Novel lipid and preparation method therefor

Abstract

The present invention relates to a novel lipid and a preparation method therefor and, more particularly, to a lipid and a preparation method therefor, in which the lipid can be ionized to form a complex with an anionic drug, allowing various molecular designs according to a target tissue or organ, and thus is useful for tissue- or organ-specific drug delivery.

Description

Novel geological features and method for manufacturing the same

[0001] The present invention relates to novel lipids and methods for producing the same, and more specifically, to lipids that are ionizable and can form complexes with anionic drugs, and allow for various molecular designs depending on the target tissue or organ, which are useful for tissue or organ-specific drug delivery, and methods for producing the same.

[0002] In the treatment of anionic drugs, including nucleic acids, safe and efficient drug delivery technologies have long been studied, and various carriers and delivery technologies have been developed. Carriers are broadly classified into viral carriers utilizing adenoviruses or retroviruses, and non-viral carriers utilizing cationic lipids and cationic polymers. Viral carriers are known to pose risks such as non-specific immune responses and face significant challenges in commercialization due to their complex production processes. Therefore, recent research trends are moving toward improving these shortcomings by utilizing non-viral carriers. Compared to viral carriers, non-viral carriers offer the advantages of fewer side effects in terms of in vivo safety and lower production costs in terms of economic feasibility.

[0003] Representative non-viral delivery vehicles used for the delivery of nucleic acid materials are lipoplexes and polyplexes. These cationic lipids or polycationic polymers have been the subject of extensive research because they stabilize anionic drugs and enhance intracellular delivery by forming complexes through electrostatic interactions (De Paula D, Bentley MV, Mahato RI, Hydrophobization and bioconjugation for enhanced siRNA delivery and targeting, RNA 13 (2007) 431-56; Gary DJ, Puri N, Won YY, Polymer-based siRNA delivery: Perspectives on the fundamental and phenomenological distinctions from polymer-based DNA delivery, J Control release 121 (2007) 64-73).

[0004] However, polycation polymers have problems with actual use due to cytotoxicity caused by multivalent cation charges, and nucleic acid-cationic lipid complexes are difficult to use in vivo due to low stability in the blood. In addition, ionic liposomes containing cationic lipids, neutral lipids, and fusogenic lipids have disadvantages such as complex synthesis methods for the cationic lipids used, cytotoxicity, and low efficiency of intracellular nucleic acid delivery.

[0005] The objective of the present invention is to provide a lipid that is ionizable to form a complex with anionic drugs and allows for various molecular designs depending on the target tissue or organ, which is useful for tissue or organ-specific drug delivery, and a method for manufacturing the same.

[0006] A first aspect of the present invention provides a lipid having a structure represented by the following chemical formula 1:

[0007] [Chemical Formula 1]

[0008]

[0009] In the above chemical formula 1,

[0010] R p and R q Each is independently a hydrogen atom (H) or a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0011] R1 and R2 are each independently substituted or unsubstituted alkylene groups, and

[0012] n is an integer greater than or equal to 0 or 1, and

[0013] Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and

[0014] R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and

[0015] R8, R 10 , R 11 and R 13Each is independently a substituted or unsubstituted divalent saturated or unsaturated hydrocarbon group, and

[0016] R9, R 12 and R 14 Each is independently a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and

[0017] L1 and L2 are each independently selected from the group consisting of -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-, arylene groups, and heteroarylene groups, wherein L' is a directly bonded, substituted or unsubstituted alkylene group or substituted or unsubstituted alkenylene group, and R' is each independently selected from the group consisting of a hydrogen atom, a substituted or unsubstituted alkyl group and a substituted or unsubstituted alkenyl group, and

[0018] X is -OH or -SH.

[0019] In one embodiment, the above R p , R q , R1~R 14 One or more of the may be further substituted with one or more moieties selected from one or more amino acids, sugars, vitamins, peptides, proteins, hormone antibodies, neurotransmitters, pharmaceutically active small molecules, endosome-degrading agents, cell membrane permeability agents, charge blockers, drugs, nucleic acids, or derivatives thereof, which are specifically suitable for target tissues or organs (e.g., liver, lungs, spleen, brain, heart, etc.).

[0020] More specifically, as mentioned above,

[0021] R p and R q is independently a hydrogen atom (H), or a substituted or unsubstituted C 1-6 It is an alkyl group, and optionally Rp and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0022] R1 and R2 are each independently substituted or unsubstituted C 1-4 It is an alkylene group, and

[0023] n is 0 or 1, and

[0024] Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and

[0025] R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and

[0026] R8, R 10 , R 11 and R 13 Each is independently substituted or unsubstituted divalent saturated or unsaturated C 2-20 It is a hydrocarbon group, and

[0027] R9, R 12 and R 14 are independently substituted or unsubstituted monovalent saturated or unsaturated C 2-20 It is a hydrocarbon group, and

[0028] L1 and L2 are each independently -C(O)O-, -OC(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 6-20 C having an arylene group and one or more heteroatoms selected from N, O, and S 3-20 Selected from the group consisting of heteroarylene groups, where L' is directly bonded, substituted, or unsubstituted C 1-13alkylene group or substituted or unsubstituted C 2-13 It is an alkenylene group, and R' are each independently hydrogen atoms, substituted or unsubstituted C. 1-18 Alkyl groups and substituted or unsubstituted C 2-18 Selected from a group composed of alkenyl groups, and

[0029] X is -OH or -SH.

[0030] More specifically, as mentioned above,

[0031] R p and R q is independently a hydrogen atom (H), or a substituted or unsubstituted C 2-4 It is an alkyl group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0032] R1 and R2 are each independently substituted or unsubstituted C 2-4 It is an alkylene group, and

[0033] n is 0, and

[0034] m is 0 and,

[0035] R3, R4 and R6 each independently (ii) , (iii) or (iv) And,

[0036] R8, R 10 , R 11 and R 13 Each is independently substituted or unsubstituted divalent saturated or unsaturated C 2-10 It is a hydrocarbon group, and

[0037] R9, R 12 and R 14 are independently substituted or unsubstituted monovalent saturated or unsaturated C 10-20 It is a hydrocarbon group, and

[0038] L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R')-, and -N(R')C(O)-, where R' is each independently a hydrogen atom, and a substituted or unsubstituted C 1-4 Selected from the group consisting of alkyl groups, and

[0039] X is -OH.

[0040] More specifically, the lipid of the present invention may have any one of the following chemical formulas A to E:

[0041]

[0042] The lipid according to the present invention is ionizable and can form complexes with anionic drugs, and is useful for specifically delivering drugs to target tissues or organs. To this end, if necessary, the lipid according to the present invention may include, in part of its structure, a moiety specifically suitable for target tissues or organs, such as amino acids, sugars, vitamins, peptides, proteins, hormones, antibodies, neurotransmitters, pharmaceutically active small molecules, endosome-degrading agents, cell membrane permeability agents, charge blockers, drugs, nucleic acids, or derivatives thereof, or may bind to such a moiety. In particular, the lipid of the present invention exhibits excellent tissue selectivity when formulated as a genetic material delivery vehicle.

