Cationic lipids and methods for producing the same

Abstract

The present invention relates to a cationic lipid and a method for producing the same, and more specifically to a cationic lipid that easily forms a complex with an anionic drug and is useful for drug delivery, and a method 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 readily form complexes with anionic drugs and are useful for drug delivery, and to methods for producing the same. [Background technology]

[0002] In the treatment of anionic drugs containing nucleic acids, safe and efficient drug delivery technologies have been studied for a long time, and various delivery bodies and technologies have been developed. Delivery bodies are mainly divided into viral delivery bodies that utilize adenoviruses and retroviruses, and non-viral delivery bodies that utilize cationic lipids and cationic polymers. Viral delivery bodies are known to be exposed to risks such as nonspecific immune reactions and have many problems for commercialization due to the complexity of their manufacturing process. Therefore, recent research is moving in the direction of improving these shortcomings by utilizing non-viral delivery bodies. Compared to viral delivery bodies, non-viral delivery bodies have advantages in terms of in vivo safety, with fewer side effects, and lower manufacturing costs.

[0003] Typical nonviral delivery bodies for nucleic acid substances include complexes of cationic lipids and nucleic acids (lipoplexes) and complexes of polycationic polymers and nucleic acids (polyplexes). Such cationic lipids and polycationic polymers have been the subject of various studies because they stabilize anionic drugs and increase their intracellular delivery by forming complexes with anionic drugs through electrostatic interactions (Non-patent documents 1 and 2).

[0004] However, polycationic polymers have cytotoxicity due to their polyvalent cation charge, posing a problem for practical application. Furthermore, conventional cationic lipids used in ionic lipid nanoparticles, which consist of cationic lipids, neutral lipids, and soluble lipids (fusogenic lipids), have drawbacks such as the complexity of their 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 [Overview of the project] [Problems that the invention aims to solve]

[0006] The object of the present invention is to provide cationic lipids with a specific structure that readily form complexes with anionic drugs and are useful for drug delivery, as well as a method for producing the same. [Means for solving the problem]

[0007] A first aspect of the present invention is to provide a lipid having the structure shown by the following formula (1).

[0008] [ka] (In the formula, M1 and M2 are each independently divalent linker groups, R1 and R2 are independently substituted or unsubstituted carbocyclic or heterocyclic groups. R3 is a hydrogen atom, or a substituted or unsubstituted organic group optionally containing one or more heteroatoms, R4 to R 11 are each independently a hydrogen atom, or a substituted or unsubstituted saturated or unsaturated hydrocarbon group, Me is a methyl group, a, b, c, and d are each independently an integer from 1 to 20.)

[0009] According to one embodiment of the present invention, in the formula (1), M1 and M2 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 (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), and may be selected from the group consisting of, where M' is a direct bond, C 1-13 alkylene or C 2-13 alkenylene, and R' are each independently a hydrogen atom, C 1-18 alkyl, and C 2-18 alkenyl and may be selected from the group consisting of.

[0010] According to one embodiment of the present invention, in the formula (1), R1 and R2 are each independently a substituted or unsubstituted C 3-20 cycloalkyl, a substituted or unsubstituted C 3-20 cycloalkenyl, a substituted or unsubstituted C 6-20 aryl, a substituted or unsubstituted C 3-20 heterocycloalkyl, a substituted or unsubstituted C 3-20 heterocycloalkenyl, and a substituted or unsubstituted C 3-20The group consisting of heteroaryls may be selected, where each of the heterocycloalkyl, heterocycloalkenyl, and heteroaryl may independently have one or more heteroatoms (e.g., 1 to 3) selected from N, O, and S.

[0011] According to one embodiment of the present invention, in 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 The group may be selected from CHQR, -CHQR, and -CQ(R)2, where each R is independently a hydrogen atom, C 1-3 Alkyl and C 2-3 Q may be selected from the group consisting of alkenyl groups; Q is a carbocyclic group, a heterocyclic group, -OR, -O(CH2) n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, -N(R)R 12 N(R)S(O)2R 12 -O(CH2) n OR, -N(R)C(=NR 13 )N(R)2, -N(R)C(=CHR 13 )N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR 13 )N(R)2, -N(OR)C(=CHR 13 )N(R)2, -C(=NR 13 )N(R)2, -C(=NR 13 ) may be selected from the group consisting of R, -C(O)N(R)OR and -C(R)N(R)2C(O)OR, where each n is an independent integer between 1 and 5, and R 12 C 3-6 Selected from the group consisting of carbocyclic groups and heterocyclic groups, R13 H, CN, NO2, C 1-6 Alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C 2-6 Alkenil, C 3-6 Selected from the group consisting of carbocyclic and heterocyclic groups, each R is independently a hydrogen atom, C 1-3 Alkyl and C 2-3 Each X is selected from the group consisting of alkenyls, and each X is independently selected from the group consisting of F, CI, BR, and I, except that R3 is -(CH2) n Q, -(CH2) n If it is CHQR, -CHQR, or -CQ(R)2, then (i) if n is 1, 2, 3, 4, or 5, Q is not -N(R)2, or (ii) if n is 1 or 2, Q is not a 5-, 6-, or 7-membered heterocycloalkyl.

