medicine, it may contain one or more components exhibiting the same or similar function as the effective ingredient of the present invention. The components exhibiting the same or similar function 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, mithramycin, irinotecan, camptothecin, novobiocin, epirubicin, dactinomycin, amsacrine, teniposide, etoposide, cisplatin, carboplatin, One or more selected from oxaliplatin, paclitaxel, docetaxel, gefitinib, erlotinib, and afatinib.

[0055] 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.

[0056]

[0057] Another example of the present invention provides a pharmaceutical composition for treating or preventing a neurodegenerative disease, comprising metal-organic framework particles loaded with protac as an active ingredient.

[0058] Another example of the present invention provides a pharmaceutical composition for treating or preventing an autoimmune disease, comprising a metal-organic framework particle loaded with a protac as an active ingredient.

[0059] In the above composition, the protac, metal-organic framework may be selected from the protac, metal-organic frameworks described above.

[0060]

[0061] The present invention will be described in more detail through the following examples. However, these examples are intended to exemplify the present invention and the scope of the present invention is not limited to these examples.

[0062]

[0063] Example

[0064] Synthesis of metal-organic frameworks (MOFs) and fabrication of protac-loaded MOFs

[0065] Device used

[0066] Scanning electron microscope (SEM): Zeiss Ultra Plus was used.

[0067] X-ray diffraction analysis: Bruker's D2 Phaser was used.

[0068] N2 adsorption / desorption isotherm: Belsorp Mini X from Microtaac was used.

[0069]

[0070] Manufacturing Example 1: Synthesis of PCN-223

[0071] 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 the solution and stirred. The mixture was then 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.

[0072] The results of measuring the obtained PCN-223 using a scanning electron microscope (Zeiss, Ultra Plus) are shown in Fig. 3.

[0073] The results of X-ray diffraction analysis (Bruker, D2 Phaser) of PCN-223 are shown in Fig. 4.

[0074] The results of N2 adsorption / desorption isotherm measurement of PCN-223 are shown in Figure 5 (Microtaac, Belsorp Mini X).

[0075] The surface area of ​​PCN-223 is 2273.6 m 2 / g, the pore size (Pore Diameter) was measured to be 1.42 nm.

[0076] The results of measuring the Zeta Potential of PCN-223 are shown in Fig. 6.

[0077]

[0078] Example 1: Preparation of PCN-223 loaded with ARV-771

[0079] PCN-223 manufactured according to Manufacturing Example 1 was placed in a vacuum oven at 150°C and pretreated 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 placed 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 obtained by drying at 80°C.

[0080] The SEM measurement results of PCN-223 loaded with ARV-771 are shown in Fig. 8.

[0081] The FTIR measurement results of PCN-223 loaded with ARV-771 are shown in Fig. 9.

[0082]

[0083] Manufacturing Example 2: Synthesis of PCN-224

[0084] 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. The mixture was then reacted at 125°C for 24 hours. The particles obtained by the reaction were washed three times with DMF and three times with ethanol, and then dried at 80°C to obtain a powder.

[0085] The results of measuring the obtained PCN-224 using a scanning electron microscope are shown in Fig. 10.

[0086] The results of X-ray diffraction analysis (Bruker, D2 Phaser) of PCN-224 are shown in Fig. 11.

[0087] The results of N2 adsorption / desorption isotherm measurement of PCN-224 are shown in Figure 12 (Microtaac, Belsorp Mini X).

[0088] The surface area of ​​PCN=224 is 1894 m2 / g, the pore size (Pore Diameter) was measured to be 1.61 nm.

[0089] The results of measuring the Zeta Potential of PCN-224 are shown in Fig. 13.

[0090]

[0091] Example 2: Preparation of PCN-224 loaded with ARV-771

[0092] PCN-224 synthesized by Manufacturing 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 obtained by drying at 80°C.

[0093] The SEM measurement results of PCN-224 loaded with ARV-771 are shown in Fig. 14.

[0094] The FTIR measurement results of PCN-224 loaded with ARV-771 are shown in Fig. 15.

[0095]

[0096] Manufacturing Example 3: Synthesis of MOF-525

[0097] 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. The mixture was then 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.

[0098] The results of measuring the obtained MOF-525 using a scanning electron microscope are shown in Fig. 16.

[0099] The results of X-ray diffraction analysis of MOF-525 are shown in Fig. 17.