[0043] In one embodiment, the “moiety specifically suited to a target tissue or organ” includes all molecules such as amino acids, sugars, vitamins, peptides, proteins, hormones, antibodies, neurotransmitters, pharmaceutically active small molecules, endosome-degrading agents, cell membrane permeability agents, charge blockers, drugs, nucleic acids, or derivatives thereof that can interact directly or indirectly with other compounds such as receptors. The sugar may include, but is not limited to, galactose, galactosamine, N-acetylgalactosamine, or combinations thereof. The hormone may include, but is not limited to, estrogen, testosterone, progesterone, glucocortisone, adrenaline, insulin, glucagon, cortisol, vitamin D, thyroid hormone, retinoic acid, growth hormone, or combinations thereof. The neurotransmitter may include growth factors such as VEGF, EGF, NGF, and PDGF; cholesterol; bile acids; It may include, but is not limited to, GABA, glutamate, acetylcholine, or combinations thereof.

[0044] In one embodiment, a “moiety specifically suitable for a target tissue or organ” may be attached to the lipid of the present invention using a linker molecule. The linker molecule may include, but is not limited to, amides, carbonyls, esters, peptides, disulfides, silanes, nucleosides, abasic nucleosides, polyethers, polyamines, polyamides, carbohydrates, lipids, polyhydrocarbons, phosphate esters, phosphoramidates, thiophosphates, alkyl phosphates, biodegradable linkers, photolabile linkers, etc.

[0045] A second aspect of the present invention provides a method for producing a lipid having a structure represented by Formula 1, comprising the step of reacting a compound of Formula a with a compound of Formula b:

[0046] [Chemical formula a]

[0047]

[0048] [Chemical formula b]

[0049]

[0050] [Chemical Formula 1]

[0051]

[0052] In the above,

[0053] Hal is a halogen atom, and

[0054] R p and R q Each is independently a hydrogen atom (H) or a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0055] R1 and R2 are each independently substituted or unsubstituted alkylene groups, and

[0056] n is an integer greater than or equal to 0 or 1, and

[0057] Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and

[0058] R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and

[0059] R8, R 10 , R 11 and R 13 Each is independently a substituted or unsubstituted divalent saturated or unsaturated hydrocarbon group, and

[0060] R9, R 12 and R 14Each is independently a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and

[0061] L1 and L2 are each independently selected from the group consisting of -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-, arylene groups, and heteroarylene groups, wherein L' is a directly bonded, substituted or unsubstituted alkylene group or substituted or unsubstituted alkenylene group, and R' is each independently selected from the group consisting of a hydrogen atom, a substituted or unsubstituted alkyl group and a substituted or unsubstituted alkenyl group, and

[0062] X is -OH or -SH.

[0063] A third aspect of the present invention provides a drug delivery composition comprising lipids of the present invention.

[0064] The lipid according to the present invention is ionizable and can form a complex with anionic drugs, and is useful for specifically delivering drugs to target tissues or organs. In particular, the lipid of the present invention has a structure in which a boronate region is introduced into the head portion of the ionizable lipid, so when formulated as a genetic material delivery vehicle, it exhibits excellent selectivity for target tissues or organs, and when sugars (such as glucose) are added to the formulation solution, it can exert a targeting function to specific receptors through the interaction between the boronate and the diol.

[0065] Figure 1 is a schematic diagram of the reaction for the synthesis process of compound A of formula performed in Example 1.

[0066] Figure 2 is a schematic diagram of the reaction for the synthesis process of compounds of formulas B and C performed in Example 2.

[0067] Figure 3 is a schematic diagram of the reaction for the synthesis process of the compound of formula D performed in Example 3.

[0068] Figure 4 is a schematic diagram of the reaction for the synthesis process of the compound of formula E performed in Example 4.

[0069] The present invention will be described in more detail below.

[0070] The lipid provided according to the first aspect of the present invention has a structure represented by the following chemical formula 1:

[0071] [Chemical Formula 1]

[0072]

[0073] In the above chemical formula 1,

[0074] R p and R q Each is independently a hydrogen atom (H) or a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0075] R1 and R2 are each independently substituted or unsubstituted alkylene groups, and

[0076] n is an integer greater than or equal to 0 or 1, and

[0077] Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and

[0078] R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and

[0079] R8, R 10 , R 11 and R 13Each is independently a substituted or unsubstituted divalent saturated or unsaturated hydrocarbon group, and

[0080] R9, R 12 and R 14 Each is independently a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and

[0081] L1 and L2 are each independently selected from the group consisting of -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-, arylene groups, and heteroarylene groups, wherein L' is a directly bonded, substituted or unsubstituted alkylene group or substituted or unsubstituted alkenylene group, and R' is each independently selected from the group consisting of a hydrogen atom, a substituted or unsubstituted alkyl group and a substituted or unsubstituted alkenyl group, and

[0082] X is -OH or -SH.

[0083] In one embodiment, the above R p , R q , R1~R 14 One or more of the may be further substituted with one or more moieties specifically suitable for a target tissue or organ (e.g., liver, lung, spleen, brain, heart, etc.), preferably one or more moieties selected from one or more amino acids, sugars, vitamins, peptides, proteins, hormones, or antibodies, or derivatives thereof.

[0084] The range of lipids of the present invention includes not only those having the structure of Chemical Formula 1, but also their cationized forms.

[0085] In this specification, the expression “substituted or unsubstituted” means that, unless otherwise specified, the group is not substituted, or -OH, a halogen atom, C 1-6 Alkyl group, C 1-6Alkoxy group, C 1-6 alkyl halide group, C 1-6 Halogenated alkoxy group, C 3-20 Cycloalkyl group, C 3-20 heterocycloalkyl group, C 6-20 aryl group or C 3-20 This means that it is substituted with one or more substituents selected from heteroaryl groups. In addition, in one embodiment, the substituent may include amino acids, sugars, vitamins, peptides, proteins, hormones, or antibodies.

[0086] In this specification, the expression that any group (e.g., heteroaryl, heterocycloalkyl, etc.) is a “hetero” group means that, unless otherwise specified, the group has one or more (e.g., 1 to 3) hetero atoms selected from N, O and S.

[0087] In this specification, the “hydrocarbon group” may be branched or unbranched, annular or non-annular, or aromatic.