[0012] According to one embodiment of the present invention, in formula (1), R4~R 11 These are, independently, hydrogen atoms and C 1-3 Alkyl and C 2-3 The group consisting of alkenyls may be selected.

[0013] According to one embodiment of the present invention, in formula (1), a, b, c, and d may each be an integer from 1 to 15, independently of each other.

[0014] More specifically, in formula (1) above, M1 and M2 may each be 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)- (where M' and R' are the same as the definitions above).

[0015] More specifically, in formula (1) above, R1 and R2 are each independently of a substituted or unsubstituted C 3-20 Cycloalkyl and substituted or unsubstituted C 3-20The heterocycloalkyl group may be selected from the group consisting of heterocycloalkyls, where the heterocycloalkyl group may have one or more heteroatoms (e.g., 1 to 3) selected from N, O, and S.

[0016] More specifically, in formula (1) above, R3 is a hydrogen atom, a substituted or unsubstituted carbon atom. 1-6 Alkyl and substituted or unsubstituted C 3-6 Carbocyclic groups (for example, C 3-6 The group consisting of cycloalkyls may be selected.

[0017] More specifically, in equation (1) above, R4~R 11 Each of these is independently a hydrogen atom or C 1-3 Alkyl is also acceptable.

[0018] More specifically, in formula (1) above, a, b, c, and d may each be an integer between 3 and 11, independently of each other.

[0019] More specifically, in formula (1) above, M1 and M2 may each be independently -C(O)O- or -OC(O)-.

[0020] More specifically, in formula (1) above, R1 and R2 are each independently of a substituted or unsubstituted C 3-6 It may also be a cycloalkyl group.

[0021] More specifically, in formula (1) above, R3 is a hydrogen atom or an unsubstituted carbon. 1-3 Alkyl is also acceptable.

[0022] More specifically, in equation (1) above, R4~R 11 This could be a hydrogen atom.

[0023] More specifically, in formula (1) above, a, b, c, and d may each be an integer between 5 and 9, independently of each other.

[0024] More specifically, the lipid may have a structure selected from the following formulas (A to O).

[0025] [ka] [ka]

[0026] A second aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1'), comprising the following steps: (1) A step of reacting the compound of formula (a) with the compound of formula (b) to obtain the compound of formula (c); (2) A step of reacting the compound of formula (c) with the compound of formula (d) to obtain the compound of formula (e); and (3) A step of reacting the compound of formula (e) with the compound of formula (f).

[0027] [ka] (In the formula, M1, R1, R3, R4, R5, R8, R9, Me, a, and b are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0028] A third aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1), comprising the following steps: (1) A step of reacting the compound of formula (a') with the compound of formula (b') to obtain the compound of formula (c'); (2) A step of reacting the compound of formula (c') with the compound of formula (d') to obtain the compound of formula (e'); and (3) A step of reacting the compound of formula (e') and the compound of formula (e) obtained in the second aspect of the present invention with the compound of formula (f).

[0029] [ka] (In the formula, M1, M2, R1~R 11Me, a, b, c, and d are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0030] A fourth aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1'), comprising the following steps: (1) A step of reacting the compound of formula (i) with the compound of formula (ii) to obtain the compound of formula (iii); (2) A step of reacting the compound of formula (iii) with the compound of formula (iv) to obtain the compound of formula (v); (3) A step of reacting the compound of formula (v) with the compound of formula (vi) to obtain the compound of formula (vii); and (4) A step of reacting the compound of formula (v) with the compound of formula (vii).

[0031] [ka] (In the formula, M1, R1, R3, R4, R5, R8, R9, Me, a, and b are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0032] A fifth aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1), comprising the following steps: (1) A step of reacting the compound of formula (i') with the compound of formula (ii') to obtain the compound of formula (iii'); (2) A step of reacting the compound of formula (iii') with the compound of formula (iv') to obtain the compound of formula (v'); (3) A step of reacting the compound of formula (v') with the compound of formula (vi) to obtain the compound of formula (vii'); and (4) A step of reacting the compound of formula (vii') with the compound of formula (v) obtained in the fourth aspect of the present invention.

[0033] [ka] (In the formula, M1, M2, R1~R11 Me, a, b, c, and d are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0034] A sixth aspect of the present invention provides a drug delivery composition containing lipids according to the present invention. [Effects of the Invention]

[0035] The lipids with a specific structure according to the present invention can readily form complexes with anionic drugs, and by utilizing these complexes, drugs can be efficiently delivered to target biological tissues. [Brief explanation of the drawing]

[0036] [Figure 1] This is the reaction scheme for the lipid synthesis process carried out in Example 1. [Figure 2] This is the reaction scheme for the lipid synthesis process performed in Example 2. [Figure 3] This is the reaction scheme for the lipid synthesis process carried out in Example 3. [Figure 4] This is the reaction scheme for the lipid synthesis process carried out in Example 4. [Figure 5] This is the reaction scheme for the lipid synthesis process carried out in Example 5. [Figure 6] This is the reaction scheme for the lipid synthesis process carried out in Example 6. [Figure 7] This is the reaction scheme for the lipid synthesis process carried out in Example 7. [Figure 8] This is the reaction scheme for the lipid synthesis process carried out in Example 8. [Figure 9] This is the reaction scheme for the lipid synthesis process carried out in Example 9. [Figure 10] This is the reaction scheme for the lipid synthesis process carried out in Example 10. [Figure 11] This is the reaction scheme for the lipid synthesis process carried out in Example 11. [Figure 12]This is the reaction scheme for the lipid synthesis process carried out in Example 12. [Figure 13] This is the reaction scheme for the lipid synthesis process carried out in Example 13. [Figure 14] This is the reaction scheme for the lipid synthesis process carried out in Example 14. [Figure 15] This is the reaction scheme for the lipid synthesis process carried out in Example 15. [Best Mode for Carrying Out the Invention]

[0037] The present invention will be described in more detail below.

