Carrier comprising metal-organic framework and lipid, and use thereof

Abstract

The present invention relates to a metal-organic framework (MOF)-lipid complex and a use thereof, the MOF-lipid complex comprising a metal-organic framework (MOF) and a lipid. It has been confirmed that the MOF-lipid complex or a carrier comprising same of the present invention can effectively deliver active ingredients including nucleic acids such as RNA, and thus can be used as a drug carrier of ingredients such as nucleic acids.

Description

Carriers comprising metal-organic frameworks and lipids and uses thereof

[0001] The present invention relates to a metal-organic framework (MOF)-lipid complex comprising a metal-organic framework (MOF) and a lipid, and uses thereof.

[0002] Even for the same drug, efficacy can vary significantly depending on the delivery method. Thus, a drug delivery system (DDS) is a drug delivery system (DDS) that efficiently delivers the required amount of medication while minimizing side effects and maximizing efficacy and effectiveness. In the pharmaceutical industry, DDS is a high-value, core technology with a high potential for success, generating economic benefits comparable to new drug development.

[0003] Recently, the development of gene therapy using gene drug delivery systems has been expanding among various drug delivery systems. For gene therapy to be successful and safe, it is crucial to deliver genes or gene regulatory factors to the target tissue. Representative examples of these genes or gene regulatory factors include mRNA, siRNA, and miRNA. Nucleic acids such as mRNA function to express specific proteins, allowing them to complement proteins deficient in genetic factors. Furthermore, the use of mRNA to express cancer markers or viral surface proteins enables the development of anticancer drugs and vaccines that activate the immune response in vivo. Furthermore, nucleic acids such as siRNA and miRNA, which can suppress the expression of specific proteins in vivo, are attracting attention as important tools for the treatment of cancer, genetic diseases, infectious diseases, and autoimmune diseases. However, direct delivery of these nucleic acids into cells is difficult and they are easily degraded by enzymes in the blood. Therefore, extensive research is being conducted to overcome this limitation.

[0004] Non-viral gene carriers, such as cationic liposomes and polymers, are attracting attention for the effective delivery of these substances. In particular, lipid nanoparticles have been actively studied as non-viral gene delivery vehicles. Lipid nanoparticles (LNPs) utilize substances existing in the body, such as phospholipids and cholesterol, and are thus particulate drug delivery vehicles with high bioavailability and affinity, as well as the ability to release and control drugs, and high stability against degradation by enzymes.

[0005] Accordingly, the inventors of the present invention confirmed that the delivery efficiency of lipid nanoparticles can be improved by applying a metal-organic framework, thereby improving the delivery efficiency, and completed the present invention.

[0006] One aspect provides a MOF-lipid complex comprising 1) a metal-organic framework (MOF); and 2) one or more lipid components.

[0007] Another aspect provides a drug delivery complex comprising the MOF-lipid complex and a drug of interest.

[0008] Another aspect provides a drug delivery composition comprising the drug delivery complex.

[0009] Another aspect is to provide a pharmaceutical composition comprising the drug delivery complex.

[0010] One aspect is to provide a MOF-lipid complex comprising 1) a metal-organic framework (MOF); and 2) one or more lipid components.

[0011] The term "metal-organic framework (MOF)" as used herein 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, MOFs not only have a very large surface area but also have an open pore structure, so they 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 medium, they have the advantage of having many active sites, which can maximize efficiency. In addition, the MOF does not easily deform at high temperatures and has a rigid skeleton, so it has excellent chemical and thermal stability.

[0012] The above metal-organic framework is made of aluminum (Al, Al 2+ , Al 3+ ), iron (Fe, Fe 2+ , Fe 3+ ), zinc (Zn, Zn 2+ , Zn 3+ ), magnesium (Mg, Mg 2+ , Mg 3+ ) and zirconium (Zr, Zr 2+ , Zr 3+ , Zr 4+) may be composed of one or more metals or metal ions thereof, and may be synthesized using the selected metal or metal ion, and specifically may be composed of the metal or metal ion and an organic ligand coordinated thereto. The metal portion (center metal) of the metal-organic framework is aluminum (Al, Al 3+ ), iron (Fe, Fe 2+ , Fe 3+ ), zinc (Zn, Zn 2+ , Zn 3+ ), magnesium (Mg, Mg 2+ , Mg 3+ ) and zirconium (Zr, Zr 4+ ) may be composed of one or more metals or metal ions thereof selected from the group consisting of:

[0013] In one embodiment, the metal-organic framework can be a magnesium-based MOF, an aluminum-based MOF, a zinc-based MOF, an iron-based MOF, and / or a zirconium-based MOF.

[0014] The above metal-organic framework is made of aluminum (Al, Al 3+ ), iron (Fe, Fe 2+ , Fe 3+ ), zinc (Zn, Zn 2+ , Zn 3+ ), magnesium (Mg, Mg 2+ , Mg 3+ ) and zirconium (Zr, Zr 4+) and one or more metals or metal ions thereof selected from the group consisting of 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)benzene, 4,4,4˝-s-triazine-2,4,4,4,4˝-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, 1,3,5-tris(4-carboxyphenyl)benzene, It may include one or more organic ligands selected from the group consisting of 1,3,5-triscarboxyphenylethynylbenzene, 2-methylimidazole,

[0015] The above metal-organic framework is UIO-66[C 48 H 28 O 32 Zr6], UIO-66-NH2[C 48 H 34 N6O 32 Zr6], UIO-66-ED[C 50 H 36 O 32 N2Zr6], UIO-67[C 84 H 52 O 32 Zr6], UIO-67-NH2[C 84 H 58 N6O 32 Zr6], UIO-67-ED[C 86 H 60 O 32 N2Zr6], PCN-128[C 108 H 76 O 30 Zr6], PCN-222[C 96 H 68 N8O 32 Zr6], PCN-223[C144 H 82 N 12 O 32 Zr6], PCN-224[C 144 H 112 N 12 O 64 Zr 15 ], MOF-525[C 144 H 88 N 12 O 32 Zr6], MOF-545[C 96 H 72 N8O 32 ], MOF-801[C 24 H 16 O 32 Zr6], MOF-808[C 24 H 16 O 32 Zr6], MOF-867[C 72 H 40 N 12 O 32 Zr6], Al-MIL-53[C8H5AlO5], Al-MIL-53-NH2[C8H6NO5Al], Al-MIL-88[C 24 H 12 Al3O 13 ], Al-MIL-88-NH2[C 24 H 15 Al3N3O 13 ], Al-MIL-100[C9H6O6Al], Al-MIL-100-NH2[C9H7NO6Al], Al-MIL-101[C 24 H 12 ClAl3O 13 ], Al-MIL-101-NH2[C 24 H 19 Al3ClN3O 15 ], Al-MIL-125[C 48 H 28 O 36 Al8], Al-MIL-125-NH2[C 48 H 34 N6O 36 Al8], Fe-MIL-53[C8H5FeO5], Fe-MIL-53-NH2[C8H6FeNO5], Fe-MIL-88[C 24 H 12Fe3O 13 ], Fe-MIL-88-NH2[C 24 H 15 Fe3N3O 13 ], Fe-MIL-100[C9H6O6Fe], Fe-MIL-100-NH2[C9H7NO6Fe], Fe-MIL-101[C 24 H 12 ClFe3O 13 ], Fe-MIL-101-NH2[C 24 H 19 Fe3ClN3O 15 ] , Fe-MIL-125[C 48 H 28 O 36 Fe8], Fe-MIL-125-NH2[C 48 H 34 N6O 36 Fe8], Mg-MOF-74[C8H4O8Mg2], Mg-MOF-1, Mg2(dobdc) [C 14 H6O6Mg2], Mg-CUK-1, MOF-5 [C 24 H 12 O 13 Zn4], ZIF-8 [C8H 12 N4Zn], IRMOF-3 [C 24 H 15 N3O 13 Zn4], Zn-BTC, Zn-MOF-74[C8H4O8Zn2] and derivatives thereof, but is not limited thereto.

