Cationic lipid and method for producing same

Abstract

The present invention relates to cationic lipids and methods for producing the same, and more specifically to cationic lipids that easily form complexes with anionic drugs and are useful for drug delivery, and methods for producing the same.

Description

[Technical Field]

[0001] The present invention relates to cationic lipids and methods for producing the same, and more specifically to cationic lipids that easily form complexes with anionic drugs and are useful for drug delivery, and methods for producing the same. [Background technology]

[0002] Safe and efficient drug delivery technologies for treatment using anionic drugs, including nucleic acids, have long been studied, and various delivery vehicles and delivery technologies have been developed. Delivery vehicles are mainly divided into viral delivery vehicles using adenoviruses, retroviruses, etc., and non-viral delivery vehicles using cationic lipids, cationic polymers, etc. Viral delivery vehicles are known to pose many challenges to commercialization due to risks such as non-specific immune responses and complex manufacturing processes. Therefore, recent research has focused on the use of non-viral delivery vehicles, and research is being conducted toward improving these drawbacks. Non-viral delivery vehicles have the advantages of fewer side effects in terms of safety in the body and lower manufacturing costs compared to viral delivery vehicles.

[0003] Representative non-viral delivery vehicles for delivering nucleic acid substances include cationic lipid-nucleic acid complexes (lipoplexes) and polycationic polymer-nucleic acid complexes (polyplexes). Such cationic lipids and polycationic polymers form complexes with anionic drugs through electrostatic interactions, stabilizing the anionic drugs and increasing their intracellular delivery, and thus have been the subject of extensive research (Non-Patent Documents 1 and 2).

[0004] However, polycationic polymers have cytotoxicity due to their multivalent cationic charge, making their practical application problematic. Furthermore, conventional cationic lipids used in ionic lipid nanoparticles, which consist of cationic lipids, neutral lipids, and fusogenic lipids, have drawbacks such as complex synthesis, cytotoxicity, and low intracellular nucleic acid delivery efficiency. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] De Paula D, Bentley MV, Mahato RI, Hydrophobization and bioconjugation for enhanced siRNA delivery and targeting, RNA 13 (2007) 431-56 [Non-patent document 2] 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 Summary of the Invention [Problem to be solved by the invention]

[0006] The first aspect of the present invention is a compound represented by the following formula (1): [ka] (In the formula, M1 and M2 each independently represent a divalent linker group, R1 and R2 are each independently a substituted or unsubstituted carbocyclic or heterocyclic group; R3 is a hydrogen atom or a substituted or unsubstituted organic group optionally containing one or more heteroatoms; R4 to R7 each independently represent a hydrogen atom or a substituted or unsubstituted saturated or unsaturated hydrocarbon group; a and b are each independently an integer of 1 to 20.

[0007] According to one embodiment of the present invention, in the formula (1), M1 and M2 each independently represent -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)-, -SS-, arylene (more specifically, C 6-20 arylene), and heteroarylene (more specifically, C having one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S). 3-20 heteroarylene), where M' is a direct bond, C 1-13 Alkylene or C 2-13 alkenylene, and each R' is independently a hydrogen atom, C 1-18 Alkyl and C 2-18 It may be selected from the group consisting of alkenyl.

[0008] According to one embodiment of the present invention, in the formula (1), R1 and R2 are each independently C 3-20 Cycloalkyl, C 3-20 Cycloalkenyl, C 6-20 Aryl, C 3-20 Heterocycloalkyl, C 3-20 Heterocycloalkenyl, and C 3-20 heteroaryl, each of which is independently unsubstituted or C 1-18 Alkyl or C 2-18 The heterocycloalkyl, heterocycloalkenyl, and heteroaryl may each independently have one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S.

[0009] According to one embodiment of the present invention, in the formula (1), R3 is a hydrogen atom, a substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Carbocyclic group, -(CH2) n Q, -(CH2) nand -CQ(R)2, wherein each R is independently selected from the group consisting of a hydrogen atom, C 1-3 Alkyl and C 2-3 alkenyl; Q may be selected from the group consisting of a carbocyclic group, a heterocyclic group, -OR, -O(CH) n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, -N(R)R 12 , N(R)S(O)2R 12 , -O(CH2) n OR, -N(R)C(=NR 13 )N(R)2, -N(R)C(=CHR 13 )N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR 13 )N(R)2, -N(OR)C(=CHR 13 )N(R)2, -C(=NR 13 )N(R)2, -C(=NR 13 )R, —C(O)N(R)OR and —C(R)N(R)C(O)OR, where each n is independently an integer from 1 to 5; R 12 is C 3-6 is selected from the group consisting of carbocyclic and heterocyclic groups; R 13 are H, CN, NO2, C 1-6 Alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C 2-6 Alkenyl, C 3-6 and each R is independently selected from the group consisting of a hydrogen atom, a C 1-3 Alkyl and C 2-3 alkenyl, and each X is independently selected from the group consisting of F, CI, BR, and I, with the proviso that R3 is -(CH2) n Q, -(CH2) nWhen CHQR, -CHQR or -CQ(R), (i) when n is 1, 2, 3, 4 or 5, Q is not -N(R), or (ii) when n is 1 or 2, Q is not a 5-, 6- or 7-membered heterocycloalkyl.

[0010] According to one embodiment of the present invention, in the formula (1), R4 to R7 are each independently a hydrogen atom, C 1-3 Alkyl and C 2-3 It may be selected from the group consisting of alkenyl.

[0011] According to one embodiment of the present invention, in the formula (1), a and b may each independently be an integer of 1-15.

[0012] More specifically, in the formula (1), M1 and M2 may each independently be selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)- and -C(O)- (wherein M' and R' are as defined above).

[0013] More specifically, in the formula (1), R1 and R2 are each independently C 3-20 Cycloalkyl and C 3-20 heterocycloalkyl, each of which is independently unsubstituted or C 1-18 Alkyl or C 2-18 It may be substituted with alkenyl, wherein the heterocycloalkyl may have one or more (eg, 1 to 3) heteroatoms selected from N, O and S.

[0014] More specifically, in the formula (1), R3 is a hydrogen atom, a substituted or unsubstituted C 1-6 Alkyl and substituted or unsubstituted C 3-6 Carbocyclic groups (e.g., C 3-6 cycloalkyl).

[0015] More specifically, in the formula (1), R4 to R7 are each independently a hydrogen atom or C 1-3 It may also be alkyl.

[0016] More specifically, in the formula (1), a and b may each independently be an integer of 3 to 13.

[0017] More specifically, in the formula (1), M1 and M2 may each independently be selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R')-, and -N(R')C(O)- (wherein R' is as defined above).

[0018] More specifically, in the formula (1), R1 and R2 are each independently a substituted or unsubstituted C 3-15 It may also be cycloalkyl.

[0019] More specifically, in the formula (1), R3 is a hydrogen atom or a substituted or unsubstituted C 1-3 It may also be alkyl.

[0020] More specifically, in the formula (1), R4 to R7 may be hydrogen atoms.

[0021] More specifically, in the formula (1), a and b may each independently be an integer of 5 to 11, and even more specifically, may be an integer of 5 to 9.

[0022] More specifically, the lipid may have a structure selected from the following formulas (A to R): [ka]

[0023] A second aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1'), the method comprising the steps of: (1) reacting a compound of formula (a) with a compound of formula (b) to obtain a compound of formula (c); and (2) reacting the compound of formula (c) with a compound of formula (d). [ka] wherein M, R, R, R, R, and a are as defined in the first aspect above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0024] A third aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1), the method comprising the steps of: (1) reacting a compound of formula (a') with a compound of formula (b') to obtain a compound of formula (c'); (2) reacting a compound of formula (c') with a compound of formula (d) to obtain a compound of formula (e); and (3) reacting a compound of formula (e) with a compound of formula (c) obtained in the second aspect of the present invention. [ka] (In the formula, M1, M2, R1 to R7, a, and b are as defined in the first aspect, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0025] A fourth aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1'), comprising the steps of: (1) reacting a compound of formula (i) with a compound of formula (ii) to obtain a compound of formula (iii); and (2) reacting a compound of formula (iii) with a compound of formula (iv). [ka] wherein M, R, R, R, R, and a are as defined in the first aspect above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0026] A fifth aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1), the method comprising the steps of: (1) reacting a compound of formula (i') with a compound of formula (ii') to obtain a compound of formula (iii'); (2) reacting a compound of formula (iii') with a compound of formula (iv) to obtain a compound of formula (v); and (3) reacting a compound of formula (v) with a compound of formula (iii) obtained in the fourth aspect of the present invention. [ka] (In the formula, M1, M2, R1 to R7, a, and b are as defined in the first aspect, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0027] A sixth aspect of the present invention provides a drug delivery composition comprising a lipid according to the present invention. [Effects of the Invention]

[0028] The lipids having a specific structure according to the present invention can easily form a complex with an anionic drug, and by utilizing this complex, the drug can be efficiently delivered to the target biological tissue. [Brief explanation of the drawings]

[0029] [Figure 1] 1 is a reaction scheme for the lipid synthesis process carried out in Example 1. [Figure 2] 1 is a reaction scheme for the lipid synthesis process carried out in Example 2. [Figure 3] 1 is a reaction scheme for the lipid synthesis process carried out in Example 3. [Figure 4] 1 is a reaction scheme for the lipid synthesis process carried out in Example 4. [Figure 5] 1 is a reaction scheme for the lipid synthesis process carried out in Example 5. [Figure 6] 1 is a reaction scheme for the lipid synthesis process carried out in Example 6. [Figure 7]1 is a reaction scheme for the lipid synthesis process carried out in Example 7. [Figure 8] 1 is a reaction scheme for the lipid synthesis process carried out in Example 8. [Figure 9] 1 is a reaction scheme for the lipid synthesis process carried out in Example 9. [Figure 10] 1 is a reaction scheme for the lipid synthesis process carried out in Example 10. [Figure 11] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 11. [Figure 12] 1 is a reaction scheme for the lipid synthesis process carried out in Example 12. [Figure 13] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 13. [Figure 14] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 14. [Figure 15] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 15. [Figure 16] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 16. [Figure 17] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 17. [Figure 18] 1 shows a reaction scheme for the lipid synthesis process carried out in Example 18. BEST MODE FOR CARRYING OUT THE INVENTION

[0030] The present invention will now be described in further detail.

