Novel ionizable lipid and lipid nanoparticle composition using same

Abstract

The present invention relates to a novel ionized lipid and a lipid nanoparticle composition using the same. The lipid nanoparticles comprising the novel ionized lipid according to the present invention exhibit excellent gene encapsulation rate and in vivo gene delivery rate, and as well as excellent gene delivery ability even upon re-administered, and thus can be advantageously utilized in the development of gene therapeutic agents for various indications.

Description

Novel ionized lipids and lipid nanoparticle compositions using the same

[0001] The present invention relates to a novel ionized lipid, a lipid nanoparticle composition using the same, and uses thereof.

[0002] Nucleic acid-based medicines are being used not only as therapeutic agents, but also as preventative agents that protect against diseases by injecting genes that can express antigens for specific diseases. Gene-based vaccines are categorized into DNA vaccines, RNA vaccines, and viral vector vaccines. Among them, RNA vaccines inject mRNA encoding an antigen into the body, causing the antigen to be expressed in the body and inducing antibody formation. RNA vaccines do not have the potential risks of infection associated with viral vector-based vaccines or genetic mutation associated with DNA vaccines. Furthermore, they can be developed quickly, drawing attention as an effective response to the COVID-19 outbreak in 2019.

[0003] However, genes are easily degraded by nucleases in the human body and are negatively charged macromolecules that are not easily delivered into cells. Therefore, a method for delivering them stably and efficiently to the desired location is needed. Various delivery techniques based on materials such as lipids, polymers, dendrimers, and inorganic metal materials have been reported as gene delivery systems. Among these, lipid nanoparticles as lipid delivery vehicles were the first siRNA new drug to receive FDA approval in 2018, patisiran (ONPATTRO). ® ) and after being applied to the development of the mRNA vaccine for COVID-19, which was approved for emergency use in 2020, lipid nanoparticle technology has been recognized for its technological value as a clinically useful delivery system for nucleic acid drugs such as siRNA and mRNA.

[0004] Current lipid nanoparticles are generally used in a form in which four components are mixed in a certain ratio: ionized lipid, phospholipid (helper lipid), cholesterol (structure-maintaining lipid), and PEG-lipid. However, as the development of gene therapy for various indications is in full swing, unmet needs have been identified in various aspects such as enhanced gene delivery ability, maintenance of gene delivery ability in specific diseases such as liver disease, enhanced stability, improved targeting characteristics, ensuring ease of storage and distribution, mitigation of side effects, cost reduction, and response to breakthrough infection. Therefore, the development of lipid nanoparticles using new ionized lipid delivery systems is continuously required.

[0005] Systemic administration of lipid nanoparticles can be utilized in the development of gene therapy. However, most lipid nanoparticles have problems with gene delivery capacity being reduced and not maintained when re-administered to maintain the therapeutic efficacy. Therefore, the development of novel ionizable lipid carriers to address this issue is needed. Local administration of lipid nanoparticles can be utilized in the development of mRNA vaccines or topical therapeutics. However, in this case, unnecessary systemic exposure of lipid nanoparticles can lead to adverse effects. Therefore, the development of ionizable lipid carriers with high local protein expression rates is required.

[0006] As gene therapy enters the pharmaceutical market in the future and continues to expand, it is expected that by securing various ionized lipids different from the existing ionized lipids and developing new lipid nanoparticle systems using them, gene delivery ability will be further improved and unmet needs for systemic or repeated administration will be addressed, thereby enabling the development of gene therapy for numerous rare diseases and various mRNA vaccines and cancer vaccines.

[0007] The purpose of the present invention is to provide a novel ionizable lipid compound.

[0008] Another object of the present invention is to provide lipid nanoparticles using the above ionized lipid compound.

[0009] Another object of the present invention is to provide medical uses of the above lipid nanoparticles.

[0010] In order to achieve the above purpose, the present invention comprises R around the central carbon as shown in the following chemical formula 1. 1 , R 2 , R 3 and (R 4 )-X coordinated compound, pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof is provided:

[0011] <Chemical Formula 1>

[0012]

[0013] In the above chemical formula 1, R 1 , R 2 and R 3 are each independently selected from (C6-C30) alkyl, alkenyl, alkynyl, (C6-C30) cycloalkyl, aryl or heteroaryl with or without a branch, and among these, at least one -CH2- at a third or more carbon position from the central carbon is -O-, -CH(OH)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OP(=O)(OR 5 )O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -SC(=S)O-, -OC(=O)S-, -OC(=S)O-, -S-, -SS-, -NR 5 -, -C(=O)NR 5 -, -NR 5 C(=O)-, -OC(=O)NR 5 -, -NR 5 C(=O)O-, -C(=S)NR 5 -, -NR 5 C(=S)-, -OC(=S)NR5 -, -NR 5 C(=S)O-, -SC(=O)NR 5 -, -SC(=S)NR 5 - and -NR 5 is substituted with any one selected from the group consisting of C(=O)S-, wherein, R 5 is independently selected from hydrogen, (C1-C20) alkyl, (C2-C20) alkenyl, alkynyl, (C3-C20) cycloalkyl, aryl or heteroaryl, and R 4 is selected from (C2-C12) alkylene, alkenylene, alkynylene, (C6-C12) cycloalkylene, arylene or heteroarylene with or without a branch, and one or more -CH2- is -O-, -CH(OH)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OP(=O)(OR 5 )O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -SC(=S)O-, -OC(=O)S-, -OC(=S)O-, -S-, -SS-, -NR 5 -, -C(=O)NR 5 -, -NR 5 C(=O)-, -OC(=O)NR 5 -, -NR 5 C(=O)O-, -C(=S)NR 5 -, -NR 5 C(=S)-, -OC(=S)NR 5 -, -NR 5 C(=S)O-, -SC(=O)NR 5 -, -SC(=S)NR 5 - and -NR 5Any one selected from the group consisting of C(=O)S- is substituted or unsubstituted, X is selected from a chain or cyclic hetero group containing hydrogen or an ionizable amine, and the cyclic hetero group is substituted or unsubstituted with (C1-C6)alkyl, hydroxy or hydroxy(C1-C6)alkyl, and n can be selected from 0 or 1.

[0014] The present invention provides a lipid nanoparticle composition comprising the above compound.

[0015] In addition, the present invention provides a composition for drug delivery comprising the lipid nanoparticle composition described above.

[0016] Lipid nanoparticles containing novel ionized lipids according to the present invention exhibit excellent gene encapsulation rates and in vivo gene transfer rates, and have been confirmed to exhibit excellent gene transfer ability even upon re-administration, and thus can be usefully utilized in the development of gene therapeutic agents for various indications.

[0017] Hereinafter, the present invention will be described in detail.

[0018]

[0019] The present invention relates to a compound of formula 1 below, wherein R is formed around a central carbon. 1 , R 2 , R 3 and (R 4 )-X coordinated compound, pharmaceutically acceptable salt, tautomer, prodrug or stereoisomer thereof is provided:

[0020] <Chemical Formula 1>

[0021]

[0022] In the above chemical formula 1, R 1 , R 2 and R 3are independently selected from (C6-C30) alkyl, alkenyl, alkynyl, (C6-C30) cycloalkyl, aryl or heteroaryl with or without a branch, and among these, at least one -CH2- at the third or more carbon positions from the central carbon is -O-, -CH(OH)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OP(=O)(OR 5 )O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -SC(=S)O-, -OC(=O)S-, -OC(=S)O-, -S-, -SS-, -NR 5 -, -C(=O)NR 5 -, -NR 5 C(=O)-, -OC(=O)NR 5 -, -NR 5 C(=O)O-, -C(=S)NR 5 -, -NR 5 C(=S)-, -OC(=S)NR 5 -, -NR 5 C(=S)O-, -SC(=O)NR 5 -, -SC(=S)NR 5 - and -NR 5 It can be substituted with any one selected from the group consisting of C(=O)S-, where R 5 may be independently selected from hydrogen, (C1-C20)alkyl, (C2-C20)alkenyl, alkynyl, (C3-C20)cycloalkyl, aryl or heteroaryl.

[0023] In the above chemical formula 1, R 4 may be selected from (C2-C12) alkylene, alkenylene, alkynylene, (C6-C12) cycloalkylene, arylene, or heteroarylene with or without a branch, and one or more -CH2- are -O-, -CH(OH)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OP(=O)(OR 5)O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -SC(=S)O-, -OC(=O)S-, -OC(=S)O-, -S-, -SS-, -NR 5 -, -C(=O)NR 5 -, -NR 5 C(=O)-, -OC(=O)NR 5 -, -NR 5 C(=O)O-, -C(=S)NR 5 -, -NR 5 C(=S)-, -OC(=S)NR 5 -, -NR 5 C(=S)O-, -SC(=O)NR 5 -, -SC(=S)NR 5 - and -NR 5 It may be substituted or unsubstituted with any one selected from the group consisting of C(=O)S-, X may be selected from a chain or cyclic hetero group containing hydrogen or an ionizable amine, and the cyclic hetero group may be substituted or unsubstituted with (C1-C6)alkyl, hydroxy or hydroxy(C1-C6)alkyl, and n may be selected from 0 or 1.