[0038] The lipid provided by the first aspect of the present invention has a structure represented by the following formula (1).

[0039] [ka] (In the formula, M1 and M2 are each independently divalent linker groups, R1 and R2 are independently substituted or unsubstituted carbocyclic or heterocyclic groups. R3 is a substituted or unsubstituted organic group containing a hydrogen atom or one or more heteroatoms. R4~R 11 Each of these is independently a hydrogen atom or a substituted or unsubstituted saturated or unsaturated hydrocarbon group. Me is a methyl group, a, b, c, and d are each independent integers between 1 and 20.

[0040] In this specification, the expression "substituted or unsubstituted" for any group means, unless otherwise specified, that the group is unsubstituted or is a hydroxyl group or C 1-6 This means that it is substituted with an alkyl group.

[0041] According to one embodiment of the present invention, in formula (1), M1 and M2 are 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-, -SS-, aliene (more specifically, C 6-20 Allylene, more specifically, C 6-10 Allylene, and heteroalylene (more specifically, C having one or more heteroatoms (e.g., 1 to 3) selected from N, O, and S) 3-20 Heteralylene, more specifically, C 3-10 The group consisting of heteroalylenes may be selected, where M' is directly bonded, C 1-13 Alkylene (more specifically, C 1-6 Alkylene) or C 2-13 Alkenylene (more specifically, C 2-6 It may also be an alkenylene, and R' independently consists of a hydrogen atom and C 1-18 Alkyl (more specifically, C 1-10 Alkyl, more specifically, C 1-6 Alkyl) and C 2-18 Alkenil (more specifically, C 2-10 Alkenyl, more specifically, C 2-6 The group consisting of alkenyls may be selected.

[0042] According to one embodiment of the present invention, in formula (1), R1 and R2 are each independently of a substituted or unsubstituted C 3-20 Cycloalkyl (more specifically, C 3-10 Cycloalkyl, more specifically, C 3-6 Cycloalkyl, substituted or unsubstituted C 3-20 Cycloalkenyl (more specifically, C 3-10 Cycloalkenyl, more specifically, C 3-6 Cycloalkenyl, substituted or unsubstituted C 6-20 Ariel (more specifically, C 6-10Aryl, more specifically C6 aryl), substituted or unsubstituted C 3-20 Heterocycloalkyl (more specifically, C 3-10 Heterocycloalkyl, more specifically, C 3-6 Heterocycloalkyl), substituted or unsubstituted C 3-20 Heterocycloalkenyl (more specifically, C 3-10 Heterocycloalkenyl, more specifically, C 3-6 Heterocycloalkenyl), and substituted or unsubstituted C 3-2 0 Heteroaryl (more specifically, C 3-10 Heteroaryl, more specifically, C 3-6 Heteroaryl), and may be selected from the group consisting of, 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.

[0043] According to one embodiment of the present invention, in the formula (1), R3 is a hydrogen atom, substituted or unsubstituted C 1-6 Alkyl, substituted or unsubstituted C 3-6 Carbocyclic group, -(CH2) n Q, -(CH2) n CHQR, -CHQR and -CQ(R)2, where each R is independently a hydrogen atom, C 1-3 Alkyl and C 2-3 May be selected from the group consisting of alkenyl; Q is a carbocyclic group, a heterocyclic group, -OR, -O(CH2) n N(R)2, -C(O)OR, -OC(O)R, -CX3, -CX2H, -CXH2, -CN, -N(R)2, -C(O)N(R)2, -N(R)C(O)R, -N(R)S(O)2R, -N(R)C(O)N(R)2, -N(R)C(S)N(R)2, -N(R)R 12 , N(R)S(O)2R 12 , -O(CH2) n OR, -N(R)C(=NR 13 )N(R)2, -N(R)C(=CHR 13)N(R)2, -OC(O)N(R)2, -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O)2R, -N(OR)C(O)OR, -N(OR)C(O)N(R)2, -N(OR)C(S)N(R)2, -N(OR)C(=NR 13 )N(R)2, -N(OR)C(=CHR 13 )N(R)2, -C(=NR 13 )N(R)2, -C(=NR 13 ) may be selected from the group consisting of R, -C(O)N(R)OR and -C(R)N(R)2C(O)OR, where each n is an independent integer between 1 and 5, and R 12 C 3-6 Selected from the group consisting of carbocyclic groups and heterocyclic groups, R 13 H, CN, NO2, C 1-6 Alkyl, -OR, -S(O)2R, -S(O)2N(R)2, C 2-6 Alkenil, C 3-6 Selected from the group consisting of carbocyclic and heterocyclic groups, each R is independently a hydrogen atom, C 1-3 Alkyl and C 2-3 Each X is selected from the group consisting of alkenyls, and each X is independently selected from the group consisting of F, CI, BR, and I, except that R3 is -(CH2) n Q, -(CH2) n If it is CHQR, -CHQR, or -CQ(R)2, then (i) if n is 1, 2, 3, 4, or 5, Q is not -N(R)2, or (ii) if n is 1 or 2, Q is not a 5-, 6-, or 7-membered heterocycloalkyl.