[0031] The lipid provided by the first aspect of the present invention is a lipid represented by the following formula (1): [ka] (In the formula, M1 and M2 each independently represent a divalent linker group, R1 and R2 are each independently a substituted or unsubstituted carbocyclic or heterocyclic group; R3 is a hydrogen atom or a substituted or unsubstituted organic group optionally containing one or more heteroatoms; R4 to R7 each independently represent a hydrogen atom or a substituted or unsubstituted saturated or unsaturated hydrocarbon group; a and b are each independently an integer of 1 to 20.

[0032] As used herein, the term "substituted or unsubstituted" of any group means that the group is not substituted or is substituted with an -OH, halogen atom, C 1-8 Alkyl groups (more specifically, C 3-7 alkyl group) or C 1-8 Halogenated alkyl groups (more specifically, C 3-7 It means that the group is substituted with one or more substituents selected from the group consisting of halogenated alkyl groups.

[0033] According to one embodiment of the present invention, in the formula (1), M1 and M2 each independently represent -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)-, -SS-, arylene (more specifically, C 6-20 Arylene, more specifically C 6-10 arylene), and heteroarylene (more specifically, C having one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S). 3-20 Heteroarylene, more specifically C 3-10 heteroarylene), where M' is a direct bond, C 1-13 Alkylene (more specifically C 1-6 alkylene) or C 2-13 Alkenylene (more specifically C 2-6 alkenylene), and each R' may independently represent a hydrogen atom, C 1-18 Alkyl (more specifically C 1-10 Alkyl, more specifically C 1-6 alkyl) and C2-18 Alkenyl (more specifically C 2-10 Alkenyl, more specifically C 2-6 alkenyl).

[0034] According to one embodiment of the present invention, in the formula (1), R and R are each independently substituted or unsubstituted C 3-20 Cycloalkyl (more specifically C 3-15 Cycloalkyl, more specifically C 6-15 cycloalkyl), substituted or unsubstituted C 3-20 Cycloalkenyl (more specifically C 3-15 Cycloalkenyl, more specifically C 6-15 cycloalkenyl), substituted or unsubstituted C 6-20 Aryl (more specifically C 6-14 aryl), substituted or unsubstituted C 3-20 Heterocycloalkyl (more specifically C 3-15 heterocycloalkyl), substituted or unsubstituted C 3-20 Heterocycloalkenyl (more specifically C 3-15 heterocycloalkenyl), and substituted or unsubstituted C 3-20 Heteroaryl (more specifically C 3-15 and heteroaryl), wherein the heterocycloalkyl, heterocycloalkenyl, and heteroaryl each independently may have one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S.

[0035] According to one embodiment of the present invention, in the formula (1), R3 is a hydrogen atom, a substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Carbocyclic group, -(CH2) n Q, -(CH2) n and -CQ(R)2, wherein each R is independently selected from the group consisting of a hydrogen atom, C 1-3 Alkyl and C 2-3alkenyl; Q may be selected from the group consisting of a carbocyclic group, a heterocyclic group, -OR, -O(CH) n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, -N(R)R 12 , N(R)S(O)2R 12 , -O(CH2) n OR, -N(R)C(=NR 13 )N(R)2, -N(R)C(=CHR 13 )N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR 13 )N(R)2, -N(OR)C(=CHR 13 )N(R)2, -C(=NR 13 )N(R)2, -C(=NR 13 )R, —C(O)N(R)OR and —C(R)N(R)C(O)OR, where each n is independently an integer from 1 to 5; R 12 is C 3-6 is selected from the group consisting of carbocyclic and heterocyclic groups; R 13 are H, CN, NO2, C 1-6 Alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C 2-6 Alkenyl, C 3-6 and each R is independently selected from the group consisting of a hydrogen atom, a C 1-3 Alkyl and C 2-3 alkenyl, and each X is independently selected from the group consisting of F, CI, BR, and I, with the proviso that R3 is -(CH2) n Q, -(CH2) n When CHQR, -CHQR or -CQ(R), (i) when n is 1, 2, 3, 4 or 5, Q is not -N(R), or (ii) when n is 1 or 2, Q is not a 5-, 6- or 7-membered heterocycloalkyl.

[0036] According to one embodiment of the present invention, in the formula (1), R4 to R7 are each independently a hydrogen atom, C 1-3 Alkyl and C 2-3 It may be selected from the group consisting of alkenyl.

[0037] According to one embodiment of the present invention, in the formula (1), a and b may each independently be an integer of 1 to 15, more specifically an integer of 3 to 13.

[0038] More specifically, in the formula (1), M1 and M2 may each independently be selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R')-, and -N(R')C(O)-, where R' is as defined above.

[0039] More specifically, in the formula (1), R1 and R2 are each independently a substituted or unsubstituted C 3-15 It may also be cycloalkyl.

[0040] More specifically, in the formula (1), R3 is a hydrogen atom or a substituted or unsubstituted C 1-3 It may also be alkyl.

[0041] More specifically, in the formula (1), R4 to R7 may be hydrogen atoms.

[0042] More specifically, in the formula (1), a and b may each independently be an integer of 5 to 11, and even more specifically, may be an integer of 5 to 9.

[0043] In one embodiment, R and R are different from each other, and R is a substituted or unsubstituted C 6-15 cycloalkyl, and R2 may be substituted or unsubstituted C 3-6 It may also be cycloalkyl.

[0044] More specifically, the lipid may have a structure selected from the following formulas (A to R): [ka]

[0045] A second aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1'), the method comprising the steps of: (1) reacting a compound of formula (a) with a compound of formula (b) to obtain a compound of formula (c); and (2) reacting the compound of formula (c) with a compound of formula (d). [ka] wherein M, R, R, R, R, and a are as defined in the first aspect above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0046] In one embodiment of the lipid production method according to the second aspect of the present invention, the reaction in step (1) can be carried out in a solvent (e.g., methylene chloride (DCM) or toluene) in the presence of a catalyst (e.g., triethylamine (TEA) or sulfuric acid) at room temperature (e.g., 20°C to 30°C) or at an elevated temperature (e.g., 40 to 150°C), and the reaction in step (2) can be carried out in a solvent (e.g., ethanol (EtOH) or dioxane) optionally in the presence of Na2CO3 or a catalyst (e.g., N,N-diisopropylethylamine (DIEA)) at an elevated temperature (e.g., 40 to 150°C), but is not limited thereto.

[0047] According to one embodiment, prior to the reaction of the compound of formula (a) with the compound of formula (b) in step (1), the compound of formula (b) may be pretreated. This pretreatment may be carried out using a chloride (e.g., oxalyl chloride) in a solvent (e.g., methylene chloride (DCM)) in the presence of dimethylformamide (DMF) at a low temperature (e.g., −10° C. to 10° C.) or room temperature (e.g., 20° C. to 30° C.), but is not limited thereto.

[0048] A third aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1), the method comprising the steps of: (1) reacting a compound of formula (a') with a compound of formula (b') to obtain a compound of formula (c'); (2) reacting a compound of formula (c') with a compound of formula (d) to obtain a compound of formula (e); and (3) reacting a compound of formula (e) with a compound of formula (c) obtained in the second aspect of the present invention. [ka] (In the formula, M1, M2, R1 to R7, a, and b are as defined in the first aspect, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0049] In one embodiment of the lipid production method according to the third aspect of the present invention, the reaction in step (1) can be carried out in a solvent (e.g., methylene chloride (DCM) or toluene) in the presence of a catalyst (e.g., triethylamine (TEA) or sulfuric acid) at room temperature (e.g., 20°C to 30°C) or at an elevated temperature (e.g., 40 to 150°C), and the reactions in steps (2) and (3) can each be independently carried out in a solvent (e.g., ethanol (EtOH)) optionally in the presence of Na2CO3 or a catalyst (e.g., N,N-diisopropylethylamine (DIEA)) at an elevated temperature (e.g., 40 to 150°C), but are not limited thereto.

[0050] According to one embodiment, prior to the reaction of the compound of formula (a') with the compound of formula (b') in step (1), the compound of formula (b') may be pretreated. This pretreatment may be carried out using a chloride (e.g., oxalyl chloride) in a solvent (e.g., methylene chloride (DCM)) in the presence of dimethylformamide (DMF) at low temperature (e.g., -10°C to 10°C) or room temperature (e.g., 20°C to 30°C), but is not limited thereto.