[0088] More specifically, as mentioned above,

[0089] R p and R q is independently a hydrogen atom (H), or a substituted or unsubstituted C 1-6 It can be an alkyl group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0090] R1 and R2 are each independently substituted or unsubstituted C 1-4 It can be an alkylene group, and

[0091] n can be 0 or 1, and

[0092] R8, R 10 , R 11 and R 13 Each is independently substituted or unsubstituted divalent saturated or unsaturated C 2-20 It can be a hydrocarbon group,

[0093] R9, R 12 and R 14 are independently substituted or unsubstituted monovalent saturated or unsaturated C 2-20 It can be a hydrocarbon group,

[0094] L1 and L2 are each independently -C(O)O-, -OC(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 6-20 C having an arylene group and one or more heteroatoms selected from N, O, and S 3-20 It can be selected from the group consisting of heteroarylene groups, where L' is directly bonded, substituted, or unsubstituted C 1-13 alkylene group or substituted or unsubstituted C 2-13 It can be an alkenylene group, and R' are each independently hydrogen atoms, substituted or unsubstituted C 1-18 Alkyl groups and substituted or unsubstituted C 2-18 It can be selected from the group composed of alkenyl groups.

[0095] More specifically, as mentioned above,

[0096] R p and R q is independently a hydrogen atom (H), or a substituted or unsubstituted C 2-4 It can be an alkyl group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and

[0097] R1 and R2 are each independently substituted or unsubstituted C 2-4 It can be an alkylene group, and

[0098] n can be 0, and

[0099] m can be 0, and

[0100] R3, R4 and R6 each independently (ii) , (iii) or (iv) It could be,

[0101] R8, R 10 , R 11 and R 13 Each is independently substituted or unsubstituted divalent saturated or unsaturated C 2-10 It can be a hydrocarbon group,

[0102] R9, R 12 and R 14 are independently substituted or unsubstituted monovalent saturated or unsaturated C 10-20 It can be a hydrocarbon group,

[0103] L1 and L2 can each be independently selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R')-, and -N(R')C(O)-, where R' is each independently a hydrogen atom, and a substituted or unsubstituted C 1-4 It can be selected from the group consisting of alkyl groups, and

[0104] X can be -OH.

[0105] More specifically, the lipid of the present invention may have any one of the following chemical formulas A to E:

[0106]

[0107] The lipid according to the present invention is ionizable and can form a complex with anionic drugs, and is useful for specifically delivering drugs to target tissues or organs. To this end, if necessary, the lipid according to the present invention may include a moiety specifically suitable for the target tissue or organ, such as an amino acid, sugar, vitamin, peptide, protein, hormone, or antibody, in part of its structure, or may bind to such a moiety. In particular, the lipid of the present invention has a structure in which a boronate region is introduced into the head portion of the ionizable lipid, so when formulated as a genetic material delivery vehicle, it exhibits excellent selectivity for target tissues or organs, and when sugars (such as glucose) are added to the formulation solution, it can exert a targeting function to a specific receptor through the interaction between the boronate and the diol.

[0108] A second aspect of the present invention provides a method for producing a lipid having a structure represented by Formula 1, comprising the step of reacting a compound of Formula a with a compound of Formula b:

[0109] [Chemical formula a]

[0110]

[0111] [Chemical formula b]

[0112]

[0113] [Chemical Formula 1]

[0114]

[0115] In the above,

[0116] Hal is a halogen atom (i.e., F, Cl, Br, or I), and

[0117] R p , R q , R1~R7, n and m are as defined in Chemical Formula 1 above.

[0118] In one embodiment of the lipid preparation method according to the second aspect of the present invention, the reaction between the compound of formula a and the compound of formula b may be carried out in a solvent (e.g., acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., K2CO3, etc.) at a temperature (e.g., 60°C to 100°C), but is not limited thereto.

[0119] The lipid according to the present invention can easily form a complex with anionic drugs, making it useful for drug delivery and useful for specifically delivering drugs to target tissues or organs. In particular, the lipid of the present invention has a structure in which a boronate site is introduced into the head portion of the ionizable lipid, so when formulated as a genetic material delivery vehicle, it exhibits excellent selectivity for target tissues or organs.

[0120] Accordingly, according to a third aspect of the present invention, a drug delivery composition comprising lipids of the present invention is provided.

[0121] In one embodiment, the drug may be selected from nucleic acids, polypeptides, viruses, or a combination thereof.

[0122] The “nucleic acid” mentioned above may be, for example, DNA, RNA, siRNA, shRNA, miRNA, mRNA, aptamers, antisense oligonucleotides, or combinations thereof, but is not limited thereto.

[0123] The above “polypeptide” may refer to a protein having activity in the body, such as an antibody or a fragment thereof, a cytokine, a hormone or an analog thereof, or a protein that can be recognized as an antigen through a series of processes in the body, including a polypeptide sequence of an antigen, an analog thereof, or a precursor thereof.

[0124] In one embodiment, the lipid of the present invention forms a complex with a drug, and this complex can be encapsulated inside a nanoparticle structure formed by an amphiphilic block copolymer.

[0125] In one embodiment, the amphiphilic block copolymer may be an AB-type block copolymer comprising a hydrophilic A block and a hydrophobic B block. The AB-type block copolymer forms core-shell type polymer nanoparticles in an aqueous solution, wherein the hydrophobic B block forms the core (inner wall) and the hydrophilic A block forms the shell (outer wall).

[0126] In one embodiment, the hydrophilic A block may be one or more selected from the group consisting of polyalkylene glycol, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, and derivatives thereof.

[0127] More specifically, the hydrophilic A block may be one or more selected from the group consisting of monomethoxypolyethylene glycol (mPEG), monoacetoxypolyethylene glycol, polyethylene glycol, copolymers of polyethylene and propylene glycol, and polyvinylpyrrolidone.

[0128] Additionally, if necessary, a functional group, a ligand capable of reaching specific tissues or cells, or a functional group capable of promoting intracellular delivery may be chemically bonded to the terminal end of the hydrophilic A block to control the in vivo distribution of the nanoparticle delivery vehicle or to increase the efficiency of the nanoparticle delivery vehicle being delivered into cells. In one embodiment, the functional group or ligand may be one or more selected from the group consisting of monosaccharides, polysaccharides, vitamins, peptides, proteins, and antibodies against cell surface receptors. More specifically, the functional group or ligand may be one or more selected from the group consisting of anisamide, vitamin B9 (folic acid), vitamin B12, vitamin A, galactose, lactose, mannose, hyaluronic acid, RGD peptide, NGR peptide, transferrin, and antibodies against transferrin receptors.

[0129] The above hydrophobic B block is a biocompatible biodegradable polymer, and in one embodiment, it may be one or more selected from the group consisting of polyester, polyanhydride, polyamino acid, polyorthoester, and polyphosphazine.

[0130] More specifically, the hydrophobic B block may be one or more selected from the group consisting of polylactide (PLA), polyglycolide, polycaprolactone, polydioxane-2-one, copolymers of polylactide and glycolide, copolymers of polylactide and polydioxane-2-one, copolymers of polylactide and polycaprolactone, and copolymers of polyglycolide and polycaprolactone.

[0131] In another embodiment, the hydrophobic B block is a biocompatible biodegradable polymer having repeating units of a structure represented by the following chemical formula e.

[0132] [Chemical formula e]

[0133]

[0134] In the above chemical formula e, R represents a branched alkylene group having 3 or more carbon atoms.

