Nanoparticle compositions for drug delivery

Abstract

The present invention relates to a composition for drug delivery and a method for preparing the same, and more particularly, to a composition for drug delivery in which a drug is encapsulated in a nanoparticle structure formed of a specific polymer and a cationic compound; and a method for preparing the same.

Description

Technical Field

[0001] This invention relates to a drug delivery composition and its preparation method, and more specifically, to a drug delivery composition and its preparation method that encapsulates a drug within a nanoparticle structure formed of a specific polymer and a cationic compound. Background Technology

[0002] In the treatment of anionic drugs, including nucleic acids, safe and effective drug delivery technologies have long been researched, and various carriers and delivery techniques have been developed. Carriers are mainly divided into viral carriers utilizing adenoviruses, retroviruses, etc., and non-viral carriers utilizing cationic lipids, cationic polymers, etc. It is well known that viral carriers face risks such as non-specific immune responses. Therefore, recent research is moving towards using non-viral carriers to mitigate these drawbacks. Although non-viral carriers are less efficient than viral carriers, they have the advantage of fewer side effects in terms of in vivo safety.

[0003] Representative non-viral vectors for delivering nucleic acid substances include complexes of cationic lipids and nucleic acids (lipoplexes) and complexes of polycationic polymers and nucleic acids (polyplexes). These cationic lipids or polycationic polymers stabilize anionic drugs by forming complexes with them through electrostatic interactions, thus increasing intracellular delivery, and have therefore been extensively studied. However, when administered intravenously in the amounts required to achieve sufficient efficacy, they cause severe toxicity, albeit less than that of viral vectors, making them unsuitable for drug use. Therefore, there is a need to develop anionic drug delivery technologies that are stable in vivo and capable of intracellular delivery to achieve adequate efficacy, while minimizing the use of potentially toxic cationic polymers or cationic lipids.

[0004] Various anionic drug delivery compositions and their preparation methods have been disclosed, in which a complex is formed through electrostatic interactions between nucleic acids and cationic lipids, and the complex is encapsulated within a nanoparticle structure of an amphiphilic block copolymer. For example, Korean Patent Publication No. 10-2017-0032858 discloses a composition for delivering anionic drugs and its preparation method, which comprises an anionic drug as an active ingredient; a cationic compound; an amphiphilic block copolymer; and polylactic acid salt, wherein the anionic drug forms a complex with the cationic compound through electrostatic interactions, and the formed complex is encapsulated within a nanoparticle structure formed of an amphiphilic block copolymer and polylactic acid salt. Existing nanoparticle drug delivery systems include those disclosed in the aforementioned patent disclosures; however, they still lack the efficiency for delivering drugs such as nucleic acids, peptides, or viruses (especially mRNA) into the body. Summary of the Invention

[0005] Technical problems to be solved The purpose of this invention is to provide a drug delivery composition and a method for preparing the same, which, compared with previously known nanoparticle drug delivery systems, has significantly improved in vivo delivery efficiency of drugs such as nucleic acids, peptides, or viruses (especially mRNA).

[0006] Technical solution A first aspect of the present invention provides a drug delivery composition comprising: an active ingredient selected from nucleic acids, peptides, viruses, or combinations thereof; an amphiphilic block copolymer containing hydrophilic and hydrophobic blocks; and a cationic compound, wherein the hydrophobic blocks are biocompatible and biodegradable polymers having repeating units represented by Formula 1. [Formula 1] , In Equation 1 above, R represents a branched alkylene group having 3 or more carbon atoms.

[0007] A second aspect of the present invention provides a method for preparing a composition for drug delivery, the method comprising the steps of: (a) preparing a solution in which an amphiphilic block copolymer comprising hydrophilic and hydrophobic blocks and a cationic compound are dissolved in an aqueous miscible organic solvent; and (b) adding an active ingredient selected from nucleic acids, peptides, viruses, or combinations thereof to the solution prepared in step (a) and mixing therewith, wherein the hydrophobic blocks are biocompatible and biodegradable polymers having repeating units represented by Formula 1 above.

[0008] Beneficial effects Compared to previously known nanoparticle drug delivery systems, the drug delivery composition according to the present invention can significantly improve the in vivo delivery efficiency of drugs such as nucleic acids, peptides or viruses (especially mRNA). Detailed Implementation

[0009] The present invention will now be described in detail.

[0010] Active ingredient The active ingredient contained in the drug delivery composition of the present invention is selected from nucleic acids, polypeptides, viruses, or combinations thereof.

[0011] "Nucleic acid" can be, for example, DNA, RNA, siRNA, shRNA, miRNA, mRNA, aptamers, antisense oligonucleotides, or combinations thereof, but is not limited to these.

[0012] "Polypeptide" can refer to a protein that is active in the body, such as an antibody or fragment thereof, a cytokine, a hormone or its analogue, or a protein that can be recognized as an antigen through a series of processes in the body, including polypeptide sequences of antigens, their analogues or precursors.

[0013] The "virus" can be an oncolytic virus, for example, one or more selected from the group consisting of adenovirus, AAV, vaccinia virus, herpes simplex virus (HSV), and vesicular stomatitis virus (VSV). In one embodiment, the oncolytic virus is an adenovirus. The adenovirus used in embodiments of the present invention contains a luciferase gene, which can be confirmed by imaging.

[0014] Viruses can express various types of therapeutic genes within an individual, not limited to specific molecular weights, proteins, biological activities, or therapeutic areas. Prophylactic viruses can induce immunity against a target disease within an individual. Compositions containing prophylactic viruses have the advantages of reducing the immune induction, targeting, or amplification of target cells by the virus itself, and reducing hyperimmune responses to the virus upon re-administration, thereby enabling effective results through multiple vaccinations.

[0015] In one implementation, the active ingredient is mRNA (messenger RNA).

[0016] mRNA can be chemically modified in its backbone, sugars, or bases, and its ends can be modified to improve its stability in the blood or reduce the immune response.

[0017] In one embodiment, based on the total dry weight of the composition, the content of the active ingredient can be 0.05% by weight or more, 0.1% by weight or more, 0.2% by weight or more, 0.3% by weight or more, 0.4% by weight or more, or 0.5% by weight or more, and can also be less than 10% by weight, less than 9% by weight, less than 8% by weight, less than 7% by weight, less than 6% by weight, less than 5% by weight, less than 4% by weight, or less than 3% by weight. If the content of the active ingredient is too low, the content of the delivery carrier will be too high compared to the drug, and therefore may cause side effects due to the delivery carrier. Conversely, if the content of the active ingredient is too high, the amount of drug not encapsulated in the nanoparticles will be too high, thereby reducing efficiency.

[0018] Amphiphilic block copolymer The drug delivery composition of the present invention comprises an amphiphilic block copolymer containing hydrophilic and hydrophobic blocks, wherein the hydrophobic blocks are biocompatible and biodegradable polymers having repeating units represented by Formula 1: [Formula 1] , In Equation 1 above, R represents a branched alkylene group having 3 or more carbon atoms.

[0019] In one embodiment, the number (degree of polymerization) of repeating units of the hydrophobic block can be, for example, more than 1, more than 2, more than 3, more than 4, more than 5, more than 6, more than 7, more than 8, more than 9, or more than 10, and can also be less than 25, less than 24, less than 23, less than 22, less than 21, or less than 20, but is not limited thereto.

[0020] In one embodiment, the amphiphilic block copolymer may be an AB-type block copolymer comprising a hydrophilic block (A) and a hydrophobic block (B). In an aqueous environment, the AB-type block copolymer forms core-shell polymer nanoparticles, wherein the hydrophobic block (B) forms the core (inner wall) and the hydrophilic block (A) forms the shell (outer wall).

[0021] In one embodiment, the hydrophilic block may be one or more selected from the group consisting of: polyalkylene glycols, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, and derivatives thereof.

[0022] More specifically, the hydrophilic block can be one or more selected from the group consisting of: monomethoxy polyethylene glycol (mPEG), monoacetoxy polyethylene glycol, polyethylene glycol, copolymers of polyethylene glycol and propylene glycol, and polyvinylpyrrolidone.

[0023] In one embodiment, the number-average molecular weight (g / mol) of the hydrophilic block can be greater than 200, greater than 500, greater than 1000, or greater than 2000, and can be less than 50000, less than 20000, less than 10000, or less than 5000, but is not limited thereto.

[0024] Furthermore, if desired, the ends of the hydrophilic block can be chemically bound to functional groups or ligands capable of reaching specific tissues or cells, or functional groups capable of promoting intracellular delivery, to control the in vivo distribution of the polymer nanoparticle carrier formed from the amphiphilic block copolymer and polylactic acid salt, or to improve the efficiency of delivering the nanoparticle carrier into cells. In one embodiment, the functional group or ligand includes any molecule capable of interacting directly or indirectly with another compound such as a receptor, including but not limited to amino acids, sugars, vitamins, peptides, proteins, hormones, antibodies, neurotransmitters, pharmaceutically active small molecules, endosomal disruptors, cell membrane permeators, charge masking agents, drugs, nucleic acids, or derivatives thereof. Sugars may include, but are not limited to, galactose, galactosamine, N-acetylgalactosamine, or combinations thereof. The hormones may include, but are not limited to, estrogen, testosterone, progesterone, glucocorticoids, adrenaline, insulin, glucagon, cortisol, vitamin D, thyroid hormones, retinoic acid, growth hormone, or combinations thereof. Neurotransmitters may include, but are not limited to, growth factors such as VEGF, EGF, NGF, and PDGF; cholesterol; bile acids; γ-aminobutyric acid (GABA); glutamate; acetylcholine; or combinations thereof. In one embodiment, a functional group or ligand may be linked to the end of a hydrophilic block via a linker molecule. The linker molecule may include, but is not limited to, amides, carbonyl groups, esters, peptides, disulfides, silanes, nucleosides, non-basic nucleosides, polyethers, polyamines, polyamides, carbohydrates, lipids, polyhydrocarbons, phosphate esters, phosphoramides, thiophosphate esters, alkyl phosphate esters, biodegradable linkers, photostable linkers, or combinations thereof.

[0025] Hydrophobic blocks are biocompatible and biodegradable polymers having repeating units represented by Formula 1 above.