[0044] According to one embodiment of the present invention, in formula (1), R4~R 11 These are, independently, hydrogen atoms and C 1-3 Alkyl and C 2-3 The group consisting of alkenyls may be selected.

[0045] According to one embodiment of the present invention, in formula (1), a, b, c, and d may each be an integer from 1 to 15, independently of each other.

[0046] More specifically, in formula (1) above, M1 and M2 may each be independently -C(O)O- or -OC(O)-.

[0047] More specifically, in formula (1) above, R1 and R2 are each independently of a substituted or unsubstituted C 3-6 It may also be a cycloalkyl group.

[0048] More specifically, in formula (1) above, R3 is a hydrogen atom or a substituted or unsubstituted carbon atom. 1-3 It may be an alkyl group, and more specifically, an unsubstituted C 1-3 C substituted with alkyl or hydroxyl groups 1-3 Alkyl is also acceptable.

[0049] More specifically, in equation (1) above, R4~R 11 This could be a hydrogen atom.

[0050] More specifically, in formula (1) above, a, b, c, and d may each be an integer between 3 and 11, and more specifically, an integer between 5 and 9.

[0051] More specifically, the lipid may have a structure selected from the following formulas (A to O).

[0052] [ka] [ka]

[0053] A second aspect of the present invention provides a method for producing a lipid having the structure represented by the following formula (1') included in formula (1), comprising the following steps: (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 (f).

[0054] [ka] (In the formula, M1, R1, R3, R4, R5, R8, R9, Me, a, and b are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0055] 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., tetrahydrofuran (THF)) under low to room temperature conditions (e.g., -75°C to 30°C), the reaction in step (2) can be carried out in a solvent (e.g., methylene chloride (DCM)) in the presence of a catalyst (e.g., 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 4-dimethylaminopyridine (DMAP), triethylamine (Et3N, TEA), or a combination thereof) under room temperature conditions (e.g., 20°C to 30°C), and the reaction in step (3) can be carried out in a solvent (e.g., ethanol (EtOH) or THF) in the presence of Na2CO3 or a catalyst (e.g., N,N-diisopropylethylamine (DIEA)) optionally under high temperature conditions (e.g., 40 to 100°C), but is not limited thereto.

[0056] A third aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1), comprising the following steps: (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') and the compound of formula (e) obtained in the second aspect of the present invention with a compound of formula (f).

[0057] [ka] (In the formula, M2, R2, R3, R6, R7, R 10 , R 11 Me, c, and d are synonymous with those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0058] 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., tetrahydrofuran (THF)) under low to room temperature conditions (e.g., -75°C to 30°C), the reaction in step (2) can be carried out in a solvent (e.g., methylene chloride (DCM)) in the presence of a catalyst (e.g., 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 4-dimethylaminopyridine (DMAP), triethylamine (Et3N, TEA), or a combination thereof) under room temperature conditions (e.g., 20°C to 30°C), and the reaction in step (3) can be carried out in a solvent (e.g., ethanol (EtOH) or THF) in the presence of Na2CO3 or a catalyst (e.g., N,N-diisopropylethylamine (DIEA)) optionally under high temperature conditions (e.g., 40 to 100°C), but is not limited thereto.

[0059] A fourth aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1'), comprising the following steps: (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); (3) reacting a compound of formula (v) with a compound of formula (vi) to obtain a compound of formula (vii); and (4) reacting a compound of formula (v) with a compound of formula (vii).

[0060] [ka] (In the formula, M1, R1, R3, R4, R5, R8, R9, Me, a, and b are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.)

[0061] 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., tetrahydrofuran (THF)) in the presence of a catalyst (e.g., sodium hydride (NaH) or lithium diisopropylamide (LDA)) under low to high temperature conditions (e.g., -20°C to 60°C), the reaction in step (2) can be carried out in a solvent (e.g., methylene chloride (DCM)) in the presence of a catalyst (e.g., 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 4-dimethylaminopyridine (DMAP), or a combination thereof) under room temperature conditions (e.g., 20°C to 30°C), the reaction in step (3) can be carried out under high temperature conditions (e.g., 40 to 70°C), and the reaction in step (4) can be carried out in a solvent (e.g., dioxane) optionally in the presence of Na2CO3 under high temperature conditions (e.g., 40 to 120°C), but is not limited thereto.

[0062] A fifth aspect of the present invention provides a method for producing a lipid having the structure represented by formula (1), comprising the following steps: (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'); (3) reacting a compound of formula (v') with a compound of formula (vi) to obtain a compound of formula (vii'); and (4) reacting a compound of formula (vii') with a compound of formula (v) obtained in the fourth aspect of the present invention.

[0063] [ka] (In the formula, M1, M2, R1, R2, R4~R 11 Me, a, b, c, and d are equivalent to those defined in formula (1) above, and each X is independently selected from the group consisting of F, CI, BR, and I.