[0016] Meanwhile, the UIO-66-ED is a metal-organic framework obtained by post-modifying UIO-66 with ethylene diamine, and has a structure represented by the following chemical formula 1.

[0017] [Chemical Formula 1]

[0018]

[0019] In one embodiment, the aluminum-based MOF may include, but is not limited to, one or more selected from the group consisting of Al-MIL-53, Al-MIL-53-NH2, Al-MIL-88, Al-MIL-88-NH2, Al-MIL-100, Al-MIL-100-NH2, Al-MIL-101, Al-MIL-101-NH2, Al-MIL-125, Al-MIL-125-NH2, and derivatives thereof.

[0020] In one embodiment, the iron-based MOF may include, but is not limited to, one or more selected from the group consisting of Fe-MIL-53, Fe-MIL-53-NH2, Fe-MIL-88, Fe-MIL-88-NH2, Fe-MIL-100, Fe-MIL-100-NH2, Fe-MIL-101, Fe-MIL-101-NH2, Fe-MIL-125, Fe-MIL-125-NH2, and derivatives thereof.

[0021] In one embodiment, the zirconium-based MOF may include, but is not limited to, one or more selected from the group consisting of UIO-66, UIO-66-ED, UIO-66-NH2, UIO-67, UIO-67-ED, UIO-67-NH2, PCN-128, PCN-222, PCN-223, PCN-224, MOF-525, MOF-545, MOF-801, MOF-808, MOF-867 and derivatives thereof.

[0022] In one embodiment, the magnesium-based MOF may include, but is not limited to, one or more selected from the group consisting of Mg-MOF-74, Mg-MOF-1, Mg2(dobdc), Mg-CUK-1, and derivatives thereof.

[0023] In one embodiment, the zinc-based MOF may include, but is not limited to, one or more selected from the group consisting of MOF-5, ZIF-8, IRMOF-3, Zn-BTC, Zn-MOF-74, and derivatives thereof.

[0024] In one embodiment, the metal-organic framework may include at least one selected from the group consisting of MOF-808, MIL-101-NH2(Al), Mg-MOF-74, UIO-67, UIO-66-ED, and MIL-88-NH2(Fe).

[0025] The one or more lipid components may include one or more selected from the group consisting of phospholipids, sterols, PEG (polyethylene glycol)-lipids, and ionizable lipids (or cationic lipids).

[0026] The phospholipids include DOPE (dioleoylphosphatidylethanolamine; 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine), DSPC (distearoylphosphatidylcholine), POPC (palmitoyloleoylphosphatidylcholine; 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine), EPC (egg) phosphatidylcholine), DOPC (dioleoylphosphatidylcholine), DPPC (dipalmitoylphosphatidylcholine), DOPG (dioleoylphosphatidylglycerol), DPPG (dipalmitoylphosphatidylglycerol), DSPE (distearoylphosphatidylethanolamine), PE (phosphatidylethanolamine, DPPE (dipalmitoylphosphatidylethanolamine), POPE (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoethanolamine), It may include, but is not limited to, one or more selected from the group consisting of DOPS (1,2-dioleoyl-sn-glycero-3-[phospho-L-serine]) and sphingomyelin.

[0027] The above ionized lipid may be a cationic lipid.

[0028] The cationic lipid may include, but is not limited to, one or more selected from the group consisting of DOTAP (1,2-dioleoyl-3-trimethylammonium propane), DOTMA (N-[1-(2,3-dioleyloxy)propyl]-N,N,N-trimethylammonium chloride), DOEPC (1,2-dioleoyl-sn-glycero-3-ethylphosphocholine), MC3 (dilinoleyl-methyl-4-dimethylaminobutyrate), and SM102 (heptadecan-9-yl 8-[2-hydroxyethyl-(6-oxo-6-undecoxyhexyl)amino]octanoate).

[0029] The above PEG-lipid (PEGylated lipid or lipid containing PEG) may include at least one selected from the group consisting of DMG (dimyristoyl glycerol)-PEG and DSPE (1,2-distearoyl-sn-glycero-3-phosphoethanolamine)-PEG, but is not limited thereto.

[0030] The above sterol may include at least one selected from the group consisting of animal sterols and derivatives such as cholesterol, cholesteryl chloride, cholesteryl octanoate, cholesteryl nonanoate, cholesteryl oleyl carbonate, and cholesteryl isostearyl carbonate, and plant sterols and derivatives such as phytosterol, campesterol, sitosterol, and stigmasterol.

[0031] In one embodiment, one or more lipid components included in the MOF-lipid complex may not include PEG-lipid. Accordingly, one or more lipid components included in the MOF-lipid complex may include one or more selected from the group consisting of phospholipids, sterols, and ionizable lipids.

[0032] In the above MOF-lipid complex, the entire lipid components (one or more lipid components) and the metal-organic framework included in the complex may be included in a mass ratio (entire lipid components:metal-organic framework) of 1:1 to 15:1, specifically 1:1 to 15:1, 1:1 to 13:1, 1:1 to 12.5:1, 1:1 to 12.3:1, 1:1 to 10:1, 1:1 to 8:1, 1:1 to 6:1, 1:1 to 5:1, 1:1 to 4:1, 1:1 to 3.5:1, 1:1 to 3:1, 1:1 to 2:1, 1:1 to 1.5:1, 1.5:1 to 15:1, 1.5:1 to 13:1, 1.5:1 to It may be included in a mass ratio of 12.5:1, 1.5:1 to 12.3:1, 1.5:1 to 10:1, 1.5:1 to 8:1, 1.5:1 to 6:1, 1.5:1 to 5:1, 1.5:1 to 4:1, 1.5:1 to 3.5:1, 1.5:1 to 3:1, or 1.5:1 to 2:1.