[0024] Preferably, the R 1 , R 2 and R 3 can be independently represented by the following chemical formula 1-A:

[0025] <Chemical Formula 1-A>

[0026]

[0027] In the above chemical formula 1-A, L may be (C2-C5)alkylene, and A may be -O-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -OC(=O)S-, -S-, -SS-, -C(=O)NR5 -, -OC(=O)NR 5 -, -C(=S)NR 5 - and -OC(=S)NR 5 - may be selected from the group consisting of, and R may be selected from (C4-C25) alkyl, alkenyl, alkynyl, cycloalkyl, aryl or heteroaryl with or without a branch, and at least one -CH2- among these may be substituted or unsubstituted with any one selected from the group consisting of -O-, -S-, and -SS-, but is not limited thereto.

[0028]

[0029] For example, the above chemical formula 1-A corresponding to the lipid tail may be selected from the following structural formulas 1 to 3, or a mixed form thereof, but is not limited thereto:

[0030] <Structural formula 1>

[0031]

[0032] <Structural formula 2>

[0033]

[0034] <Structural formula 3>

[0035]

[0036]

[0037] In the above chemical formula 1, the R 4 may be selected from alkylene, alkenylene or alkynylene of (C2-C6), and at least one of them -CH2- is -NR 5 -, -C(=O)NR 5 -, -OC(=O)NR 5 -, -C(=S)NR 5 -, -OC(=S)NR 5 -, -SC(=O)NR 5 -, and -SC(=S)NR 5 - may be substituted or unsubstituted with any one selected from the group consisting of, wherein, R 5may be independently selected from hydrogen or (C1-C6)alkyl, but is not limited thereto.

[0038] In the above chemical formula 1, X may be selected from amino substituted with one or more (C1-C4)alkyl; or a cyclic hetero group selected from the group consisting of piperidine, piperazine, and imidazole, and the cyclic hetero group may be substituted or unsubstituted with (C1-C4)alkyl, hydroxy, or hydroxy(C1-C4)alkyl, but is not limited thereto.

[0039]

[0040] For example, the following X and R corresponding to the polar head structure 4 The structure represented by the chemical formula 1-B may be selected from the structural formulas 4 or 5 below, or a mixed form thereof, but is not limited thereto:

[0041] <Chemical Formula 1-B>

[0042]

[0043] <Structural formula 4>

[0044]

[0045] <Structural formula 5>

[0046]

[0047]

[0048] More preferably, the present invention may include compounds represented by the following chemical formulas:

[0049] <Chemical Formula 1-1>

[0050]

[0051] Compound 1: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, Molecular formula: C 48 H 84 N2O7, molecular weight: 801.21]

[0052]

[0053] <Chemical Formula 1-2>

[0054]

[0055] Compound 2: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(5-(dimethylamino)pentanamido)heptanedioate, Molecular formula: C 47 H 84 N2O7, molecular weight: 789.20]

[0056]

[0057] <Chemical Formula 1-3>

[0058]

[0059] Compound 3: Di((Z)-dodec-5-en-1-yl) 4-(((4-(dimethylamino)butoxy)carbonyl)amino)-4-(3-oxo-3-(((Z)-tridec-6-en-1-yl)oxy)propyl)heptanedioate, Molecular formula: C 54 H98 N2O8, molecular weight: 903.38]

[0060]

[0061] <Chemical Formula 1-4>

[0062]

[0063] Compound 4: Di((Z)-dodec-5-en-1-yl) 4-(3-(((Z)-dodec-5-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, Molecular formula: C 54 H 96 N2O7, molecular weight: 885.37]

[0064]

[0065] <Chemical Formula 1-5>

[0066]

[0067] Compound 5: Di((Z)-tetradec-9-en-1-yl) 4-(3-oxo-3-(((Z)-tetradec-9-en-1-yl)oxy)propyl)-4-(3-piperidin-1-yl)propanamido)heptanedioate, Molecular formula: C 60 H 108 N2O7, molecular weight: 969.53]

[0068]

[0069] <Chemical Formula 1-6>

[0070]

[0071] Compound 6: Di((Z)-oct-3-en-1-yl) 4-(3(((Z)-oct-3-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, Molecular formula: C 42 H 72 N2O7, molecular weight: 717.05]

[0072]

[0073] <Chemical Formula 1-7>

[0074]

[0075] Compound 7: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-methylpiperidine-4-carboxamido)heptanedioate, Molecular formula: C 47 H 82 N2O7, molecular weight: 787.18]

[0076]

[0077] <Chemical Formula 1-8>

[0078]

[0079] Compound 8: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-methylpiperidine-3-carboxamido)heptanedioate, Molecular formula: C 47 H 82N2O7, molecular weight: 787.18]

[0080]

[0081] <Chemical Formula 1-9>

[0082]

[0083] Compound 9: Bis(2-(heptylthio)ethyl) 4-(3-(2-(heptylthio)ethoxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, molecular formula: C 45 H 84 N2O7S3, molecular weight: 861.35]

[0084]

[0085] <Chemical Formula 1-10>

[0086]

[0087] Compound 10: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-(2-hydroxyethyl)piperidine-4-carboxamido)heptanedioate, Molecular formula: C 48 H 84 N2O8, molecular weight: 817.21]

[0088]

[0089] <Chemical Formula 1-11>

[0090]

[0091] Compound 11: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(4-piperidin-1-yl)butanamido)heptanedioate, Molecular formula: C 49 H 86 N2O7, molecular weight: 815.23]

[0092]

[0093] <Chemical Formula 1-12>

[0094]

[0095] Compound 12: 4-(3-(piperidin-1-yl)propanamido)-4-(3-(((Z)-tetradec-9-enoyl)oxy)propyl)heptane-1,7-diyl(9Z,9'Z)-bis(tetradec-9-enoate)[4-(3-(piperidin-1-yl)propanamido)-4-(3-(((Z)-tetradec-9-enoyl)oxy)propyl)heptane-1,7-diyl(9Z,9'Z)-bis(tetradec-9-enoate), Molecular formula: C 60 H 108 N2O7, molecular weight: 969.53]

[0096]

[0097] <Chemical Formula 1-13>

[0098]

[0099] Compound 13: Di((Z)-dec-4-en-1-yl) 4-(3-(1H-imidazol-1-yl)propanamido)-4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)heptanedioate, Molecular formula: C 46H 77 N3O7, molecular weight: 784.14]

[0100]

[0101] <Chemical Formula 1-14>

[0102]

[0103] Compound 14: Bis(3-(hexylundecyl) 4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, molecular formula: C 69 H 132 N2O7, molecular weight: 1101.82]

[0104]

[0105] <Chemical Formula 1-15>

[0106]

[0107] Compound 15: Bis(3-(hexylundecyl) 4-(3-oxo-3-(undecyloxy)propyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, molecular formula: C 63 H 120 N2O7, molecular weight: 1017.66]

[0108]

[0109] <Chemical Formula 1-16>

[0110]

[0111] Compound 16: Bis(3-(hexylundecyl) 4-(((3-(dimethylamino)propoxy)carbonyl)amino)-4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)heptanedioate, Molecular formula: C 67 H 130 N2O8, molecular weight: 1091.78]

[0112]

[0113] <Chemical Formula 1-17>

[0114]

[0115] Compound 17: Bis(3-(hexylundecyl) 4-(((3-(diethylamino)propoxy)carbonyl)amino)-4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)heptanedioate, Molecular formula: C 69 H 134 N2O8, molecular weight: 1119.84]

[0116]

[0117] <Chemical Formula 1-18>

[0118]

[0119] Compound 18: Bis(3-hexylundecyl) 4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)-4-(1-(2-hydroxyethyl)piperidine-4-carboxamido)heptanedioate, Molecular formula: C 69 H132 N2O8, molecular weight: 1117.82]

[0120]

[0121] <Chemical Formula 1-19>

[0122]

[0123] Compound 19: 4-(3-((2-hexyldecanoyl)oxy)propyl)-4-(3-(piperidin-1-yl)propanamido)heptane-1,7-diyl bis(2-hexyldecanoate)[4-(3-((2-hexyldecanoyl)oxy)propyl)-4-(3-(piperidin-1-yl)propanamido)heptane-1,7-diyl bis(2-hexyldecanoate), Molecular formula: C 66 H 126 N2O7, molecular weight: 1059.74]

[0124]

[0125] <Chemical Formula 1-20>

[0126]

[0127] Compound 20: Bis(3-hexylundecyl) 4-(3-(1H-imidazol-1-yl)propanamido)-4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)heptanedioate, Molecular formula: C 67 H 125 N3O7, molecular weight: 1084.75]

[0128]

[0129] <Chemical Formula 1-21>

[0130]

[0131] Compound 21: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(3-(4-methylpiperazin-1-yl)propanamido)heptanedioate, Molecular formula: C 48 H 85 N3O7, molecular weight: 816.22]