[0064] 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., tetrahydrofuran (THF)) in the presence of a catalyst (e.g., sodium hydride (NaH) or lithium diisopropylamide (LDA)) under low to high temperature conditions (e.g., -20°C to 60°C), the reaction in step (2) can be carried out in a solvent (e.g., methylene chloride (DCM)) in the presence of a catalyst (e.g., 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI), 4-dimethylaminopyridine (DMAP), or a combination thereof) under room temperature conditions (e.g., 20°C to 30°C), the reaction in step (3) can be carried out under high temperature conditions (e.g., 40 to 70°C), and the reaction in step (4) can be carried out in a solvent (e.g., dioxane) optionally in the presence of Na2CO3 under high temperature conditions (e.g., 40 to 120°C), but is not limited thereto.

[0065] Lipids having the structure represented by formula (1) of the present invention are useful for drug delivery because they can easily form complexes with anionic drugs.

[0066] Accordingly, according to a sixth aspect of the present invention, a drug delivery composition is provided which comprises a lipid having the structure represented by formula (1) of the present invention.

[0067] In one embodiment, the drug may be selected from nucleic acids, polypeptides, viruses, or combinations thereof.

[0068] The aforementioned "nucleic acid" may, but is not limited to, DNA, RNA, siRNA, shRNA, miRNA, mRNA, aptamers, antisense oligonucleotides, or combinations thereof.

[0069] The term "polypeptide" may refer to proteins that are active in the body, such as antibodies or fragments thereof, cytokines, hormones or their analogues, or polypeptide sequences of antigens, their analogues or precursors, and that can be recognized as antigens in the body through a series of processes.

[0070] In one embodiment, the lipid of the present invention forms a complex with a drug, and this complex is encapsulated within a nanoparticle structure formed by an amphiphilic block copolymer.

[0071] In one embodiment, the amphiphilic block copolymer may be an AB-type block copolymer containing hydrophilic A blocks and hydrophobic B blocks. In an aqueous environment, the AB-type block copolymer forms core-shell type polymer nanoparticles in which the hydrophobic B blocks form the core (inner wall) and the hydrophilic A blocks form the shell (outer wall).

[0072] 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.

[0073] 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.

[0074] Furthermore, if necessary, by chemically attaching a functional group, ligand, or functional group capable of promoting intracellular delivery to the end of the hydrophilic A block, it is possible to control the in vivo distribution of the high molecular weight nanoparticle deliverer formed from the amphiphilic block copolymer and the polylactic acid salt, or to increase the intracellular delivery efficiency of the nanoparticle deliverer. 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.

[0075] The 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 polyphosphatidine.

[0076] More specifically, the hydrophobic B block may be one or more selected from the group consisting of polylactide (PLA), polyglycolide, polycaprolactone, polydioxan-2-one, copolymer of polylactide and glycoside, copolymer of polylactide and polydioxan-2-one, copolymer of polylactide and polycaprolactone, and copolymer of polyglycolide and polycaprolactone.

[0077] 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 terminal end of the hydrophobic B block in order to enhance its hydrophobicity and improve the stability of the nanoparticles.

[0078] The present invention will be described in more detail below with reference to examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited in any way by them. Examples

[0079] Example 1 The compound of formula (A) below was prepared according to the synthesis scheme shown in Figure 1.

[0080] [ka]

[0081] (1) Synthesis of 1-cyclopropylnonan-1-ol In a 2000 mL three-necked round-bottom flask (RBF), cyclopropanecarboxyl (35.0 g, 499 mmol, 1.00 eq) and tetrahydrofuran (THF) (700 mL) were added under a nitrogen atmosphere. After cooling to -65 °C, magnesium octyl bromide (2 M, 375 mL, 1.50 eq) was added, and the mixture was stirred at -65 °C for 2 hours. After heating the reactor to 15 °C, the mixture was poured into a saturated aqueous solution of NH4Cl (500 mL) to separate the organic layer from the aqueous layer. The aqueous layer was extracted with ethyl acetate ( Depositphotos) (450 mL) (150 mL each, 3 times). The resulting organic layer was collected, concentrated under vacuum, and purified using a silica column with petroleum ether: Depositphotos = 50:1 → 0:1 to obtain 1-cyclopropylnonan-1-ol (87.5 g, 73.1%). 1H NMR (400 MHz, CHLOROFORM-d): δ 2.93 - 2.81 (m, 1H), 1.61 (br d, 2H), 1.52 - 1.27 (m, 12H), 0.95 - 0.86 (m, 4H), 0.60 - 0.45 (m, 2H), 0.34 - 0.19 (m, 2H)

[0082] (2) Synthesis of 1-cyclopropylnonyl 8-bromooctanoate 1-Cyclopropylnonan-1-ol (30.0 g, 163 mmol, 1.00 eq), 8-bromooctanoic acid (72.6 g, 326 mmol, 2.00 eq), methylene chloride (DCM) (300 mL), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (31.2 g, 163 mmol, 1.00 eq), and 4-dimethylaminopyridine (DMAP) (19.9 g, 163 mmol, 1.00 eq) were added to a 1000 mL three-necked RBF and stirred at 25 °C for 16 hours. After vacuum concentration of the mixture in the reactor, silica powder was added and the mixture was purified using a silica column with petroleum ether:siRNA = 10:1 → 50:1 to obtain 1-cyclopropylnonyl 8-bromooctanoate (22.8 g, 36.0%). 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.29 (td, 1H), 3.59 - 3.31 (m, 2H), 2.32 (t, 2H), 1.94 - 1.75 (m, 2H), 1.73 - 1.60 (m, 4H), 1.49 - 1.25 (m, 18H), 1.03 - 0.93 (m, 1H), 0.90 (t, 3H), 0.61 - 0.24 (m, 4H)