[0026] In one embodiment, the number of carbon atoms in R in Formula 1 above can be, for example, 3 or more, 4 or more, 5 or more, 6 or more, or 7 or more, and can also be 20 or less, 19 or less, 18 or less, 17 or less, 16 or less, 15 or less, 14 or less, or 13 or less, but is not limited thereto.

[0027] In one embodiment, R in Formula 1 above may represent a branched alkylene with 3 to 20 carbon atoms, more specifically, a branched alkylene with 3 to 17 carbon atoms, even more specifically, a branched alkylene with 3 to 15 carbon atoms, or even more specifically, a branched alkylene with 3 to 13 carbon atoms, but is not limited thereto.

[0028] In one embodiment, the hydrophobic block can be, but is not limited to, a biocompatible and biodegradable polymer having repeating units selected from, but not limited to, the following structures: , , , , , , , , , , , , , , , , .

[0029] In one embodiment, the repeating unit represented by Formula 1 above can be obtained by ring-opening polymerization of a lactone compound.

[0030] In one embodiment, the number-average molecular weight (g / mol) of the hydrophobic block can be 80 or more, 100 or more, 150 or more, 200 or more, 500 or more, 1000 or more, or 1700 or more, and can also be less than 50,000, less than 20,000, less than 10,000, or less than 6,000, but is not limited thereto.

[0031] For example, the number-average molecular weight combinations of hydrophilic-hydrophobic blocks can be 2000-6000, 2000-4000, 2000-3000, 2000-1700, 2000-1300, 2000-1000, 2000-800, 2000-500, etc., but are not limited to these.

[0032] Furthermore, in one embodiment, in order to increase the hydrophobicity of the hydrophobic block and thereby improve the stability of the nanoparticles, the hydrophobic block can be modified by chemically binding the hydroxyl groups at the end of the hydrophobic block with tocopherol, cholesterol, or fatty acids having 10 to 24 carbon atoms.

[0033] In one embodiment, the ratio of hydrophilic blocks to hydrophobic blocks in the amphiphilic block copolymer allows the hydrophilic block content to be from 25% to 95% by weight, specifically from 40% to 90% by weight, and more specifically from 50% to 80% by weight, based on the total weight of the copolymer. If the ratio of hydrophilic blocks based on the total weight of the copolymer is less than 25% by weight, the polymer has low solubility in water, making it difficult to form nanoparticles. Therefore, to ensure sufficient water solubility of the copolymer for nanoparticle formation, a hydrophilic block ratio of 25% by weight or more is preferred. Conversely, if the ratio of hydrophilic blocks based on the total weight of the copolymer is greater than 95% by weight, the hydrophilicity becomes excessive, reducing the stability of the polymer nanoparticles and making it difficult to use the copolymer as a solubilizing composition for complexes containing active ingredients. Therefore, considering the stability of the nanoparticles, a hydrophilic block ratio of 95% by weight or less is preferred.

[0034] In one embodiment, based on the total dry weight of the composition, the content of the amphiphilic block copolymer in the drug delivery composition of the present invention can be 5% by weight or more, 6% by weight or more, 7% by weight or more, 10% by weight or more, 12% by weight or more, 15% by weight or more, 17% by weight or more, or 18% by weight or more, and can also be less than 90% by weight, less than 80% by weight, less than 70% by weight, less than 60% by weight, less than 50% by weight, less than 40% by weight, or less than 30% by weight. If the polymer content is too low, the size of the nanoparticles will become too large, and therefore the stability of the nanoparticles may decrease, and the loss rate during filter sterilization may increase. Conversely, if the polymer content is too high, there is a concern that the amount of active ingredient that can be incorporated will become too low.

[0035] According to one embodiment, to the extent that the objectives of the invention can be achieved, the hydrophobic block may further include additional hydrophobic repeating units in addition to the repeating units having the structure represented by Formula 1 above.

[0036] In one embodiment, the additional hydrophobic repeating unit may be one or more selected from the group consisting of polyesters, polyanhydrides, polyamino acids, polyorthoesters, and polyphosphazenes. More specifically, the additional hydrophobic repeating unit may be one or more selected from the group consisting of polylactic acid (PLA), polyglycolic acid, polydioxane-2-one, copolymers of lactide and glycolide, and copolymers of lactide and dioxane-2-one, but is not limited thereto.

[0037] Cationic compound The drug delivery composition of the present invention comprises a cationic compound.

[0038] In one embodiment, the cationic compound may be a cationic lipid, a cationic polymer, or a combination thereof, and more specifically, it may be a cationic lipid.

[0039] For example, cationic lipids can be one or more combinations selected from the group consisting of: N,N-diolenoyl-N,N-dimethylammonium chloride (DODAC), N,N-distearate-N,N-dimethylammonium bromide (DDAB), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N,N-dimethyl-(2,3-dioleoyloxy)propylamine (DODMA), N,N,N-trimethyl-(2,3-dioleoyloxy)propylamine (DOTMA), 1,2-diacyl-3-trimethylammonium propane (TAP), 1,2-diacyl-3-dimethylammonium propane (DA). P), 3β-[N-(N',N',N'-trimethylaminoethyl)carbamoyl]cholesterol (TC-cholesterol), 3β-[N-(N',N'-dimethylaminoethyl)carbamoyl]cholesterol (DC-cholesterol), 3β-[N-(N'-monomethylaminoethyl)carbamoyl]cholesterol (MC-cholesterol), 3β-[N-(aminoethyl)carbamoyl]cholesterol (AC-cholesterol), cholesterol oxypropyl-1-amine (COPA), N-(N'-aminoethyl)carbamoylpropionyl tocopherol (AC-tocopherol), and N-(N'-methylaminoethyl)carbamoylpropionyl tocopherol (MC-tocopherol). More specifically, the cationic lipid can be one or more selected from the group consisting of: 3β-[N-(N',N',N'-trimethylaminoethyl)carbamoyl]cholesterol (TC-cholesterol), 3β-[N-(N',N'-dimethylaminoethyl)carbamoyl]cholesterol (DC-cholesterol), 3β-[N-(N'-monomethylaminoethyl)carbamoyl]cholesterol (MC-cholesterol), 3β-[N-(aminoethyl)carbamoyl]cholesterol (AC-cholesterol), N-(1-(2,3-dioleoyloxy)propyl)-N,N,N-trimethylammonium chloride (DOTAP), N,N-dimethyl-(2,3-dioleoyloxy)propylamine (DODMA), and N,N,N-trimethyl-(2,3-dioleoyloxy)propylamine (DOTMA).

[0040] In one embodiment, the cationic lipid may be a lipid having a structure represented by the following formula 2, or an ionized form thereof: [Equation 2] , In Equation 2 above, M1 and M2 are each independently a divalent linker group. R1 and R2 are each independently a substituted or unsubstituted carbocyclic group or a heterocyclic group. R3 is a hydrogen atom or an organic group, optionally containing one or more heteroatoms, whether substituted or unsubstituted. R4 to R 11 Each is an independent hydrogen atom, or a substituted or unsubstituted saturated or unsaturated hydrocarbon group. Me is a methyl group, and a, b, c, and d are each independent integers from 1 to 20.

[0041] In Formula 2 above, the expression "substituted or unsubstituted" for any group indicates that, unless otherwise stated, the group is not substituted, or is substituted by a hydroxyl group or C. 1-6 Alkyl substitution.

[0042] According to one implementation scheme, in Equation 2 above, M1 and M2 can each be independently selected from the following groups: -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, aryl (more specifically C 6-20 Alpha-aryl, and more specifically C 6-10 (aryl), and heteroaryl (more specifically C) 3-20 Hybrid aryl, and more specifically C 3-10 A heteroaryl group having one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S, wherein M' can be a direct bond, C, or N. 1-13 Alkylene (more specifically C) 1-6 (alkylene) or C 2-13 alkenyl (more specifically C) 2-6 (alkenyl), and each R' can independently be selected from the group consisting of: hydrogen atoms, C 1-18 Alkyl (more specifically C) 1-10 Alkyl groups, and more specifically C14 groups. 1-6 alkyl) and C 2-18 alkenyl (more specifically C) 2-10 Alkenyl, and more specifically C 2-6 (alkenyl).

[0043] According to one implementation scheme, in Equation 2 above, R1 and R2 can each be independently selected from the group consisting of: substituted or unsubstituted C. 3-20 cycloalkyl (more specifically C) 3-10 cycloalkyl, and more specifically C 3-6 cycloalkyl), substituted or unsubstituted C3-20 Cycloalkenyl (more specifically C) 3-10 Cycloalkenyl, and more specifically C 3-6 Cycloalkenyl), substituted or unsubstituted C 6-20 Aryl (more specifically C) 6-10 Aryl (more specifically C6 aryl), substituted or unsubstituted C 3-20 Heterocyclic alkyl groups (more specifically C14) 3-10 Heterocyclic alkyl groups, and more specifically C146 ... 3-6 Heterocyclic alkyl), substituted or unsubstituted C 3-20 Heterocyclic alkenyl (more specifically C) 3-10 Heterocyclic alkenyl groups, and more specifically C 3-6 Heterocyclic alkenyl groups, and substituted or unsubstituted C groups. 3-20 heteroaryl (more specifically C 3-10 heteroaryl, and more specifically C 3-6 (heteroaryl), wherein each heterocyclic alkyl, heterocyclic alkenyl and heteroaryl group may independently have one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0044] According to one implementation scheme, in Equation 2 above, R3 can be selected from the group consisting of: hydrogen atoms, substituted or unsubstituted C atoms. 1-6 Alkyl, substituted or unsubstituted C 3-6 Carbocyclic group, -(CH2) n Q、-(CH2) n CHQR, -CHQR, and -CQ(R)2, where each R can be independently selected from the group consisting of: hydrogen atoms, C ... 1-3 Alkyl and C 2-3 Alkenyl group; Q can be selected from the following groups: carbocyclic group, heterocyclic group, -OR, -O(CH2). n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, -N(R)R 12 、N(R)S(O)2R 12 -O(CH2) n OR, -N(R)C(=NR) 13 )N(R)2、-N(R)C(=CHR 13)N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR 13 )N(R)2、-N(OR)C(=CHR 13 )N(R)2、-C(=NR 13 )N(R)2、-C(=NR 13 R, -C(O)N(R)OR, and -C(R)N(R)2C(O)OR, where each n is an independent integer from 1 to 5; R 12 Choose from the following groups: C 3-6 Carbocyclic groups and heterocyclic groups; R 13 Choose from the following groups: H, CN, NO2, C 1-6 Alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C 2-6 alkenyl, C 3-6 Carbocyclic and heterocyclic groups; each R is independently selected from the group consisting of: hydrogen atom, C 1-3 Alkyl and C 2-3 Alkenyl; each X is independently chosen from the group consisting of: F, CI, Br, and I, provided that R3 is -(CH2). n Q、-(CH2) n When it is CHQR, -CHQR or -CQ(R)2, (i) if n is 1, 2, 3, 4 or 5, then Q is not -N(R)2, or (ii) if n is 1 or 2, then Q is not a 5-, 6- or 7-membered heterocyclic alkyl group.