[0033] In one embodiment, the MOF-lipid complex comprises a metal-organic framework and one or more lipid components, wherein the one or more lipid components comprise an ionizable lipid, a phospholipid, and a sterol, and may not comprise a PEG-lipid. Specifically, in the MOF-lipid complex, based on 100 parts by weight of the total lipid components (one or more lipid components) included in the MOF-lipid complex, the ionized lipid is 40 to 85 parts by weight, 40 to 80 parts by weight, 40 to 75 parts by weight, 40 to 70 parts by weight, 40 to 65 parts by weight, 40 to 63 parts by weight, 50 to 85 parts by weight, 50 to 80 parts by weight, 50 to 75 parts by weight, 50 to 70 parts by weight, 50 to 65 parts by weight, 50 to 63 parts by weight, 55 to 85 parts by weight, 55 to 80 parts by weight, 55 to 75 parts by weight, 55 to 70 parts by weight, 55 to 65 parts by weight, 55 to 63 parts by weight, 60 to 85 parts by weight, 60 to 80 parts by weight, 60 to 75 parts by weight, 60 to 70 parts by weight, 60 to 65 parts by weight or 60 to 63 parts by weight, the phospholipid may be included in 5 to 40 parts by weight, 5 to 30 parts by weight, 5 to 25 parts by weight, 5 to 20 parts by weight, 5 to 15 parts by weight, 5 to 13 parts by weight, 10 to 40 parts by weight, 10 to 30 parts by weight, 10 to 25 parts by weight, 10 to 20 parts by weight, 10 to 15 parts by weight or 10 to 13 parts by weight, the sterol may be included in 10 to 50 parts by weight, 10 to 40 parts by weight, 10 to 35 parts by weight, 10 to 30 parts by weight, 10 to 25 parts by weight, 15 to 50 parts by weight, 15 to 40 parts by weight, 15 to It may be included in an amount of 35 parts by weight, 15 to 30 parts by weight, 15 to 25 parts by weight, 20 to 50 parts by weight, 20 to 40 parts by weight, 20 to 35 parts by weight, 20 to 30 parts by weight, or 20 to 25 parts by weight.

[0034] In one embodiment, the MOF-lipid complex comprises a metal-organic framework and one or more lipid components, wherein the one or more lipid components comprise an ionizable lipid and a sterol, and may not comprise a PEG-lipid. Specifically, in the MOF-lipid complex, based on 100 parts by weight of the total lipid components (one or more lipid components) included in the MOF-lipid complex, the ionized lipid is 50 to 90 parts by weight, 50 to 85 parts by weight, 50 to 80 parts by weight, 50 to 77 parts by weight, 60 to 90 parts by weight, 60 to 85 parts by weight, 60 to 80 parts by weight, 60 to 77 parts by weight, 70 to 90 parts by weight, 70 to 85 parts by weight, 70 to 80 parts by weight, 70 to 77 parts by weight, 70 to 90 parts by weight, 70 to 85 parts by weight, 70 to 80 parts by weight, 70 to 77 parts by weight, 75 to 90 parts by weight, 75 to 85 parts by weight, 75 to 80 parts by weight, or 75 to 77 The sterol may be included in an amount of 10 to 50 parts by weight, 10 to 40 parts by weight, 10 to 35 parts by weight, 10 to 30 parts by weight, 10 to 25 parts by weight, 15 to 50 parts by weight, 15 to 40 parts by weight, 15 to 35 parts by weight, 15 to 30 parts by weight, 15 to 25 parts by weight, 20 to 50 parts by weight, 20 to 40 parts by weight, 20 to 35 parts by weight, 20 to 30 parts by weight, or 20 to 25 parts by weight.

[0035] In one embodiment, the MOF-lipid complex comprises a metal-organic framework and one or more lipid components, wherein the one or more lipid components comprise an ionizable lipid and a phospholipid, and may not comprise a PEG-lipid. Specifically, in the MOF-lipid complex, the ionized lipid may be included in an amount of 60 to 95 parts by weight, 60 to 92 parts by weight, 60 to 90 parts by weight, 60 to 88 parts by weight, 70 to 95 parts by weight, 70 to 92 parts by weight, 70 to 90 parts by weight, 70 to 88 parts by weight, 80 to 95 parts by weight, 80 to 92 parts by weight, 80 to 90 parts by weight, 80 to 88 parts by weight, 85 to 95 parts by weight, 85 to 92 parts by weight, 85 to 90 parts by weight, or 85 to 88 parts by weight, based on 100 parts by weight of the total lipid component (one or more lipid components) included in the MOF-lipid complex, and the phospholipid may be included in an amount of 5 to 40 parts by weight, 5 to 30 parts by weight, 5 to 25 parts by weight, 5 to It may be included in an amount of 20 parts by weight, 5 to 15 parts by weight, 5 to 13 parts by weight, 10 to 40 parts by weight, 10 to 30 parts by weight, 10 to 25 parts by weight, 10 to 20 parts by weight, 10 to 15 parts by weight, or 10 to 13 parts by weight.

[0036] In one embodiment, the MOF-lipid complex may comprise a metal-organic framework and an ionizable lipid, and may not comprise a PEG-lipid.

[0037] The complex may be intended to deliver an active ingredient or bioactive substance capable of exhibiting pharmacological activity within a cell and / or in vivo. Specifically, the delivery system may be intended to deliver a desired drug. Furthermore, the active ingredient or bioactive substance may comprise a nucleic acid.

[0038]

[0039] Another aspect is to provide 1) the MOF-lipid complex; and 2) a drug delivery complex comprising a desired drug. The same parts described above also apply to the complex.

[0040] The term "drug delivery complex", "drug carrier" or "drug delivery system (DDS)" in this specification refers to a series of technologies or substances that control the delivery and release of pharmacologically active substances into cells, tissues, organs and systems to achieve optimal efficacy by utilizing various physicochemical technologies, and refers to a technology that optimizes drug treatment by designing a formulation to efficiently deliver the required amount of drugs while minimizing the side effects of existing drugs and maximizing their efficacy and effectiveness. The use of the drug delivery system technology has various advantages, such as being able to effectively deliver therapeutic substances, increasing the amount of drugs reaching the target site, enabling more effective treatment, and reducing side effects by preventing delivery to areas other than the target site.

[0041] The above complex may be for delivering a desired drug into a living body and / or into a cell.

[0042] The above drug may be mutually bonded to the MOF and / or lipid within the complex, or may be mixed with each other without being mutually bonded to form a non-covalent complex, and may be applied without limitation as long as it can improve drug delivery efficiency without inhibiting the pharmacological activity of the drug.

[0043] The drug is loaded into the delivery complex, and the drug may be bound or linked to a lipid component or may be encapsulated therein.

[0044] The above drug is a substance capable of exhibiting pharmacological activity within a cell and / or in a living body, and may include at least one selected from the group consisting of nucleic acids, proteins and compounds, and may specifically be at least one selected from the group consisting of nucleic acids, peptides, proteins and protein-nucleic acid structures, and more specifically may be a nucleic acid.

[0045] The nucleic acid may include at least one selected from the group consisting of single-stranded siRNA, double-stranded siRNA, rRNA, RNA, DNA, cDNA, plasmid, aptamer, mRNA, tRNA, lncRNA, piRNA, circRNA, saRNA, antisense oligonucleotide, shRNA, miRNA, ribozyme, PNA, microRNA, ASO, gapmer, antagomir, agomir, and DNAzyme.

[0046] The above drug may include a therapeutic agent and / or a vaccine.

[0047] The above vaccine is for preventing or treating bacterial and / or viral infection or infectious disease, and the vaccine may be selected from the group consisting of coronavirus vaccine, Japanese encephalitis vaccine, Hemophilus influenzae type B vaccine, MERS vaccine, Zika vaccine, Pseudomonas aeruginosa vaccine, cancer vaccine, tuberculosis vaccine, anthrax vaccine, HAV vaccine, HBV vaccine, HCV vaccine, HIV vaccine, herpes simplex vaccine, meningitis vaccine, diphtheria vaccine, pertussis vaccine, tetanus vaccine, varicella vaccine, multidrug-resistant bacteria vaccine, enterococcus vaccine, staphylococcus vaccine, Klebsiella pneumoniae vaccine, Acinetobacter vaccine, Enterobacter vaccine, Helicobacter vaccine, malaria vaccine, dengue fever vaccine, tsutsugamushi vaccine, severe fever with thrombocytopenia syndrome vaccine, SARS vaccine, Ebola vaccine, influenza vaccine, COVID-19 vaccine, and pneumococcal vaccine, but It is not limited.