[0051] A fourth aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1'), comprising the steps of: (1) reacting a compound of formula (i) with a compound of formula (ii) to obtain a compound of formula (iii); and (2) reacting a compound of formula (iii) with a compound of formula (iv). [ka] wherein M, R, R, R, R, and a are as defined in the first aspect above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0052] In one embodiment of the lipid production method according to the fourth aspect of the present invention, the reaction in step (1) can be carried out in a solvent (e.g., methylene chloride (DCM) or toluene) in the presence of a catalyst (e.g., triethylamine (TEA) or sulfuric acid) at room temperature (e.g., 20°C to 30°C) or at an elevated temperature (e.g., 40 to 150°C), and the reaction in step (2) can be carried out in a solvent (e.g., ethanol (EtOH) or dioxane) optionally in the presence of Na2CO3 or a catalyst (e.g., N,N-diisopropylethylamine (DIEA)) at an elevated temperature (e.g., 40 to 150°C), but is not limited thereto.

[0053] A fifth aspect of the present invention provides a method for producing a lipid having a structure represented by formula (1), the method comprising the steps of: (1) reacting a compound of formula (i') with a compound of formula (ii') to obtain a compound of formula (iii'); (2) reacting a compound of formula (iii') with a compound of formula (iv) to obtain a compound of formula (v); and (3) reacting a compound of formula (v) with a compound of formula (iii) obtained in the fourth aspect of the present invention. [ka] (In the formula, M1, M2, R1 to R7, a, and b are as defined in the first aspect, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0054] In one embodiment of the lipid production method according to the fifth aspect of the present invention, the reaction in step (1) can be carried out in a solvent (e.g., methylene chloride (DCM) or toluene) in the presence of a catalyst (e.g., triethylamine (TEA) or sulfuric acid) at room temperature (e.g., 20°C to 30°C) or at an elevated temperature (e.g., 40 to 150°C), and the reactions in steps (2) and (3) can each be independently carried out in a solvent (e.g., ethanol (EtOH)) optionally in the presence of Na2CO3 or a catalyst (e.g., N,N-diisopropylethylamine (DIEA)) at an elevated temperature (e.g., 40 to 150°C), but are not limited thereto.

[0055] The lipid having the structure represented by formula (1) of the present invention can easily form a complex with an anionic drug, and is therefore useful for drug delivery.

[0056] Therefore, according to a sixth aspect of the present invention, there is provided a drug delivery composition comprising a lipid having a structure represented by formula (1) of the present invention.

[0057] In one embodiment, the drug may be selected from a nucleic acid, a polypeptide, a virus, or a combination thereof.

[0058] The "nucleic acid" may be, for example, but is not limited to, DNA, RNA, siRNA, shRNA, miRNA, mRNA, an aptamer, an antisense oligonucleotide, or a combination thereof.

[0059] The term "polypeptide" may refer to a protein that has 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 the polypeptide sequence of an antigen, an analog thereof, or a precursor thereof.

[0060] In one embodiment, the lipids of the present invention are complexed with a drug, and the complex is encapsulated within the nanoparticle structure formed by the amphiphilic block copolymer.

[0061] In one embodiment, the amphiphilic block copolymer may be an AB type block copolymer comprising a hydrophilic A block and a hydrophobic B block, which in an aqueous environment forms core-shell polymeric nanoparticles in which the hydrophobic B block forms the core (inner wall) and the hydrophilic A block forms the shell (outer wall).

[0062] 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.

[0063] More specifically, the hydrophilic A block may be one or more selected from the group consisting of monomethoxypolyethylene glycol (mPEG), monoacetoxypolyethylene glycol, polyethylene glycol, a copolymer of polyethylene and propylene glycol, and polyvinylpyrrolidone.

[0064] Furthermore, if necessary, functional groups, ligands, or functional groups capable of targeting specific tissues or cells, or functional groups capable of promoting intracellular delivery, can be chemically bonded to the ends of the hydrophilic A block to control the biodistribution of polymeric nanoparticle delivery vehicles formed from amphiphilic block copolymers and polylactic acid salts or to enhance the intracellular delivery efficiency of the nanoparticle delivery vehicles. 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, antibodies against transferrin receptors, and the like.

[0065] The hydrophobic B block is a biocompatible, biodegradable polymer, which in one embodiment may be one or more selected from the group consisting of polyesters, polyanhydrides, polyamino acids, polyorthoesters, and polyphosphazines.

[0066] More specifically, the hydrophobic B block may be one or more selected from the group consisting of polylactide (PLA), polyglycolide, polycaprolactone, polydioxane-2-one, a copolymer of polylactide and glycolide, a copolymer of polylactide and polydioxane-2-one, a copolymer of polylactide and polycaprolactone, and a copolymer of polyglycolide and polycaprolactone.

[0067] In one embodiment, in order to increase the hydrophobicity of the hydrophobic B block and thereby improve the stability of the nanoparticles, the hydrophobic B block may be modified by chemically bonding tocopherol, cholesterol, or a fatty acid having 10 to 24 carbon atoms to the hydroxy group at the end of the hydrophobic B block.

[0068] The present invention will be described in more detail below with reference to examples. However, these examples are for the purpose of illustrating the present invention and are not intended to limit the scope of the present invention in any way.

[0069] Example Example 1 1-1. The compound of formula (A) below was produced according to the synthesis scheme shown in FIG. [ka]

[0070] Synthesis of 1-2,4-pentylcyclohexyl 8-bromooctanoate A 250 mL three-necked round-bottom flask (RBF) was charged with 8-bromooctanoic acid (2.00 g, 8.96 mmol, 1.00 eq), methylene chloride (DCM) (40 mL), and dimethylformamide (DMF) (0.5 mL), and oxalyl chloride (2.28 g, 17.9 mmol, 2.00 eq) was added at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at 25 °C for 4 h under a nitrogen atmosphere, after which 4-pentylcyclohexan-1-ol (2.29 g, 13.5 mmol, 1.50 eq) and triethylamine (TEA) (1.36 g, 13.5 mmol, 1.50 eq) were added, and the mixture was stirred at 25 °C under a nitrogen atmosphere for an additional 12 h. The reaction mixture was concentrated in vacuo, and the concentrated residue was purified on a silica column with petroleum ether:ethyl acetate (EtOAc) = 1:0 → 50:1 to give 4-pentylcyclohexyl 8-bromooctanoate (2.46 g, 6.55 mmol, yield 73.1%) as a pale yellow oil.

[0071] 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.76 - 0.86 (m, 3 H) 0.88 - 1.94 (m, 28 H) 2.21 (dt, 2 H) 3.33 (td, 2 H) 4.52 - 4.66 (m, 1 H) 4.87 - 4.95 (m, 1 H)

[0072] Synthesis of 1-3,4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate 4-Pentylcyclohexyl 8-bromooctanoate (2.46 g, 6.55 mmol, 1.00 eq), 2-aminoethan-1-ol (2.00 g, 32.8 mmol, 5.00 eq), and ethanol (EtOH) (50 mL) were added to a 100 mL three-neck RBF and stirred under a nitrogen atmosphere at 80° C. for 16 hours. The reaction mixture was concentrated in vacuo, and the concentrated residue was purified on a silica column with petroleum ether:EtOAc = 1:0 → 50:1 to give 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate (2.00 g, 5.62 mmol, 85.8% yield) as a yellow solid.

[0073] 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.76 - 0.86 (m, 3 H) 0.88 - 1.94 (m, 27 H) 2.32 (t, 2 H) 2.51 (t, 2 H) 2.71 (t, 2H), 3.54 (d, 2 H) 3.91 - 4.00 (m, 1 H) 4.11 - 4.31 (m, 1 H)

[0074] 1-4. Synthesis of cyclopentadecyl 8-bromooctanoate To a 250 mL three-neck RBF, cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 eq), 8-bromooctanoic acid (4.93 g, 22.1 mmol, 1 eq), sulfuric acid (HSO) (217 mg, 2.21 mmol, 0.10 eq), and toluene (100 mL) were added, and the mixture was stirred under a nitrogen atmosphere at 120 °C for 16 h. Subsequently, after evaporation of the solvent, the residue was purified on a silica column with petroleum ether: EtOAc = 1:0 → 50:1 to give cyclopentadecyl 8-bromooctanoate (2.60 g, 6.03 mmol, 27.3% yield) as a colorless oil.

[0075] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.89 (quin, 1H), 3.41 (dt, 2H), 2.28 (t, 2H), 1.85 (quin, 2H), 1.73 - 1.16 (m, 36H)

[0076] 1-5. Synthesis of Compound of Formula (A) A 100 mL three-neck RBF was charged with cyclopentadecyl 8-bromooctanoate (1.60 g, 3.71 mmol, 1.00 eq), 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate (1.32 g, 3.71 mmol, 1.00 eq), N,N-diisopropylethylamine (DIEA) (527 mg, 4.08 mmol, 1.10 eq), and EtOH (30 mL) and stirred at 80 °C for 48 h. The solvent was evaporated, and the residue was purified on a silica column with petroleum ether: EtOAc (10:1 → 1:1). The product was then purified again by prep-HPLC (folic acid conditions). After washing with aqueous NaHCO (300 mL), the organic layer was concentrated and extracted with DCM (200 mL × 2). The organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated to give the compound of formula (A) (0.240 g, 340 μmol, 9.16% yield) as a yellow oil.

[0077] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.91 (br s, 1H), 4.82 (quin, 1H), 3.46 (br t, 2H), 2.51 (br d, 2H), 2.37 (br t, 4H), 2.21 (td, 4H), 1.58 - 1.42 (m, 14H), 1.33 - 1.17 (m, 52H), 0.83 - 0.80 (m, 3H).