[0132]

[0133] <Chemical Formula 1-22>

[0134]

[0135] Compound 22: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(piperidin-1-yl)heptanedioate, Molecular formula: C 45 H 79 NO6, molecular weight: 730.13]

[0136]

[0137] <Chemical Formula 1-23>

[0138]

[0139] Compound 23: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(4-(hydroxymethyl)piperidin-1-yl)heptanedioate, Molecular formula: C 46 H 81NO7, molecular weight: 760.15]

[0140]

[0141] <Chemical Formula 1-24>

[0142]

[0143] Compound 24: Di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(4-methylpiperazin-1-yl)heptanedioate, Molecular formula: C 45 H 80 N2O6, molecular weight: 745.14]

[0144]

[0145] Most preferably, the compound is (1) di((z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (3) di((z)-dodec-5-en-yl) 4-(((4-(dimethylamino)butoxy)carbonyl)amino)-4-(3-oxo-3-(((Z)-tridec-6-en-1-yl)oxy)propyl)heptanedioate, (4) di((z)-dodec-5-en-1-yl) 4-(3-(((Z)-dodec-5-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (5) di((z)-tetradec-9-en-1-yl) 4-(3-oxo-3-(((Z)-tetradec-9-en-1-yl)oxy)propyl)-4-(3-piperidin-1-yl)propanamido)heptanedioate, (6) di((z)-oct-3-en-1-yl) 4-(3(((Z)-oct-3-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (7) di((z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-methylpiperidine-4-carboxamido)heptanedioate, (8) di((z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-methylpiperidine-3-carboxamido)heptanedioate, (14) bis(3-(hexylundecyl) 4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, and (18) bis(3-hexylundecyl) It may be at least one selected from the group consisting of 4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)-4-(1-(2-hydroxyethyl)piperidine-4-carboxamido)heptanedioate, but is not limited thereto.

[0146]

[0147] The above compound may include a pharmaceutically acceptable salt form exhibiting the same or similar activity.

[0148] As used herein, “pharmaceutically acceptable” means that the compound or salt thereof is not toxic to cells or humans exposed to it, and thus has a safety and efficacy profile suitable for administration to humans.

[0149] The above salt may be used in the form of either a pharmaceutically acceptable basic salt or an acid salt. The basic salt may be used in the form of either an organic basic salt or an inorganic basic salt, and may be selected from the group consisting of sodium salt, potassium salt, calcium salt, lithium salt, magnesium salt, cesium salt, aminium salt, ammonium salt, triethylaminium salt, and pyridinium salt. The acid salt is useful as an acid addition salt formed by a free acid. Inorganic acids and organic acids can be used as free acids, and inorganic acids such as hydrochloric acid, hydrobromic acid, sulfuric acid, sulfurous acid, phosphoric acid, diphosphoric acid, nitric acid, etc. can be used, and organic acids such as citric acid, acetic acid, maleic acid, malic acid, fumaric acid, gluconic acid, methanesulfonic acid, benzenesulfonic acid, camphorsulfonic acid, oxalic acid, malonic acid, glutaric acid, acetic acid, glycolic acid, succinic acid, tartaric acid, 4-toluenesulfonic acid, galacturonic acid, embonic acid, glutamic acid, citric acid, aspartic acid, stearic acid, etc. can be used, but are not limited thereto, and all salts formed using various inorganic acids and organic acids commonly used in the art can be included.

[0150] In addition, the compound may include not only the above salts, but also all salts, hydrates, solvates, derivatives, etc. that can be prepared by conventional methods. Addition salts can be prepared by conventional methods, and can be prepared by dissolving in a water-miscible organic solvent such as acetone, methanol, ethanol, or acetonitrile, adding an excess amount of organic base, or adding an aqueous base solution of an inorganic base, and then precipitating or crystallizing. Alternatively, the addition salt can be obtained by evaporating the solvent or excess base from the mixture and then drying, or by suction filtration of the precipitated salt.

[0151]

[0152] The present invention provides a lipid nanoparticle composition comprising the above compound.

[0153] The above compound can act as an ionizable lipid.

[0154] The lipid nanoparticle composition comprising the above compound has excellent safety and stability, and can function as a delivery vehicle so that a gene containing RNA, DNA, or a mixture thereof as an active ingredient can effectively exhibit its effect within a cell. Accordingly, the lipid nanoparticles comprising the novel ionizable lipid compound exhibit excellent gene encapsulation rates and in vivo gene delivery rates, and on the other hand, exhibit excellent gene delivery functions even upon re-administration, and thus can be usefully utilized as a drug delivery composition.

[0155] The lipid nanoparticle composition may further comprise one or more selected from the group consisting of neutral lipids, steroids, and polymerized lipids.

[0156] The above neutral lipid may be a phospholipid or a glycolipid. Specifically, the phospholipids include dioleoylphosphatidylethanolamine (DOPE), distearoylphosphatidylcholine (DSPC), palmitoyloleoylphosphatidylcholine (POPC), egg phosphatidylcholine (EPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), distearoylphosphatidylethanolamine (DSPE), It may be at least one selected from the group consisting of phosphatidylethanolamine (PE) and dipalmitoylphosphatidylethanolamine, and the glycolipid may be at least one selected from the group consisting of glucosylceramide, galactosylceramide, glucosylsphingosine, galactosylsphingosine, and phosphoglycoceramide, but is not limited thereto.

[0157] The above steroid may be at least one selected from the group consisting of cholesterol, bile acid derivatives and cholic acid derivatives, but is not limited thereto.

[0158] The above polymer lipid may be a pegylated lipid having a structure in which a water-soluble polymer and a lipid are combined, and the pegylated lipid may be at least one selected from the group consisting of PEG bound to dialkyloxypropyl (PEG-DAA); PEG bound to diacylglycerol (PEG-DAG) such as PEG-c-DOMG and PEG-DMG; PEG bound to phospholipids such as phosphatidylethanolamine (PEG-DLPE, PEG-DMPE and PEG-DSPE); PEG bound to ceramide (PEG-CER); PEG bound to cholesterol or a derivative thereof; PEG-modified phosphatidic acid; PEG-modified dialkylamine; and PEG-modified dialkylglycerol, but is not limited thereto. In addition, the pegylated lipid may be a functionalized PEG in which a functional group is combined on a side that is not combined with lipid. The functional group that can be used at this time may be at least one selected from the group consisting of succinyl, carboxylic acid, maleimide, n-hydroxysuccinimide, amine, biotin, cyanuric, and folate, but is not limited thereto.

[0159]

[0160] The above ionized lipids can be prepared into lipid nanoparticles, including neutral lipids, steroids, polymerized lipids, etc.

[0161] In order to have a composition suitable for manufacturing lipid nanoparticles, the lipid nanoparticle composition, excluding the active ingredient, may contain 20 to 65 mol% of ionized lipids, 2.5 to 30 mol% of neutral lipids, 20 to 60 mol% of steroids, and 0.5 to 5 mol% of polymerized lipids. For example, ionized lipid:neutral lipid:steroid:polymer polymerized lipid is 60:5:33.5:1.5, 50:10:38.5:1.5, 40:15:43.5:1.5, 30:20:48.5:1.5, 25:25:48.5:1.5, 59.5:5:33.5:2.0, 49.5:10:38.5:2.0, 39.5:15:43.5:2.0, 29.5:20:48.5:2.0, 24.5:25:48.5:2.0, 59:5:33.5:2.5, 49:10:38.5:2.5, 39:15:43.5:2.5, It can be composed in one mol% ratio selected from the group consisting of 29:20:48.5:2.5, 24:25:48.5:2.5, 58.5:5:33.5:3.0, 48.5:10:38.5:3.0, 38.5:15:43.5:3.0, 28.5:20:48.5:3.0, 23.5:25:48.5:3.0, 58:5:33.5:3.5, 48:10:38.5:3.5, 38:15:43.5:3.5, 28:20:48.5:3.5, and 23:25:48.5:3.5.

[0162]

[0163] The lipid nanoparticle composition may further comprise a prophylactic or therapeutic agent.

[0164] The agent may be a gene comprising RNA, DNA or a mixture thereof composed of single strands or double strands, and specifically may be at least one selected from the group consisting of small interfering ribonucleic acid (siRNA), ribosomal ribonucleic acid (rRNA), ribonucleic acid (RNA), deoxyribonucleic acid (DNA), complementary deoxyribonucleic acid (cDNA), aptamer, messenger ribonucleic acid (mRNA), transfer ribonucleic acid (tRNA), antisense oligonucleotide, small hairpin ribonucleic acid (shRNA), micro ribonucleic acid (miRNA), asymmetric interfering ribonucleic acid (aiRNA), dicer-substrate ribonucleic acid (dsRNA), ribozyme, peptide nucleic acid (PNA), deoxyribozyme (DNAzyme) and guide ribonucleic acid for gene correction (sgRNA), but is not limited thereto. The active ingredient of the above composition may be an anionic biopolymer-drug conjugate such as various anionic peptides, protein drugs, protein-nucleic acid structures, or hyaluronic acid-peptide conjugates, hyaluronic acid-protein conjugates, antibodies, etc.