[0083] (3) Synthesis of compound (A) In a 100 mL three-necked flask, methylamine hydrochloride (173 mg, 2.57 mmol, 1.00 eq), ethanol (EtOH) (30 mL), N,N-diisopropylethylamine (DIEA) (1.66 g, 12.8 mmol, 5.00 eq), and 1-cyclopropylnonyl 8-bromooctanoate (3.00 g, 7.70 mmol, 3.00 eq) were added in sequence, and the mixture was stirred at 80°C for 72 hours. After vacuum concentration of the mixture in the reactor, silica powder was added, and the mixture was purified using a silica column with petroleum ether:siRNA = 10:1 → 1:1 to obtain the compound of formula (A) (660 mg, 38.9%). 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.29 (td, 2H), 2.32 (br t, 8H), 2.22 (s, 3H), 1.74 - 1.60 (m, 8H), 1.54 - 1.42 (m, 4H), 1.39 - 1.23 (m, 36H), 1.02 - 0.87 (m, 8H), 0.61 - 0.23 (m, 8H)

[0084] Example 2 The compound of formula (B) below was prepared according to the synthesis scheme shown in Figure 2.

[0085] [ka]

[0086] (1) Synthesis of 1-cyclopropylheptan-1-ol In a 2000 mL three-necked RBF, cyclopropanecarboxyl (27.0 g, 385 mmol, 1.00 eq) and THF (500 mL) were added under a nitrogen atmosphere. After cooling to -65°C, magnesium hexylbromide (1 M, 500 mL, 1.30 eq) was slowly added. The mixture was stirred at -65°C for 3 hours, and then the reactor temperature was slowly raised to 25°C. Subsequently, the mixture was poured into a saturated aqueous solution of NH4Cl (500 mL) to separate the organic layer from the aqueous layer. The aqueous layer was extracted with ethyl acetate (500 mL each, 3 times). The organic layer was collected and concentrated under vacuum. The residue after concentration was purified using a silica column with petroleum ether:ethyl acetate = 50:1 → 10:1 to obtain 1-cyclopropylheptan-1-ol (46.0 g, 294 mmol, yield 76.4%) as a colorless oil. 1 H NMR (400 MHz, CHLOROFORM-d): δ 2.86 (td, 1H), 1.67 - 1.38 (m, 6H), 1.36 - 1.27 (m, 6H), 0.91 - 0.87 (m, 3H), 0.59 - 0.42 (m, 2H), 0.32 - 0.17 (m, 2H)

[0087] (2) Synthesis of 1-cyclopropylheptyl 6-bromohexanoate 1-Cyclopropylheptan-1-ol (10.0 g, 63.9 mmol, 1.00 eq), 6-bromohexanoic acid (12.5 g, 63.9 mmol, 1.00 eq), DCM (100 mL), EDCI (15.9 g, 83.2 mmol, 1.30 eq), and DMAP (10.2 g, 83.2 mmol, 1.30 eq) were added to a 500 mL three-necked RBF and stirred at 25 °C for 16 hours. After vacuum concentration of the reactor mixture, it was purified using a silica column with petroleum ether:siRNA = 50:1 → 10:1 to obtain 1-cyclopropylheptyl 6-bromohexanoate (7.50 g, 22.5 mmol, 35.1%). 1H NMR: (400 MHz, CHLOROFORM-d): δ 4.28 (td, 1H), 3.55 (t, 1H), 3.42 (t, 1H), 2.39 - 2.26 (m, 2H), 1.97 - 1.76 (m, 2H), 1.71 - 1.60 (m, 4H), 1.57 - 1.42 (m, 3H), 1.33 - 1.28 (m, 6H), 0.91 - 0.86 (m, 5H), 0.63 - 0.51 (m, 1H), 0.50 - 0.41 (m, 1H), 0.40 - 0.32 (m, 1H), 0.30 - 0.22 (m, 1H)

[0088] (3) Synthesis of compound (B) 100 mL of three-necked RBF was mixed with 1-cyclopropylheptyl 6-bromohexanoate (500 mg, 1.00 eq), and under a nitrogen atmosphere, methylamine in THF (2 M, 2 g, 42.8 eq) was added, followed by stirring at 50°C for 16 hours. After stirring, 20 mL of aqueous sodium carbonate (Na2CO3) solution was added to the reaction mixture, and the mixture was stirred for 2 hours to adjust the pH to 8. The mixture was then extracted using DCM (30 mL x 3 times), and the organic layer was stored. The stored organic layer was concentrated under vacuum and purified using a silica column with DCM:methanol = 10:1 to obtain the compound of formula (B) (68 mg, 8.56%) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ 4.27 (td, 2H), 2.43 - 2.16 (m, 10H), 1.75 - 1.62 (m, 8H), 1.55 - 1.43 (m, 5H), 1.40 - 1.20 (m, 22H), 1.00 - 0.91 (m, 2H), 0.90 - 0.85 (m, 4H), 0.62 - 0.50 (m, 2H), 0.49 - 0.33 (m, 4H), 0.30 - 0.21 (m, 2H)

[0089] Example 3 The compound of formula (C) below was prepared according to the synthesis scheme shown in Figure 3.