[0078] Example 2 2-1. The compound of formula (B) below was produced according to the synthesis scheme shown in FIG. [ka]

[0079] 2-2. Synthesis of cyclopentadecyl 6-bromohexanoate A 100 mL three-neck RBF was charged with cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 eq), 6-bromohexanoic acid (6.46 g, 33.1 mmol, 1.50 eq), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (5.08 g, 26.5 mmol, 1.20 eq), 4-dimethylaminopyridine (DMAP) (540 mg, 4.42 mmol, 0.20 eq), triethanolamine (TEA) (4.47 g, 44.2 mmol, 6.15 mL, 2.00 eq), and DCM (50 mL). The mixture was purged with nitrogen gas three times. The resulting mixture was stirred under nitrogen at 25 °C for 16 h. The reaction was quenched with 200 mL of water at 20 °C and extracted with 600 mL of DCM (3 x 200 mL). The organic layer was collected, dried over NaSO, filtered, and concentrated under vacuum. The residue was purified on a silica column with petroleum ether: EtOAc = 100:1 → 1:1 to give cyclopentadecyl 6-bromohexanoate (2.20 g, 5.45 mmol, 24.7% yield) as a yellow oil.

[0080] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.83 (quin, 1H), 3.50 - 3.30 (m, 2H), 2.23 (t, 2H), 1.87 - 1.68 (m, 2H), 1.63 - 1.37 (m, 10H), 1.26 (br s, 22H)

[0081] Synthesis of 2-3,4-propylcyclohexyl 6-bromohexanoate To a 250 mL three-necked RBF was added 4-propylcyclohexan-1-ol (10.0 g, 70.3 mmol, 1.00 eq), 6-bromohexanoic acid (16.5 g, 84.4 mmol, 1.20 eq), EDCI (20.2 g, 105 mmol, 1.50 eq), DMAP (8.59 g, 70.3 mmol, 1.00 eq), and TEA (7.11 g, 70.3 mmol, 9.79 mL, 1.00 eq) in DCM (100 mL), and the mixture was purged with nitrogen three times and then stirred under nitrogen atmosphere at 25 °C for 16 h. The reaction mixture was neutralized by adding 100 mL of water at 20°C and extracted with 600 mL of DCM (200 mL × 3 times). The organic layer was then collected and concentrated in vacuo. The concentrated residue was purified on a silica column with petroleum ether: EtOAc = 100:1 → 10:1 to give 4-propylcyclohexyl 6-bromohexanoate (6.80 g, 21.3 mmol, yield 30.3%) as a yellow oil.

[0082] 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.76 - 0.86 (m, 3 H) 0.88 - 1.94 (m, 19 H) 2.21 (dt, 2 H) 3.33 (td, 2 H) 4.52 - 4.66 (m, 1 H)

[0083] Synthesis of 2-4,4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate To a 250 mL three-neck RBF was added 4-propylcyclohexyl 6-bromohexanoate (3.00 g, 9.40 mmol, 1.00 eq), 2-aminoethan-1-ol (2.87 g, 47.0 mmol, 2.84 mL, 5.00 eq), and EtOH (60 mL). The mixture was purged with nitrogen three times and then stirred under a nitrogen atmosphere at 5 °C for 16 h. The reaction mixture was cooled to room temperature and concentrated to remove the solvent. The residue was purified by column chromatography (SiO, DCM:MeOH = 100:1 → 1:1) to give 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate (0.70 g, 2.34 mmol, 24.9% yield) as a yellow oil.

[0084] 1 H NMR (400 MHz, CHLOROFORM-d): δ 5.04 - 4.60 (m, 1H), 3.70 - 3.61 (m, 2H), 2.79 (t, 2H), 2.64 (dt, 2H), 2.35 - 2.24 (m, 2H), 2.01 - 1.94 (m, 2H), 1.87 - 1.74 (m, 2H), 1.65 (qd, 2H), 1.56 - 1.48 (m, 4H), 1.41 - 1.16 (m, 9H), 1.07 - 0.94 (m, 1H), 0.89 (dt, 3H)

[0085] 2-5. Synthesis of Compound of Formula (B) To a 50 mL three-neck RBF, 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate (371 mg, 1.24 mmol, 1.00 eq), 4-propylcyclohexyl 6-bromohexanoate (0.50 g, 1.24 mmol, 1.00 eq), N,N-diisopropylethylamine (DIEA) (800 mg, 6.20 mmol, 1.08 mL, 5 eq) were added along with 1,4-dioxane (10 mL). The mixture was purged with nitrogen three times and then stirred at 95 °C under a nitrogen atmosphere for 16 h. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO, DCM:MeOH = 100:1 → 10:1) to give compound of formula (B) (0.21 g, 337.64 μmol, 27.2% yield) as a yellow oil.

[0086] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.94 - 4.52 (m, 2H), 3.79 (br s, 2H), 3.06 - 2.63 (m, 6H), 2.29 - 2.17 (m, 4H), 1.93 - 1.81 (m, 1H), 1.79 - 1.37 (m, 17H), 1.36 - 1.09 (m, 34H), 0.99 - 0.86 (m, 1H), 0.85 - 0.76 (m, 3H)

[0087] Example 3 3-1. A compound of the following formula (C) was produced according to the synthesis scheme shown in FIG. [ka]

[0088] 3-2. Synthesis of cyclopentadecyl 10-bromodecanoate To a 100 mL three-neck RBF, cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 eq), 10-bromodecanoic acid (8.32 g, 33.1 mmol, 1.50 eq), DMAP (540 mg, 4.42 mmol, 0.20 eq), EDCI (5.08 g, 26.5 mmol, 1.20 eq), and TEA (4.47 g, 44.2 mmol, 2.00 eq) were added along with DCM (50 mL). The mixture was purged with nitrogen three times and then stirred at 50 °C under a nitrogen atmosphere for 16 h. The reaction mixture was cooled to 20 °C and quenched with water. The mixture was then extracted with 600 mL of DCM (3 x 200 mL). The organic layer was collected, dried over Na2SO4, filtered, and the filtrate was concentrated. The resulting residue was purified by column chromatography (SiO 2 , petroleum ether: EtOAc = 100:1 → 1:1) to afford cyclopentadecyl 10-bromodecanoate (2.60 g, 5.66 mmol, 25.6% yield) as a yellow oil.

[0089] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.90 (quin, 1H), 3.58 - 3.36 (m, 2H), 2.27 (t, 2H), 1.90 - 1.72 (m, 2H), 1.64 - 1.53 (m, 6H), 1.43 - 1.28 (m, 34H)

[0090] Synthesis of 3-3,4-heptylcyclohexan-1-ol LiAlH4 (2.50 M in THF, 61.1 mL, 1.20 eq) was added to a 500 mL three-neck RBF along with THF (250 mL). The mixture was cooled to 0 °C, and a solution of 4-heptylcyclohexan-1-one (25.0 g, 127 mmol, 1.00 eq) in THF (250 mL) was added slowly over 20 min under a nitrogen atmosphere. The temperature of the mixture was then raised to 25 °C and stirred under nitrogen for 3 h. The reaction mixture was cooled to 0 °C, and water (120 mL) was added under a nitrogen atmosphere, taking care to avoid excessive foaming. 15% aqueous NaOH (12 mL) was then slowly added to the reaction mixture at 0 °C. After 5 min, water (36 mL) was added at the same temperature. The temperature was then slowly raised to 25 °C, and the mixture was stirred for 15 min. The resulting mixture was filtered, and the filter cake was concentrated in vacuo to give 4-heptylcyclohexan-1-ol (50.0 g, 252 mmol, 98.9% yield) as a colorless powder.

[0091] 1 H NMR (400 MHz, CHLOROFORM-d): δ 3.47 (tt, H), 1.95 - 1.84 (m, 2H), 1.75 - 1.65 (m, 2H), 1.42 (s, 1H), 1.24 - 1.14 (m, 14H), 1.11 - 1.08 (m, 2H), 0.83 - 0.80 (m, 3H)

[0092] Synthesis of 3-4,4-heptylcyclohexyl 10-bromodecanoate To a 250 mL three-necked RBF was added 4-heptylcyclohexan-1-ol (10.0 g, 50.4 mmol, 1.00 eq), 10-bromodecanoic acid (15.2 g, 60.5 mmol, 1.20 eq), EDCI (14.5 g, 75.6 mmol, 1.50 eq), DMAP (6.16 g, 50.4 mmol, 1.00 eq), TEA (5.10 g, 50.4 mmol, 1.00 eq), and DCM (100 mL). The mixture was purged with nitrogen three times and then stirred under a nitrogen atmosphere at 25° C. for 16 h. The reaction mixture was neutralized by adding 200 mL of water at 20°C and extracted with 600 mL of DCM (200 mL × 3 times). The organic layer was collected and concentrated in vacuo, and the concentrated residue was purified by column chromatography (SiO, petroleum ether: EtOAc = 100:1 → 1:1) to give 4-heptylcyclohexyl 10-bromodecanoate (6.60 g, 15.3 mmol, yield 30.3%) as a white solid.