[0135] In one embodiment, the number of carbon atoms of R in the above formula e may be, for example, 3 or more, 4 or more, 5 or more, 6 or more, or 7 or more, and may 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.

[0136] In one embodiment, in the above formula e, R may represent a branched alkylene group having 3 to 20 carbon atoms, more specifically a branched alkylene group having 3 to 17 carbon atoms, even more specifically a branched alkylene group having 3 to 15 carbon atoms, and even more specifically a branched alkylene group having 3 to 13 carbon atoms, but is not limited thereto.

[0137] In one embodiment, the hydrophobic B block may be a biocompatible biodegradable polymer having repeating units of a structure selected from the following, but is not limited thereto.

[0138]

[0139] In one embodiment, the repeating unit of the structure represented by the above formula e can be obtained by ring-opening polymerization of a lactone compound.

[0140] In addition, in one embodiment, the hydrophobic B block may be modified by chemically bonding tocopherol, cholesterol, or a fatty acid having 10 to 24 carbon atoms to the hydroxyl group at the end of the hydrophobic B block in order to increase the hydrophobicity of the hydrophobic B block and improve the stability of the nanoparticle.

[0141] The present invention will be explained in more detail below based on the following examples, but these are for the purpose of explaining the invention and do not limit the scope of the invention in any way.

[0142] [Example]

[0143] Example 1

[0144] A compound of the following chemical formula A (compound A) was prepared according to the synthesis outline shown in Fig. 1.

[0145] [Chemical Formula A]

[0146]

[0147] (1) Synthesis of 2-octyldodecyl 6-((tert-butoxycarbonyl)amino)hexanoate

[0148]

[0149] 6-((tert-butoxycarbonyl)amino)hexanoic acid (17 g, 73.5 mmol, 1.70 eq) and dichloromethane (DCM) (170 mL) were mixed at 25°C. Triethylamine (TEA) (6.13 g, 60.5 mmol, 1.40 eq) was added to this mixture at 25°C. Subsequently, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (15.7 g, 82.1 mmol, 1.90 eq) was added at the same temperature. The mixture was stirred at 25°C for 30 minutes. Subsequently, 4-dimethylaminopyridine (DMAP) (7.39 g, 60.5 mmol, 1.40 eq) was added, followed by the addition of 2-octyldodecan-1-ol (12.9 g, 43.2 mmol, 1.00 eq) to the mixture. The mixture was stirred at 25°C for 16 hours under a nitrogen (N2) atmosphere. The residue was diluted with water (100 mL) and extracted three times with DCM (100 mL), and the bound organic layer was washed twice with water (50 mL). After drying with Na2SO4 and filtering, the residue was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (dichloromethane (DCM) / methanol (MeOH) = 100 / 1 to 0 / 1) to obtain 2-octyldodecyl 6-((tert-butoxycarbonyl)amino)hexanoate in the form of a yellow oil (14 g, 27.3 mmol, yield 63.2%).

[0150] 1HNMR (400 MHz, CHLOROFORM-d)δ0.84 - 0.91 (m, 6 H) 1.24 - 1.26 (m, 7 H) 1.42 - 1.46 (m, 9 H) 1.62 (br s, 4 H) 2.05 (s, 11 H) 2.29 (s, 2 H) 3.08 - 3.14 (m, 2 H) 3.96 (d, 2 H) 4.46 - 4.62 (m, 1 H)

[0151] (2) Synthesis of 2-octyldodecyl 6-aminohexanoate

[0152]

[0153] 2-octyldodecyl 6-((tert-butoxycarbonyl)amino)hexanoate (6 g, 11.7 mmol, 1.00 eq) and DCM (60 mL) were mixed at 25°C. Trifluoroacetic acid (TFA) (60 mL, 117.23 mmol, 10 V) was added to this mixture at 25°C. The mixture was stirred at 25°C for 2 hours. After the reaction was complete, the reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1 to 0 / 1). Finally, 2-octyldodecyl 6-aminohexanoate (3 g, 7.29 mmol, yield 62.1%) in the form of a yellow oil was obtained.

[0154] 1 HNMR (400 MHz, CHLOROFORM-d)δ0.89 (t, 6 H) 1.16 - 1.46 (m, 36 H) 1.57 - 1.70 (m, 5 H) 2.32 (t, 2 H) 2.82 - 2.93 (m, 2 H) 3.97 (d, 2 H)

[0155] (3) Synthesis of bis(2-octyldodecyl) 6,6'-azanediyldihexanoate

[0156]

[0157] 2-octyldodecyl 6-aminohexanoate (3 g, 7.29 mmol, 1.00 eq) and acetonitrile (ACN) (30 mL) were mixed at 25°C. To this mixture, 2-octyldodecyl 6-bromohexanoate (3.47 g, 7.29 mmol, 1.00 eq) was added at 25°C, followed by the addition of K2CO3 (1.01 g, 7.29 mmol, 1.00 eq) at the same temperature. The mixture was stirred at 60°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1 to 0 / 1). Finally, bis(2-octyldodecyl) 6,6'-azandyldihexanoate (1.20 g, 1.49 mmol, yield 20.4%) in the form of a yellow oil was obtained.

[0158] 1 HNMR (400 MHz, CHLOROFORM-d) δ0.89 (t, 12 H) 1.19 - 1.40 (m, 70 H) 1.49 - 1.69 (m, 11 H) 2.25 - 2.39 (m, 4 H) 2.52 - 2.67 (m, 4 H) 3.97 (d, 4 H).

[0159] (4) Synthesis of Compound A

[0160]

[0161] Bis(2-octyldodecyl) 6,6'-azandyldihexanoate (3 g, 3.72 mmol, 1.00 eq) and ACN (30 mL) were mixed at 25°C. To the mixture, 2-(4-bromobutyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (978 mg, 3.72 mmol, 1.00 eq) was added at 25°C, followed by the addition of K2CO3 (771 mg, 5.58 mmol, 1.50 eq) at the same temperature. The mixture was stirred at 60°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 10 / 1 to 0 / 1). Finally, compound A in the form of a yellow oil (1 g, 1.01 mmol, yield 27.2%) was obtained.

[0162] 1 HNMR (400 MHz, CHLOROFORM-d)δ0.79 (br t, 2 H) 0.87 - 0.94 (m, 12 H) 1.26 (br d, 80 H) 1.38 - 1.47 (m, 8 H) 1.59 - 1.66 (m, 6 H) 2.31 (br t, 10 H) 3.97 (d, 4H)

[0163] Example 2

[0164] Compounds of the following chemical formulas B and C (compound B and compound C, respectively) were prepared according to the synthesis outline shown in Figure 2.