[0045] According to one implementation scheme, in Equation 2 above, R4 to R 11 Each can be independently selected from the following groups: hydrogen atom, C... 1-3 Alkyl and C 2-3 Alkenyl group.

[0046] According to one implementation scheme, in Equation 2 above, a, b, c, and d can each be an integer from 1 to 15 independently.

[0047] More specifically, in Equation 2 above, M1 and M2 can each be independently -C(O)O- or -OC(O)-.

[0048] More specifically, in Equation 2 above, R1 and R2 can each independently be substituted or unsubstituted C. 3-6 Cycloalkyl.

[0049] More specifically, in Equation 2 above, R3 can be a hydrogen atom or a substituted or unsubstituted C atom. 1-3 Alkyl groups, or more specifically, unsubstituted C4 groups. 1-3 alkyl or hydroxy substituted C 1-3 alkyl.

[0050] More specifically, in Equation 2 above, R4 to R 11 It can be a hydrogen atom.

[0051] More specifically, in Equation 2 above, a, b, c, and d can each be an integer from 3 to 11, or even more specifically, an integer from 5 to 9.

[0052] More specifically, the lipids of Formula 2 above can be lipids having a structure selected from Formulas 2-A to 2-O:

[0053] In other embodiments, the cationic lipid may be a lipid having the structure shown in Formula 3, or an ionized form thereof: [Formula 3] ; ;and ; In each of the structures shown in Equation 3 above, At least two (more specifically, 2 to 7) of the R groups are Rx, and the other R groups are Ry, wherein Each Rx is selected independently , , and In , each of a, b, and c is an independent integer from 2 to 20, R1 is a substituted or unsubstituted, saturated or unsaturated divalent hydrocarbon group, and R2 is a substituted or unsubstituted, unsaturated monovalent hydrocarbon group. Indicates substituted or unsubstituted methylene, and Each Ry is independently H, or a substituted or unsubstituted alkyl group, wherein two Ry groups that are not H can be linked together with the nitrogen atom to which they are attached to form a ring structure; and Each L is independently a substituted or unsubstituted alkylene group, and may optionally have an ether bond (-O-), a thioether bond (-S-), or a disulfide bond (-SS-) in its structure.

[0054] In Formula 3 above, unless otherwise stated, "substituted or unsubstituted" for any group means that the group is not substituted, or is substituted by one or more atoms selected from -OH, halogen atoms, C1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C 3-20 cycloalkyl, C 3-20 Heterocyclic alkyl, C 6-20 Aryl or C 3-20 Substituent substitution of heteroaryl groups.

[0055] According to one implementation scheme, in Equation 3 above, each Rx is independently selected from... , , and Where a, b, and c can each be an integer from 2 to 20 or from 2 to 15, and R1 can be a substituted or unsubstituted saturated or unsaturated divalent C. 1-12 The hydrocarbon group, R2, can be a substituted or unsubstituted unsaturated monovalent carbon. 2-24 hydrocarbon group, and Indicates whether the methylene group is substituted or unsubstituted.

[0056] According to one implementation scheme, in Equation 3 above, a, b, and c can each be an integer from 2 to 15 independently, more specifically, they can each be an integer from 3 to 12 independently. Even more specifically, they can each be an integer from 5 to 7 independently, and b and c can each be an integer from 3 to 11 independently, but are not limited thereto.

[0057] According to one implementation scheme, in Equation 3 above, each Ry can be independently H or C. 1-20 Alkyl group, wherein the alkyl group may be independently unsubstituted or may be substituted by one or more of the following: -OH, C 1-20 Alkyl, C 1-20 Alkoxy, -NH2, -NH(C) 1-20 alkyl), -N(C) 1-20 alkyl)2, optionally substituted C 3-20 Carbocyclic groups (e.g., C) 3-20 cycloalkyl or C 6-20 aryl) and optionally substituted C 3-20 Heterocyclic groups (e.g., C) 3-20 Heterocyclic alkyl or C 3-20 (Heteroaryl), wherein the heterocyclic group may have one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S; and two Ry groups that are not H may be linked to the nitrogen atom to which they are attached to form a ring structure optionally having one or more heteroatoms selected from N and O. Furthermore, in the above, the alkyl or alkoxy group may more specifically be C 1-10 Alkyl or alkoxy, and more specifically C 1-6Alkyl or alkoxy, but not limited to these.

[0058] According to one implementation scheme, in Equation 3 above, each L can be independently represented as C. 1-20 Alkylene (more specifically C) 1-10 Alkylene, and more specifically C 1-6 Alkylenes), wherein each may be independently unsubstituted or may be substituted by one or more of the following: -OH, C 1-20 Alkyl, C 1-20 Alkoxy, -NH2, -NH(C) 1-20 alkyl), -N(C) 1-20 alkyl)2, optionally substituted C 3-20 Carbocyclic groups (e.g., C) 3-20 cycloalkyl or C 6-20 aryl), and optionally substituted C 3-20 Heterocyclic groups (e.g., C) 3-20 Heterocyclic alkyl or C 3-20 (Heteroaryl), wherein the heterocyclic group may have one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S. Furthermore, in the above, the alkyl or alkoxy group may more specifically be C. 1-10 Alkyl or alkoxy, and more specifically C 1-6 Alkyl or alkoxy, but not limited to these.

[0059] More specifically, in Equation 3 above, each Ry can be independently H or C. 1-10 Alkyl groups, wherein each alkyl group may be independently unsubstituted or may be substituted with one or more of the following: -OH, C 1-10 Alkyl, C 1-10 Alkoxy, -NH2, -NH(C) 1-10 alkyl), -N(C) 1-10 alkyl)2, optionally substituted C 3-10 Carbocyclic groups and optionally substituted C 3-10 A heterocyclic group, wherein the heterocyclic group may have one or more (e.g., 1 to 3) heteroatoms selected from N, O and S; and two Ry groups that are not H may be connected to the nitrogen atom to which they are attached to form a ring structure having one or more heteroatoms selected from N and O.

[0060] More specifically, in Equation 3 above, each L can be independently represented by C. 1-10 Alkylenes, each of which may be independently unsubstituted or may be substituted with one or more of the following: -OH, C 1-10 Alkyl, C 1-10 Alkoxy, -NH2, -NH(C) 1-10 alkyl), -N(C)1-10 alkyl)2, optionally substituted C 3-10 Carbocyclic groups and optionally substituted C 3-10 Heterocyclic groups, wherein the heterocyclic groups may have one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0061] More specifically, in Equation 3 above, each Rx is independently selected from... , and Where a, b, and c can each be an integer from 3 to 12 independently, and R1 can be a substituted or unsubstituted C. 1-12 Alkylene, substituted or unsubstituted C 2-12 alkenyl or substituted or unsubstituted C 2-12 The alkynyl group, R2 can be a substituted or unsubstituted C. 2-24 alkenyl or substituted or unsubstituted C 2-24 alkynyl group, and Indicates whether the methylene group is substituted or unsubstituted.

[0062] More specifically, in Equation 3 above, each Ry can be independently H or C. 1-6 Alkyl groups, wherein the alkyl group may be independently unsubstituted or may be substituted by one or more groups selected from -OH and -NH2, and two Ry groups that are not H may be linked to the nitrogen atom to which they are attached to form a ring structure having one or more heteroatoms selected from N and O.

[0063] More specifically, in Equation 3 above, each L can independently be unsubstituted C. 1-6 Alkylene.

[0064] Specifically, the lipids of Formula 3 above can have structures selected from the following: In each of the above structures, R1 to R7 are each independently selected from... , , and Where a, b, and c are each an independent integer from 2 to 20, R1 is a substituted or unsubstituted saturated or unsaturated divalent hydrocarbon group, and R2 is a substituted or unsubstituted unsaturated monovalent hydrocarbon group. Indicates whether the methylene group is substituted or unsubstituted.

[0065] More specifically, the lipid of Formula 3 above can be a lipid having a structure selected from Formulas 3-A to 3-V:

[0066] In another embodiment, the cationic lipid may be a lipid having a structure represented by the following formula 4, or an ionized form thereof: [Formula 4] , In Equation 4 above, M1 and M2 are each independently a divalent linker group. R1 and R2 are each independently a substituted or unsubstituted carbocyclic group or a heterocyclic group. R3 is a hydrogen atom or an organic group, optionally containing one or more heteroatoms, whether substituted or unsubstituted. R4 through R7 are each independently a hydrogen atom or a substituted or unsubstituted saturated or unsaturated hydrocarbon group, and a and b are each an independent integer from 1 to 20.

[0067] In Formula 4 above, the expression "substituted or unsubstituted" for any group indicates that, unless otherwise stated, the group is not substituted, or is selected from -OH, halogen atoms, C 1-8 Alkyl (more specifically, C) 3-7 alkyl) or C 1-8 Haloalkyl (more specifically, C) 3-7 One or more substituents of a haloalkyl group are used for substitution.

[0068] According to one implementation scheme, in Equation 4 above, M1 and M2 can each be independently selected from the following groups: -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, aryl (more specifically C 6-20 Alpha-aryl, and more specifically C 6-10 (aryl), and heteroaryl (more specifically C) 3-20 Hybrid aryl, and more specifically C 3-10 A heteroaryl group having one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S, wherein M' can be a direct bond, C, or N. 1-13 Alkylene (more specifically C) 1-6 (alkylene) or C 2-13 alkenyl (more specifically C) 2-6 (alkenyl), and each R' can independently be selected from the group consisting of: hydrogen atoms, C 1-18Alkyl (more specifically C) 1-10 Alkyl groups, and more specifically C14 groups. 1-6 alkyl) and C 2-18 alkenyl (more specifically C) 2-10 Alkenyl, and more specifically C 2-6 (alkenyl).