[0165]

[0166] The lipid nanoparticle composition including ionized lipids, neutral lipids, steroids, polymerized lipids, etc. of the present invention can be prepared as a lipid solution by dissolving in a solvent miscible with ethanol or water. Separately, the active ingredient can be prepared as an active ingredient solution by dissolving in a citric acid or acetic acid buffer having a pH of 4.0±1.0. In addition, the lipid solution and the active ingredient solution can be mixed at a volume ratio of 1:3 at a flow rate of about 10 to 15 mL / min using a microfluidic mixing device (Benchtop Nanoassemblr, Precision Nanosystems) to prepare lipid nanoparticles.

[0167]

[0168] In addition, the present invention provides a drug delivery composition comprising the lipid nanoparticle composition and a preventive or therapeutic agent.

[0169] The above drug delivery composition can be administered to mammals, including humans, via various routes, including parenteral administration. Parenteral administration can be applied intravenously, subcutaneously, intraperitoneally, or locally. The dosage varies depending on the patient's condition and weight, the extent of the disease, the drug form, the route and time of administration, but can be appropriately selected by those skilled in the art. When formulating the drug delivery composition according to an example, it is manufactured using diluents or excipients such as commonly used fillers, bulking agents, lyophilizing agents, binders, wetting agents, disintegrating agents, and surfactants.

[0170] Formulations for parenteral administration include sterile aqueous solutions, non-aqueous solutions, suspensions, emulsions, lyophilized preparations, and suppositories. Non-aqueous solutions and suspensions can include propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable esters such as ethyl oleate. Suppository bases include witepsol, macrogol, Tween 61, cocoa butter, laurin, glycerol, and gelatin.

[0171] The drug delivery composition of the present invention can be administered containing a pharmaceutically effective amount of a preventive or therapeutic agent. The effective dosage level of the preventive or therapeutic agent can be determined based on the type and severity of the patient's disease, the activity and sensitivity of the drug, the time of administration, the route and excretion rate, the duration of treatment, factors including concomitant medications, and other factors well known in the medical field. In one embodiment, the composition can be administered as an individual therapeutic agent or in combination with another therapeutic agent, can be administered sequentially or simultaneously with conventional therapeutic agents, and can be administered singly or in multiple doses. It is important to administer an amount that achieves the maximum effect with the minimum amount without causing side effects, taking all of the above factors into consideration, and this can be easily determined by those skilled in the art. For example, the composition can be administered at 0.001 to 10 mg / kg, 0.01 to 5 mg / kg, or 0.1 to 1 mg / kg.

[0172] Hereinafter, to aid understanding of the present invention, examples will be given in detail. However, the following examples are intended only to illustrate the scope of the present invention and are not intended to limit its scope. These examples are provided to more fully explain the present invention to those of average skill in the art.

[0173]

[0174] <Example 1> Preparation of compound

[0175] 1-1. Preparation of compound 1

[0176] Compound 1 was prepared as shown in the following reaction scheme 1.

[0177] [Reaction Formula 1]

[0178]

[0179] Specifically, compound 1-a (1.57 g, 10.0 mmol) was dissolved in dichloromethane (29 mL, hereinafter DCM) at room temperature, and then compound 1-b (4.16 g, 10.0 mmol) and 4-dimethylaminopyridine (122 mg, 1.00 mmol, hereinafter DMAP), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride (2.88 g, 15.0 mmol, hereinafter EDCI) were added, followed by stirring at room temperature for 16 hours. After adding purified water (29 mL), stirring was performed, the organic layer was separated by standing, dried over sodium sulfate (hereinafter Na2SO4), and filtered. The solution was purified by column chromatography [methanol (hereinafter MeOH): DCM = 0:10 → 1:9] to obtain compound 1-c. Compound 1-c (3.56 g, 6.42 mmol) was dissolved in DCM (18 mL) at room temperature, trifluoroacetic acid (18 mL, hereinafter referred to as TFA) was slowly added, and the mixture was stirred at room temperature for 3 hours, concentrated under reduced pressure, and azeotropically concentrated with DCM 4 to 5 times to obtain compound 1-d. Compound 1-d (1.07 g, 2.14 mmol) was dissolved in N-methyl-2-pyrrolidone (32 mL, hereinafter referred to as NMP) at room temperature, and compound 1-e (2.68 g, 17.2 mmol), DMAP (3.14 g, 25.7 mmol), and EDCI (4.93 g, 25.7 mmol) were added, and the external temperature was raised to 40°C and stirred for 16 hours. Purified water (32 mL) was added, extracted with methyl-tert-butyl ether (32 mL, methyl-tert-butyl ether, hereinafter referred to as MTBE), washed with saturated sodium chloride solution (32 mL*2 times, hereinafter referred to as brine), dried with Na2SO4, filtered, concentrated under reduced pressure, dissolved in 10 mL of DCM, and column purified (MeOH: DCM = 0:10 → 1:9) to obtain compound 1.

[0180] 1 H NMR: (400 MHz, CDCl3) δ H8.67 (1H, s), 5.3-5.44 (6H, m), 4.04-4.07 (6H, m), 2.52-2.53 (4H, m), 2.26-2.31 (6H, m), 1.98-2.12 (16H, m), 1.56-1.71 (12H, m), 1.12-1.36 (24H, m), 0.87-0.90 (9H, m)

[0181]

[0182] 1-2. Preparation of compound 3

[0183] Compound 3 was prepared according to the following reaction scheme 2.

[0184] [Reaction Formula 2]

[0185]

[0186] Compound 3-a (831 mg, 2.00 mmol) was dissolved in diethyl ether (42 mL, hereinafter referred to as Et2O) at room temperature, triethylamine (697 μL, hereinafter referred to as TEA) and triphosgene (386 mg, 1.30 mmol, triphosgene) were added, and the mixture was stirred under nitrogen for 2 hours to obtain compound 3-b. Compound 3-c (533 μL, 4.00 mmol) was added, and the mixture was stirred under nitrogen for 16 hours at room temperature. After washing with saturated sodium bicarbonate solution (42 mL, hereinafter referred to as sat.aq, NaHCO3), the organic layer was dried over Na2SO4, concentrated under reduced pressure, dissolved in DCM (10 mL), and purified by column chromatography (MeOH: DCM = 0:10 → 2:8) to obtain compound 3-d. Compound 3-d (332 mg, 0.594 mmol) was dissolved in 1-4-dioxane (1.5 mL, hereinafter referred to as Dioxane) at room temperature, 4.0 M hydrochloric acid solution (1.49 mL, 5.91 mmol, in Dioxane) was added, and the mixture was stirred at room temperature for 16 hours. The mixture was concentrated under reduced pressure and azeotropically concentrated with DCM 4 to 5 times to obtain compound 3-e. Compound 3-e (254 mg, 0.594 mmol) was dissolved in NMP (7.6 mL) at room temperature, and compound 3-f (548 g, 2.97 mmol), DMAP (333 mg, 25.7 mmol), and EDCI (570 mg, 2.97 mmol) were added, and the temperature was raised to 45°C and stirred for 16 hours. After extraction with purified water (7.6 mL) and MTBE (7.6 mL), washing with brine (7.6 mL*2 times), drying with Na2SO4, filtering and concentrating under reduced pressure, dissolving in DCM (3 mL) and column purification (MeOH: DCM = 0:10 → 1:9) was performed to obtain compound 3.

[0187] 1 H NMR: (400 MHz, CDCl3) δ H5.30-5.41 (6H, m), 4.02-4.07 (8H, m), 2.41-2.52 (2H, m), 2.28-2.31 (6H, m), 1.96-2.07 (16H, m), 1.62-1.66 (14H, m), 1.39-1.42 (4H, m), 1.25-1.33 (26H, m), 0.84-0.90 (9H, m)

[0188]

[0189] 1-3. Preparation of compound 4

[0190] Compound 4 was prepared according to the following reaction scheme 3.

[0191] [Reaction Formula 3]

[0192]

[0193] Compound 4-a (1.57 g, 10.0 mmol) was dissolved in DCM (29 mL) at room temperature, and compound 4-b (4.16 g, 10.0 mmol), DMAP (122 mg, 1.00 mmol), and EDCI (2.88 g, 15.0 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After adding purified water (29 mL), stirring was performed, the organic layer was separated, dried over Na2SO4, and filtered. The solution was purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 4-c. Compound 4-c (3.56 g, 6.42 mmol) was dissolved in DCM (18 mL) at room temperature, and TFA (18 mL) was slowly added. The mixture was stirred at room temperature for 3 hours, concentrated under reduced pressure, and azeotropically concentrated with DCM 4 to 5 times to obtain compound 4-d. Compound 4-d (400 mg, 0.799 mmol) was dissolved in NMP (12 mL) at room temperature, then compound 4-e (954 mg, 5.18 mmol), DMAP (632 mg, 5.18 mmol), and EDCI (992 mg, 5.18 mmol) were added, the temperature was raised to 40°C, and the mixture was stirred for 16 hours. Purified water (12 mL) was added, extracted with MTBE (12 mL), washed with brine (12 mL*2 times), dried over Na2SO4, filtered, and concentrated under reduced pressure. Then, the mixture was dissolved in DCM (10 mL) and purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 4.