[0165] [Chemical Formula B]

[0166]

[0167] [Chemical Formula C]

[0168]

[0169] (1) Synthesis of 2-octyldodecyl 6-((tert-butoxycarbonyl)amino)hexanoate

[0170]

[0171] 6-((tert-butoxycarbonyl)amino)hexanoic acid (17.0 g, 73.5 mmol, 1.70 eq) was added to DCM (170 mL) at 25°C. TEA (6.13 g, 60.5 mmol, 1.40 eq) was added to the mixture at 25°C, followed by the addition of EDCI (15.7 g, 82.1 mmol, 1.90 eq) to the mixture at 25°C. The mixture was stirred at 25°C for 30 minutes. Subsequently, DMAP (7.39 g, 60.5 mmol, 1.40 eq) was added to the mixture at 25°C, followed by the addition of 2-octyldodecan-1-ol (12.9 g, 43.2 mmol, 1.00 eq). The mixture was stirred at 25°C for 16 hours under a nitrogen atmosphere. The residue was diluted with water (100 mL) and extracted with DCM (100 mL x 3). The bound organic layer was washed with water (50.0 mL x 2), dried with Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 100 / 1 to 0 / 1). Finally, 2-octyldodecyl 6-((tert-butoxycarbonyl)amino)hexanoate (14.0 g, 27.3 mmol, yield 63.2%) in the form of a yellow oil was obtained.

[0172] 1 HNMR (400 MHz, CHLOROFORM-d)δ4.46 - 4.62 (m, 1H), 3.96 (d, 2H), 3.08 - 3.14 (m, 2H), 2.29 (s, 2H), 2.05 (s, 11H), 1.62 (br s, 4H), 1.42 - 1.46 (m, 9H), 1.24 - 1.26 (m, 7H), 0.84 - 0.91 (m, 6H)

[0173] (2) Synthesis of 2-octyldodecyl 6-aminohexanoate

[0174]

[0175] 2-octyldodecyl 6-((tert-butoxycarbonyl)amino)hexanoate (6.00 g, 11.7 mmol, 1.0 eq) was added to DCM (60.0 mL) at 25°C. TFA (60.0 mL, 117 mmol, 10 V) was added to the mixture at 25°C. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 100 / 1 to 10 / 1). Finally, 2-octyldodecyl 6-aminohexanoate (3.00 g, 7.29 mmol, yield 62.1%) in the form of a yellow oil was obtained.

[0176] 1 HNMR (400 MHz, CHLOROFORM-d)δ3.97 (d, 2H), 2.82 - 2.93 (m, 2H), 2.32 (t, 2H), 1.57 - 1.70 (m, 5H), 1.16 - 1.46 (m, 36H), 0.89 (t, 6H)

[0177] (3) Synthesis of bis(2-octyldodecyl) 6,6'-azandyldihexanoate

[0178]

[0179] 2-octyldodecyl 6-aminohexanoate (3.00 g, 7.29 mmol, 1.0 eq) was added to ACN (30.0 mL) at 25°C. 2-octyldodecyl 6-bromohexanoate (3.47 g, 7.29 mmol, 1.0 eq) was added to the mixture at 25°C. Subsequently, K2CO3 (1.01 g, 7.29 mmol, 1.0 eq) was added to the mixture at 25°C. The mixture was stirred at 60°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 100 / 1 to 10 / 1). Finally, bis(2-octyldodecyl) 6,6'-azandyldihexanoate (1.20 g, 1.49 mmol, yield 20.4%) in the form of a yellow oil was obtained.

[0180] 1 HNMR (400 MHz, CHLOROFORM-d)δ3.97 (d, 4 H), 2.52 - 2.67 (m, 4 H), 2.25 - 2.39 (m, 4 H), 1.49 - 1.69 (m, 11 H), 1.19 - 1.40 (m, 70 H), 0.89 (t, 12H)

[0181] (4) Synthesis of Compound B

[0182]

[0183] Bis(2-octyldodecyl) 6,6'-azandyldihexanoate (2.00 g, 2.48 mmol, 1.00 eq) was added to ACN (20.0 mL) at 25°C. 2-(3-bromopropyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (617 mg, 2.48 mmol, 1.00 eq) was added to the mixture. Subsequently, K2CO3 (514 mg, 3.72 mmol, 1.50 eq) was added to the mixture. The mixture was stirred at 60°C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM / MeOH = 100 / 1 to 1 / 10). Finally, compound B in the form of a yellow oil (1.00 g, 1.03 mmol, yield 41.3%) was obtained.

[0184] 1 HNMR (400 MHz, CHLOROFORM-d)δ3.97 (d, 2H), 3.63 (br t, 2H), 2.25 - 2.39 (m, 2H), 1.58 - 1.81 (m, 15H), 1.25 - 1.40 (m, 64H), 0.70 - 1.01 (m, 13H)

[0185] (5) Synthesis of Compound C

[0186]

[0187] Compound B (1.00 g, 1.03 mmol, 1.00 eq) was added to MeOH (10.0 mL) at 25°C. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated with DCM (2.00 mL × 3), washed with water (10.0 mL), dried with Na2SO4, filtered, and then concentrated under reduced pressure to obtain the residue. Finally, compound C (0.20 g, 224 μmol, yield 21.8%) in the form of a yellow oil was obtained.

[0188] 1 HNMR (400 MHz, CHLOROFORM-d)δ3.97 (d, 4H), 2.47 (br d, 6H), 2.31 (t, 4H), 1.66 - 1.72 (m, 2H), 1.54 (br s, 10H), 1.20 - 1.32 (m, 68H), 0.93 - 1.00 (m, 2H), 0.89 (t, 12H)

[0189] Example 3

[0190] A compound of the following chemical formula D (compound D) was prepared according to the synthesis outline shown in Fig. 3.

[0191] [Chemical Formula D]

[0192]

[0193] (1) Synthesis of heptadecan-9-yl 8-((tert-butoxycarbonyl)amino)octanoate

[0194]

[0195] To a mixture in which heptadecan-9-ol (20.1 g, 65.5 mmol, 1.00 eq) and 8-((tert-butoxycarbonyl)amino)octanoic acid (17.0 g, 65.5 mmol, 1.00 eq) were dissolved in DCM (201 mL), EDCI (23.8 g, 124 mmol, 1.90 eq), TEA (9.29 g, 91.7 mmol, 1.40 eq), and DMAP (11.2 g, 91.7 mmol, 1.40 eq) were added at 25°C. The mixture was stirred at 25°C for 16 hours. The reaction mixture was separated into layers in DCM (50.0 mL) and H2O (100 mL). The organic layer was separated, and the aqueous layer was further extracted with DCM (50.0 mL × 2) and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 10 / 1). Finally, heptadecan-9-yl 8-((tert-butoxycarbonyl)amino)octanoate (23.8 g, 47.8 mmol, yield 72.9%) in the form of a yellow solid was obtained.