[0069] According to one implementation scheme, in Equation 4 above, R1 and R2 can each be independently selected from the group consisting of: substituted or unsubstituted C. 3-20 cycloalkyl (more specifically C) 3-15 cycloalkyl, and more specifically C 6-15 cycloalkyl), substituted or unsubstituted C 3-20 Cycloalkenyl (more specifically C) 3-15 Cycloalkenyl, and more specifically C 6-15 Cycloalkenyl), substituted or unsubstituted C 6-20 Aryl (more specifically C) 6-14 aryl), substituted or unsubstituted C 3-20 Heterocyclic alkyl groups (more specifically C14) 3-15 Heterocyclic alkyl), substituted or unsubstituted C 3-20 Heterocyclic alkenyl (more specifically C) 3-15 Heterocyclic alkenyl groups, and substituted or unsubstituted C groups 3-20 heteroaryl (more specifically C 3-15 (heteroaryl), wherein each heterocyclic alkyl, heterocyclic alkenyl and heteroaryl group may independently have one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0070] According to one implementation scheme, in Equation 4 above, R3 can be selected from the group consisting of: hydrogen atoms, substituted or unsubstituted C atoms. 1-6 Alkyl, substituted or unsubstituted C 3-6 Carbocyclic group, -(CH2) n Q、-(CH2) n CHQR, -CHQR, and -CQ(R)2, where each R can be independently selected from the group consisting of: hydrogen atoms, C ... 1-3 Alkyl and C 2-3 Alkenyl group; Q can be selected from the following groups: carbocyclic group, heterocyclic group, -OR, -O(CH2). n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, -N(R)R 12 、N(R)S(O)2R12 -O(CH2) n OR, -N(R)C(=NR) 13 )N(R)2、-N(R)C(=CHR 13 )N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR 13 )N(R)2、-N(OR)C(=CHR 13 )N(R)2、-C(=NR 13 )N(R)2、-C(=NR 13 R, -C(O)N(R)OR, and -C(R)N(R)2C(O)OR, where each n is an independent integer from 1 to 5; R 12 Choose from the following groups: C 3-6 Carbocyclic groups and heterocyclic groups; R 13 Choose from the following groups: H, CN, NO2, C 1-6 Alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C 2-6 alkenyl, C 3-6 Carbocyclic and heterocyclic groups; each R is independently selected from the group consisting of: hydrogen atom, C 1-3 Alkyl and C 2-3 Alkenyl; each X is independently selected from the group consisting of: F, CI, Br, and I, provided that R3 is -(CH2). n Q、-(CH2) n When it is CHQR, -CHQR or -CQ(R)2, (i) if n is 1, 2, 3, 4 or 5, then Q is not -N(R)2, or (ii) if n is 1 or 2, then Q is not a 5-, 6- or 7-membered heterocyclic alkyl group.

[0071] According to one implementation scheme, in Equation 4 above, R4 to R7 can each be independently selected from the following groups: hydrogen atoms, C atoms, etc. 1-3 Alkyl and C 2-3 Alkenyl group.

[0072] According to one implementation scheme, in Equation 4 above, a and b can each be an integer from 1 to 15, or even more specifically, an integer from 3 to 13.

[0073] More specifically, in Equation 4 above, M1 and M2 can each be independently selected from the following groups: -C(O)O-, -OC(O)-, -C(O)N(R')- and -N(R')C(O)-, where R' is the same as defined above.

[0074] More specifically, in Equation 4 above, R1 and R2 can each independently be substituted or unsubstituted C. 3-15 Cycloalkyl.

[0075] More specifically, in Equation 4 above, R3 can be a hydrogen atom or a substituted or unsubstituted C atom. 1-3 alkyl.

[0076] More specifically, in Equation 4 above, R4 to R7 can be hydrogen atoms.

[0077] More specifically, in Equation 4 above, a and b can each be an integer from 5 to 11, or even more specifically, an integer from 5 to 9.

[0078] More specifically, the lipids of Formula 4 above can be lipids having a structure selected from Formulas 4-A to 4-R:

[0079] In another embodiment, the cationic lipid may be a lipid having the structure represented by Formula 5: [Formula 5] , In Equation 5 above, R1 is a substituted or unsubstituted alkylene, alkenylene, or ynylene group. R2, R3, and R4 are each independently substituted or unsubstituted alkylene, alkenylene, or ynylene groups. R5, R6, and R7 are each independently a substituted or unsubstituted saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl, alkenyl, ynyl, or carbocyclic group, or independently a -R group. 10 -(L4) n -R 11 , Each R 10 Independently, it is a substituted or unsubstituted alkylene group. Each R 11 Independently, it can be a substituted or unsubstituted saturated or unsaturated monovalent hydrocarbon group. L1, L2, L3, and L4 are each independently selected from the following groups: -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, ynyl, aryl, and heteroaryl, wherein L' is a direct bond, alkylene, alkenyl, or ynylene, and each R' is independently selected from the following groups: hydrogen atom, alkyl, alkenyl, and ynyl. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion.

[0080] In Formula 5 above, the expression "substituted or unsubstituted" for any group indicates that, unless otherwise stated, the group is not substituted, or is selected from -OH, halogen atoms, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 3-20 cycloalkyl, C 3-20 Heterocyclic alkyl, C 6-20 Aryl or C 3-20 One or more substituents of the heteroaryl group are substituted.

[0081] In Formula 5 above, "hetero-" for any group (e.g., heteroaryl, heterocycloalkyl, etc.) means that, unless otherwise stated, the group has one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0082] In Equation 5 above, the "monovalent hydrocarbon group" can be branched or unbranched, cyclic or acyclic or aromatic.

[0083] According to one implementation plan, in Equation 5 above, R1 can be substituted or unsubstituted C. 1-6 Alkylene, C 2-6 imide or C 2-6 Ethyne-1, R2, R3, and R4 can each independently be substituted or unsubstituted C. 3-12 Alkylene, C 3-12 imide or C 3-12 Imyethynyl, R5, R6, and R7 can each independently be substituted or unsubstituted saturated or unsaturated monovalent C. 3-20 hydrocarbon group, R8 and R9 can be independently substituted or unsubstituted C.1-6 Alkyl, C 2-6 alkenyl, C 2-6 alkynyl or C 3-6 The carbocyclic group, or each group can be independently -R 10 -(L4) n -R 11 , Each R 10 It can be independently substituted or unsubstituted C 3-12 Alkylene Each R 11 The unit valence C can be independently substituted or unsubstituted saturated or unsaturated. 3-20 hydrocarbon group, L1, L2, L3, and L4 can each be independently selected from the following groups: -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, C 2-6 imidene group, C 2-6 Ethyne group, C 6-20 Aromatic and C 3-20 Hybrid aryl groups, where L' can be a direct bond, C 1-13 Alkylene, C 2-13 imide or C 2-13 The ynylene group, and each R' can independently be selected from the group consisting of: hydrogen atoms, C atoms, and so on. 1-18 Alkyl, C 2-18 alkenyl and C 2-18 alkynyl group, n is 0 or 1, and X - It can be a pharmaceutically acceptable monovalent anion of inorganic or organic acids.

[0084] More specifically, in Equation 5 above, R1 can be substituted or unsubstituted C. 3-4 Alkylene, C 3-4 imide or C 3-4 Imyethynyl, R2, R3, and R4 can each independently be substituted or unsubstituted C. 6-8 Alkylene, C 6-8 imide or C 6-8 Imyethynyl, R5, R6, and R7 can each independently be substituted or unsubstituted saturated or unsaturated monovalent C. 5-15 hydrocarbon group, R8 and R9 can be independently substituted or unsubstituted C. 1-2 Alkyl, C 2-3 alkenyl or C 2-3 The alkynyl group, or each group can be independently -R 10 -(L4) n -R 11 , Each R 10 It can be independently substituted or unsubstituted C 6-8 Alkylene Each R 11 The unit valence C can be independently substituted or unsubstituted saturated or unsaturated. 5-15 hydrocarbon group, L1, L2, L3, and L4 can each be independently selected from the following groups: -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -P(O)(OR')O-, -SS-, C 2-5 imide and C 2-5 The ynylene group, wherein each R' can be independently selected from the group consisting of: hydrogen atoms, C atoms, and so on. 1-6 Alkyl, C 2-6 alkenyl and C 2-6 alkynyl group, n is 0 or 1, and X - It can be a halide anion (F - Cl - ,Br - I - ), nitrate anion (NO3) - ), benzoate anion (C6H5COO) - ), mesylate anion, acetate anion (CH3COO) - (i.e., AcO) - (or trihaloacetate anion)

[0085] Even more specifically, in Equation 5 above, R1 can be substituted or unsubstituted C. 3-4 Alkylene R2, R3, and R4 can each independently be substituted or unsubstituted C. 6-8 Alkylene R5, R6, and R7 can each independently be substituted or unsubstituted saturated or unsaturated monovalent C. 5-15 hydrocarbon group, R8 and R9 can be independently substituted or unsubstituted C. 1-2 Alkyl groups, or each can be independently -R 10 -(L4) n-R 11 , Each R 10 It can be independently substituted or unsubstituted C 6-8 Alkylene Each R 11 The unit valence C can be independently substituted or unsubstituted saturated or unsaturated. 5-15 hydrocarbon group, L1, L2, L3, and L4 can each be independently selected from the following groups: -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -P(O)(OR')O-, -SS-, and C 2-5 The subalkenyl group, wherein each R' can be independently selected from the group consisting of hydrogen atoms and C atoms. 1-6 alkyl, n is 0 or 1, and X - It can be Cl - ,Br - or acetate anion (CH3COO) - (i.e., AcO) - ).