[0194] 1 H NMR: (400 MHz, CDCl3) δ H 5.29-5.42 (6H, m), 4.03-4.07 (6H, m),2.45-2.54 (H6, m), 2.26-2.3 (6H, m), 1.99-2.08 (12H, m), 1.57-1.65 (18H, m), 1.38-1.44 (6H, m), 1.24-1.36 (26H, m), 0.87-0.90 (9H, m)

[0195]

[0196] 1-4. Preparation of compound 5

[0197] Compound 5 was prepared as shown in the following reaction scheme 4.

[0198] [Reaction Formula 4]

[0199]

[0200] Compound 5-a (1.57 g, 10.0 mmol) was dissolved in DCM (29 mL) at room temperature. Compound 5-b (4.16 g, 10.0 mmol), DMAP (122 mg, 1.00 mmol), and EDCI (2.88 g, 15.0 mmol) were added, and the mixture was stirred at room temperature for 16 hours. Purified water (29 mL) was added, stirred, allowed to stand, and the organic layer was separated, dried over Na2SO4, and filtered. The solution was purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 5-c. Compound 5-c (3.56 g, 6.42 mmol) was dissolved in DCM (18 mL) at room temperature, TFA (18 mL) was slowly added, and the mixture was stirred at room temperature for 3 hours. The mixture was concentrated under reduced pressure and azeotropically concentrated with DCM 4 to 5 times to obtain compound 5-d. Compound 5-d (400 mg, 0.799 mmol) was dissolved in NMP (12 mL) at room temperature, then compound 5-e (1.10 g, 5.18 mmol), DMAP (632 mg, 5.18 mmol), and EDCI (992 mg, 5.18 mmol) were added, the temperature was raised to 40°C, and the mixture was stirred for 16 hours. 12 mL of purified water was added, extracted with MTBE (12 mL), washed with brine (12 mL*2 times), dried over Na2SO4, filtered, and concentrated under reduced pressure. Then, the mixture was dissolved in DCM (10 mL) and purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 5.

[0201] 1 H NMR: (400 MHz, CDCl3) δ H8.63 (1H, s), 5.33-5.39 (6H, m), 4.02-4.06 (6H, m), 2.45-2.56 (2H, m), 2.25-2.29 (8H, m), 1.99-2.03 (12H, m), 1.48-1.59 (12H, m), 1.24-1.34 (52H, m), 0.87-0.90 (9H, m)

[0202]

[0203] 1-5. Preparation of compound 6

[0204] Compound 6 was prepared as shown in the following reaction scheme 5.

[0205] [Reaction Formula 5]

[0206]

[0207] Compound 6-a (1.57 g, 10.0 mmol) was dissolved in DCM (29 mL) at room temperature, and compound 6-b (4.16 g, 10.0 mmol), DMAP (122 mg, 1.00 mmol), and EDCI (2.88 g, 15.0 mmol) were added, and the mixture was stirred at room temperature for 16 hours. After adding purified water (29 mL), stirring was performed, the organic layer was separated, dried over Na2SO4, and filtered. The solution was purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 6-c. Compound 6-c (3.56 g, 6.42 mmol) was dissolved in DCM (18 mL) at room temperature, and TFA (18 mL) was slowly added. The mixture was stirred at room temperature for 3 hours, concentrated under reduced pressure, and azeotropically concentrated with DCM 4 to 5 times to obtain compound 6-d. Compound 6-d (500 mg, 0.999 mmol) was dissolved in NMP (15 mL) at room temperature, then compound 6-e (1.03 g, 7.99 mmol), DMAP (1.47 g, 12.0 mmol), and EDCI (2.30 g, 12.0 mmol) were added, the temperature was raised to 40°C, and the mixture was stirred for 16 hours. Purified water (15 mL) was added, extracted with MTBE (15 mL), washed with brine (15 mL*2 times), dried over Na2SO4, filtered, and concentrated under reduced pressure. Then, the mixture was dissolved in DCM (10 mL) and purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 6.

[0208] 1 H NMR: (400 MHz, CDCl3) δ H 8.66 (1H, s), 5.29-5.53 (6H, m), 2.25-2.56 (24H, m), 1.99-2.07 (6H, m), 1.59-1.62 (8H, m), 1.49 (2H, s) 1.24-1.36 (10H, m), 0.87-0.90 (9H, m)

[0209]

[0210] 1-6. Preparation of compound 7

[0211] Compound 7 was prepared as shown in the following reaction scheme 6.

[0212] [Reaction Formula 6]

[0213]

[0214] Compound 7-a (1.43 g, 10.0 mmol) was dissolved in DCM (29 mL) at room temperature, and compound 7-b (4.16 g, 10.0 mmol), DMAP (122 mg, 1.00 mmol), and EDCI (2.88 g, 15.0 mmol) were added. The mixture was stirred at room temperature for 16 hours. Purified water (29 mL) was added, stirred for 10 minutes, allowed to stand, and the organic layer was separated, dried over Na2SO4, filtered, and the solution was purified by column chromatography (methanol: DCM = 0:10 → 1:9) to obtain compound 7-c. Compound 7-c (3.56 g, 6.58 mmol) was dissolved in 18 mL of DCM at room temperature, TFA (18 mL) was slowly added, and the mixture was stirred at room temperature for 3 hours, concentrated under reduced pressure, and azeotropically concentrated with DCM 4 to 5 times to obtain compound 7-d. Compound 7-d (730 mg, 1.50 mmol) was dissolved in NMP (21 mL) at room temperature, and compound 7-e (1.88 g, 12.0 mmol), DMAP (2.20 g, 18.0 mmol), and EDCI (3.45 g, 18.0 mmol) were added, and the mixture was heated to 40°C and stirred for 16 hours. Add 21 mL of purified water, extract with MTBE (21 mL), wash with brine (21 mL*2 times), dry with Na2SO4, filter and concentrate under reduced pressure, then dissolve in 10 mL of DCM and column purify (MeOH: DCM = 0:10 → 1:9) to obtain compound 7.

[0215] 1 H NMR: (400 MHz, CDCl3) δ H5.78 (1H, s), 5.31-5.43 (6H, m), 4.04-4.07 (6H, t), 2.89-2.92 (3H, m), 2.26-2.3 (9H, m), 1.98-2.12 (14H, m), 1.64-1.84 (16H, m), 1.23-1.38 (18H, m), 0.87-0.90 (9H, t)

[0216]

[0217] 1-7. Preparation of compound 8

[0218] Compound 8 was prepared as shown in the following reaction scheme 7.

[0219] [Reaction Formula 7]

[0220]

[0221] Compound 8-a (1.80 g, 10.0 mmol) was dissolved in DCM (36 mL) at room temperature, and compound 8-b (4.16 g, 10.0 mmol), DMAP (122 mg, 1.00 mmol), and EDCI (2.88 g, 15.0 mmol) were added, and the mixture was stirred at room temperature for 16 hours. 29 mL of purified water was added, stirred, allowed to settle, and the organic layer was separated, dried over Na2SO4, and filtered. The solution was purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 8-c. Compound 8-c (4.87 g, 9.01 mmol) was dissolved in DCM (24 mL) at room temperature, and TFA (24 mL) was slowly added. The mixture was stirred at room temperature for 3 hours, concentrated under reduced pressure, and azeotropically concentrated with DCM to obtain compound 8-d. Compound 8-d (671 mg, 1.43 mmol) was dissolved in NMP (17 mL) at room temperature, then compound 8-e (2.33 g, 14.3 mmol), DMAP (2.62 g, 21.4 mmol), and EDCI (4.11 g, 21.4 mmol) were added, the temperature was raised to 40°C, and the mixture was stirred for 16 hours. Purified water (17 mL) was added, extracted with MTBE (17 mL), washed with brine (17 mL*2 times), dried over Na2SO4, filtered, and concentrated under reduced pressure. Then, the mixture was dissolved in DCM (10 mL) and purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 8.

[0222] 1 H NMR: (400 MHz, CDCl3) δ H 5.29-5.44 (6H, m), 4.04-4.08 (6H, t), 2.62-2.68 (2H, m), 2.25-2.31 (6H, m), 1.98-2.12 (14H, m), 1.58-1.72 (16H, m), 1.23-1.38 (20H, m), 0.87-0.90 (9H, m)

[0223]

[0224] 1-8. Preparation of compound 14

[0225] Compound 14 was prepared as shown in the following reaction scheme 8.