[0196] 1 HNMR (400 MHz, CHLOROFORM-d)δ4.87 (quin, 1 H), 4.50 (br s, 1 H), 3.10 (br d, 2 H), 2.28 (t, 2 H), 1.59 - 1.66 (m, 2 H), 1.47 - 1.58 (m, 6 H), 1.45 (s, 9 H), 1.21 - 1.35 (m, 30 H), 0.80 - 0.93 (m, 6 H)

[0197] (2) Synthesis of heptadecan-9-yl 8-aminooctanoate

[0198]

[0199] TFA (238 mL) was added to a mixture of heptadecan-9-yl 8-((tert-butoxycarbonyl)amino)octanoate (23.8 g, 47.8 mmol, 1.00 eq) dissolved in DCM (238 mL) at 25°C. The mixture was stirred at 25°C for 4 hours. The pH was adjusted to 8 using saturated Na2CO3 at 20°C for 0.5 hours, and the aqueous layer was extracted with DCM (200 mL × 3). The organic layer was concentrated under reduced pressure to obtain the final product. Finally, heptadecan-9-yl 8-aminooctanoate (19.0 g, 47.7 mmol, yield 99.9%) in the form of a yellow oil was obtained.

[0200] 1 HNMR (400 MHz, CHLOROFORM-d)δ6.25 (br s, 2 H), 4.86 (quin, 1 H), 2.87 (t, 2 H), 2.27 (t, 2 H), 1.61 (br d, 4 H), 1.51 (br d, 4 H), 1.33 (br s, 6 H), 1.20 - 1.31 (m, 24 H), 0.88 (t, 6 H)

[0201] (3) Synthesis of undecan-3-yl 8-bromooctanoate

[0202]

[0203] 8-bromooctanoic acid (48.6 g, 218 mmol, 1.50 eq) was added at 25°C to a mixture in which undecan-3-ol (25.0 g, 145 mmol, 1.00 eq) was dissolved in toluene (250 mL). Subsequently, H2SO4 (2.13 g, 21.8 mmol, 0.15 eq) was added at 25°C and purged three times with nitrogen (N2). The mixture was stirred at 120°C for 16 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 100 / 1 to 1 / 100). Finally, undecane-3-yl 8-bromooctanoate in the form of a yellow oil (40.0 g, 106 mmol, yield 73.1%) was obtained.

[0204] 1 HNMR (400 MHz, CHLOROFORM-d)δ4.82 (t, 1H), 3.41 (t, 2H), 2.30 (t, 2H), 1.86 (quin, 2H), 1.68 - 1.59 (m, 2H), 1.59 - 1.55 (m, 2H), 1.54 - 1.50 (m, 2H), 1.48 - 1.41 (m, 2H), 1.35 (td, 4H), 1.27 (br s, 12H), 0.94 - 0.83 (m, 6H)

[0205] (4) Synthesis of heptadecan-9-yl 8-((8-oxo-8-(undecan-3-yloxy)octyl)amino)octanoate

[0206]

[0207] K2CO3 (1.74 g, 12.5 mmol, 1.00 eq) was added at 25°C to a mixture of heptadecan-9-yl 8-aminooctanoate (5.00 g, 12.5 mmol, 1.00 eq) and undecane-3-yl 8-bromooctanoate (4.75 g, 12.5 mmol, 1.00 eq) dissolved in acetonitrile (ACN, 50.0 mL). The mixture was stirred at 60°C for 16 hours. The reaction mixture was separated into DCM (50.0 mL) and water (100 mL). The organic layer was separated, and the aqueous layer was extracted with DCM (50 mL x 2) to bind the organic layer. The residue was then concentrated under reduced pressure. This residue was purified by silica gel column chromatography (DCM / MeOH=10 / 1) to obtain residue 2. Residue 2 was further purified by prep-HPLC (Column: PHS-Phenyl-Hexyl 250*50mm*7um; Mobile phase: [water (TFA)-ACN]; Gradient: 50%-80% B, for 25 min). The purified product was adjusted to pH 10 with saturated K2CO3 (30.0 mL), concentrated under reduced pressure to remove ACN, extracted the aqueous layer with DCM (30.0 mL x 3), and concentrated the organic layer to obtain the final product. Finally, colorless oil-form heptadecan-9-yl 8-((8-oxo-8-(undecan-3-yloxy)octyl)amino)octanoate (2.00 g, 2.88 mmol, yield 22.9%) was obtained.

[0208] 1 HNMR (400 MHz, CHLOROFORM-d)δ4.78 - 4.94 (m, 2 H), 2.58 (t, 4 H), 2.24 - 2.33 (m, 4 H), 1.63 - 1.67 (m, 2 H), 1.46 - 1.54 (m, 10 H), 1.22 - 1.36 (m, 52 H), 0.80 - 0.97 (m, 12 H)

[0209] (5) Synthesis of Compound D

[0210]

[0211] K2CO3 (298 mg, 2.16 mmol, 1.50 eq) was added to a mixture of heptadecan-9-yl 8-((8-oxo-8-(undecan-3-yloxy)octyl)amino)octanoate (1.00 g, 1.44 mmol, 1.00 eq) and 2-(3-bromopropyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (358 mg, 1.44 mmol, 1.00 eq) dissolved in acetonitrile (ACN, 10 mL). The mixture was stirred at 80°C for 16 hours. The reaction was completed cleanly according to TLC. After filtering the reaction mixture, the residue was obtained by concentrating under reduced pressure. This residue was purified by silica gel column chromatography (DCM / MeOH=10 / 1). Finally, compound D (0.15 g, 187.49 μmol, yield 13.0%, purity 97.5%) was obtained in the form of a light yellow oil.

[0212] 1 HNMR (400 MHz, CHLOROFORM-d)δ7.73 - 8.30 (m, 2 H), 4.78 - 4.91 (m, 2 H), 2.46 (br s, 6 H), 2.29 (td, 4 H), 1.58 - 1.69 (m, 8 H), 1.47 - 1.55 (m, 10 H), 1.24 - 1.34 (m, 48 H), 0.85 - 0.99 (m, 14 H)

[0213] Example 4

[0214] A compound of the following chemical formula E (compound E) was prepared according to the synthesis outline shown in Fig. 4.

[0215] [Chemical Formula E]

[0216]

[0217] (1) Synthesis of heptadecan-9-yl 8-((tert-butoxycarbonyl)amino)octanoate

[0218]

[0219] Heptadecan-9-ol (20.2 g, 78.7 mmol, 1.00 eq) and 8-((tert-butoxycarbonyl)amino)octosan (17.0 g, 65.6 mmol, 0.80 eq) were dissolved in dichloromethane (DCM, 80.0 mL). To this mixture, EDCI (23.9 g, 125 mmol, 1.90 eq), TEA (9.29 g, 91.8 mmol, 1.40 eq), and DMAP (11.2 g, 91.8 mmol, 1.40 eq) were added at 25°C. The mixture was stirred at 25°C for 16 hours. The reaction mixture was partitioned into dichloromethane (50.0 mL) and water (100 mL). The organic phase was separated, and the aqueous phase was further extracted with DCM (50.0 mL x 2). The organic layers were combined and concentrated under reduced pressure to obtain the residue. This residue was purified by silica gel column chromatography (petroleum ether / ethyl acetate = 20 / 1 to 10 / 1). Finally, heptadecan-9-yl 8-((tert-butoxycarbonyl)amino)octanoate (23.8 g, 47.8 mmol, yield 72.9%) was obtained in the form of a pale yellow solid.