[0086] More specifically, cationic lipids can be lipids having a structure selected from the following formulas 5-A to 5-Q:

[0087] In another embodiment, the cationic lipid may be a lipid having a structure represented by the following formula 6, or an ionized form thereof: [Formula 6] , In Equation 6 above, R1 is a substituted or unsubstituted alkyl, alkenyl, or alkynyl group. R2 is a substituted or unsubstituted alkylene, alkenylene, or ynylene group. R3 is an unsubstituted alkylene group. R is a hydrogen atom (H) or Where R4 is a substituted or unsubstituted alkyl, alkenyl, or ynyl group, and R5 is a substituted or unsubstituted alkylene, alkenyl, or ynylene group, and This indicates the connection point with the nitrogen atom.

[0088] In Formula 6 above, the expression "substituted or unsubstituted" for any group indicates that, unless otherwise stated, the group is not substituted, or is selected from -OH, halogen atoms, C1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 3-20 cycloalkyl, C 3-20 Heterocyclic alkyl, C 6-20 Aryl or C 3-20 One or more substituents of the heteroaryl group are substituted.

[0089] In Formula 6 above, each of "alkyl", "alkenyl", "alkynyl", "alkylene", "alkenyl", and "alkynylene" can be independently branched or unbranched, or cyclic or acyclic.

[0090] According to one implementation plan, in Equation 6 above, R1 and R4 can each be independently substituted or unsubstituted C. 1-30 Alkyl, substituted or unsubstituted C 2-30 alkenyl or substituted or unsubstituted C 2-30 alkynyl group, R2 and R5 can be independently substituted or unsubstituted C. 1-15 Alkylene, substituted or unsubstituted C 2-15 alkenyl or substituted or unsubstituted C 2-15 alkyne group, and R3 can be unsubstituted C 2-9 Alkylene.

[0091] More specifically, in Equation 6 above, R1 and R4 can each be independently substituted or unsubstituted C. 1-20 Alkyl, substituted or unsubstituted C 2-20 alkenyl or substituted or unsubstituted C 2-20 alkynyl group, R2 and R5 can be independently substituted or unsubstituted C. 1-12 Alkylene, substituted or unsubstituted C 2-12 alkenyl or substituted or unsubstituted C 2-12 alkyne group, and R3 can be unsubstituted C 2-7 Alkylene.

[0092] More specifically, in Equation 6 above, R1 and R4 can each be independently substituted or unsubstituted C. 5-20 Alkyl, substituted or unsubstituted C 5-20 alkenyl or substituted or unsubstituted C 5-20 alkynyl group, R2 and R5 can be independently substituted or unsubstituted C. 3-12 Alkylene, substituted or unsubstituted C3-12 alkenyl or substituted or unsubstituted C 3-12 alkyne group, and R3 can be unsubstituted C 2-5 Alkylene.

[0093] More specifically, the cationic lipids of Formula 6 above can be lipids having a structure selected from Formulas 6-A to 6-L:

[0094] In another embodiment, the cationic lipid may be a lipid having the structure represented by Formula 7: [Formula 7] , In Equation 7 above, R1, R2, and R3 are substituted or unsubstituted alkyl groups. R4 is a substituted or unsubstituted divalent hydrocarbon group. R5, R6, R7, and R8 are each independently a substituted or unsubstituted saturated or unsaturated divalent hydrocarbon group. R9 and R 10 Each is an independent monovalent hydrocarbon group, either substituted or unsubstituted, saturated or unsaturated. L1, L2, L3, and L4 are each independently selected from the following groups: -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, and -S(O)2-, where L' is a direct bond, alkylene, alkenyl, or ynylene, and each R' is independently selected from the following groups: hydrogen atom, alkyl, alkenyl, and ynylene. X - It is a monovalent anion.

[0095] In Formula 7 above, the expression "substituted or unsubstituted" for any group indicates that, unless otherwise stated, the group is not substituted, or is selected from -OH, halogen atoms, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 3-20 cycloalkyl, C 3-20 Heterocyclic alkyl, C 6-20 Aryl or C 3-20 One or more substituents of the heteroaryl group are substituted.

[0096] In Formula 7 above, the expression “hetero-” for any group (e.g., heteroaryl, heterocycloalkyl, etc.) indicates that, unless otherwise stated, the group has one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0097] In Equation 7 above, "monovalent hydrocarbon group" and "divalent hydrocarbon group" can be branched or unbranched, cyclic or acyclic or aromatic.

[0098] In Formula 7 above, each of "alkyl", "alkenyl", "alkynyl", "alkylene", "alkenyl", and "alkynylene" can be independently branched or unbranched, or cyclic or acyclic.

[0099] According to one implementation plan, in Equation 7 above, R1, R2, and R3 are each C1 with or without substitution. 1-6 alkyl, R4 is a divalent C that is either substituted or unsubstituted. 2-6 hydrocarbon group, R5, R6, R7, and R8 are each independently a substituted or unsubstituted saturated or unsaturated divalent carbon. 2-20 hydrocarbon group, R9 and R 10 Each is independently priced as either substituted or unsubstituted saturated or unsaturated. 2-20 hydrocarbon group, L1, L2, L3, and L4 are each independently selected from the following groups: -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, and -S(O)2-, where L' is a direct bond, C 1-13 Alkylene, C 2-13 imide or C 2-13 The ynylene group, and each R' is independently selected from the group consisting of: hydrogen atom, C... 1-18 Alkyl, C 2-18 alkenyl and C 2-18 alkynyl group, and X - It can be a monovalent anion of inorganic or organic acids.

[0100] More specifically, in Equation 7 above, R1, R2, and R3 are each C1 with or without substitution. 1-3 alkyl, R4 is a divalent C that is either substituted or unsubstituted. 2-4 Alkylene R5, R6, R7, and R8 are each independently substituted or unsubstituted C. 2-13 Alkylene or C 2-13 alkenyl, R9 and R 10 Each is independently either substituted or unsubstituted C 6-20 Alkyl, C 6-20 alkenyl or C 6-20 alkynyl group, L1, L2, L3, and L4 are each independently selected from the following groups: -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, and -P(O)(OR')O-, where each R' is independently selected from the following groups: hydrogen atom and C. 1-6 Alkyl, and X - It can be F - Cl - ,Br - I - Nitrate anion, benzoate anion, methanesulfonate anion, acetate anion (CH3COO) - (i.e., AcO) - ) or trihaloacetate anion (CF3COO) - ).

[0101] More specifically, the cationic lipids of Formula 7 above can be lipids having structures selected from Formulas 7-A and 7-B:

[0102] In one embodiment, the cationic polymer may be selected from the group consisting of: chitosan, ethylene glycol chitosan, protamine, polylysine, polyarginine, polyamidoamine (PAMAM), polyethyleneimine, dextran, hyaluronic acid, albumin, branched-chain polyethyleneimine (PEI), polyamines, and polyethyleneamine (PVAm). More specifically, the cationic polymer may be one or more selected from the group consisting of: polyethyleneimine (PEI), polyamines, and polyethyleneamine (PVAm).

[0103] In one embodiment, based on the total dry weight of the composition, the content of the cationic compound in the drug delivery composition of the present invention can be 5% by weight or more, 10% by weight or more, 15% by weight or more, 20% by weight or more, 25% by weight or more, 30% by weight or more, or 35% by weight or more, and can also be 95% by weight or less, 90% by weight or less, 85% by weight or less, 80% by weight or less, 75% by weight or less, 70% by weight or less, or 65% by weight or less. If the content of the cationic compound is too low, it may be insufficient to form nanoparticles. Conversely, if the content of the cationic compound is too high, the size of the nanoparticles will become too large, and therefore the stability of the nanoparticles may decrease, and the loss rate during filter sterilization may increase.

[0104] In the drug delivery composition of the present invention, the active ingredient is maintained in a state encapsulated within a nanoparticle structure formed of a polymer component comprising an amphiphilic block copolymer and a cationic compound, thereby improving stability in blood or body fluids.

[0105] In one embodiment, the particle size of the nanoparticles can be defined by the Z-average value. For example, the particle size of the nanoparticles can be less than 800 nm, less than 600 nm, less than 500 nm, less than 400 nm, less than 300 nm, less than 200 nm, or less than 150 nm, and can also be greater than 10 nm, greater than 50 nm, or greater than 100 nm. In another embodiment, the particle size of the nanoparticles is defined by the Z-average value, which can be, for example, 10 to 800 nm, 20 to 600 nm, 30 to 500 nm, 50 to 400 nm, or 80 to 300 nm.

[0106] In one embodiment, based on 1 part by weight of the cationic compound, the relative amount of the polymer component comprising the amphiphilic block copolymer to the cationic compound may be more than 0.01 parts by weight, more than 0.02 parts by weight, more than 0.03 parts by weight, more than 0.04 parts by weight, or more than 0.05 parts by weight, and may be less than 50 parts by weight, less than 49 parts by weight, less than 47 parts by weight, less than 45 parts by weight, less than 43 parts by weight, less than 41 parts by weight, less than 40 parts by weight, less than 39 parts by weight, or less than 37 parts by weight, but is not limited thereto.

[0107] Optional additive components In one embodiment, to improve the in vivo delivery efficiency of the active ingredient, the drug delivery composition of the present invention may further contain fusion-promoting lipids.

[0108] In one implementation, the fusion-promoting lipid may be one or more combinations selected from the group consisting of phospholipids, PEGylated lipids (PEGylated lipids), cholesterol, and tocopherol.

[0109] Specifically, phospholipids can be one or more selected from the group consisting of phosphatidylethanolamine (PE), phosphatidylcholine (PC), and phosphatidic acid. Phosphatidylethanolamine (PE), phosphatidylcholine (PC), and phosphatidic acid can be combined with one or two C... 10-24 The form in which fatty acids are bound. Cholesterol and tocopherol include their respective analogues, derivatives, and metabolites.

[0110] Specifically, PEG lipids refer to polyethylene glycol (PEG) modified lipids, which are PEG derivatives, such as DMG or DSPE, with attached lipid moieties. PEG lipids can be used to improve the circulation time of active ingredients encapsulated in lipid nanoparticles and reduce non-specific uptake. PEG lipids can be one or more selected from the group consisting of: PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide, PEG-modified dialkylamine, PEG-modified diacylglycerol, and PEG-modified dialkylglycerol, as well as combinations thereof. For example, PEG lipids include 1,2-dimyristoyl-sn-glycerol methoxy polyethylene glycol (PEG-DMG), 1,2-distearyl-sn-glycerol-3-phosphoethanolamine-N-[amino(polyethylene glycol)] (PEG-DSPE), PEG distearylglycerol (PEG-DSG), PEG dipalmitoyl, PEG dioleoyl, PEG distearyl, PEG diacyl glycosamide (PEG-DAG), PEG dipalmitoylphosphatidylethanolamine (PEG-DPPE), or PEG-1,2-dimyristoylpropyl-3-amine (PEG-c-DMA).