[0226] [Reaction Formula 8]

[0227]

[0228] Compound 14-a (157 mg, 1.00 mmol) was dissolved in DCM (3.2 mL) at room temperature, and compound 14-b (416 mg, 1.00 mmol), DMAP (12.2 mg, 0.10 mmol), and EDCI (288 mg, 1.50 mmol) were added. The mixture was stirred at room temperature for 16 hours. Purified water (3.2 mL) was added, stirred, and allowed to stand to separate the organic layer. The solution was dried over Na2SO4 and filtered. The resulting solution was purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 14-c. Compound 14-c (442 mg, 0.797 mmol) was dissolved in DCM (4.4 mL) at room temperature, TFA (4.4 mL) was slowly added, and the mixture was stirred at room temperature for 3 hours, concentrated under reduced pressure, and azeotropically concentrated with DCM 4 to 5 times to obtain compound 14-d. Compound 14-d (175 mg, 0.414 mmol) was dissolved in NMP (5.3 mL) at room temperature, and compound 14-e (603 mg, 1.45 mmol), DMAP (253 mg, 2.07 mmol), and 2,4,6-tripropyl-1,3,5,2,4,6-trioxatriphosphorinane-2,4,6-trioxide solution (1.32 g, 2.07 mmol, 50 wt% in DCM) were added, and the temperature was raised to 100°C and stirred for 16 hours. Purified water (5.3 mL) was added, extracted with EA (11 mL), washed with brine (5.3 mL*2 times), dried with Na2SO4, filtered, concentrated under reduced pressure, dissolved in DCM (10 mL), and column purified (MeOH: DCM = 0:10 → 1:9) to obtain compound 14.

[0229] 1H NMR: (400 MHz, CDCl3) δ H 4.04-4.07 (6H, m), 3.70-3.74 (1H, m), 2.25-2.28 (6H, m), 2.01-2.03 (6H, m), 1.56-1.57 (18H, m), 1.23-1.26 (78H, m), 0.87-0.90 (18H, m)

[0230]

[0231] 1-9. Preparation of compound 18

[0232] Compound 18 was prepared as shown in the following reaction scheme 9.

[0233] [Reaction Formula 9]

[0234]

[0235] Compound 18-a (901 mg, 3.42 mmol) was dissolved in DCM (18 mL) at room temperature, and compound 18-b (1.42 g, 3.42 mmol), DMAP (41.8 mg, 0.342 mmol), and EDCI (983 mg, 5.13 mmol) were added. The mixture was stirred at room temperature for 16 hours. Purified water (18 mL) was added, stirred, and allowed to stand to separate the organic layer. The solution was dried over Na2SO4 and filtered. The resulting solution was purified by column chromatography (MeOH: DCM = 0:10 → 1:9) to obtain compound 18-c. Compound 18-c (1.12 g, 1.83 mmol) was dissolved in EA (5 mL) at room temperature, hydrochloric acid solution (4.34 g, 18.3 mmol, 15.4 wt% in EA) was added, and the mixture was stirred at room temperature for 16 hours and concentrated under reduced pressure to obtain compound 18-d. Compound 18-d (892 mg, 1.81 mmol) was dissolved in NMP (29 mL) at room temperature, and compound 18-e (2.78 g, 10.8 mmol), DMAP (1.99 g, 16.6 mmol), and EDCI (3.12 g, 16.6 mmol) were added, and the temperature was raised to 85°C and stirred for 16 hours. After cooling, purified water (29 mL) was added, extracted with EA (58 mL), washed with brine (29 mL x 2), dried with Na2SO4, filtered and concentrated under reduced pressure, dissolved in DCM (10 mL), and column purified (MeOH : DCM = 0:10 → 1:9) to obtain compound 18-f. Compound 18-f (185 mg, 0.153 mmol) was dissolved in MeOH (3.7 mL) at room temperature, and palladium-on-carbon catalyst (18.5 mg, 5 wt%) was added, followed by replacement with hydrogen gas and stirring for 16 hours. The mixture was filtered with celite and washed with MeOH (3.7 mL). The filtered solution was concentrated under reduced pressure and dried to obtain compound 18.

[0236] 1 H NMR: (400 MHz, CDCl3) δ H4.05-4.11 (6H, m), 3.65-3.75 (4H, m), 3.04 (2H, s), 2.62 (2H, s), 2.26-2.30 (6H, m), 2.01-2.05 (6H, m), 1.54-1.69 (10H, m), 1.23-1.32 (78H, m), 0.87-0.90 (18H, m)

[0237]

[0238] <Example 2> Preparation of lipid nanoparticles

[0239] Lipid nanoparticles encapsulating Firefly Luciferase mRNA (RNA Gene, Korea) were prepared using the compound (ionized lipid) prepared in Example 1 above, dioleoylphosphatidylethanolamine (DOPE) or distearoylphosphatidylcholine (DSPC) (neutral lipid), cholesterol (steroid), and PEG 2000-DMG (polymerized lipid). Ionized lipids, DOPE / DSPC, cholesterol, and PEG 2000-DMG were prepared in a molar % ratio of 50:10:38.5:1.5 (composition indicated as A) or 49:10:38.5:2.5 (composition indicated as B) and dissolved in ethanol at a concentration of 4.8 mg / mL to prepare a lipid solution. Separately, Firefly Luciferase mRNA was dissolved in a citric acid buffer solution at pH 4.0±1.0 or an acetate buffer solution at pH 5.0±1.0 (composition indicated as *) at a weight ratio of 1:16 mRNA:ionized lipid to prepare an active ingredient solution. Then, lipid nanoparticles were prepared by passing the mixture through a microfluidic mixing device (Benchtop Nanoassemblr, Precision Nanosystems) at a flow rate of approximately 15 mL / min so that the lipid solution:active ingredient solution had a volume ratio of 1:3. The manufactured lipid nanoparticles were diluted with Tris buffer containing 8.7% sucrose and dialyzed using a dialysis centrifuge tube to ensure that the ethanol content was less than 1%, and the final concentration was manufactured to be 0.2 mg / mL based on mRNA.

[0240] For comparison, lipid nanoparticles encapsulating Firefly Luciferase mRNA (RNA Gene, Korea) were prepared using SM-102 ionizable lipid, distearoylphosphatidylcholine (DSPC) (neutral lipid), cholesterol (steroid), and DMG-PEG2000 (polymeric lipid). SM-102, DSPC, cholesterol, and DMG-PEG2000 were prepared in a molar % ratio of 50:10:38.5:1.5, the same as in the SpikeVax example, and used as lipid nanoparticles for comparison.

[0241]

[0242] <Analysis Example 1> Confirmation of lipid nanoparticle size and polydispersity

[0243] The size and surface charge of lipid nanoparticles containing the compound of Example 1 were measured. To measure the size of lipid nanoparticles, the concentration of mRNA contained in each lipid nanoparticle was diluted with PBS to 1 μg / mL, and the diameter and polydispersity index (PDI) of lipid nanoparticles (LNPs) were measured using dynamic light scattering (DLS) on a Malvern Zetasizer Nano (Malvern Instruments, UK).

[0244] As a result, as shown in Table 1, all lipid nanoparticles exhibited a size of approximately 140 nm or less and a good polydispersity of less than 0.2.

[0245] Compound No. Particle Size (nm) PDI Compound 1 A9 10.06 Compound 1 B7 9 0.04 Compound 3 A1 0.06 Compound 3 B6 4 0.08 Compound 4 A8 4 0.04 Compound 4 B6 4 0.05 Compound 5 A8 8 0.05 Compound 5 B7 2 0.06 Compound 7 B5 5 0.09 Compound 8 B5 9 0.06 Compound 14 A1 3 9 0.14 Compound 14 B1 1 2 0.17 Compound 18 A1 0 3 0.04 Compound 18 B7 2 0.05

[0246]

[0247] <Analysis Example 2> Confirming Gene Encapsulation Rate

[0248] To confirm the gene encapsulation rate of lipid nanoparticles including the compound of Example 1 above, Quant-iT TM Ribogreen TM RNA Reagent and Kit were used. 5 μL of ribosomal RNA standard (100 μg / mL in TE buffer) was taken and diluted with 245 μL TE buffer or 0.4% Triton-TE buffer to prepare a stock solution for the calibration curve. 2, 5, 10, 25, and 50 μL of the stock solution were diluted with TE buffer or 0.4% Triton-TE buffer to make a total volume of 100 μL, and then added to Quant-iTTM Ribogreen. TMAfter adding 100 μL of RNA Reagent and mixing, the fluorescence intensity (excitation 475 nm and emission 500-550 nm) was measured to create a calibration curve. 5 μL of the nanoparticle solution prepared in Example 2 was diluted with 245 μL of TE buffer (10 mM Tris-HCl, 1 mM EDTA, pH 7.5 in DEPC-treated water). 50 μL of TE buffer or 2% Triton-TE buffer was added to 50 μL of the diluted nanoparticle solution, and incubated at 37°C for about 10 minutes. 40 μL of each culture medium was then placed in a microplate, and 60 μL of TE buffer and Quant-iT were added. TM Ribogreen TM 100 μL of RNA reagent was sequentially added, mixed, and the fluorescence intensity was measured. The measured fluorescence intensity was applied to the calibration curve to calculate the gene content in TE buffer (TO) and in 2% Triton-TE buffer (T2). Then, the gene inclusion rate was calculated using Equation 1 below. The gene inclusion rate was confirmed and is shown in Table 2. As a result, all samples showed good gene inclusion rates.