[0220] 1 H NMR (400 MHz, CHLOROFORM-d)δ4.87 (quin, 1H), 4.50 (br s, 1H), 3.10 (br d, 2H), 2.28 (t, 2H), 1.59 - 1.66 (m, 2H), 1.47 - 1.58 (m, 6H), 1.45 (s, 9H), 1.21 - 1.35 (m, 30H), 0.80 - 0.93 (m, 6 H)

[0221] (2) Synthesis of heptadecan-9-yl 8-aminooctanoate

[0222]

[0223] Heptadecan-9-yl 8-((tert-butoxycarbonyl)amino)octanoate (23.8 g, 47.8 mmol, 1.00 eq) was dissolved in dichloromethane (DCM, 238 mL), and TFA (307 g, 2.69 mol, 238 mL) was added at 25°C. The mixture was stirred at 25°C for 4 hours. The pH of the reaction mixture was adjusted to 8 using saturated Na2CO3 (30.0 mL) at 20°C for 0.5 hours, and the aqueous phase was extracted with dichloromethane (30.0 mL x 3). The organic phase was concentrated under reduced pressure to obtain the product. Without purification, heptadecan-9-yl 8-aminooctanoate (19.0 g, 47.8 mmol, yield 99.9%) was obtained as a light yellow oil.

[0224] 1 H NMR (400 MHz, CHLOROFORM-d)δ6.25 (br s, 2H) 4.86 (quin, 1H) 2.87 (t, 2H) 2.27 (t, 2H) 1.61 (br d, 4H) 1.51 (br d, 4H) 1.33 (br s, 6H) 1.20 - 1.31 (m, 24H) 0.88 (t, 6H)

[0225] (3) Synthesis of heptadecan-9-yl 8-bromooctanoate

[0226]

[0227] Heptadecan-9-ol (50.0 g, 195 mmol, 1.00 eq) and 8-bromooctanic acid (47.9 g, 214 mmol, 1.10 eq) were dissolved in toluene (500 mL), and H2SO4 (1.91 g, 19.5 mmol, 0.10 eq) was added. The mixture was stirred at 110°C for 16 hours. The mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (SiO₂, petroleum ether / ethyl acetate = 20 / 1 to 10 / 1). Finally, heptadecan-9-yl 8-bromooctanoate (50.0 g, 108 mmol, yield 55.5%) was obtained in the form of a pale yellow oil.

[0228] 1 H NMR (400 MHz, CHLOROFORM-d)δ4.87 (quin, 1H), 3.40 (t, 2H), 2.29 (t, 2H), 1.86 (quin, 2H), 1.50 - 1.52 (m, 2H), 1.35 - 1.36 (m, 4H), 1.33 - 1.34 (m, 2H), 1.23 - 1.33 (m, 28H), 0.82 - 0.93 (m, 6H)

[0229] (4) Synthesis of di(heptadecan-9-yl) 8,8'-azanediyldioctanoate

[0230]

[0231] Heptadecan-9-yl 8-aminooctanoate (5.00 g, 12.6 mmol, 1.00 eq), heptadecan-9-yl 8-bromooctanoate (5.80 g, 12.6 mmol, 1.00 eq), and K2CO3 (1.74 g, 12.6 mmol, 1.00 eq) were mixed with acetonitrile (ACN, 50.0 mL), followed by degassing and purging three times with nitrogen (N2). The mixture was then stirred at 60°C for 16 hours under a nitrogen atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to remove the solvent. The residue was extracted with saturated K2CO3 and dichloromethane (DCM, 100 mL × 3), and the organic layer was concentrated under reduced pressure to obtain the residue. The residue was dissolved in acetonitrile (50.0 mL), and then water (60.0 mL) and excess K2CO3 were added, and the mixture was stirred at 25°C for 16 hours. The mixture was filtered, and the acetonitrile was removed by concentration under reduced pressure. The aqueous layer was extracted with dichloromethane (100 mL × 3), and the organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (SiO₂, DCM / MeOH = 100 / 1 to 10 / 1). Finally, di(heptadecan-9-yl) 8,8'-azandiyldioctanoate (1.20 g, 1.54 mmol, yield 12.3%) was obtained in the form of a light yellow oil.

[0232] 1 H NMR (400 MHz, CHLOROFORM-d)δ4.87 (t, 2H), 2.60 (t, 4H), 2.28 (t, 4H), 1.57 - 1.67 (m, 6H), 1.51 (br d, 12H), 1.23 - 1.33 (m, 58H), 0.82 - 0.93 (m, 12H)

[0233] (5) Synthesis of Compound E

[0234]

[0235] Di(heptadecan-9-yl) 8,8'-azandiyldioctanoate (1.20 g, 1.54 mmol, 1.00 eq), 2-(3-bromopropyl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (384 mg, 1.54 mmol, 1.00 eq), and K2CO3 (320 mg, 2.31 mmol, 1.50 eq) were mixed with acetonitrile (ACN, 12 mL) and degassed and purged three times with nitrogen (N2). Subsequently, the mixture was stirred at 60°C for 16 hours under a nitrogen atmosphere. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (SiO₂, DCM / MeOH = 20 / 1 to 10 / 1). The purified product was adjusted to pH 10 with saturated K2CO3 (30.0 mL), and the mixture was stirred at 20°C for 5 hours. Afterward, acetonitrile was removed by concentration under reduced pressure, the aqueous layer was extracted with dichloromethane (30.0 mL × 3), and the organic layer was concentrated under reduced pressure to obtain the final product. Finally, compound E (0.12 g, 139 μmol, yield 9.01%) was obtained in the form of a light yellow oil.

[0236] 1 H NMR (400 MHz, CHLOROFORM-d)δ7.69 - 8.24 (m, 2H), 4.87 (quin, 2H), 2.46 (br s, 6H), 2.28 (t, 4H), 1.44 - 1.71 (m, 26H), 1.24 - 1.32 (m, 52H), 0.93 - 0.98 (m, 2H), 0.89 (t, 12H)

[0237] [Example of preparation of a drug delivery composition]

[0238] 1. Preparation of raw materials

[0239] As shown in the table below, the substances required for formulation preparation were dissolved in each dilution solvent to the required concentrations. When dissolving, the substances were brought to room temperature before adding the solvent and dissolving them.

[0240]

[0241] 2. Mixing of raw materials

[0242] The required amounts of raw materials were taken and mixed according to the ratio of E:DOPE:cholesterol:DMG-PEG = 50:10:38.5:1.5, with the NP ratio (amine group of lipid: phosphate group of mRNA) set to 6. Ethanol was added to the ethanol layer so that the sum of all raw materials was within 12.5 mM, and the aqueous phase and ethanol phase were mixed while maintaining a volume ratio of 3:1. Buffer exchange was performed as follows to lower the total ethanol content after mixing: the mixture was concentrated by centrifuging at 4,000 rpm using an Amicon-Ultra tube filter (Merk Millipore, UFC505096 or UFC805024, pore size: 50K or 100K, volume 0.5 mL or 4 mL or 15 mL), and then the process of diluting with PBS and centrifuging to concentrate was repeated to perform buffer exchange.