[0048] The above drug (specifically, a nucleic acid) may form a complex with the MOF.

[0049] In the drug delivery complex, the MOF and the drug (specifically, nucleic acid) may be included in a mass ratio (MOF:drug) of 3:1 to 50:1, specifically 3:1 to 50:1, 3:1 to 40:1, 3:1 to 35:1, 3:1 to 30:1, 3:1 to 26:1, 3:1 to 25:1, 3:1 to 20:1, 3:1 to 15:1, 3:1 to 14:1, 3:1 to 13:1, 3:1 to 10:1, 3:1 to 8:1, 3:1 to 5:1, 3:1 to 4:1, 5:1 to 50:1, 5:1 to 40:1, 5:1 to 35:1, 5:1 to 30:1, It may be included in a mass ratio of 5:1 to 26:1, 5:1 to 25:1, 5:1 to 20:1, 5:1 to 15:1, 5:1 to 14:1, 5:1 to 13:1, 5:1 to 10:1, 5:1 to 8:1, 10:1 to 50:1, 10:1 to 40:1, 10:1 to 35:1, 10:1 to 30:1, 10:1 to 26:1, 10:1 to 25:1, 10:1 to 20:1, 10:1 to 15:1, 10:1 to 14:1, or 10:1 to 13:1.

[0050]

[0051] Another aspect provides a drug delivery composition comprising the drug delivery complex. The same aspects described above also apply to the composition.

[0052]

[0053] Another aspect is the use of the drug delivery complex for drug delivery. The same principles described above apply to this use.

[0054]

[0055] Another aspect provides a method for delivering a desired drug to a subject, comprising administering the drug delivery complex to the subject. The same aspects described above also apply to the method.

[0056] In one embodiment, the method may be for delivering a desired drug to a target cell, tissue, organ and / or system of an individual using the drug delivery complex of the present disclosure.

[0057]

[0058] Another aspect provides a pharmaceutical composition comprising the drug delivery complex. The same principles as described above apply to the composition.

[0059] The above composition may be a composition for a vaccine.

[0060] 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 the adaptability of the subject of application (prescription). Any carrier commonly used in the art and pharmaceutically acceptable for the pharmaceutical composition may be used. 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 can be formulated and used in the form of oral formulations such as powders, granules, tablets, capsules, suspensions, emulsions, syrups, aerosols, external preparations, suppositories, or sterile injection solutions, each according to a conventional method.

[0061] When formulating the above pharmaceutical composition, it may be prepared using a diluent or excipient such as a generally used filler, bulking agent, binder, wetting agent, disintegrant, or surfactant, but may not be limited thereto.

[0062] When the above pharmaceutical composition is manufactured into an oral dosage form, it can be manufactured into a dosage form such as powder, granules, tablets, pills, dragees, capsules, liquids, gels, syrups, suspensions, wafers, etc., using a suitable carrier according to a method known in the art. At this time, examples of suitable pharmaceutically acceptable carriers include sugars such as lactose, glucose, sucrose, dextrose, sorbitol, mannitol, xylitol, etc.; starches such as corn starch, potato starch, and wheat starch; celluloses such as cellulose, methylcellulose, ethylcellulose, sodium carboxymethylcellulose, and hydroxypropylmethylcellulose; polyvinyl pyrrolidone, water, methylhydroxybenzoate, propylhydroxybenzoate, magnesium stearate, mineral oil, malt, gelatin, talc, polyols, and vegetable oils. In case of formulation, the formulation may include diluents and / or excipients such as fillers, bulking agents, binders, wetting agents, disintegrants, and surfactants, as needed.

[0063] When the above pharmaceutical composition is prepared as a parenteral dosage form, it can be formulated in the form of injections, transdermal administration, nasal inhalation, and suppositories using a suitable carrier according to a method known in the art. When formulated as an injection, suitable carriers include sterile water, ethanol, polyols such as glycerol or propylene glycol, or mixtures thereof, and preferably, Ringer's solution, phosphate buffered saline (PBS) containing triethanolamine, sterile water for injection, and isotonic solutions such as 5% dextrose can be used. When formulated as a transdermal dosage form, it can be formulated in the form of ointments, creams, lotions, gels, external solutions, pastes, liniments, aerosols, etc. In the case of nasal inhalation, it can be formulated in the form of an aerosol spray using a suitable propellant such as dichlorofluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane, or carbon dioxide, and in the case of formulating it as a suppository, the base can be witepsol, tween 61, polyethylene glycol, cacao butter, laurin butter, polyoxyethylene sorbitan fatty acid ester, polyoxyethylene stearate, sorbitan fatty acid ester, etc.

[0064] The pharmaceutical composition may be administered in a pharmaceutically effective amount. The term "pharmaceutically effective amount" refers to an amount sufficient to treat or prevent a disease at a reasonable benefit / risk ratio applicable to medical treatment or prevention, and the effective dosage level may be determined based on the severity of the disease, the activity of the drug, the patient's age, weight, health, sex, the patient's sensitivity to the drug, the time of administration of the composition of the present invention used, the route of administration and the excretion rate, the treatment period, the drug used in combination with or concurrently with the composition of the present invention used, and other factors well known in the medical field. The pharmaceutical composition of the present invention may be administered alone or in combination with a component known to exhibit a known therapeutic effect. It is important to take all of the above factors into consideration and administer an amount that can achieve the maximum effect with the minimum amount without side effects.

[0065] The dosage of the pharmaceutical composition may be determined by a person skilled in the art in consideration of the intended use, the degree of toxicity of the disease, the patient's age, weight, sex, medical history, or the type of substance used as the active ingredient. For example, the pharmaceutical composition of the present invention may be administered at about 0.1 ng to about 1,000 mg / kg, preferably 1 ng to about 100 mg / kg, per adult, and the frequency of administration of the composition of the present disclosure is not particularly limited thereto, but may be administered once a day or administered in divided doses several times. The dosage or frequency of administration does not limit the scope of the present disclosure in any way.

[0066] It has been confirmed that the MOF-lipid complex of the present invention or a carrier comprising the same can effectively deliver an active ingredient including a nucleic acid such as RNA, and thus can be used as a drug carrier for nucleic acids, etc.

[0067] Figure 1A shows the results of scanning electron microscopy observation of MOF-808; B shows the results of adsorption / desorption isotherm and specific surface area / pore measurement; and C shows the results of X-ray diffraction analysis.

[0068] Figure 2A is a drawing showing the results of scanning electron microscopy observation of MIL-101-NH2(Al); B is a drawing showing the results of adsorption / desorption isotherm and specific surface area / pore measurement; and C is a drawing showing the results of X-ray diffraction analysis.

[0069] Figure 3A shows the results of scanning electron microscopy observation of Mg-MOF-74; B shows the results of adsorption / desorption isotherm and specific surface area / pore measurement; and C shows the results of X-ray diffraction analysis.