[0249] [Formula 1]

[0250]

[0251]

[0252] As a result, as shown in Table 2 below, it was confirmed that all compounds showed a good gene encapsulation rate of 85% or more.

[0253] Compound number Gene inclusion rate (%) Compound 1 A87.2 Compound 1 B91.7 Compound 3 A93.3 Compound 3 B96.0 Compound 4 A90.4 Compound 4 B88.5 Compound 5 A92.9 Compound 5 B93.5 Compound 7 B93.5 Compound 8 B94.6 Compound 14 A95.8 Compound 14 B95.9 Compound 18 A95.9 Compound 18 B95.1

[0254]

[0255] <Analysis Example 3> Confirmation of lipid nanoparticle gene delivery efficiency

[0256] 3-1. Confirmation of intracellular gene transfer efficiency

[0257] In order to confirm the intracellular gene delivery efficiency of lipid nanoparticles including the compound of Example 1, the reporter gene FLuc mRNA was encapsulated in lipid nanoparticles and its intracellular expression efficiency was evaluated. Since lipid nanoparticles mediate the low-density lipoprotein receptor (LDL receptor), a cell surface receptor, as a cellular uptake mechanism, HeLa cell line (Korea Cell Line Bank, Seoul), a human cervical cancer cell line with high expression of LDL receptor on the cell surface, was used, and cell culture was performed using MEM (minimum essential medium) medium (Gibco) containing 10% fetal bovine serum (Gibco, NY, USA) and penicillin / streptomycin (Gibco) at 37°C and 5% CO2 conditions. Before lipid nanoparticle treatment, HeLa cell line was seeded at 2 × 10 per well. 4 Cells were cultured in 96-well plates, and after stabilization for 16 to 24 hours, lipid nanoparticles at a concentration of 0.25 μg / mL based on the encapsulated mRNA were diluted in cell culture medium and treated to the cells.

[0258] To confirm the expression of intracellular luciferase, 6 hours after lipid nanoparticle treatment, the culture medium in the wells was removed, washed with PBS, and 100 μL of Glo-Lysis Buffer (Promega, WI, USA) was added to lyse the cells at room temperature for 5 minutes. Next, 50 μL of each cell lysate and Steady-Glo™ Luciferase Assay solution (Promega) were added at a 1:1 ratio to a 96-well white plate, and after incubation at room temperature for 5 minutes, the luminescence value was detected using a GloMax Discover microplate reader (Promega). A calibration curve was created using recombinant luciferase (Promega) as a standard, and the luciferase protein concentration (pg / mL) of the cell lysate was derived from this.

[0259] As a result, as shown in Table 3 below, it was confirmed that lipid nanoparticles including the compound of Example 1 exhibited good gene delivery efficacy into cells.

[0260] Compound number Intracellular luciferase expression level (mean ± SD, pg / mL) Compound 1 B2 56,690 ± 2,228 Compound 3 A10,777 ± 936 Compound 3 B6,744 ± 619 Compound 4 A168,042 ± 11,196 Compound 4 B75,952 ± 6,333 Compound 5 A3 36,723 ± 18,075 Compound 5 B171,157 ± 29,254 Compound 7 B2,701 ± 138 Compound 8 B109,640 ± 11,923 Compound 18 A7,566 ± 662 Compound 18 B4,471 ± 1,181

[0261]

[0262] 3-2. Confirmation of in vivo gene transfer efficiency 1

[0263] In order to confirm the in vivo gene transfer efficiency of lipid nanoparticles including the compound of Example 1, lipid nanoparticles were intravenously injected once at a dose of 1 mg / kg based on the luciferase gene loaded into 7-week-old C57BL / 6 mice, and after 4 hours, the mice were sacrificed and their livers were removed.

[0264] To determine the level of luciferase expression in liver tissue samples, the excised liver tissue was transferred to a tube containing 3 mm metal beads, 500 μL of Glo-Lysis Buffer (Promega) per 50 mg was added, and the tissue was homogenized using a bead homogenizer. The homogenate was centrifuged at 1,000 g for 10 minutes, and the supernatant was diluted to an appropriate ratio using Glo-Lysis Buffer. The diluted tissue homogenate and Steady-Glo Luciferase Assay (Promega) solution were each transferred to a 96-well white plate at a 1:1 ratio, and reacted at room temperature for 5 minutes. Next, the luminescence value was detected using a GloMax Discover microplate reader (Promega), and a calibration curve was generated using recombinant luciferase (Promega) as a standard, through which the luciferase expression level of the sample (ng / g tissue) was derived.

[0265] As a result, as shown in Table 4 below, the lipid nanoparticles containing the compound of Example 1 exhibited high levels of luciferase expression in liver tissue upon intravenous injection, thereby confirming their function as a gene delivery vehicle with excellent delivery efficacy in the body through systemic administration. In addition, in an in vivo test, composition B showed similar or higher delivery efficacy compared to composition A.

[0266] Compound number: Luciferase expression in the liver after intravenous injection (Mean ± SD, ng / g tissue) Compound 1 A90,263 ± 7,749 Compound 1 B105,111 ± 3,985 Compound 1 B *148,472 ± 19,655 Compound 4 A66,632 ± 5,590 Compound 5 A52,549 ± 12,859 Compound 5 B69,984 ± 14,910 Compound 14 A1,211 ± 144 Compound 14 B1,469 ± 664

[0267]

[0268] 3-3. Confirmation of in vivo gene transfer efficiency 2

[0269] To determine the in vivo gene transfer efficiency of repeated administration of lipid nanoparticles, 7-week-old C57BL / 6 mice were intravenously injected with lipid nanoparticles loaded with EPO mRNA (TriLink BioTechnologies, CA, USA) at a dose of 0.5 mg / kg per gene. Blood samples were collected 6 hours after lipid nanoparticle administration, and plasma was separated by anticoagulation.

[0270] To determine the concentration of EPO protein expressed in plasma, an EPO enzyme-linked immunosorbent assay (ELISA) kit (R&D Systems, MN, USA) was used for analysis according to the manufacturer's method. More specifically, plasma samples were diluted 1:5000 using the specimen diluent included in the kit, mixed with 100 μL of assay diluent in a 1:1 ratio in microwells, and incubated for 2 hours at room temperature. After removing all the solution in each well, 200 μL of EPO conjugate was added, and incubated for another 2 hours at room temperature. After washing four times with 300 μL of washing buffer, 200 μL of substrate solution was added, and incubated for 20 minutes. To terminate the reaction, 100 μL of stop solution was added, and the absorbance was measured at wavelengths of 450 nm and 600 nm using a microplate reader (Promega). A calibration curve was prepared using the standard material provided in the kit, and the EPO protein concentration (ng / mL) of each sample was calculated within the defined range of the standard material.

[0271] As a result, as shown in Table 5 below, the lipid nanoparticles including the compound of Example 1 showed a higher level of EPO expression in the blood compared to the lipid nanoparticles including the ionized lipid Lipid 5 used as a control during repeated intravenous injections, and when administered again, it was confirmed that, unlike the Lipid 5 lipid nanoparticles, the lipid nanoparticles including the compound of Example 1 maintained a high level of EPO expression well. Considering that Lipid 5 is known to be a carrier that maintains a good gene delivery ability during repeated administration among carriers to date, it can be confirmed that the compound of Example 1 has characteristics that are advantageous for repeated administration.

[0272] Compound number EPO expression in blood after intravenous injection (Mean ± SD, ng / mL) 1st dose 2nd dose Compound 1 B 6,841 ± 87 46,441 ± 609 Lipid 52,321 ± 40 28 14 ± 321

[0273]

[0274] While specific aspects of the present invention have been described in detail above, it should be apparent to those skilled in the art that these specific descriptions merely represent preferred embodiments and are not intended to limit the scope of the present invention. In other words, the substantial scope of the present invention is defined by the appended claims and their equivalents.