[0243] The specific process sequence is as follows.

[0244] 1) Two autoclaved tubes were prepared (tubes (A), (B)).

[0245] 2) Each compound of chemical formulas A, C, D, and E, with moles calculated according to experimental conditions, and DOPE, cholesterol, and DMG-PEG were added sequentially to Tube (A), and mixed by vortexing after addition.

[0246] 3) Ethanol was added as needed in the ethanol phase so that the total of all raw materials was within 6.25-12.5 mM.

[0247] 4) mRNA and 20 mM sodium acetate buffer (pH 4.6) were mixed in Tube (B). The ratio was calculated so that the volume of the aqueous phase was three times the volume of the ethanol phase.

[0248] 5) Mixing of tube (A) and tube (B) was performed using a Microfluidics (Ignite, Precision Nanosystem) instrument. The Microfluidics operating conditions were a Flow Rate Ratio (FRR) of C:R=3:1 and a Total Flow Rate (TRR) of 12 mL / min.

[0249] 6) The resulting mixture from step 5) was centrifuged at 4,000 rpm using an Amicon-Ultra tube filter (50K) to concentrate it, then diluted with PBS and centrifuged to concentrate it again to remove excess ethanol, and then concentrated to a final x mg / mL (theoretical concentration).

[0250] 7) Once concentrated to the desired concentration, it was sterilized using a 0.22 µm pore size filter.

[0251] 3. Evaluation of Physical Properties of the Formulation

[0252] 1) For the prepared formulations, particle characteristics (i.e., zeta-average particle size, polydispersity index (PDI), and zeta-potential) were determined using a particle size analyzer (Dynamic Light Scattering, DLS), and the results are shown in Table 1 below.

[0253] 2) For the prepared formulations, mRNA encapsulation efficiency was confirmed through a Ribo-green assay, and the results are shown in Table 1 below.

[0254] [Table 1]

[0255]

Claims

1. Lipids having a structure represented by the following chemical formula 1: [Chemical Formula 1] In the above chemical formula 1, R p and R q Each is independently a hydrogen atom (H) or a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and R1 and R2 are each independently substituted or unsubstituted alkylene groups, and n is an integer greater than or equal to 0 or 1, and Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and R8, R 10 , R 11 and R 13 Each is independently a substituted or unsubstituted divalent saturated or unsaturated hydrocarbon group, and R9, R 12 and R 14 Each is independently a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and L1 and L2 are each independently selected from the group consisting of -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-, arylene groups, and heteroarylene groups, wherein L' is a directly bonded, substituted or unsubstituted alkylene group or substituted or unsubstituted alkenylene group, and R' is each independently selected from the group consisting of a hydrogen atom, a substituted or unsubstituted alkyl group and a substituted or unsubstituted alkenyl group, and X is -OH or -SH.

2. In paragraph 1, the above R p , R q , R1~R 14 A lipid in which one or more of the following are further substituted with one or more moieties selected from amino acids, sugars, vitamins, peptides, proteins, hormones or antibodies, or derivatives thereof.

3. In Paragraph 1, R p and R q is independently a hydrogen atom (H), or a substituted or unsubstituted C 1-6 It is an alkyl group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and R1 and R2 are each independently substituted or unsubstituted C 1-4 It is an alkylene group, and n is 0 or 1, and Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and R8, R 10 , R 11 and R 13 Each is independently substituted or unsubstituted divalent saturated or unsaturated C 2-20 It is a hydrocarbon group, and R9, R 12 and R 14 are independently substituted or unsubstituted monovalent saturated or unsaturated C 2-20 It is a hydrocarbon group, and L1 and L2 are each independently -C(O)O-, -OC(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 6-20 C having an arylene group and one or more heteroatoms selected from N, O, and S 3-20 Selected from the group consisting of heteroarylene groups, where L' is directly bonded, substituted, or unsubstituted C 1-13 alkylene group or substituted or unsubstituted C 2-13 It is an alkenylene group, and R' are each independently hydrogen atoms, substituted or unsubstituted C. 1-18 Alkyl groups and substituted or unsubstituted C 2-18 Selected from a group composed of alkenyl groups, and X is -OH or -SH, Geology.

4. In Paragraph 1, R p and R q is independently a hydrogen atom (H), or a substituted or unsubstituted C 2-4 It is an alkyl group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and R1 and R2 are each independently substituted or unsubstituted C 2-4 It is an alkylene group, and n is 0, and m is 0 and, R3, R4 and R6 each independently (ii) , (iii) or (iv) And, R8, R 10 , R 11 and R 13 Each is independently substituted or unsubstituted divalent saturated or unsaturated C 2-10 It is a hydrocarbon group, and R9, R 12 and R 14 are independently substituted or unsubstituted monovalent saturated or unsaturated C 10-20 It is a hydrocarbon group, and L1 and L2 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R')-, and -N(R')C(O)-, where R' is each independently a hydrogen atom, and a substituted or unsubstituted C 1-4 Selected from the group consisting of alkyl groups, and X is -OH, Geology.

5. In claim 1, a lipid having any one of the following chemical formulas A to E having a structure:

6. A method for preparing a lipid having a structure represented by Chemical Formula 1, comprising the step of reacting a compound of Chemical Formula a with a compound of Chemical Formula b: [Chemical formula a] [Chemical formula b] [Chemical Formula 1] As mentioned above, Hal is a halogen atom, and R p and R q Each is independently a hydrogen atom (H) or a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and optionally R p and R q They can bond and form rings together with the oxygen atoms to which they are attached, and R1 and R2 are each independently substituted or unsubstituted alkylene groups, and n is an integer greater than or equal to 0 or 1, and Each m is independently 0 or 1, and when m is 1, the N with R attached pointed to by m carries a univalent positive charge (+1), and R3 to R7 are each independently (i) a hydrogen atom (H), (ii) , (iii) or (iv) However, at least (n+2) of R3 to R7 present in the molecule are any one of (ii) to (iv), and R8, R 10 , R 11 and R 13 Each is independently a substituted or unsubstituted divalent saturated or unsaturated hydrocarbon group, and R9, R 12 and R 14 Each is independently a substituted or unsubstituted monovalent saturated or unsaturated hydrocarbon group, and L1 and L2 are each independently selected from the group consisting of -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-, arylene groups, and heteroarylene groups, wherein L' is a directly bonded, substituted or unsubstituted alkylene group or substituted or unsubstituted alkenylene group, and R' is each independently selected from the group consisting of a hydrogen atom, a substituted or unsubstituted alkyl group and a substituted or unsubstituted alkenyl group, and X is -OH or -SH.

7. A drug delivery composition comprising a lipid according to any one of claims 1 to 5.

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