[0093] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.59 (tt, 1H), 3.46 (t, 1H), 3.33 (t, 1H), 2.19 (t, 2H), 1.87 (br dd, 2H), 1.82 - 1.67 (m, 4H), 1.58 - 1.50 (m, 2H), 1.38 - 1.32 (m, 2H), 1.21 (br d, 23H), 0.97 - 0.86 (m, 2H), 0.81 (t, 3H)

[0094] Synthesis of 3-5,4-heptylcyclohexyl 10-((2-hydroxyethyl)amino)decanoate To a 250 mL three-neck RBF, 4-heptylcyclohexyl 10-bromodecanoate (2.30 g, 5.33 mmol, 1.00 eq) and 2-aminoethan-1-ol (1.63 g, 26.6 mmol, 5 eq) were added along with EtOH (60 mL). The mixture was purged with nitrogen three times and then stirred under a nitrogen atmosphere at 95 °C for 16 h. The reaction mixture was concentrated in vacuo, and the residue was purified by column chromatography (SiO, DCM:MeOH = 100:1 → 1:1) to give 4-heptylcyclohexyl 10-((2-hydroxyethyl)amino)decanoate (1.30 g, 3.16 mmol, 59.2% yield) as a white solid.

[0095] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.66 (tt, 1H), 3.83 - 3.73 (m, 2H), 2.96 - 2.88 (m, 2H), 2.81 - 2.73 (m, 2H), 2.26 (t, 2H), 1.95 (br dd, 2H), 1.78 (br d, 2H), 1.62 (td, 4H), 1.28 (br d, 24H), 1.18 (br d, 2H), 1.04 - 0.96 (m, 2H), 0.89 (t, 3H)

[0096] 3-6. Synthesis of Compound of Formula (C) To a 50 mL three-neck RBF, 4-heptylcyclohexyl 10-((2-hydroxyethyl)amino)decanoate (0.80 g, 1.94 mmol, 1.00 eq), cyclopentadecyl 10-bromodecanoate (893 mg, 1.94 mmol, 1.00 eq), DIEA (1.26 g, 9.72 mmol, 1.69 mL, 5.00 eq) and EtOH (3 mL) were added. The mixture was purged with nitrogen three times and then stirred at 95 °C under a nitrogen atmosphere for 16 h. The reaction mixture was cooled to room temperature, concentrated in vacuo, and the residue was purified by chromatography (SiO, DCM:MeOH = 100:1 → 10:1) to give compound of formula (C) (0.35 g, 447 μmol, 22.9% yield) as a white solid.

[0097] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.96 - 4.61 (m, 2H), 3.77 - 3.52 (m, 2H), 2.84 - 2.37 (m, 6H), 2.27 (dt, 4H), 2.01 - 1.92 (m, 2H), 1.79 (br d, 2H), 1.68 - 1.46 (m, 18H), 1.37 - 1.25 (m, 56H), 1.00 (br d, 1H), 0.89 (br t, 3H)

[0098] Example 4 4-1. A compound of the following formula (D) was produced according to the synthesis scheme shown in FIG. [ka]

[0099] 4-2. Synthesis of (E)-2-(hept-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane To a 500 mL three-necked RBF was added hept-1-yne (64.4 g, 670 mmol, 1.00 eq), TEA (EtN, 6.78 g, 670 mmol, 0.10 eq), 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (90.0 g, 703 mmol, 1.05 eq), zirconium chloride; cyclopentane (17.9 g, 670 mmol, 0.10 eq), and the mixture was purged with nitrogen three times and then stirred under a nitrogen atmosphere at 60 °C for 16 h. The reaction mixture was cooled to room temperature, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1) to give (E)-2-(hept-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (102 g, 455 mmol, 67.9% yield) as a yellow oil.

[0100] 1H NMR: (400 MHz, CHLOROFORM-d): δ 6.64 (td, 1H), 5.43 (td, 1H), 2.19 - 2.12 (m, 2H), 1.44 - 1.39 (m, 2H), 1.30 (br d, 4H), 1.27 (s, 12H), 0.90 - 0.87 (m, 3H)

[0101] Synthesis of 4-3,4,4,5,5-tetramethyl-2-((1R,2R)-2-pentylcyclopropyl)-1,3,2-dioxaborolane Diethylzinc (1M, 178.5 mL, 2.00 eq) was added to a 1000 mL three-neck RBF under a nitrogen atmosphere along with purified, distilled DCM (80 mL). A solution of trifluoroacetic acid (TFA) (20.4 g, 178 mmol, 2.00 eq) in DCM (40 mL) was added portionwise at 0 °C. A solution of diiodomethane (47.8 g, 178 mmol, 2.00 eq) in DCM (40 mL) was added with stirring at 0 °C for 30 min. The reaction mixture was stirred for an additional 30 min, after which a solution of (E)-2-(hept-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (20.0 g, 89.2 mmol, 1.00 eq) in DCM (40 mL) was added at 0 °C, and the mixture was stirred under a nitrogen atmosphere at 25 °C for 2 h. The reaction mixture was neutralized with water and extracted with DCM (1000 mL x 3). The organic layer was collected, dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo. The residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1 → 1:100) to give 4,4,5,5-tetramethyl-2-((1R,2R)-2-pentylcyclopropyl)-1,3,2-dioxaborolane (14.0 g, 58.8 mmol, 65.9% yield) as a yellow oil.

[0102] 1H NMR (400 MHz, CHLOROFORM-d): δ 1.42 - 1.37 (m, 2H), 1.32 - 1.26 (m, 6H), 1.26 - 1.24 (m, 2H), 1.22 (s, 10H), 0.89 (br t, 4H), 0.67 (dt, 1H), 0.43 - 0.33 (m, 1H), -0.42 (td, 1H)

[0103] 4-4. Synthesis of (1R,2R)-2-pentylcyclopropan-1-ol To a 500 mL three-necked RBF was added 4,4,5,5-tetramethyl-2-((1R,2R)-2-pentylcyclopropyl)-1,3,2-dioxaborolane (8.00 g, 33.6 mmol, 1.00 eq) and THF (160 mL). To this mixture was added NaOH (2.69 g, 67.2 mmol, 2.00 eq) slowly at 0 °C, and then HO (7.87 g, 69.4 mmol, 30% purity, 2.07 eq) was added at 0 °C. The mixture was stirred at 25 °C for 16 h. The reaction mixture was then poured into ice-water containing Na2SO3 (2 eq) to quench the reaction, extracted with DCM (30 mL x 3). The organic layer was collected, dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1 → 1:100) to give (1R,2R)-2-pentylcyclopropan-1-ol (2.50 g, 19.5 mmol, 58.1% yield) as a yellow oil.

[0104] 1 H NMR (400 MHz, CHLOROFORM-d): δ 3.20 (td, 1H), 1.84 - 1.67 (m, 1H), 1.41 - 1.35 (m, 2H), 1.33 - 1.26 (m, 4H), 1.23 - 1.06 (m, 2H), 0.94 - 0.87 (m, 4H), 0.68 (ddd, 1H), 0.31 (q, 1H)

[0105] 4-5. Synthesis of (1R,2R)-2-pentylcyclopropyl 8-bromooctanoate To a 100 mL three-neck RBF, (1R,2R)-2-pentylcyclopropan-1-ol (2.50 g, 19.5 mmol, 1.00 eq) was added along with DCM (25 mL), 8-bromooctanoic acid (5.22 g, 23.4 mmol, 1.20 eq), EDCI (4.49 g, 23.4 mmol, 1.20 eq), DMAP (476 mg, 3.90 mmol, 0.20 eq), and EtN (3.95 g, 39.00 mmol, 2.00 eq). The mixture was purged with nitrogen three times and then stirred under nitrogen atmosphere at 25 °C for 16 h. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1 → 1:100) to give (1R,2R)-2-pentylcyclopropyl 8-bromooctanoate (3.50 g, 10.5 mmol, 53.9% yield) as a yellow oil.

[0106] 1 H NMR (400 MHz, CHLOROFORM-d): δ 3.85 - 3.79 (m, 1H), 3.41 (t, 2H), 2.26 (t, 2H), 1.86 (quin, 2H), 1.65 - 1.58 (m, 2H), 1.46 - 1.38 (m, 4H), 1.36 - 1.27 (m, 10H), 1.00 (ddd, 1H), 0.89 (br t, 3H), 0.78 (ddd, 1H), 0.52 (q, 1H)

[0107] 4-6. Synthesis of cyclohexyl 8-bromooctanoate 8-Bromooctanoic acid (15.0 g, 67.2 mmol, 1.00 eq) was added to a 1000 mL three-neck RBF together with DCM (150 mL). Trifluoroacetic anhydride (TFAA) (14.1 g, 67.2 mmol, 1.00 eq) was added under a nitrogen atmosphere and stirred at 0-25°C for 2.5 h. Cyclohexanol (33.7 g, 336 mmol, 35.1 mL, 5.00 eq) was added to the mixture at 0°C. After stirring at 25°C for 16 h under a nitrogen atmosphere, the reaction mixture was neutralized with 100 mL of water at 20°C and extracted with 600 mL of DCM (3 x 200 mL). The organic layer was collected, dried over Na2SO4, and filtered. The filtrate was concentrated in vacuo, and the residue was purified by silica column chromatography (SiO2, petroleum ether: EtOAc = 100:1 → 10:1) to give cyclohexyl 8-bromooctanoate (7.10 g, 23.3 mmol, 34.6% yield) as a colorless oil.

[0108] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.77 (br d, 1H), 3.42 (t, 2H), 2.30 (t, 2H), 1.93 - 1.82 (m, 4H), 1.74 (br dd, 2H), 1.67 - 1.55 (m, 4H), 1.47 - 1.31 (m, 10H)

[0109] 4-7. Synthesis of cyclohexyl 8-((2-hydroxyethyl)amino)octanoate To a 250 mL three-neck RBF, cyclohexyl 8-bromooctanoate (3.00 g, 9.83 mmol, 1.00 eq), 2-aminoethan-1-ol (3.00 g, 49.1 mmol, 5.00 eq), Na2CO3 (1.04 g, 9.83 mmol, 1.00 eq) and 1,4-dioxane (60 mL) were added. The mixture was purged with nitrogen three times and then stirred at 80 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the residue was purified by silica column chromatography (DCM:MeOH = 100:1 → 10:1) to give cyclohexyl 8-((2-hydroxyethyl)amino)octanoate (2.80 g, 9.81 mmol, 99.8% yield) as a yellow oil.