Claims

1. R around the central carbon as in the chemical formula 1 below 1 , R 2 , R 3 and (R 4 )-X is selected from the group consisting of a compound, a pharmaceutically acceptable salt, a tautomer, a prodrug or a stereoisomer thereof: <Chemical Formula 1> In the above chemical formula 1, R 1 , R 2 and R 3 are each independently selected from (C6-C30) alkyl, alkenyl, alkynyl, (C6-C30) cycloalkyl, aryl or heteroaryl with or without a branch, and among these, at least one -CH2- at a third or more carbon position from the central carbon is -O-, -CH(OH)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OP(=O)(OR 5 )O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -SC(=S)O-, -OC(=O)S-, -OC(=S)O-, -S-, -SS-, -NR 5 -, -C(=O)NR 5 -, -NR 5 C(=O)-, -OC(=O)NR 5 -, -NR 5 C(=O)O-, -C(=S)NR 5 -, -NR 5 C(=S)-, -OC(=S)NR 5 -, -NR 5 C(=S)O-, -SC(=O)NR 5 -, -SC(=S)NR 5 - and -NR 5 is substituted with one selected from the group consisting of C(=O)S-, wherein, R 5 is independently selected from hydrogen, (C1-C20) alkyl, (C2-C20) alkenyl, alkynyl, (C3-C20) cycloalkyl, aryl or heteroaryl, R 4 is selected from (C2-C12) alkylene, alkenylene, alkynylene, (C6-C12) cycloalkylene, aryl or heteroarylene with or without a branch, and one or more -CH2- is -O-, -CH(OH)-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OP(=O)(OR 5 )O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -SC(=S)O-, -OC(=O)S-, -OC(=S)O-, -S-, -SS-, -NR 5 -, -C(=O)NR 5 -, -NR 5 C(=O)-, -OC(=O)NR 5 -, -NR 5 C(=O)O-, -C(=S)NR 5 -, -NR 5 C(=S)-, -OC(=S)NR 5 -, -NR 5 C(=S)O-, -SC(=O)NR 5 -, -SC(=S)NR 5 - and -NR 5 Substituted or unsubstituted with any one selected from the group consisting of C(=O)S-, X is selected from a chain or cyclic hetero group containing hydrogen or an ionizable amine, wherein the cyclic hetero group is unsubstituted or substituted with (C1-C6)alkyl, hydroxy or hydroxy(C1-C6)alkyl, n is chosen from 0 or 1.

2. In paragraph 1, The above R 1 , R 2 and R 3 are each independently a compound represented by the following chemical formula 1-A: <Chemical Formula 1-A> In the above chemical formula 1-A, L is (C2-C5)alkylene, A is -O-, -C(=O)-, -OC(=O)-, -C(=O)O-, -OC(=O)O-, -OC(=S)-, -C(=S)O-, -SC(=O)-, -C(=O)S-, -SC(=S)-, -C(=S)S-, -SC(=O)O-, -OC(=O)S-, -S-, -SS-, -C(=O)NR 5 -, -OC(=O)NR 5 -, -C(=S)NR 5 - and -OC(=S)NR 5 - is selected from the group consisting of, R is selected from (C4-C25) alkyl, alkenyl, alkynyl, cycloalkyl, aryl or heteroaryl with or without branching, wherein at least one -CH2- among these is substituted or unsubstituted with any one selected from the group consisting of -O-, -S-, and -SS-.

3. In paragraph 1, The above R 4 is selected from alkylene, alkenylene or alkynylene of (C2-C6), and at least one of them -CH2- is -NR 5 -, -C(=O)NR 5 -, -OC(=O)NR 5 -, -C(=S)NR 5 -, -OC(=S)NR 5 -, -SC(=O)NR 5 -, and -SC(=S)NR 5 - is substituted or unsubstituted with any one selected from the group consisting of, wherein, R 5 A compound characterized in that it is independently selected from hydrogen or (C1-C6)alkyl.

4. In paragraph 1, A compound wherein X is selected from amino substituted with one or more (C1-C4)alkyl; or a cyclic hetero group selected from the group consisting of piperidine, piperazine, and imidazole, wherein the cyclic hetero group is substituted or unsubstituted with (C1-C4)alkyl, hydroxy, or hydroxy(C1-C4)alkyl.

5. In paragraph 1, The above compound is, (1) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (2) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(5-(dimethylamino)pentanamido)heptanedioate, (3) di((Z)-dodec-5-en-1-yl) 4-(((4-(dimethylamino)butoxy)carbonyl)amino)-4-(3-oxo-3-(((Z)-tridec-6-en-1-yl)oxy)propyl)heptanedioate, (4) di((Z)-dodec-5-en-1-yl) 4-(3-(((Z)-dodec-5-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (5) di((Z)-tetradec-9-en-1-yl) 4-(3-oxo-3-(((Z)-tetradec-9-en-1-yl)oxy)propyl)-4-(3-piperidin-1-yl)propanamido)heptanedioate, (6) di((z)-oct-3-en-1-yl) 4-(3(((Z)-oct-3-en-1-yl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (7) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-methylpiperidine-4-carboxamido)heptanedioate, (8) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-methylpiperidine-3-carboxamido)heptanedioate, (9) bis(2-(heptylthio)ethyl) 4-(3-(2-(heptylthio)ethoxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (10) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(1-(2-hydroxyethyl)piperidine-4-carboxamido)heptanedioate, (11) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(4-piperidin-1-yl)butanamido)heptanedioate, (12) 4-(3-(piperidin-1-yl)propanamido)-4-(3-(((Z)-tetradec-9-enoyl)oxy)propyl)heptane-1,7-diyl(9Z,9'Z)-bis(tetradec-9-enoate)], (13) di((Z)-dec-4-en-1-yl) 4-(3-(1H-imidazol-1-yl)propanamido)-4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)heptanedioate, (14) bis(3-(hexylundecyl) 4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (15) bis(3-(hexylundecyl) 4-(3-oxo-3-(undecyloxy)propyl)-4-(3-(piperidin-1-yl)propanamido)heptanedioate, (16) bis(3-(hexylundecyl) 4-(((3-(dimethylamino)propoxy)carbonyl)amino)-4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)heptanedioate, (17) bis(3-(hexylundecyl) 4-(((3-(diethylamino)propoxy)carbonyl)amino)-4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)heptanedioate, (18) bis(3-hexylundecyl) 4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)-4-(1-(2-hydroxyethyl)piperidine-4-carboxamido)heptanedioate, (19) 4-(3-((2-hexyldecanoyl)oxy)propyl)-4-(3-(piperidin-1-yl)propanamido)heptane-1,7-diyl bis(2-hexyldecanoate)[4-(3-((2-hexyldecanoyl)oxy)propyl)-4-(3-(piperidin-1-yl)propanamido)heptane-1,7-diyl bis(2-hexyldecanoate)], (20) bis(3-hexylundecyl) 4-(3-(1H-imidazol-1-yl)propanamido)-4-(3-((3-hexylundecyl)oxy)-3-oxopropyl)heptanedioate, (21) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(3-(4-methylpiperazin-1-yl)propanamido)heptanedioate, (22) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(piperidin-1-yl)heptanedioate, (23) di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(4-(hydroxymethyl)piperidin-1-yl)heptanedioate, and (24) A compound characterized by comprising at least one selected from the group consisting of di((Z)-dec-4-en-1-yl) 4-(3-(((Z)-dec-4-en-1-yl)oxy)-3-oxopropyl)-4-(4-methylpiperazin-1-yl)heptanedioate.

6. A lipid nanoparticle composition comprising a compound according to any one of claims 1 to 5.

7. In paragraph 6, The above compound is, A lipid nanoparticle composition characterized by being an ionized lipid.

8. In paragraph 7, The above composition, A lipid nanoparticle composition characterized in that it further comprises at least one selected from the group consisting of neutral lipids, steroids, and polymerized lipids.

9. In paragraph 8, The above neutral lipids are, A lipid nanoparticle composition characterized by being a phospholipid or a glycolipid.

10. In paragraph 8, The above steroids are, A lipid nanoparticle composition characterized by comprising at least one selected from the group consisting of cholesterol, bile acid derivatives, and cholic acid derivatives.

11. In paragraph 8, The above polymer polymerized lipid is, A lipid nanoparticle composition characterized in that the PEGylated lipid comprises at least one 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.

12. In paragraph 8, The above composition, A lipid nanoparticle composition comprising 20 to 65 mol% of ionized lipid, 2.5 to 30 mol% of neutral lipid, 25 to 60 mol% of steroid, and 0.5 to 5 mol% of polymerized lipid.

13. In paragraph 8, The above composition, A lipid nanoparticle composition characterized in that it further comprises a preventive or therapeutic agent.

14. In paragraph 13, The above preventive or therapeutic agent, A lipid nanoparticle composition characterized by comprising at least one selected from the group consisting of small interfering ribonucleic acid (siRNA), ribosomal ribonucleic acid (rRNA), ribonucleic acid (RNA), deoxyribonucleic acid (DNA), complementary deoxyribonucleic acid (cDNA), aptamer, messenger ribonucleic acid (mRNA), transfer ribonucleic acid (tRNA), antisense oligonucleotide, small hairpin ribonucleic acid (shRNA), micro ribonucleic acid (miRNA), asymmetric interfering ribonucleic acid (aiRNA), dicer-substrate ribonucleic acid (dsRNA), ribozyme, peptide nucleic acid (PNA), deoxyribozyme (DNAzyme), guide ribonucleic acid for gene correction (sgRNA), and mixtures thereof.

15. A composition for drug delivery comprising a lipid nanoparticle composition according to claim 13.

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