[0070] Figure 4A is a drawing showing the results of scanning electron microscope observation of UIO-67; B is a drawing showing the results of adsorption / desorption isotherm and specific surface area / pore measurement; and C is a drawing showing the results of X-ray diffraction analysis.

[0071] Figure 5A is a drawing showing the results of scanning electron microscopy observation of UIO-66-ED; B is a drawing showing the results of adsorption / desorption isotherm and specific surface area / pore measurement; and C is a drawing showing the results of X-ray diffraction analysis.

[0072] Figure 6A is a drawing showing the results of scanning electron microscopy observation of MIL-88-NH2(Fe); B is a drawing showing the results of adsorption / desorption isotherm and specific surface area / pore measurement; and C is a drawing showing the results of X-ray diffraction analysis.

[0073] Figure 7 is a diagram showing the cytotoxicity results of metal-organic frameworks, where A represents MOF-808, B represents MIL-101-NH2(Al), C represents Mg-MOF-74, D represents UIO-67, E represents UIO-66-ED, and F represents MIL-88-NH2(Fe).

[0074] Figure 8 is a drawing showing the results of confirming the formation of a complex between a metal-organic framework and mRNA.

[0075] Figure 9 is a drawing showing the results of confirming the RNA delivery efficiency of a lipid carrier including MOF-808 as an example of a metal-organic framework.

[0076] Figure 10 is a drawing showing the results of confirming the RNA delivery efficiency of a lipid delivery system including UIO-66-ED as an example of a metal-organic framework.

[0077] Figure 11 is a diagram showing the results of confirming the RNA delivery efficiency of lipid carriers with various metal-organic framework content ratios.

[0078] The following examples and experimental examples are provided for more detailed explanation. However, these examples and experimental examples are provided for illustrative purposes only and the scope of the present invention is not limited to these examples and experimental examples.

[0079]

[0080] Experimental Example 1: Synthesis of a Metal-Organic Framework and Evaluation of Its Physicochemical Properties

[0081] Various types of metal-organic frameworks (MOFs) used in the present invention were produced / synthesized using the following methods, and their physicochemical properties were evaluated.

[0082]

[0083] 1-1: MOF-808

[0084] MOF-808 (C), a zirconium (Zr)-based metal-organic framework 24 H 16 O 32 To synthesize Zr6)(CAS No. 1579984-19-2), the following experiments were performed.

[0085] 1) Add 420 mg of trimesic acid (1,3,5-Benezenetricarboxylic acid) and 1.165 g of zirconium chloride to 64 ml of DMF and stir.

[0086] 2) Allow to react at 120℃ for 24 hours.

[0087] 4) The synthesized particles were washed three times each using DMF and ethanol, and dried at 80°C to obtain a powder.

[0088] Next, to confirm the physicochemical properties of the synthesized metal-organic framework, the following experiments were performed.

[0089] First, to measure with a scanning electron microscope (SEM), a 2-3 mg powder sample was placed in an Eppen tube, 100 μl of ethanol was added, and bath sonication was performed. After that, 10 μl was spotted on a silicon wafer and dried in an oven at 80°C. The dried silicon wafer was attached to an SEM mount using carbon tape, and then moved to the SEM equipment to measure images (Company name: Zeiss / Model name: ULTRA PLUS).

[0090] 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 2-30℃ range (Company name: Bruker / Model name: D2 phaser).

[0091] Next, for N2 adsorption isotherm and BET specific surface area-pore measurement, about 40 mg of completely dried sample was prepared and carefully placed into 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 measured in advance was subtracted to calculate the weight of the intact sample. The glass sampler was attached to the N2 adsorption-desorption device and measurements were performed (Company name: MICROTAAC / Model name: BELSORP MINI X).

[0092] Next, to measure the zeta potential, 3 mg of particles were weighed and placed in a vial, 6 mL of distilled water was added, and the particles were uniformly dispersed by ultrasonic treatment for 30 seconds. The sample, prepared at a concentration of 500 ug / mL, was injected into a zeta measurement cell, filled, and placed in a measuring device to measure the zeta potential (Company name: Malvern Panalytical / Model name: Zetasizer Nano).

[0093] As a result of confirming the physicochemical properties of the above-mentioned synthesized metal-organic framework, the zeta potential was +32.17 mV, and the results of SEM, X-ray diffraction, N2 adsorption isotherm, and BET specific surface area-pore measurement are shown in Fig. 1.

[0094]

[0095] 1-2: MIL-101-NH2(Al)

[0096] MIL-101-NH2(Al) (C), a metal-organic framework based on aluminum (Al) 24 H 19 Al3ClN3O 15 )(CAS No. 1404201-64-4), the following experiments were performed.

[0097] 1) Add 272 mg of 2-aminoterephthalic acid and 724 mg of aluminum chloride (AlCl3) to 60 ml of DMF and stir.

[0098] 2) React at 110℃ for 16 hours.

[0099] 4) The synthesized particles were washed three times each using DMF and ethanol, and dried at 80°C to obtain a powder.

[0100] Next, the physicochemical properties of the synthesized metal-organic framework were confirmed, and the zeta potential was +12.23 mV, and the SEM, X-ray diffraction, N2 adsorption isotherm, and BET specific surface area-pore measurement results are shown in Fig. 2.

[0101]

[0102] 1-3: Mg-MOF-74

[0103] To synthesize a magnesium (Mg)-based metal-organic framework, Mg-MOF-74 (CAS No. 1174268-74-6), the following experiments were performed.

[0104] 1) Dissolve 155 mg of 2,5-dihydroxyterephthalic acid and 665 mg of magnesium nitrate hexahydrate (MgNO3) in 62 ml of DMF.

[0105] 2) Allow to react at 125℃ for 24 hours.

[0106] 4) The synthesized particles were washed three times each using DMF and ethanol, and dried at 80°C to obtain a powder.

[0107] Next, the physicochemical properties of the synthesized metal-organic framework were confirmed, and the zeta potential was -23.7 mV, and the SEM, X-ray diffraction, N2 adsorption isotherm, and BET surface area-pore measurement results are shown in Fig. 3.

[0108]

[0109] 1-4: UIO-67

[0110] UIO-67 (C), a zirconium (Zr)-based metal-organic framework 84 H 52 O 32 To synthesize Zr6)(CAS No. 1072413-83-2), the following experiments were performed.

[0111] 1) Dissolve 240 mg of BPDC (Biphenyl-4,4-dicarboxylic acid) and 230 mg of ZrCl4 in 80 ml of DMF.

[0112] 2) React at 95℃ for 17 hours.

[0113] 4) The synthesized particles were washed three times each using DMF and ethanol, and dried at 80°C to obtain a powder.

[0114] Next, the physicochemical properties of the synthesized metal-organic framework were confirmed, and the zeta potential was -26.36 mV, and the SEM, X-ray diffraction, N2 adsorption isotherm, and BET specific surface area-pore measurement results are shown in Fig. 4.

[0115]

[0116] 1-5: UIO-66-ED

[0117] UIO-66-ED (C), a zirconium (Zr)-based metal-organic framework 50 H 36 O 32 In order to synthesize N2Zr6, the following experiments were performed. Meanwhile, the UIO-66-ED is a metal-organic framework obtained by post-modifying UIO-66 with ethylene diamine, and has a structure represented by the following chemical formula 1.