[0090] [ka]

[0091] (1) Synthesis of 1-cyclopropylundecane-1-ol To a 2000 mL three-necked RBF, cyclopropanecarboxyl (27.0 g, 385 mmol, 1.00 eq) and THF (270 mL) were added. The mixture was purged three times with nitrogen and cooled to -60°C. Then, magnesium decylbromide (1 M, 501 mL, 1.30 eq) was added, and the mixture was stirred under a nitrogen atmosphere at -60°C for 16 hours. The reactor was heated to 25°C and poured into a saturated aqueous solution of NH4Cl (200 mL) to separate the organic layer from the aqueous layer. The aqueous layer was extracted with siRNA (100 mL each, four times), and the resulting organic layer was collected, concentrated under vacuum, and purified using a silica column with petroleum ether:siRNA = 10:1 to obtain 1-cyclopropylundecane-1-ol (56.0 g, 68.5%). 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.17 - 0.31 (m, 2 H) 0.43 - 0.57 (m, 2 H) 0.88 (t, 4 H) 1.26 (br s, 14 H) 1.37 - 1.49 (m, 2 H) 1.54 - 1.64 (m, 3 H) 2.85 (dt, 1 H)

[0092] (2) Synthesis of 1-cyclopropylundecyl8-bromooctanoate 1-Cyclopropylundecane-1-ol (24.0 g, 113 mmol, 1.00 eq), 8-bromooctanoic acid (32.8 g, 147 mmol, 1.30 eq), DCM (300 mL), EDCI (26.0 g, 136 mmol, 1.20 eq), and DMAP (16.6 g, 136 mmol, 1.20 eq) were added to a 1000 mL three-necked RBF and purged three times with nitrogen. The mixture was stirred at 25°C for 16 hours, then poured into water (200 mL) to separate the organic and aqueous layers. The aqueous layer was extracted with DCM (200 mL each, three times). The extracted organic layer was dried over Na2SO4, the dried mixture was filtered, and the mixture was concentrated under vacuum. The concentrated residue was purified using a silica column with a ratio of petroleum ether:dimethyl = 10:1 → 1:1 to obtain 1-cyclopropyl undecyl 8-bromooctanoate (11.0 g, 23.3%). 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.22 - 0.30 (m, 1 H) 0.33 - 0.40 (m, 1 H) 0.42 - 0.49 (m, 1 H) 0.50 - 0.58 (m, 1 H) 0.85 - 1.00 (m, 4 H) 1.20 - 1.38 (m, 20 H) 1.41 - 1.49 (m, 2 H) 1.58 - 1.68 (m, 4 H) 1.73 - 1.91 (m, 2 H) 2.25 - 2.37 (m, 2 H) 3.35 - 3.56 (m, 2 H) 4.21 - 4.35 (m, 1 H)

[0093] (3) Synthesis of the compound of formula (C) 100 mL of three-necked RBF was mixed with 1-cyclopropylundecyl 8-bromooctanoate (3.00 g, 7.19 mmol, 1.00 eq) and methylamine solution (2 M in THF, 10.3 g, 331 mmol, 46.0 eq), and stirred at 50°C for 16 hours. The mixture in the reactor was vacuum concentrated, and then aqueous NaHCO3 solution was added to adjust the pH. Subsequently, the mixture was extracted using DCM, and the extracted organic layer was vacuum concentrated. The residue after concentration was purified using a silica column with DCM:methanol = 10:1 → 1:1 to obtain the compound of formula (C) (2.00 g, 63.0%) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ 0.21 - 0.30 (m, 2 H) 0.37 (dq, 2 H) 0.42 - 0.49 (m, 2 H) 0.50 - 0.58 (m, 2 H) 0.84 - 0.98 (m, 8 H) 1.24 - 1.36 (m, 44 H) 1.51 (br s, 4 H) 1.59 - 1.68 (m, 8 H) 2.23 - 2.33 (m, 7 H) 2.34 - 2.44 (m, 4 H) 4.21 - 4.33 (m, 2 H)

[0094] Example 4 The compound of formula (D) below was prepared according to the synthesis scheme shown in Figure 4.