[0111] Specifically, the fusion-promoting lipids can be one or more combinations selected from the group consisting of: dilauroyl phosphatidylethanolamine, dimyristoyl phosphatidylethanolamine, dipalmitoyl phosphatidylethanolamine, distearyl phosphatidylethanolamine, dioleoyl phosphatidylethanolamine, dilinoleoyl phosphatidylethanolamine, 1-palmitoyl-2-oleoyl phosphatidylethanolamine, 1,2-diphytanyl-3-sn-phosphatidylethanolamine, dilauroyl phosphatidylcholine, dimyristoyl phosphatidylcholine, dipalmitoyl phosphatidylethanolamine, and dipalmitoyl phosphatidylethanolamine. Acylphosphatidylcholine, distearylphosphatidylcholine, dioleoylphosphatidylcholine, dilinoleoylphosphatidylcholine, 1-palmitoyl-2-oleoylphosphatidylcholine, 1,2-diphydanyl-3-sn-phosphatidylcholine, dilauroylphosphatidyl, dimyristoylphosphatidyl, dipalmitoylphosphatidyl, distearylphosphatidyl, dioleoylphosphatidyl, dilinoleoylphosphatidyl, 1-palmitoyl-2-oleoylphosphatidyl, 1,2-diphydanyl-3-sn-phosphatidyl, cholesterol, and tocopherol.

[0112] More specifically, the fusion-promoting lipids may be one or more combinations selected from the group consisting of: dioleoylphosphatidylethanolamine (DOPE), 1,2-dipalmitoyl-sn-glycerol-3-phosphocholine (DPPC), distearate phosphatidylcholine (DSPC), 1,2-dioleoyl-sn-glycerol-3-phosphocholine (DOPC), 1-stearoyl-sn-glycerol-3-phosphocholine (18:0 lysophosphatidylcholine (LysoPC)), 1-(9Z-octadecenoyl)-sn-glycerol-3-phosphocholine (18:1 LysoPC) 1-Palmitoyl-2-oleoyl-sn-glycerol-3-phosphate choline, 1-stearoyl-2-oleoyl-sn-glycerol-3-phosphate choline (SOPC), 1,2-dioleoyl-sn-glycerol-3-phosphate choline (DOPC), 1-oleoyl-2-hydroxy-sn-glycerol-3-phosphate ethanolamine (18:1 lysophosphatidylethanolamine (Lyso PE)), 1-stearoyl-2-oleoyl-sn-glycerol-3-phosphate ethanolamine (18:0-18:1 PE), 1-stearoyl-2-linoleoyl-sn-glycerol-3-phosphate ethanolamine (18:0-18:2 PE), 1,2-dilinoleoyl-sn-glycerol-3-phosphate ethanolamine (18:2 PE), 1-stearoyl-2-hydroxy-sn-glycerol-3-phosphate-(1'-racemic glycerol)(18:0 lysophosphatidylglycerol (Lyso PG)), 1-oleoyl-2-hydroxy-sn-glycerol-3-phosphate-(1'-racemic glycerol)(18:1 Lyso PG), 1,2-dipalmitoyl-sn-glycerol-3-phosphate ethanolamine (DPPE), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), 1,2-dioleoyl-sn-glycerol-3-phosphate (18PA), cholesterol, and tocopherol.

[0113] In one embodiment of the invention, the fusion-promoting lipid may be distearylphosphatidylcholine, cholesterol, or a combination thereof.

[0114] In one embodiment, based on the total dry weight of the composition, the content of the fusion-promoting lipids can be more than 1% by weight, more than 2% by weight, more than 3% by weight, more than 4% by weight, or more than 5% by weight, and can also be less than 40% by weight, less than 35% by weight, less than 30% by weight, less than 25% by weight, or less than 20% by weight.

[0115] In one embodiment, based on 1 part by weight of the cationic compound, the relative amount of the fusion-promoting lipid to the cationic compound can be 0.05 parts by weight or more, 0.06 parts by weight or more, 0.07 parts by weight or more, 0.08 parts by weight or more, 0.09 parts by weight or more, or 0.1 parts by weight or more, and can be 6 parts by weight or less, 5.5 parts by weight or less, 5 parts by weight or less, 4.5 parts by weight or less, 4 parts by weight or less, or 3.8 parts by weight or less, but is not limited thereto.

[0116] In one embodiment, when phospholipids are used as fusion-promoting lipids, the relative amount of phospholipids based on 1 part by weight of the cationic compound can be 0.03 parts by weight or more, 0.04 parts by weight or more, 0.05 parts by weight or more, or 0.06 parts by weight or more, or 4 parts by weight or less, 3.9 parts by weight or less, 3.7 parts by weight or less, 3.5 parts by weight or less, 3.3 parts by weight or less, 3.1 parts by weight or less, 3 parts by weight or less, 2.9 parts by weight or less, or 2.7 parts by weight or less, but is not limited thereto.

[0117] In one embodiment, when cholesterol is used as the fusion-promoting lipid, the relative amount of cholesterol based on 1 part by weight of the cationic compound can be more than 0.02 parts by weight, more than 0.03 parts by weight, or more than 0.04 parts by weight, and can also be less than 2 parts by weight, less than 1.9 parts by weight, less than 1.7 parts by weight, less than 1.5 parts by weight, less than 1.3 parts by weight, or less than 1.1 parts by weight, but is not limited thereto.

[0118] Furthermore, in one embodiment, the drug delivery composition of the present invention may also contain one or more additive components (hereinafter referred to as "optional additive components") that are typically included in drug delivery compositions.

[0119] In one embodiment, the optional additive component may be, for example, one or more selected from the group consisting of: pH adjusters (e.g., acidifiers, alkalizers, buffers), tension modifiers, fillers (e.g., sugars, polyols, amino acids, polymers, proteins, etc.), wetting agents, solubilizers, surfactants, antioxidants, antibacterial agents, chelating agents, complexing agents, etc., but not limited thereto.

[0120] In one embodiment, the pH adjuster may be one or more selected from the group consisting of: acetate, citrate, tartrate, histidine, glutamate, phosphate, tris(hydroxymethyl)aminomethane (Tris), glycine, bicarbonate, succinate, sulfate, nitrate, etc., but not limited thereto.

[0121] In one embodiment, the tension modifier may be one or more selected from the group consisting of: mannitol, sorbitol, lactose, glucose, trehalose, sodium chloride, potassium chloride, glycerol, glycerin, propylene glycol, etc., but not limited thereto.

[0122] In one embodiment, the filler may be one or more selected from the group consisting of: sugars and polyols, including sucrose, trehalose, glucose, lactose, sorbitol, mannitol, glycerol, etc.; amino acids, including arginine, aspartic acid, glutamic acid, lysine, proline, glycine, histidine, methionine, alanine, etc.; polymers and proteins, including gelatin, polyvinylpyrrolidone (PVP), lactide-co-glycolic acid (PLGA), polyethylene glycol (PEG), dextran, cyclodextrin or derivatives thereof, starch derivatives, hydroxylamine sulfate (HAS), bovine serum albumin (BSA), etc.; or combinations thereof, but not limited thereto.

[0123] In one embodiment, the wetting agent and / or solubilizer may be one or more selected from the group consisting of: lecithin, PEG 300, PEG 600, PEG 1000, polyoxyethylene dodecyl ethers (e.g., Brij 30, Brij 35, Brij 56, Brij 76, Brij 97), polypropylene glycol (PPG) 2000, glucoside alkyl ethers, polyoxyethylene glycol octylphenol ethers, polyoxyethylene glycol alkylphenol ethers, glyceryl alkyl esters, polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 80, sorbitan monolaurate (Span 20), sorbitan monooleate (Span 80), sorbitan trioleate (Span 20). 85), cocamide monoethanolamine (MEA), cocamide diethanolamine (DEA), dodecyl dimethylamine oxide, poloxamer, polyvinylpyrrolidone K25, polyvinyl alcohol, oligolactic acid, sodium dioctyl sulfosuccinate, diethylene glycol monolaurate, triethanolamine oleate, sodium oleate, potassium oleate, ethyl oleate, oleic acid, ethyl laurate, sodium dodecyl sulfate, etc., but not limited to these.

[0124] In one embodiment, the antioxidant may be one or more selected from the group consisting of: tocopherol (vitamin E), α-tocopherol, α-tocopherol succinate, ascorbic acid, ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), monothioglycerol, potassium metabisulfite, propionic acid, propyl gallate, sodium ascorbate, sodium bisulfite, sodium metabisulfite, sodium sulfite, histamine, methionine, glutathione, poly(ethylamine), etc., but not limited thereto.

[0125] In one embodiment, the antimicrobial agent may be one or more selected from the group consisting of: benzalkonium chloride, benzyl chloride, benzyl alcohol, bromonitrile glycol, bromophthalic acid trimethylammonium, cetylpyridine chloride, chlorhexidine, chlorobutanol, chlorocresol, xyloxyphenol, cresol, m-cresol, ethanol, glycerin, hexoterine, imidazoline, phenol, phenoxyethanol, phenethyl alcohol, phenylmercuric nitrate, propylene glycol, mercuric sulfate, etc., but not limited thereto.

[0126] In one embodiment, the chelating agent may be one or more selected from the group consisting of: ethylenediaminetetraacetic acid (EDTA), disodium edetate, dipotassium edetate, edetate, fumaric acid, malic acid, phosphoric acid, sodium edetate, tartaric acid, diethylenetriaminepentaacetic acid (DPTA), citric acid, hexaphosphate, mercaptoacetic acid, zinc, etc., but not limited thereto.

[0127] When optional additive components are used in the drug delivery composition of the present invention, the content of each additive may be, for example, more than 0.01% by weight, more than 0.05% by weight, or more than 0.1% by weight based on the total dry weight of the composition, and may be less than 10% by weight, less than 5% by weight, or less than 1% by weight, but is not limited thereto.