[0118] [Chemical Formula 1]

[0119]

[0120] 1) Dissolve 1.36 g of Benzoic acid, 11.2 g of BDC (Benzene-1,4-dicarboxylic acid), and 2.12 g of ZrCl4 (Zirconium(IV) chloride) in 250 ml of DMF.

[0121] 2) Allow to react at 120℃ for 24 hours.

[0122] 3) Wash the synthesized particles three times each using DMF and ethanol, and dry them at 80°C to obtain a powder.

[0123] 4) Remove the residual solution in the pores of the obtained powder in a vacuum environment at 150℃ for 8 hours.

[0124] 5) After removing the residual solution, suspend 200 mg of powder in 30 ml of toluene, and then add 500 ul of ethylenediamine.

[0125] 6) Put the above solution into a round flask and connect the flask to a reflux condenser.

[0126] 7) Stir at 100℃ for 6 hours.

[0127] 8) The synthesized particles are washed three times each using toluene and ethanol, and dried at 80°C to obtain a powder.

[0128] Next, the physicochemical properties of the synthesized metal-organic framework were confirmed, and the zeta potential was -20.44 mV, and the SEM, X-ray diffraction, N2 adsorption isotherm, and BET specific surface area-pore measurement results are shown in Fig. 5.

[0129]

[0130] 1-6: MIL-88-NH2(Fe)

[0131] MIL-88-NH2(Fe) (C), an iron (Fe)-based metal-organic framework 24 H 15 Fe3N3O13 )(CAS No. 1341134-09-5), the following experiments were performed.

[0132] 1) Add 232 mg of 2-aminoterephthalic acid and 374 mg of iron chloride hexahydrate (FeCl3) to 30 ml of DMF and stir.

[0133] 2) Seal and react at 120℃ for 24 hours.

[0134] 4) The synthesized particles were washed three times each using DMF and ethanol, and dried at 80°C to obtain a powder.

[0135] Next, the physicochemical properties of the synthesized metal-organic framework were confirmed, and the zeta potential was +12.0 mV, and the SEM, X-ray diffraction, N2 adsorption isotherm, and BET specific surface area-pore measurement results are shown in Fig. 6.

[0136]

[0137] Experimental Example 2: Cytotoxicity Evaluation of Metal-Organic Frameworks

[0138] To evaluate the cytotoxicity of the metal-organic framework produced in Experimental Example 1 above, the following experiment was performed.

[0139] Specifically, to evaluate the cytotoxicity of the metal-organic framework manufactured / synthesized in Experimental Example 1 above, mouse fibroblasts (L929) were first seeded in a 96-well plate at 1X10 per well. 4Cells were added and cultured for 24 hours in a 37℃, 5% CO2 incubator. After 24 hours, the existing medium in each well was removed, and the metal-organic framework particles to be tested were suspended in the medium at a concentration of 100, 50, 25, 10, or 1 ug / ml to make a test solution, and 100 ul was added to each well. The solution was cultured for 24 hours in a 37℃, 5% CO2 incubator, and the medium was removed. Next, the plate and cells were washed three times with PBS to remove particles attached to the surface. Next, 10% EZ-cytox medium was added to each well and reacted in a 37℃ incubator for 1 hour, and the absorbance at 450 nm and 600 nm was measured using a microplate reader to compare the cell viability with the control group.

[0140] As a result of the above cytotoxicity experiment, it was confirmed that all of the metal-organic frameworks produced in the above experimental examples did not exhibit cytotoxicity (Fig. 7).

[0141]

[0142] Experimental Example 3: Confirmation of the Formation of a Metal-Organic Framework and RNA Complex

[0143] To confirm whether the metal-organic framework produced in Experimental Example 1 above and RNA can form a complex, the following experiment was performed.

[0144] First, 1.0 g of agarose was added to 100 ml of 1x TAE buffer and melted at high temperature to make a 1% agarose gel. Then, 7 ul of Dyne StainingSTAR (EtBr substitute) was added to the agarose gel solution and mixed sufficiently, and the solution was poured into an electrophoresis gel forming mold and solidified. The solidified agarose gel was placed in an electrophoresis device, and 1x TAE buffer was added so that the gel was sufficiently submerged. The MOF manufactured in Experimental Example 1 was suspended in DW at a concentration of 10 mg / ml, and then diluted to concentrations of 5 mg / ml, 2.5 mg / ml, 1.25 mg / ml, 0.625 mg / ml, and 0.312 mg / ml to prepare (for MOFs that require ligand measurement, the ligand is suspended in DW at a concentration of 1 mg / ml). 200 ng / ul mRNA solution was prepared by adding 80 ul of 10 mM NaOAC to 20 ul of mRNA (1 ug / ul), and 1 ul of mRNA solution and 10 ul of MOF solution were mixed and incubated at room temperature for 10 minutes to induce the formation of mRNA-MOF complexes. In addition, mRNA control was prepared by mixing 1 ul of mRNA solution and 10 ul of DI. After mixing 5 ul of DNA ladder (50 bp) and 1 ul of DNA loading buffer, 5 ul was loaded onto an agarose gel, and 5 ul of mRNA control and 5 ul of mRNA-MOF complex were mixed in 1 ul of DNA loading buffer, respectively, and 5 ul was loaded onto an agarose gel.

[0145] As a result of the above experiment, it was confirmed that the metal-organic framework fabricated above can form a complex with RNA (Fig. 8).

[0146]

[0147] Experimental Example 4: Method for preparing a lipid carrier comprising a metal-organic framework

[0148] In order to manufacture a lipid-based carrier including the metal-organic framework manufactured in Experimental Example 1, the following experiments were performed.

[0149] First, mRNA-loaded lipid nanoparticles were manufactured using the following method.

[0150] 1) Prepare ethanol stocks with concentrations of 40 mM DOTAP (an example of ionized lipid), 25 mM DSPC (an example of phospholipid), 25 mM cholesterol (an example of sterol), and 6.25 mM DMG-PEG (an example of PEG-lipid).

[0151] 2) Take four types of lipids in the amounts shown in Table 1 below, mix them, and add EtOH so that the final volume becomes 25 ul. This is called Lipid-Mix.

[0152] 3) Prepare an mRNA solution with a concentration of 51.23 ug / ml using 10 mM NaOAc buffer.

[0153] 4) Add 25 ul of Lipid-Mix to 75 ul of mRNA solution, vortex for about 30 seconds to prepare mRNA-LNP solution, and incubate at room temperature for 10 minutes.

[0154] 5) Add 1.9 ml of PBS and 100 μl of mRNA-LNP solution to a centrifugal filter (10 kDa), centrifuge at 3000 xg at 4°C for 20 minutes, and wash away uncomplexed substances. Repeat this process twice.

[0155] 6) The solution filtered on the filter was taken to obtain lipid nanoparticles loaded with mRNA, which were named DOTAP-LNP (Example 1, lipid nanoparticles not containing MOF).

[0156] Components used in the manufacture of the delivery system DOTAP (ug) DSPC (ug) Cholesterol (ug) DMG-PEG (ug) mRNA (ug) 187.32 19.75 37.229.4 13.84

[0157]

[0158] Next, a lipid carrier including a metal-organic framework based on the lipid nanoparticles manufactured above was manufactured by the following method.