[0100] The results of N2 adsorption / desorption isotherm measurement of MOF-525 are shown in Figure 18 (Microtaac, Belsorp Mini X).

[0101] The surface area of ​​MOF-525 is 1356.3 m 2 / g, the pore size (Pore Diameter) was measured to be 1.68 nm.

[0102] The Zeta Potential measurement results of MOF-525 are shown in Fig. 19.

[0103]

[0104] Example 3: Preparation of MOF-525 loaded with ARV-771

[0105] MOF-525 synthesized by Manufacturing 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 obtained by drying in an oven at 80°C.

[0106] The SEM measurement results of MOF-525 loaded with ARV-771 are shown in Fig. 20.

[0107] The FTIR measurement results of MOF-525 loaded with ARV-771 are shown in Fig. 21.

[0108]

[0109] Manufacturing Example 4: Synthesis of MOF-545

[0110] 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. 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.

[0111] The results of measuring the obtained MOF-545 using a scanning electron microscope are shown in Fig. 22.

[0112] The results of X-ray diffraction analysis of MOF-545 are shown in Fig. 23.

[0113] The results of the N2 adsorption / desorption isotherm measurement of MOF-545 are shown in Figure 24.

[0114] The surface area of ​​MOF-545 is 1082.2 m 2 / g, the pore size (Pore Diameter) was measured to be 3.09 nm.

[0115] The Zeta Potential measurement results of MOF-545 are shown in Fig. 25.

[0116]

[0117] Example 4: Preparation of MOF-545 loaded with ARV-771

[0118] MOF-545 synthesized in 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 containing ARV-771 dissolved in DMSO at a concentration of 1 mg / ml was prepared, 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.

[0119] The SEM measurement results of MOF-545 loaded with ARV-771 are shown in Fig. 26.

[0120] The FTIR measurement results of MOF-545 loaded with ARV-771 are shown in Fig. 27.

[0121]

[0122] Manufacturing Example 5: Synthesis of UIO-67

[0123] 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.

[0124] The results of measuring the obtained UIO-67 using a scanning electron microscope are shown in Fig. 28.

[0125] The results of X-ray diffraction analysis of UIO-67 are shown in Fig. 29.

[0126] The results of the N2 adsorption / desorption isotherm measurement of UIO-67 are shown in Figure 30.

[0127] The surface area of ​​UIO-67 is 2192.8 m 2 / g, the pore size (Pore Diameter) was measured to be 1.88 nm.

[0128] The results of measuring the Zeta Potential of UIO-67 are shown in Fig. 31.

[0129]

[0130] Example 5: Manufacturing of UIO-67 loaded with ARV-771

[0131] UIO-67 manufactured by Manufacturing 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.

[0132] The SEM measurement results of UIO-67 equipped with ARV-771 are shown in Fig. 32.

[0133] The FTIR measurement results of UIO-67 equipped with ARV-771 are shown in Fig. 33.

[0134]

[0135] Experimental Example 1: Cytotoxicity Test Method

[0136] (1) Mouse fibroblasts (L929) were placed in a 96-well plate at a density of 11,000 cells per well and cultured in a 37°C, 5% CO2 incubator for 24 hours.

[0137] (2) MOF suspensions were prepared at concentrations of 100, 50, 25, 10, and 1 ug / ml using cell culture medium.

[0138] (3) The existing medium of each well plate cell was removed, and a suspension was prepared by diluting the metal-organic framework particles to be tested in the medium at concentrations of 100, 50, 25, 10, and 1 ug / ml, and 100 ul was added to each well.

[0139] (4) Cultured for 24 hours in a 37°C, 5% CO2 incubator.

[0140] (5) The suspension was removed from the well and washed with PBS.

[0141] (6) 100 ul of 10% Ez-Cytox medium was added to each well.

[0142] (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)

[0143] The cytotoxicity test results of PCN-223, PCN-224, MOF-525, MOF-545, and UIO-67 are shown in Figures 34 to 38, respectively.

[0144]

[0145] Experimental Example 2: Measuring drug loading

[0146] (1) 10 mg of ARV-771 loaded MOF was weighed and placed in an amber vial.

[0147] (2) 20 ml of PBS (pH7.4) was added to the measured sample.

[0148] (3) Incubated for 72 hours in a shaking incubator (RPM 100) at 37.5 degrees Celsius.

[0149] (4) After 72 hours, the vial was removed and treated with ultrasonic equipment for 6 hours to completely decompose it.