[0128] Composition and its preparation method The drug delivery composition according to the present invention can be administered via routes of administration such as intravascular, intramuscular, mucosal, subcutaneous, intradermal, oral, bone, transdermal, or local tissue, and can be formulated into various oral or parenteral formulations suitable for such routes of administration. Examples of oral formulations include various formulations such as tablets, capsules, powder formulations, liquid formulations, etc., and examples of parenteral formulations include various formulations such as eye drops, injectable formulations, etc., and in one embodiment, the composition can be an injectable formulation. For example, when the composition of the present invention is freeze-dried, it can be reconstituted with distilled water for injection, 0.9% physiological saline, 5% glucose aqueous solution, etc., to prepare an injectable formulation.

[0129] The present invention also provides a method for preparing a drug delivery composition, comprising the steps of: (a) preparing a solution in which an amphiphilic block copolymer comprising hydrophilic and hydrophobic blocks and a cationic compound are dissolved in an aqueous miscible organic solvent; and (b) adding an active ingredient selected from nucleic acids, peptides, viruses, or combinations thereof to the solution prepared in step (a) and mixing therewith, wherein the hydrophobic blocks are biocompatible and biodegradable polymers having repeating units represented by Formula 1 above.

[0130] In one embodiment, the water-miscible organic solvent in step (a) may be ethanol.

[0131] In one implementation, step (a) can be carried out in solution under acidic conditions.

[0132] In one embodiment, step (b) may include: (b-1) preparing a buffer solution containing the active ingredient; and (b-2) adding the buffer solution of the active ingredient prepared in step (b-1) to the solution prepared in step (a) and mixing them.

[0133] In one embodiment, the mixing ratio of the buffer solution of the active ingredient prepared in step (b-1) to the solution prepared in step (a) can be a volume ratio of 1:1 to 1:5, more specifically, it can be 1:2 to 1:4.

[0134] In another embodiment, step (b) may include: (b-1) adding the active ingredient to the solution prepared in step (a); and (b-2) adding a buffer solution to the resulting mixture from step (b-1) and mixing it therewith.

[0135] In one embodiment, the method for preparing a drug delivery composition may further include adding a buffer solution, water for injection, or a combination thereof to the resulting mixture from step (b) for pH adjustment.

[0136] In another embodiment, the method for preparing a drug delivery composition may further include the steps of removing the solvent from the resulting mixture in step (b), then adding a freeze-drying adjuvant thereto and freeze-drying the resulting mixture.

[0137] Adding freeze-drying aids helps the freeze-dried composition maintain its cake-like shape or helps the composition dissolve uniformly within a short time during the reconstitution process after freeze-drying. Specifically, freeze-drying aids can be one or more selected from the group consisting of sugars, amino acids, polymers, and proteins; for example, freeze-drying aids can be one or more selected from the group consisting of lactose, mannitol, sorbitol, and sucrose. Based on the total dry weight of the freeze-dried composition, the content of the freeze-drying aid can be from 1% to 90% by weight, more specifically from 10% to 60% by weight.

[0138] The present invention will now be explained in more detail with reference to the following embodiments. However, the embodiments are for illustrative purposes only, and the scope of the invention is not limited in any way.

[0139] [Example] Cationic lipid preparation example 1 Compound 2-A of formula 2 is prepared as follows.

[0140] [Equation 2-A] (1) Synthesis of 1-cyclopropylnonane-1-ol In a 2000 mL three-necked round-bottom flask (RBF), cyclopropaneformaldehyde (35.0 g, 499 mmol, 1.00 equivalent) and tetrahydrofuran (THF) (700 mL) were added under nitrogen atmosphere. The mixture was cooled to -65 °C, and then octylmagnesium bromide (2 M, 375 mL, 1.50 equivalent) was added. The mixture was stirred at -65 °C for 2 hours. The reactor was heated to 15 °C, and the reaction mixture was poured into a saturated aqueous solution of NH4Cl (500 mL). The organic and aqueous layers were separated. The aqueous layer was extracted with ethyl acetate (EtOAc) (450 mL) three times (150 mL each time). The organic layer was collected, concentrated under vacuum, and purified by silica gel column chromatography (petroleum ether:EtOAc = 50:1 → 0:1) to give 1-cyclopropylnonan-1-ol (87.5 g, 73.1%).

[0141] 1 H NMR (400 MHz, chloroform-) d ): δ 2.93 - 2.81 (m, 1H), 1.61 (br d, 2H), 1.52- 1.27 (m, 12H), 0.95 - 0.86 (m, 4H), 0.60 - 0.45 (m, 2H), 0.34 - 0.19 (m,2H).

[0142] (2) Synthesis of 1-cyclopropylnonyl 8-bromooctanoic acid In 1000 mL of three-necked RBF, cyclopropylnonan-1-ol (30.0 g, 163 mmol, 1.00 equivalent), 8-bromooctanoic acid (72.6 g, 326 mmol, 2.00 equivalent), dichloromethane (DCM) (300 mL), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (31.2 g, 163 mmol, 1.00 equivalent), and 4-dimethylaminopyridine (DMAP) (19.9 g, 163 mmol, 1.00 equivalent) were added, and the mixture was stirred at 25 °C for 16 hours. The reaction mixture was concentrated under vacuum, and after the addition of silica powder, it was purified by silica gel column chromatography with petroleum ether:EtOAc = 10:1 → 50:1 to give 1-cyclopropylnonyl 8-bromooctanoic acid (22.8 g, 36.0%).

[0143] 1 H NMR (400 MHz, chloroform-) d ): δ 4.29 (td, 1H), 3.59 - 3.31 (m, 2H), 2.32(t, 2H), 1.94 - 1.75 (m, 2H), 1.73 - 1.60 (m, 4H), 1.49 - 1.25 (m, 18H), 1.03- 0.93 (m, 1H), 0.90 (t, 3H), 0.61 - 0.24 (m, 4H).

[0144] (3) Synthesis of compound 2-A In a 100 mL three-necked flask, methylamine hydrochloride (173 mg, 2.57 mmol, 1.00 equivalent), ethanol (EtOH) (30 mL), N,N-diisopropylethylamine (DIEA) (1.66 g, 12.8 mmol, 5.00 equivalent), and 1-cyclopropylnonyl 8-bromooctanoate (3.00 g, 7.70 mmol, 3.00 equivalent) were added sequentially, and the mixture was stirred at 80 °C for 72 hours. The reaction mixture was concentrated under vacuum, and after the addition of silica powder, it was purified by silica gel column chromatography using petroleum ether:EtOAc = 10:1 → 1:1 to give compound 2-A (660 mg, 38.9%).

[0145] 1 H NMR (400 MHz, chloroform-) d ): δ4.29 (td, 2H), 2.32 (br t, 8H), 2.22 (s,3H), 1.74 - 1.60 (m, 8H), 1.54 - 1.42 (m, 4H), 1.39 - 1.23 (m, 36H), 1.02 -0.87 (m, 8H), 0.61 - 0.23 (m, 8H).

[0146] Cationic lipid preparation example 2 Compound 4-A was prepared as follows.

[0147] [Equation 4-A] (1) Synthesis of 4-pentylcyclohexyl 8-bromooctanoate In a 250 mL three-necked round-bottom flask (RBF), 8-bromooctanoic acid (2.00 g, 8.96 mmol, 1.00 equivalent), dichloromethane (DCM) (40 mL), and dimethylformamide (DMF) (0.5 mL) were added together, and oxalyl chloride (2.28 g, 17.9 mmol, 2.00 equivalent) was added at 0 °C under nitrogen. The resulting mixture was stirred at 25 °C for 4 hours under nitrogen, then 4-pentylcyclohexane-1-ol (2.29 g, 13.5 mmol, 1.50 equivalent) and triethylamine (TEA) (1.36 g, 13.5 mmol, 1.50 equivalent) were added, and the mixture was stirred again at 25 °C under nitrogen for 12 hours. The reaction mixture was concentrated under vacuum, and the concentrated residue was purified by silica gel column chromatography. The petroleum ether:ethyl acetate (EtOAc) ratio was 1:0 → 50:1 to give 4-pentylcyclohexyl 8-bromooctanoate (2.46 g, 6.55 mmol, 73.1% yield).

[0148] 1 H NMR (400 MHz, chloroform-) d ): δ 0.76 - 0.86 (m, 3 H) 0.88 - 1.94 (m, 28 H)2.21 (dt, 2 H) 3.33 (td, 2 H) 4.52 - 4.66 (m, 1 H) 4.87 - 4.95 (m, 1 H).

[0149] (2) Synthesis of 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoic acid In 100 mL of three-necked RBF, 4-pentylcyclohexyl 8-bromooctanoate (2.46 g, 6.55 mmol, 1.00 equivalent), 2-aminoethanol (2.00 g, 32.8 mmol, 5.00 equivalent), and ethanol (EtOH) (50 mL) were added, and the mixture was stirred at 80 °C for 16 hours under nitrogen. The reaction mixture was concentrated under vacuum, and the concentrated residue was purified by silica gel column chromatography using petroleum ether:EtOAc = 1:0 → 50:1 to give 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate (2.00 g, 5.62 mmol, 85.8% yield) as a yellow solid.

[0150] 1 H NMR (400 MHz, chloroform-) d ): δ 0.76 - 0.86 (m, 3 H) 0.88 - 1.94 (m, 27 H)2.32 (t, 2 H) 2.51 (t, 2 H) 2.71 (t, 2H), 3.54 (d, 2 H) 3.91 - 4.00 (m, 1 H)4.11 - 4.31 (m, 1H).

[0151] (3) Synthesis of cyclopentadecanyl 8-bromooctanoate In 250 mL of three-necked RBF, cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 equivalent), 8-bromooctanoic acid (4.93 g, 22.1 mmol, 1 equivalent), sulfuric acid (H₂SO₄) (217 mg, 2.21 mmol, 0.10 equivalent), and toluene (100 mL) were added, and the mixture was stirred at 120 °C for 16 hours under nitrogen atmosphere. After evaporation of the solvent, the residue was purified by silica gel column chromatography using petroleum ether:EtOAc = 1:0 → 50:1 to give a colorless oily cyclopentadecanol 8-bromooctanoic acid (2.60 g, 6.03 mmol, 27.3% yield).