[0159] 1) Prepare ethanol stocks with concentrations of 80 mM DOTAP (an example of ionized lipid), 25 mM DSPC (an example of phospholipid), 25 mM cholesterol (an example of sterol), and 6.25 mM DMG-PEG (an example of PEG-lipid).

[0160] 2) For each group, take the lipid components in the amount of lipid listed in Table 2 below, mix them, and add EtOH so that the final volume becomes 25 ul. This is called Lipid-Mix.

[0161] 3) Prepare an mRNA solution with a concentration of 51.23 ug / ml using 10 mM NaOAc buffer.

[0162] 4) Suspend MOF in EtOH to prepare a 20 mg / ml MOF suspension.

[0163] 5) Mix 25 ul of Lipid-Mix, 2.5 ul of MOF suspension, and 75 ul of mRNA solution, then vortex for about 30 seconds to prepare a MOF-Lipid-mRNA solution, and incubate at room temperature for 10 minutes.

[0164] 6) Add 1.9 ml of PBS and 100 μl of MOF-Lipid-mRNA solution to a centrifugal filter (10 kDa), centrifuge at 3000 xg at 4 °C for 20 minutes, and wash away uncomplexed substances. Repeat this process twice.

[0165] 7) The solution filtered on the filter was taken to obtain a lipid carrier containing a metal-organic framework loaded with mRNA, which was named MOF-Lipid-mRNA complex.

[0166] Components used in the manufacture of group carriers MOF (ug) DOTAP (ug) DSPC (ug) Cholesterol (ug) DMG-PEG (ug) mRNA (ug) 150 87.32 19.75 37.229.413.84 250 107.07-37.229.413.84 350 124.5 319.75-9.4 13.84 450 96.73 19.75 37.22-3.84 550 144.29--9.4 13.84 650 116.48-37.22-3.84 750 133.94 19.75--3.84 850 153.7---3.84

[0167]

[0168] Experimental Example 5: Confirmation of RNA delivery efficiency of lipid carriers containing metal-organic frameworks.

[0169] In order to confirm the RNA delivery efficiency of the lipid carrier (MOF-LNP) containing the metal-organic framework manufactured in Experimental Example 4 above, the following experiment was performed.

[0170]

[0171] 5.1: Confirmation of RNA delivery efficiency according to lipid components of lipid carriers containing metal-organic frameworks

[0172] In relation to the lipid delivery vehicle including the metal-organic framework manufactured in Experimental Example 4 above, the following experiment was performed to compare and analyze the RNA delivery efficiency according to the presence or absence of each lipid component.

[0173] Specifically, HEK293 (human embryonic kidney cell) cells were seeded in 24-well at about 30,000 cells / well and cultured for 24 hours at 37°C, 5% CO2. Thereafter, the cell medium was replaced with 500 μl of Opti-MEM, and the complex volume that would contain 0.1 μg of mRNA contained in the LNP (Example 1) manufactured in Experimental Example 4 and the lipid carrier (LNP-MOF) including various lipid combinations and metal-organic frameworks was calculated, and each volume was added to each well to treat the cells, and incubated at 37°C for 16 hours. After 16 hours, Opti-MEM was removed, 200 ul of lysis buffer was added to each well, incubated for 15 minutes, 50 ul of luciferin and 50 ul of the above sample were added to a 96-well white plate, and luminescence was measured using a microplate reader.

[0174] In the lipid carriers containing the above metal-organic frameworks, MOF-808 or UIO-66-ED were used as examples of the metal-organic frameworks, and the efficiency of delivery of the loaded mRNA was analyzed by confirming the protein expression level of cells according to the use of the carriers of each group. The protein expression level was set to '1' when mRNA was introduced using the carrier of Example 1, which is a lipid nanoparticle that does not contain a metal-organic framework (composed of DOTAP, DSPC, cholesterol, and DMG-PEG - only MOF was excluded from Group 1), and the expression level was expressed based on this.

[0175] As a result, for lipid carriers containing zirconium-based MOF-808 as an example of a metal-organic framework, it was confirmed that the groups excluding PEG-lipid (groups 4, 6, 7, and 8) had significantly higher protein expression levels compared to the other groups (Table 3 and Fig. 9).

[0176] Example Group Composition Protein Expression Level (Compared to Example 1) MOF / mRNA wt ratio MOFDOTAPDSPCCholesterol DMG-PEG2 2 MOF-80 8 OXOO 2.68 13.03 3 OOXO 0.48 13.044 OOX 2 7.85 13.055 OXXO 5.63 13.066 OOX 11.67 13.077 OOX 2 8.31 13.088 OXX 5 0.58 13.0

[0177]

[0178] In addition, in the case of lipid carriers containing zirconium-based UIO-66-ED as another example of metal-organic frameworks, it was confirmed that the groups in which PEG-lipid was excluded (groups 4, 6, 7 and 8) had significantly higher protein expression levels compared to the other groups (Table 4 and Fig. 10).

[0179] Example Group Composition Protein Expression Level (Compared to Example 1) MOF / mRNA wt ratio MOFDOTAPDSPCCholesterol DMG-PEG9 1 UIO-66-ED OOOO 1.03 13.010 2 OX OO 2.05 13.01 13 OO X 0.02 13.01 24 OO X 10.88 13.01 35 OO X 0.11 13.01 46 OX OX 3.98 13.01 57 OO X 5.44 13.01 68 OX XX 4.08 13.0

[0180]

[0181] Based on the above results, it can be seen that in the case of a lipid carrier including a metal-organic framework, the RNA delivery efficiency is significantly improved when PEG-lipid, which is one of the components of the lipid nanoparticle, is excluded.

[0182]

[0183] 5.2: Confirmation of RNA delivery efficiency of lipid carriers containing various types of metal-organic frameworks and excluding PEG-lipids

[0184] In order to confirm whether the RNA delivery efficiency of a lipid carrier containing a metal-organic framework and excluding PEG-lipid, as confirmed in Experimental Example 5.1 above, shows the same trend when using other metal-organic frameworks, the following experiment was performed.

[0185] Specifically, lipid carriers of groups 4, 6, 7 and 8 excluding PEG-lipid were prepared using aluminum-based MIL-101-NH2(AL), magnesium-based Mg-MOF-74, zirconium-based UIO-66 or iron-based MIL-88-NH2(Fe) as metal-organic frameworks, and the RNA delivery efficiency of the carriers was confirmed by the method described in Experimental Example 5.1. Meanwhile, the protein expression level when the carrier of Example 1, which is a lipid nanoparticle without a metal-organic framework, was used was set to '1', and the expression level was expressed based on this.

[0186] The above analysis results confirmed that even when using various types of metal-organic frameworks, the RNA delivery efficiency of the groups excluding PEG-lipid (groups 4, 6, 7, and 8) was significantly increased compared to the control group (Tables 5 to 8).