[0150] (5) The sample, which was completely decomposed and from which the drug was released, was centrifuged at 14,000 RPM for 10 minutes and the supernatant was collected.

[0151] (6) The total amount of drug was quantitatively measured using HPLC from the obtained supernatant.

[0152] The results of analyzing the payload of ARV-771 are shown in Table 1 below.

[0153] Sample nameARV-771 Payload (ug / mg)ARV-771 loaded PCN-22323.61ARV-771 loaded PCN-22429.14ARV-771 loaded MOF-52532.53ARV-771 loaded MOF-54521.44ARV-771 loaded UIO-6728.37

[0154]

[0155] Experimental Example 3: Evaluation of the Cancer Cell Killing Ability of ARV-771-Loaded MOF

[0156] To evaluate the drug delivery efficacy of MOF loaded with PROTAC drugs, the cancer cell killing ability was evaluated using MC38, a representative cancer cell line.

[0157] Specifically, as examples of ARV-771-loaded metal-organic frameworks, five types of metal-organic frameworks manufactured / synthesized in the above examples were applied, and a control group containing only ARV-771 was also applied for testing.

[0158] 100 ul 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, the control group containing only ARV-771 and the 5 MOF experimental groups loaded with ARV-771 were applied at the same drug concentration standard of 2 ug / ml, and the concentration was applied equally per well of 96-well standard at 200 ng / well, N=8. Culture was performed in a CO2 incubator for 24 hours. After 24 hours, the growth rate of cancer cells was evaluated compared to the untreated group by analyzing the absorbance at 450 nm and 600 nm using a Microplate reader using EZ-Cytox reagent.

[0159] As a result of the cancer cell killing ability evaluation, the cancer cell survival rate in the ARV-771-only treatment group used as a comparison group for each ARV-771-loaded metal-organic framework was 79.2%, 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 than when ARV-771 was used alone.

[0160] Table 2 below shows the results of killing ability of ARV-771 loaded MOF MC38 colon cancer cell line.

[0161] MC38 survival rate (%) compared to the untreated group ARV-771 (2 ug / ml) 86.5 ARV-771 loaded PCN-22347.1 ARV-771 loaded PCN-22458.3 ARV-771 loaded MOF-52539.6 ARV-771 loaded MOF-54545.1 ARV-771 loaded UIO-6734.5

[0162]

[0163] The assignment information is as follows:

[0164] [Project ID] 1345370811

[0165] [Project Number] 2021RIS-001

[0166] [Ministry Name] Ministry of Education (P13)

[0167] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0168] [Research Project Name] Local Government-University Cooperation-Based Regional Innovation Project

[0169] [Research Project Name] Local Government-University Cooperation-Based Regional Innovation Project (Chungbuk Regional Innovation Platform)

[0170] [Name of Project Performing Organization] (Chungbuk Regional Innovation Platform) Chungbuk National University

[0171] [Research Period] March 1, 2024 - February 28, 2025

[0172] The present invention relates to a drug delivery system comprising metal-organic framework particles loaded with Protac and a pharmaceutical composition comprising the same.

Claims

1. A drug delivery system comprising metal-organic framework particles loaded with Protac.

2. In paragraph 1, The above metal-organic framework is a drug delivery system formed by the bonding of an organic ligand and a metal ion.

3. In paragraph 2, A drug delivery system wherein the organic ligand has a carboxyl or amine group at both terminals.

4. In paragraph 2, The above organic ligand is a drug delivery system comprising a porphyrin ligand.

5. In paragraph 1, The above metal-organic frameworks are 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-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, Fe-MIL-100, A drug delivery system comprising at least one metal-organic framework selected from the group consisting of Fe-MIL-101, Fe-MIL-101-NH2, Fe-MIL-125 and Fe-MIL-125-NH2.

6. In paragraph 1, A drug delivery system, wherein the metal-organic framework comprises one or more metals or metal ions selected from the group consisting of aluminum (Al), iron (Fe), and zirconium (Zr).

7. A pharmaceutical composition for treating or preventing cancer, comprising metal-organic framework particles loaded with protac as an active ingredient.

8. In paragraph 7, A pharmaceutical composition wherein the cancer is 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.

9. A pharmaceutical composition for treating or preventing neurodegenerative diseases, comprising metal-organic framework particles loaded with protac as an active ingredient.

10. A pharmaceutical composition for treating or preventing an autoimmune disease, comprising a metal-organic framework particle loaded with a protac as an active ingredient.

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