[0110] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.77 (td, 1H), 3.75 - 3.67 (m, 2H), 2.91 - 2.80 (m, 5H), 2.68 (t, 2H), 2.29 (t, 2H), 1.88 - 1.82 (m, 2H), 1.77 - 1.70 (m, 2H), 1.66 - 1.60 (m, 2H), 1.55(br d, 2H), 1.46 - 1.28 (m, 12H)

[0111] 4-8. Synthesis of Compound of Formula (D) To a 50 mL three-neck RBF, cyclohexyl 8-((2-hydroxyethyl)amino)octanoate (942 mg, 3.30 mmol, 1.10 eq) and EtOH (10 mL) were added, followed by (1R,2R)-2-pentylcyclopropyl 8-bromooctanoate (1.00 g, 3.00 mmol, 1.00 eq) and DIEA (1.94 g, 15.00 mmol, 5.00 eq). The mixture was stirred at 90 °C under a nitrogen atmosphere for 16 h. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the residue was purified by silica column chromatography (DCM:MeOH = 10:1 → 1:100) to give compound (D) (0.15 g, 279 μmol, 9.30% yield) as a yellow oil.

[0112] 1 H NMR (400 MHz, CHLOROFORM-d): δ 5.23 - 4.94 (m, 2H), 4.63 - 4.56 (m, 1H), 4.12 (br s, 2H), 3.68 - 3.57 (m, 2H), 3.56 - 3.37 (m, 4H), 2.40 - 2.27 (m, 2H), 2.04 (br d, 6H), 1.91 - 1.76 (m, 2H), 1.74 - 1.55 (m, 17H), 1.54 - 1.46 (m, 2H), 1.43 - 1.25 (m, 5H), 1.21 (s, 2H), 0.90 (d, 6H)

[0113] Example 5 5-1. The compound of formula (E) below was produced according to the synthesis scheme shown in FIG. [ka]

[0114] 5-2. Synthesis of cyclopentadecanecarbonitrile Cyclopentadecanone (25.0 g, 111 mmol, 1.00 eq), potassium; 2-methylpropan-2-olate (25.0 g, 223 mmol, 2.00 eq), 2-methylpropan-2-ol (250 mL) were added to a 2000 mL three-necked RBF together with THF (500 mL). The mixture was cooled to 0°C, and then 1-(isocyanomethylsulfonyl)-4-methyl-benzene (32.6 g, 167 mmol, 1.50 eq) was slowly added thereto at 0°C for 1 hour, followed by stirring at 20°C for 11 hours. The reaction mixture was diluted with HO (400 mL) and extracted with EtOAc (400 mL). The organic layer was washed with HO (400 mL), dried over NaSO, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica column chromatography (petroleum ether: EtOAc = 1:0 → 5:1) to give cyclopentadecanecarbonitrile (20 g, 84.96 mmol, 76.25% yield) as a colorless oil.

[0115] 1 H NMR (400 MHz, CHLOROFORM-d): δ 1.33 (br s, 20 H) 1.44 - 1.53 (m, 4 H) 1.67 (q, 4 H) 2.59 (quin, 1 H)

[0116] 5-3. Synthesis of cyclopentadecanecarboxylic acid A 250 mL three-neck RBF was charged with cyclopentadecanecarbonitrile (14.0 g, 59.5 mmol, 1.00 eq), KOH (6.00 M, 69.4 mL, 7.00 eq), and EtOH (70 mL). The mixture was purged with nitrogen three times and stirred under a nitrogen atmosphere at 100 °C for 16 h. The reaction mixture was concentrated in vacuo, and the residue was acidified to pH 4 with 4 M aqueous hydrochloric acid (50 mL) and extracted with EtOAc (50 mL x 3). The organic layer was collected, dried over Na2SO4, filtered, and the filtrate was concentrated in vacuo. The residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1 → 1:1) to give cyclopentadecanecarboxylic acid (1.20 g, 4.72 mmol, 7.93% yield) as a white solid.

[0117] 1 H NMR (400 MHz, CHLOROFORM-d): δ ppm 1.27 - 1.46 (m, 24 H) 1.55 - 1.72 (m, 4 H) 2.44 (quin, 1 H) 10.15 - 11.75 (m, 1 H)

[0118] Synthesis of 5-4,7-bromoheptyl cyclopentadecanecarboxylate A 50 mL three-neck RBF was charged with cyclopentadecanecarboxylic acid (1.20 g, 4.72 mmol, 1.10 eq) and toluene (12 mL). 7-Bromoheptan-1-ol (837 mg, 4.29 mmol, 1.00 eq) was added, followed by H2SO4 (84.1 mg, 858 μmol, 45.7 μL, 0.20 eq). The mixture was purged with nitrogen three times and stirred under nitrogen atmosphere at 120 °C for 16 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1) to give 7-bromoheptyl cyclopentadecanecarboxylate (0.90 g, 2.09 mmol, 48.6% yield) as a colorless oil.

[0119] 1 H NMR (400 MHz, CHLOROFORM-d): δ 1.29 - 1.40 (m, 30 H) 1.56 - 1.64 (m, 6 H) 1.87 (quin2 H) 2.34 - 2.46 (m, 1 H) 3.41 (t, 2 H) 4.07 (t, 2 H)

[0120] Synthesis of 5,5,7-bromoheptyl 4-pentylcyclohexane-1-carboxylate 4-Pentylcyclohexane-1-carboxylic acid (4.47 g, 22.9 mmol, 1.00 eq) and toluene (50 mL) were added to a 250 mL three-neck RBF. 7-Bromoheptan-1-ol (5.00 g, 25.2 mmol, 1.00 eq) was added, followed by H2SO4 (450 mg, 4.58 mmol, 244 μL, 0.20 eq). The mixture was purged with nitrogen three times and stirred under nitrogen atmosphere at 120 °C for 16 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica column chromatography (petroleum ether: EtOAc = 10:1) to give 7-bromoheptyl 4-pentylcyclohexane-1-carboxylate (7.00 g, 18.7 mmol, 81.4% yield) as a colorless oil.

[0121] 1H NMR (400 MHz, CHLOROFORM-d): δ 0.86 - 0.90 (m, 3 H) 1.18 - 1.32 (m, 10 H) 1.32 - 1.40 (m, 5 H) 1.41 - 1.59 (m, 6 H) 1.59 - 1.68 (m, 2 H) 1.81 - 1.90 (m, 2 H) 1.91 - 2.02 (m, 2 H) 2.46 - 2.54 (m, 1 H) 3.41 (t, 2 H) 4.02 - 4.12 (m, 2 H)

[0122] Synthesis of 5-6,7-((2-hydroxyethyl)amino)heptyl 4-pentylcyclohexane-1-carboxylate To a 250 mL three-neck RBF, 7-bromoheptyl 4-pentylcyclohexane-1-carboxylate (3.00 g, 7.99 mmol, 1.00 eq), 2-aminoethan-1-ol (2.44 g, 40.0 mmol, 2.41 mL, 5.00 eq), and 1,4-dioxane (90 mL) were added. The mixture was purged with nitrogen three times and stirred under nitrogen atmosphere at 100 °C for 16 h. The reaction mixture was concentrated in vacuo, and the residue was purified by silica column chromatography (DCM:MeOH = 10:1) to give 7-((2-hydroxyethyl)amino)heptyl 4-pentylcyclohexane-1-carboxylate (1.70 g, 4.78 mmol, 59.8% yield) as a yellow oil.

[0123] 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.88 (t, 3 H) 1.19 - 1.28 (m, 8 H) 1.34 (br s, 6 H) 1.41 - 1.71 (m, 8 H) 1.84 - 2.01 (m, 6 H) 2.42 - 2.54 (m, 1 H) 2.63 (t, 2 H) 2.74 - 2.86 (m, 2 H) 3.50 - 3.79 (m, 2 H) 3.99 - 4.13 (m, 2 H)

[0124] 5-7. Synthesis of Compound of Formula (E) A 50 mL three-neck RBF was charged with 7-((2-hydroxyethyl)amino)heptyl 4-pentylcyclohexane-1-carboxylate (90 mg, 209 μmol, 1.00 eq) and 1,4-dioxane (5 mL). To this was added 7-bromoheptylcyclopentadecanecarboxylate (74.2 mg, 209 μmol, 1.00 eq) and DIEA (135 mg, 1.04 mmol, 5.00 eq). The mixture was purged with nitrogen three times and stirred at 105 °C under a nitrogen atmosphere for 16 h. The reaction mixture was cooled to 25 °C, quenched with water (10 mL), and extracted with 30 mL of EtOAc (3 x 10 mL). The organic layer was collected, dried over NaSO, filtered, and the filtrate was concentrated in vacuo. The concentrated residue was purified by silica column chromatography (DCM:MeOH=10:1) to give the compound of formula (E) (70 mg, 99.1 μmol, 47.5% yield) as a yellow oil.