[0187] Example Group Composition Protein Expression Level (Compared to Example 1) MOF / mRNA wt ratio MOFDOTAPDSPCCholesterol DMG-PEG174MIL-101-NH2(AL) OOOX70.3613.0186OXOX18.4713.0197OOXX5.5413.0208OXXX56.2513.0

[0188] Example Group Composition Protein Expression Level (Compared to Example 1) MOF / mRNA wt ratio MOFDOTAPDSPCCholesterol DMG-PEG2 14 Mg-MOF-74 OOOX 11.49 13.0 226 OXOX 5.32 13.0 237 OOXX 30.31 13.0 248 OXXX 47.16 13.0

[0189] Example Group Composition Protein Expression Level (Compared to Example 1) MOF / mRNA wt ratio MOFDOTAPDSPCCholesterol DMG-PEG254UIO-67OOOX8.3313.0266OXOX7.5213.0277OOXX2.7613.0288OXXX5.2513.0

[0190] Example Group Composition Protein Expression Level (Compared to Example 1) MOF / mRNA wt ratio MOFDOTAPDSPCCholesterol DMG-PEG294MIL-88-NH2(Fe) OOOX8.6813.0306OXOX40.4213.0317OOXX19.5613.0328OXXX141.5113.0

[0191]

[0192] Based on the above results, it can be seen that the lipid carrier containing a metal-organic framework has significantly improved RNA delivery efficiency compared to lipid nanoparticles not containing a metal-organic framework when PEG-lipid, which is one of the components of the lipid nanoparticle, is excluded.

[0193] 5.3: Confirmation of RNA delivery efficiency of lipid carriers according to metal-organic framework content ratio

[0194] In relation to lipid carriers containing metal-organic frameworks, the following experiments were performed to determine the RNA delivery efficiency of lipid carriers according to the metal-organic framework content ratio.

[0195] Specifically, a lipid carrier including a metal-organic framework was manufactured based on the method for manufacturing group 8 of Experimental Example 4, and the MOF suspension used in the above 'Step 4' was used at various concentrations (2.5 mg / ml, 5 mg / ml, 20 mg / ml, 40 mg / ml, 60 mg / ml, and 80 mg / ml) to control the content ratio of the metal-organic framework, and MOF-808 was used as an example of the metal-organic framework (Table 9).

[0196] Example Group Components used in the manufacture of the delivery system Protein expression level (compared to Example 1) MOF / mRNA wt ratio Lipid / MOF wt ratioMOF-808(ug)DOTAP(ug)DSPCCholesterolDMG-PEGmRNA338200153.7XXX3.848.7853.50.77348150153.7XXX3.8412.0539.11.02358100 153.7XXX3.8411.5226.01.5436850153.7XXX3.8412.0513.03.0737812.5153.7XXX3.8412.333.312.303886.25153.7XXX3.846.941.624.59

[0197]

[0198] The RNA delivery efficiency of the lipid carrier comprising the metal-organic framework manufactured above was confirmed, and it was confirmed that the RNA delivery efficiency was excellent when MOF suspensions were used at concentrations of 5 mg / ml (12.5 ug), 20 mg / ml (50 ug), 40 mg / ml (100 ug), and 60 mg / ml (150 ug) (Fig. 11 and Table 9). Therefore, it can be seen that the RNA delivery efficiency of the lipid carrier can be further improved when it is contained in a specific ratio of the metal-organic framework.

[0199]

[0200] The foregoing description of the present invention is provided for illustrative purposes only. Those skilled in the art will readily appreciate that the present invention can be readily modified into other specific forms without altering the technical spirit or essential characteristics of the present invention. Therefore, the embodiments described above should be understood as illustrative in all respects and not restrictive.

[0201]

[0202] <Assignment Information>

[0203] [National Research and Development Project 1 that supported this invention]

[0204] [Project ID] 1425180328

[0205] [Project Number] RS-2023-00280608

[0206] [Ministry Name] Ministry of SMEs and Startups

[0207] [Name of Project Management (Specialist) Agency] Small and Medium Business Technology Information Promotion Agency

[0208] [Research Project Name] Startup Growth Technology Development Project

[0209] [Research Project Name] Development of Next-Generation Innovative New Drugs Based on Metal-Organic Frameworks (MOFs) Using Effective Delivery Technology for Proteolysis-Targeting Chimeras (PROTACs)

[0210] [Name of the project performing organization] Mediac Co., Ltd.

[0211] Research Period: September 1, 2023 - July 31, 2024

[0212] [Contribution rate] 50%

[0213]

[0214] [National Research and Development Project 2 that supported this invention]

[0215] [Project ID] 1345370811

[0216] [Project Number] 2021RIS-001

[0217] [Ministry Name] Ministry of Education (P13)

[0218] [Name of Project Management (Specialist) Institution] National Research Foundation of Korea

[0219] [Research Project Name] Local Government-University Cooperation-Based Regional Innovation Project

[0220] [Research Project Name] Local Government-University Cooperation-Based Regional Innovation Project

[0221] [Project Implementation Organization Name] (Chungbuk Regional Innovation Platform) Chungbuk National University

[0222] [Research Period] March 1, 2024 - February 28, 2025

[0223] [Contribution rate] 50%

Claims

1. 1) Metal-organic frameworks (MOFs); and 2) One or more lipid components A MOF-lipid complex comprising:

2. A MOF-lipid complex according to claim 1, wherein the metal-organic framework comprises one or more metals or metal ions selected from the group consisting of aluminum (Al), iron (Fe), zinc (Zn), magnesium (Mg), and zirconium (Zr).

3. In claim 1, the metal-organic framework is UIO-66, UIO-66-ED, UIO-66-NH2, UIO-67, UIO-67-ED, UIO-67-NH2, PCN-128, PCN-222, PCN-223, PCN-224, MOF-525, MOF-545, Mg-MOF-74, 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-100-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, Fe-MIL-100-NH2, Fe-MIL-101, Fe-MIL-101-NH2, A MOF-lipid complex comprising at least one selected from the group consisting of Fe-MIL-125, Fe-MIL-125-NH2, Mg-MOF-74, Mg-MOF-1, Mg2(dobdc), Mg-CUK-1, MOF-5, ZIF-8, IRMOF-3, Zn-BTC, and Zn-MOF-74.

4. A MOF-lipid complex according to claim 1, wherein the at least one lipid component comprises at least one selected from the group consisting of phospholipids, sterols, PEG (polyethylene glycol)-lipids, and ionizable lipids.

5. A MOF-lipid complex according to claim 1, wherein the one or more lipid components do not include PEG-lipid.

6. A MOF-lipid complex according to claim 1, wherein the at least one lipid component and the metal-organic framework are included in a mass ratio of 1:1 to 15:

1.

7. A MOF-lipid complex according to claim 1, wherein the complex is for delivering a desired drug. 8.1) A MOF-lipid complex according to any one of claims 1 to 7; and 2) Target drug A drug delivery complex comprising:

9. A drug delivery complex according to claim 8, wherein the drug is a nucleic acid, a protein, or a compound.

10. A drug delivery complex according to claim 9, wherein the drug is at least one selected from the group consisting of single-stranded siRNA, double-stranded siRNA, rRNA, RNA, DNA, cDNA, plasmid, aptamer, mRNA, tRNA, lncRNA, piRNA, circRNA, saRNA, antisense oligonucleotide, shRNA, miRNA, ribozyme, PNA, microRNA, ASO, gapmer, aptamer, antagomir, agomir, and DNAzyme.

11. A drug delivery complex according to claim 8, wherein the drug is capable of forming a complex with the MOF.

12. A drug delivery complex according to claim 8, wherein the drug is encapsulated inside the MOF-lipid complex.

13. A drug delivery complex according to claim 8, wherein the MOF and the drug are included in a mass ratio of 3:1 to 40:

1.

14. A composition for drug delivery comprising the drug delivery complex of claim 8.

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