[0152] 1 H NMR (400 MHz, chloroform-) d ): δ 4.89 (quin, 1H), 3.41 (dt, 2H), 2.28 (t,2H), 1.85 (quin, 2H), 1.73 - 1.16 (m, 36H).

[0153] (4) Synthesis of compound 4-A In 100 mL of three-necked RBF, cyclopentadecanyl 8-bromooctanoate (1.60 g, 3.71 mmol, 1.00 equivalent), 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate (1.32 g, 3.71 mmol, 1.00 equivalent), N,N-diisopropylethylamine (DIEA) (527 mg, 4.08 mmol, 1.10 equivalent), and ethanol (30 mL) were added, and the mixture was stirred at 80 °C for 48 hours. Subsequently, after solvent evaporation, the residue was purified using a silica gel column with petroleum ether:EtOAc = 10:1 → 1:1, and further purified by preparative HPLC (folic acid conditions), washed with NaHCO3 aqueous solution (300 mL), and the organic layer was concentrated and extracted with DCM (200 mL x 2). The organic layer was dried with anhydrous Na2SO4 and filtered. The filtrate was concentrated to give a yellow oily compound of formula 4-A (0.240 g, 340 μmol, 9.16% yield).

[0154] 1 H NMR (400 MHz, chloroform-) d ): δ 4.91 (br s, 1H), 4.82 (quin, 1H), 3.46 (brt, 2H), 2.51 (br d, 2H), 2.37 (br t, 4H), 2.21 (td, 4H), 1.58 - 1.42 (m,14H), 1.33 - 1.17 (m, 52H), 0.83 - 0.80 (m, 3H).

[0155] Amphiphilic block copolymer (mPEG-PNL) preparation example 1 The following amphiphilic block copolymers were prepared.

[0156] Monomethoxy polyethylene glycol (monomethoxyPEG) (5.00 g, 2.5 mmol, 1.0 equivalent) was placed in a 100 mL double-necked round-bottom flask (RBF) and dried at 120 °C for 2 hours. δ-nonanolactone and 1,5,7-triazabicyclo[4.4.0]dec-5-ene (TBD), which had been vacuum-dried at room temperature for 2 hours, were sequentially added to a polymerization reactor containing monomethoxyPEG, and the reaction mixture was stirred at 50 °C for 1 hour. After the reaction was complete, the reaction mixture was added dropwise to cold diethyl ether to induce a first precipitation. The precipitate was collected by centrifugation. The precipitate was dissolved in dichloromethane (DCM), and the resulting solution was added dropwise to cold diethyl ether to induce a second precipitation. The precipitate was collected by centrifugation and dissolved in DCM, and then cold hexane was added dropwise to induce a third precipitation. The precipitate was collected by centrifugation and dissolved in DCM, and then cold hexane was added dropwise to induce a fourth precipitation, thereby obtaining an amphiphilic block copolymer.

[0157] 1 H-NMR (400 MHz, CDCl3) δ 4.88-4.87 (CH-O-CO, m), 4.22-4.20 (CH2-O-CO,m), 3.83-3.54 (CH-OH, O-CH2-CH2-O, m), 3.34 (O-CH3, s), 2.34-2.28 (O-CO-CH2,m), 1.72-1.25 (CH2-CH2-CH2-CH3, m), 0.91-0.87 (CH2-CH3, m).

[0158] Comparative Example 1 The drug delivery composition was prepared according to the composition shown in Table 1 below. An ethanol solution was prepared by sequentially mixing a solution of lipid 5 (11.4 mg of 8-[(2-hydroxyethyl)[8-(nonoxy)-8-oxooctyl]amino]-octanoic acid, 1-octylnonyl ester) in 570 μL, a solution of DSPC (distearylphosphatidylcholine) (2.5 mg) in 250 μL, a solution of cholesterol (4.8 mg) in 480 μL, and a solution of DMG-PEG (1,2-dimyristoyl-rac-glycerol-3-methoxypolyethylene glycol-2000) (1.2 mg) in 120 μL, and thoroughly mixing by vortexing. An aqueous solution of the active ingredient was prepared by mixing 1 mg of luciferase mRNA with 20 mM sodium acetate buffer (pH 4.6). The aqueous solution of the active ingredient was then added to the ethanol solution to make the volume ratio of the aqueous phase to the ethanol phase 3:1. After mixing, the mixture was centrifuged at 4000 rpm using an Amicon Ultra centrifuge filter and concentrated to one-third of its initial volume. Then, PBS (phosphate-buffered saline) was added to restore the mixture to its initial volume, and centrifuged again at 4000 rpm to concentrate the mixture to one-third of its initial volume. This process was repeated six times to remove ethanol, exchanging the buffer with PBS while concentrating the composition. After concentration to the desired concentration, the resulting composition was sterilized by filtration through a 0.22 μm filter.

[0159] [Table 1]

[0160] Examples 1 to 18 The drug delivery composition was prepared according to the composition shown in Table 2 below. The lipid compound of formula 2-A prepared in Cationic Lipid Preparation Example 1 or the lipid compound of formula 4-A prepared in Cationic Lipid Preparation Example 2 was dissolved in ethanol at a concentration of 20 mg / mL. DOPE (1,2-dioleoyl-sn-glycerol-3-phosphate ethanolamine), cholesterol, and mPEG-PNL (monomethoxy polyethylene glycol polynonolactone) prepared in Amphiphilic Block Copolymer Preparation Example 1 were dissolved in ethanol at concentrations of 10 mg / mL, 10 mg / mL, and 25 mg / mL, respectively. For each of the above components, volumes corresponding to the amounts shown in Table 2 were taken from their respective solutions and combined. The resulting mixtures were thoroughly mixed by vortexing to prepare an ethanol solution. An aqueous solution of the active ingredient was prepared by mixing 1 mg of luciferase mRNA with 20 mM sodium acetate buffer (pH 4.6). The aqueous solution of the active ingredient was then added to the ethanol solution to make the volume ratio of the aqueous phase to the ethanol phase 3:1. After mixing, vortex the resulting mixture for approximately 5 seconds to ensure thorough mixing, then centrifuge at 4000 rpm using an Amicon Ultra centrifuge filter to concentrate the mixture to one-third of its initial volume. Add PBS (phosphate-buffered saline) to restore the mixture to its initial volume, and then centrifuge again at 4000 rpm to concentrate the mixture to one-third of its initial volume. Repeat this process six times to remove ethanol, exchanging the buffer with PBS while concentrating the composition. After concentrating to the desired concentration, sterilize the resulting composition by filtering through a 0.22 μm filter.

[0161] [Table 2] (Unit: mg)

[0162] (3) Evaluation of formulation performance For each formulation of Comparative Example 1 and Examples 1 to 18, particle characteristics were evaluated using a particle size analyzer based on dynamic light scattering (DLS), and the results are shown in Table 3 below.

[0163] [Table 3]

[0164] (4) Administration of the composition The formulations of Comparative Example 1 and Examples 1 to 10, 13, 14, 16 and 18 were prepared at a concentration of 10 μg / mL and administered intravenously at a dose of 2 μg mRNA per mouse. Four hours after administration, fluorescein was prepared dissolved in sterile water at a concentration of 15 μg / μL and administered intraperitoneally at a dose of 3 mg per 20 g mouse. Fifteen minutes after intraperitoneal injection of fluorescein, protein expression in individual organs was measured using a bioluminescence imaging system, and the results are shown in Table 4 below.

[0165] As can be seen from Table 4, the drug delivery formulation according to the present invention exhibits excellent efficiency in selectively delivering the drug to the liver after intravenous administration.

Claims

1. A drug delivery composition comprising: The active ingredient is selected from nucleic acids, polypeptides, viruses, or combinations thereof; An amphiphilic block copolymer comprising hydrophilic and hydrophobic blocks; and cationic compounds, The hydrophobic block is a biocompatible, biodegradable polymer having repeating units represented by Formula 1: [Formula 1] ， In Equation 1 above, R represents a branched alkylene group having 3 or more carbon atoms.

2. The drug delivery composition according to claim 1, wherein, The hydrophilic block is selected from one or more of the group consisting of: polyalkylene glycols, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, and derivatives thereof.

3. The drug delivery composition according to claim 1, wherein, In Formula 1, R represents a branched alkylene group having 3 to 20 carbon atoms.

4. The drug delivery composition according to claim 1, wherein, The hydrophobic block is a biocompatible, biodegradable polymer having repeating units selected from the following structures: 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 、 。 5. The drug delivery composition according to claim 1, wherein, The repeating unit represented by Formula 1 is obtained by ring-opening polymerization of lactone compounds.

6. The drug delivery composition according to claim 1, wherein, The cationic compound is a cationic lipid, a cationic polymer, or a combination thereof.

7. The drug delivery composition according to claim 1, wherein, The active ingredient is mRNA.

8. The drug delivery composition according to any one of claims 1 to 7, wherein the composition further comprises a fusion-promoting lipid.

9. The drug delivery composition according to claim 8, wherein the fusion-promoting lipid is one or more combinations selected from the group consisting of phospholipids, PEG lipids, cholesterol, and tocopherol.

10. A method for preparing a composition for drug delivery, comprising the following steps: (a) Prepare a solution in which an amphiphilic block copolymer comprising hydrophilic and hydrophobic blocks and a cationic compound are dissolved in a water-miscible organic solvent; and (b) Add an active ingredient selected from nucleic acids, peptides, viruses, or combinations thereof to the solution prepared in step (a), and mix them. The hydrophobic block is a biocompatible, biodegradable polymer having repeating units represented by Formula 1: [Formula 1] ， In Equation 1 above, R represents a branched alkylene group having 3 or more carbon atoms.

11. The method for preparing a drug delivery composition according to claim 10, wherein the water-miscible organic solvent in step (a) is ethanol.

12. The method for preparing a drug delivery composition according to claim 10, wherein step (b) comprises: (b-1) The step of preparing a buffer solution containing the active ingredient; and (b-2) The step of adding the buffer solution of the active ingredient prepared in step (b-1) to the solution prepared in step (a) and mixing them.

13. The method for preparing a drug delivery composition according to claim 10, wherein step (b) comprises: (b-1) The step of adding the active ingredient to the solution prepared in step (a); and (b-2) The step of adding a buffer solution to the mixture obtained in step (b-1) and mixing it.

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