[0125] 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.88 (t, 3 H) 1.23 - 1.40 (m, 46 H) 1.45 - 1.50 (m, 4 H) 1.52 - 1.66 (m, 12 H) 1.72 - 2.05 (m, 4 H) 2.37 - 2.55 (m, 6 H) 2.63 (br t, 2 H) 3.57 (br t, 2 H) 4.02 - 4.09 (m, 4 H)

[0126] Example 6 6-1. A compound of the following formula (F) was produced according to the synthesis scheme shown in FIG. [ka]

[0127] Synthesis of 6-2-cyclopentadecyl 10-bromodecanoate It was synthesized by the method described in Example 3-2.

[0128] Synthesis of 6-3,4-propylcyclohexyl 6-bromohexanoate It was synthesized by the method described in Example 2-3.

[0129] Synthesis of 6-4,4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate It was synthesized by the method described in Examples 2-4.

[0130] 6-5. Synthesis of Compound of Formula (F) To a 50 mL three-neck RBF, 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate (0.10 g, 1.00 eq), cyclopentadecyl 10-bromodecanoate (153 mg, 1.00 eq), DIEA (44 mg, 1.00 eq), and EtOH (5 mL) were added. The mixture was purged with nitrogen three times and stirred at 95 °C under a nitrogen atmosphere for 16 h. The reaction mixture was cooled to room temperature and concentrated in vacuo. The residue was purified by silica column chromatography (DCM:MeOH = 100:1 → 10:1) to give compound (F) (70 mg, 30.9% yield) as a yellow oil.

[0131] 1 H NMR (400 MHz, CHLOROFORM-d): δ 5.03 - 4.62 (m, 2H), 4.01 - 3.72 (m, 2H), 3.10 - 2.75 (m, 6H), 2.35 - 2.25 (m, 4H), 1.95 (br d, 1H), 1.59 (br s, 18H), 1.40 - 1.17 (m, 42H), 1.05 - 0.96 (m, 1H), 0.92 - 0.87 (m, 3H)

[0132] Example 7 According to the synthesis scheme shown in FIG. 7, a compound of the following formula (G) was produced in the same manner as in Example 1, except that 3-pentylcyclopentan-1-ol was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0133] 1 H NMR (400 MHz, CHLOROFORM-d): δ 5.03 - 4.77 (m, 2H), 4.09 - 3.88 (m, 2H), 3.04 - 2.88 (br, 6H), 2.41 - 2.31 (m, 4H), 1.89 - 1.56 (br, 14H), 1.51 - 0.96 (br, 53H), 0.88 (t, 3H)

[0134] Example 8 According to the synthesis scheme shown in FIG. 8, the compound of the following formula (H) was produced in the same manner as in Example 1, except that 6-bromohexanoic acid was used instead of 8-bromooctanoic acid and 3-pentylcyclopentan-1-ol was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0135] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.68 (m, 1H), 4.48 (m, 1H), 3.91 (m, 2H), 3.02 - 2.89 (br, 6H), 2.35 - 2.29 (m, 4H), 1.92 - 1.61 (m, 14H), 1.58 - 1.01 (m, 45H), 0.90 (t, 3H)

[0136] Example 9 According to the synthesis scheme shown in FIG. 9 , the compound of the following formula (I) was produced in the same manner as in Example 1, except that 10-bromodecanoic acid was used instead of 8-bromooctanoic acid and 3-pentylcyclopentan-1-ol was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0137] 1H NMR (400 MHz, CHLOROFORM-d): δ 4.70 (m, 1H), 4.47 (m, 1H), 3.90 (m, 2H), 2.99 - 2.79 (br, 6H), 2.31 - 2.26 (m, 4H), 1.92 - 1.61 (m, 14H), 1.58 - 1.01 (m, 61H), 0.85 (t, 3H)

[0138] Example 10 According to the synthesis scheme shown in FIG. 10, a compound of the following formula (J) was produced in the same manner as in Example 1, except that cyclohexanol was used instead of cyclopentadecanol and 3-pentylcyclopentan-1-ol was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0139] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.68 - 4.61 (m, 2H), 3.94 (m, 2H), 3.12 - 2.94 (br, 6H), 2.44 (m, 4H), 1.91 - 1.10 (m 45H), 0.88 (t, 3H)

[0140] Example 11 According to the synthesis scheme shown in FIG. 11, a compound of the following formula (K) was produced in the same manner as in Example 1, except that aminocyclopentadecane was used instead of cyclopentadecanol. [ka]

[0141] 1H NMR (400 MHz, CHLOROFORM-d): δ 8.10 (br, 1H), 4.67 - 4.63 (m, 1H), 3.95 - 3.88 (m, 2H), 3.64 - 3.59 (m, 1H), 3.11 - 2.89 (br, 6H), 2.31 - 2.19 (m, 4H), 1.95 - 1.25 (m, 65H), 0.88 (t, 3H)

[0142] Example 12 According to the synthesis scheme shown in FIG. 12, a compound of the following formula (L) was produced in the same manner as in Example 1, except that 1-amino-4-pentylcyclohexane was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0143] 1 H NMR (400 MHz, CHLOROFORM-d): δ 8.12 (br, 1H), 4.50 - 4.43 (m, 1H), 3.88 - 3.76 (m, 2H), 3.49 - 3.40 (m, 2H), 3.36 - 3.33 (m, 1H), 3.03 - 2.90 (br, 6H), 2.31 - 2.90 (m, 4H), 1.92 - 1.20 (m, 65H), 0.90 (t, 3H)

[0144] Example 13 According to the synthesis scheme shown in FIG. 13, a compound of the following formula (M) was produced in the same manner as in Example 1, except that aminocyclopentadecane was used instead of cyclopentadecanol and 1-amino-4-pentylcyclohexane was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0145] 1H NMR (400 MHz, CHLOROFORM-d): δ 8.10 (br, 2H), 3.84 - 3.78 (m, 2H), 3.69 - 3.61 (m, 2H), 3.11 - 2.98 (br, 6H), 2.33 - 2.19 (m, 4H), 1.90 - 1.22 (m, 65H), 0.91 (t, 3H)

[0146] Example 14 According to the synthesis scheme shown in FIG. 14, the compound of the following formula (N) was produced in the same manner as in Example 1, except that (1R,2R)-2-pentylcyclopropan-1-ol produced in Example 4-4 was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0147] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.56 - 4.45 (m, 1H), 3.56 - 3.52 (m, 2H), 3.42 (br, 1H), 3.11 - 3.05 (br, 4H), 2.77 - 2.70 (br, 2H), 2.44 - 2.38 (m, 4H), 1.91 - 1.21 (m, 57H), 0.9 (t, 3H), 0.62 - 0.55 (m, 2H)

[0148] Example 15 According to the synthesis scheme shown in FIG. 15, a compound of the following formula (O) was produced in the same manner as in Example 1, except that methylamine (CH3-NH2) was used instead of 2-aminoethan-1-ol. [ka]

[0149] 1H NMR (400 MHz, CHLOROFORM-d): δ 4.41 - 4.38 (m, 1H), 4.22 - 4.18 (m, 1H), 3.00 - 2.82 (m, 4H), 2.75 (s, 3H), 2.42 - 2.39 (m, 4H), 1.92 - 1.20 (m, 65H), 0.90 (t, 3H)

[0150] Example 16 According to the synthesis scheme shown in FIG. 16, the compound of the following formula (P) was produced in the same manner as in Example 1, except that methylamine (CH—NH) was used instead of 2-aminoethan-1-ol, 10-bromodecanoic acid was used instead of 8-bromooctanoic acid, and 4-heptylcyclohexan-1-ol was used instead of 4-pentylcyclohexan-1-ol. [ka]

[0151] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.43 - 4.40 (m, 1H), 4.21 - 4.18 (m, 1H), 3.12 - 2.92 (m, 4H), 2.80 (s, 3H), 2.42 - 2.39 (m, 4H), 1.93 - 1.19 (m, 73H), 0.90 (t, 3H)

[0152] Example 17 According to the synthesis scheme shown in FIG. 17, a compound of the following formula (Q) was produced in the same manner as in Example 1, except that cyclodecaneol was used instead of cyclopentadecanol. [ka]

[0153] 1H NMR (400 MHz, CHLOROFORM-d): δ 4.48 - 4.41 (m, 1H), 4.19 - 4.17 (m, 1H), 3.54 - 3.49 (m, 2H), 3.02 - 2.92 (br, 4H), 2.78 - 2.71 (br, 2H), 2.35 - 2.31 (m, 4H), 1.91 - 1.19 (m, 55H), 0.88 (t, 3H)

[0154] Example 18 According to the synthesis scheme shown in FIG. 18, a compound of the following formula (R) was produced in the same manner as in Example 1, except that cyclodecaneol was used instead of cyclopentadecanol and methylamine (CH—NH) was used instead of 2-aminoethan-1-ol. [ka]

[0155] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.48 - 4.41 (m, 1H), 4.19 - 4.17 (m, 1H), 3.01 - 2.93 (br, 4H), 2.75 (br, 3H), 2.34 - 2.30 (m, 4H), 1.90 - 1.21 (m, 55H), 0.90 (t, 3H)

[0156] [Production Example of Drug Delivery Composition] 1. Preparation of Raw Materials The raw materials required for the preparation of the formulation were dissolved in each dilution solvent according to the table below to prepare the required concentration. During dissolution, the raw materials were kept at room temperature and the solvent was added to dissolve them. [Table 1]

[0157] 2.Mixing of raw materials The required amounts of raw materials were mixed to achieve an NP ratio (lipid amine group:mRNA phosphate group) of 6, with each compound of formula (A-F):DOPE:cholesterol:DMG-PEG = 50:10:38.5:1.5. Ethanol was added to the ethanol layer to maintain a total molecular weight of all raw materials within 12.5 mM, and the aqueous and ethanol phases were mixed while maintaining a volume ratio of 3:1. After mixing, buffer exchange was performed to reduce the total ethanol content as follows: the mixed solution was concentrated by centrifugation at 4,000 rpm to 1 / 3 using an Amicon-Ultra tube filter (Merck Millipore, UFC505096 or UFC805024, pore size: 50K, volume: 15 mL). It was then diluted with 3x the volume of PBS and concentrated by centrifugation. This process was repeated six times to perform buffer exchange.

[0158] More specifically, the steps are as follows: 1) Two autoclaved tubes were prepared (tubes (A) and (B)). 2) Each compound of formulae (A to F), DSPC, cholesterol, and DMG-PEG were added to tube (A) in the molar amounts calculated according to the experimental conditions, and mixed using a brutex mixer. 3) In the ethanol phase, ethanol was added as necessary so that the total molecular weight of all raw materials was within 12.5 mM. 4) In tube (B), mRNA was mixed with 20 mM sodium acetate buffer (pH 4.6) (prepared by diluting the mRNA to 20 mM with 3 M sodium acetate buffer and titrating it to pH 4.6 with 1 M hydrochloric acid). The aqueous phase was added so that the total volume was three times the ethanol phase. 5) Tube (A) and tube (B) were mixed using a microfluidics device (Ignite, Precision Nanosystems). The microfluidics operating conditions were FRR (flow ratio) C:R = 3:1, TRR (total flow rate) 12 mL / min. 6) The mixture obtained in step 5) was centrifuged at 4,000 rpm using an Amicon-Ultra tube filter (50K) to a 1 / 3 level, then diluted 3 times with PBS and concentrated by centrifugation. This process was repeated 6 times to concentrate to the desired volume.

[0159] 3. Evaluation of the physical properties of the formulation 1) The particle characteristics (i.e., zeta-average particle size (Z-average), polydispersity index (PDI), and zeta potential) of the manufactured formulations were determined using a particle size analyzer (dynamic light scattering, DLS), and the results are shown in Table 1 below. 2) The mRNA encapsulation efficiency of the produced preparation was confirmed by RiboGreen assay, and the results are shown in Table 1 below. [Table 2]

Claims

1. The following formula (1) 【Chemistry 1】 (In the formula, M 1 and M 2 are each independently a divalent linker group, R 1 and R 2 are each independently a substituted or unsubstituted carbocyclic or heterocyclic group; R 3 is a hydrogen atom or a substituted or unsubstituted organic group optionally containing one or more heteroatoms, R 4 ~R 7 are each independently a hydrogen atom or a substituted or unsubstituted saturated or unsaturated hydrocarbon group, a and b are each independently an integer of 1 to 20.

2. M 1 and M 2 are each independently —C(O)O—, —OC(O)—, —OC(O)-M′-C(O)O—, —C(O)N(R′)—, —N(R′)C(O)—, —C(O)—, —C(S)—, —C(S)S—, —SC(S)—, —CH(OH)—, —P(O)(OR′)O—, —S(O) 2 -, -S-S-, arylene, and heteroarylene, wherein M' is a direct bond, C 1-13 Alkylene or C 2-13 alkenylene, and each R' is independently a hydrogen atom, C 1-18 Alkyl and C 2-18 alkenyl, R 1 and R 2 are each independently C 3-20 Cycloalkyl, C 3-20 Cycloalkenyl, C 6-20 Aryl, C 3-20 Heterocycloalkyl, C 3-20 heterocycloalkenyl, and C 3-20 heteroaryl, each of which is independently unsubstituted or C 1-18 Alkyl or C 2-18 substituted with alkenyl, R 3 represents a hydrogen atom, a substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Carbocyclic group, -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR and -CQ(R) 2 wherein each R is independently selected from the group consisting of a hydrogen atom, C 1-3 Alkyl and C 2-3 alkenyl; Q is a carbocyclic group, a heterocyclic group, —OR, —O(CH 2 ) n N (R) 2 , -C(O)OR, -OC(O)R, -CX 3 , -CX 2 H, -CXH 2 , -CN, -N(R) 2 , -C(O)N(R) 2 , -N(R)C(O)R, -N(R)S(O) 2 R, -N(R)C(O)N(R) 2 , -N(R)C(S)N(R) 2 , -N(R)R 12 , N(R)S(O) 2 R 12 , -O(CH 2 ) n OR, -N(R)C(=NR 13 ) N (R) 2 , -N(R)C(=CHR 13 ) N (R) 2 , -OC(O)N(R) 2 , -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O) 2 R, -N(OR)C(O)OR, -N(OR)C(O)N(R) 2 , -N(OR)C(S)N(R) 2 , -N(OR)C(=NR 13 ) N (R) 2 , -N(OR)C(=CHR 13 ) N (R) 2 , -C(=NR 13 ) N (R) 2 , -C(=NR 13 )R, -C(O)N(R)OR and -C(R)N(R) 2 C(O)OR, where each n is independently an integer from 1 to 5; 12 is C 3-6 selected from the group consisting of carbocyclic and heterocyclic groups; R 13 H, CN, NO 2 , C 1-6 Alkyl, —OR, —S(O) 2 R, -S(O) 2 N (R) 2 , C 2-6 Alkenyl, C 3-6 and each R is independently selected from the group consisting of a hydrogen atom, a C 1-3 Alkyl and C 2-3 alkenyl, and each X is independently selected from the group consisting of F, CI, BR, and I, with the proviso that R 3 But -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR or -CQ(R) 2 (i) when n is 1, 2, 3, 4 or 5, Q is -N(R) 2 or (ii) when n is 1 or 2, Q is not a 5-, 6-, or 7-membered heterocycloalkyl; R 4 ~R 7 are each independently a hydrogen atom, C 1-3 Alkyl and C 2-3 alkenyl, The lipid according to claim 1, wherein a and b are each independently an integer of 1 to 15.

3. M 1 and M 2 are each independently selected from the group consisting of —C(O)O—, —OC(O)—, —OC(O)-M′-C(O)O—, —C(O)N(R′)—, —N(R′)C(O)— and —C(O)—, wherein M′ and R′ are as defined in claim 2; R 1 and R 2 are each independently C 3-20 Cycloalkyl and C 3-20 heterocycloalkyl, each of which is independently unsubstituted or selected from the group consisting of C 1-18 Alkyl or C 2-18 substituted with alkenyl, R 3 represents a hydrogen atom, a substituted or unsubstituted C 1-6 Alkyl and substituted or unsubstituted C 3-6 is selected from the group consisting of carbocyclic groups; R 4 ~R 7 are each independently a hydrogen atom or C 1-3 is alkyl, The lipid according to claim 2, wherein a and b are each independently an integer of 3 to 13.

4. M 1 and M 2 are each independently selected from the group consisting of —C(O)O—, —OC(O)—, —C(O)N(R′)— and —N(R′)C(O)—, wherein each R′ is independently a hydrogen atom, 1-18 Alkyl and C 2-18 alkenyl, R 1 and R 2 are each independently substituted or unsubstituted C 3-15 is cycloalkyl, R 3 represents a hydrogen atom or a substituted or unsubstituted C 1-3 is alkyl, R 4 ~R 7 is a hydrogen atom, The lipid according to claim 3, wherein a and b are each independently an integer of 5 to 11.

5. The lipid according to claim 4, wherein the lipid has a structure selected from the following formulas (A to R): 【Chemistry 2】

6. A method for producing a lipid having a structure represented by formula (1'), comprising the steps of: (1) reacting a compound of formula (a) with a compound of formula (b) to obtain a compound of formula (c); and (2) reacting the compound of formula (c) with the compound of formula (d) 【Transformation 3】 (In the formula, M 1 , R 1 , R 3 , R 4 , R 5 and a are as defined in claim 1, and each X is independently selected from the group consisting of F, CI, BR, and I.

7. A method for producing a lipid having a structure represented by formula (1), comprising the steps of: (1) reacting a compound of formula (a') with a compound of formula (b') to obtain a compound of formula (c'); (2) reacting a compound of formula (c') with a compound of formula (d) to obtain a compound of formula (e); and (3) reacting the compound of formula (e) with the compound of formula (c) obtained in claim 6 【Chemistry 4】 (In the formula, M 1 , M 2 , R 1 ~R 7 , a, and b are as defined in claim 1, and each X is independently selected from the group consisting of F, Cl, Br, and I.

8. A method for producing a lipid having a structure represented by formula (1'), comprising the steps of: (1) reacting a compound of formula (i) with a compound of formula (ii) to obtain a compound of formula (iii); and (2) reacting the compound of formula (iii) with the compound of formula (iv) 【Transformation 5】 (In the formula, M 1 , R 1 , R 3 , R 4 , R 5 and a are as defined in claim 1, and each X is independently selected from the group consisting of F, CI, BR, and I.

9. A method for producing a lipid having a structure represented by formula (1), comprising the steps of: (1) reacting a compound of formula (i') with a compound of formula (ii') to obtain a compound of formula (iii'); (2) reacting the compound of formula (iii') with a compound of formula (iv) to obtain a compound of formula (v); and (3) reacting the compound of formula (v) with the compound of formula (iii) obtained in claim 8 【Transformation 6】 (In the formula, M 1 , M 2 , R 1 ~R 7 , a, and b are as defined in claim 1, and each X is independently selected from the group consisting of F, CI, BR, and I.

10. A drug delivery composition comprising the lipid according to any one of claims 1 to 5.

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