[0095] [ka]

[0096] (1) Synthesis of 1-cyclohexylnonan-1-ol To a 1000 mL three-necked RBF, cyclohexanecarbaldehyde (43.0 g, 383 mmol, 1.00 eq) and THF (430 mL) were added, the mixture was purged three times with nitrogen, cooled to -65°C, and then magnesium octyl bromide (1 M in THF, 498 mL, 1.30 eq) was added. The mixture was stirred under a nitrogen atmosphere at -65°C for 1 hour. After heating the reactor to 25°C, it was poured into a saturated aqueous solution of NH4Cl (700 mL) to separate the organic and aqueous layers. The aqueous layer was extracted with HCl (400 mL each, three times), the organic layer was collected and concentrated under vacuum, and purified using a silica column with petroleum ether:HCl = 10:1 → 1:10 to obtain 1-cyclohexylnonan-1-ol (12.0 g, 53.0 mmol, 13.8%) as a colorless oil. 1 HNMR (400 MHz, CHLOROFORM-d): δ 3.41 - 3.31 (m, 1H), 1.84 - 1.73 (m, 3H), 1.70 - 1.63 (m, 2H), 1.48 (br d, 3H), 1.36 - 1.21 (m, 15H), 1.18 - 1.00 (m, 3H), 0.94 - 0.84 (m, 3H)

[0097] (2) Synthesis of 1-cyclohexylnonyl 8-bromooctanoate 1-Cyclohexylnonan-1-ol (7.00 g, 30.9 mmol, 1.00 eq) was added to a 250 mL three-necked RBF, followed by the addition of DCM (70 mL). Then, 8-bromooctanoic acid (8.28 g, 37.1 mmol, 1.20 eq), EDCI (7.11 g, 37.1 mmol, 1.20 eq), DMAP (755 mg, 6.18 mmol, 0.20 eq), and Et3N (6.26 g, 61.8 mmol, 2.00 eq) were added and mixed. The mixture was stirred at 25°C for 16 hours and purged three times with nitrogen. The reaction mixture was filtered through a Celite plug, and the filtrate was concentrated under vacuum. The concentrated residue was purified using a silica column with a petroleum ether:dimethyl ratio of 10:1 → 1:100 to obtain 1-cyclohexylnonyl 8-bromooctanoate (3.50 g, 8.11 mmol, 26.2%) as a yellow oily substance. 1HNMR (400 MHz, CHLOROFORM-d): δ 4.07 (t, 2H), 3.41 (t, 2H), 2.30 (t, 2H), 1.91 - 1.80 (m, 2H), 1.67 - 1.58 (m, 4H), 1.47 - 1.41 (m, 2H), 1.37 - 1.26 (m, 16H), 0.91 - 0.87 (m, 3H)

[0098] (3) Synthesis of compound (D) 1.50 g, 3.48 mmol, 1.00 eq of 1-cyclohexylnonyl 8-bromooctanoate was added to a 100 mL three-necked RBF, and methylamine in THF (CH3NH2 in THF) (15.6 g, 151 mmol, purity 30%, 43.5 eq) was added. The mixture was purged three times with nitrogen and stirred at 50°C for 16 hours. After stirring, the reaction mixture was filtered, and the filtrate was concentrated under vacuum. The residue after concentration was purified using a silica column with DCM:MeOH = 100:1 → 10:1 to obtain the compound of formula (D) (0.13 g, 178 μmol, yield 5.11%) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d) δ 4.81 - 4.69 (m, 2H), 2.30 (br t, 8H), 2.22 (br s, 3H), 1.76 - 1.71 (m, 4H), 1.69 - 1.60 (m, 12H), 1.53 - 1.43 (m, 10H), 1.35 - 1.22 (m, 40H), 1.05 - 0.97 (m, 4H), 0.88 (br t, 6H)

[0099] Example 5 The compound of formula (E) below was prepared according to the synthesis scheme shown in Figure 5.

[0100] [ka]

[0101] (1) Synthesis of 1-cyclopropylnonan-1-ol To a 1000 mL three-necked RBF, cyclopropanecarboxyl (46.0 g, 656 mmol, 1.00 eq) was added, followed by THF. Under a nitrogen atmosphere, magnesium octyl bromide (2.00 M, 492 mL, 1.50 eq) was added. The mixture was stirred at -65°C for 3 hours, then 700 mL of saturated NH4Cl aqueous solution was added at 15°C to separate the organic and aqueous layers. The aqueous layer was further extracted with siRNA (200 mL x 3 times). The organic layer was collected, concentrated under vacuum, and purified using a silica column with petroleum ether:siRNA = 100:1 to obtain 1-cyclopropylnonan-1-ol (59.5 g, 49.2%) as a colorless oil. 1 H NMR (400 MHz, CHLOROFORM-d): δ 2.63 (td, 1H), 1.47 - 1.31 (m, 5H), 1.29 - 1.09 (m, 6H), 0.73 - 0.61 (m, 6H), 0.37 - 0.19 (m, 3H), 0.10 - 0.06 (m, 3H)

[0102] (2) Synthesis of 1-cyclopropylnonyl 8-bromooctanoate To a 1000 mL three-necked RBF, DCM (600 mL), 1-cyclopropylnonan-1-ol (59.5 g, 325 mmol, 1.00 eq), 8-bromooctanoic acid (94.4 g, 423 mmol, 1.30 eq), EDCI (93.6 g, 488 mmol, 1.50 eq), DMAP (39.8 g, 326 mmol, 1.00 eq), and TEA (32.9 g, 326 mmol, 45.3 mL, 1.00 eq) were added and stirred at 25°C for 16 hours under a nitrogen atmosphere. The reaction mixture was concentrated under vacuum to obtain the residue. The resulting residue was purified using a silica column with a petroleum ether:dimethyl ratio of 100:1 → 1:1 to obtain 1-cyclopropylnonyl 8-bromooctanoate (21.0 g, 53.9 mmol, 16.6% yield) as a pale yellow oily substance. 1H NMR (400 MHz, CHLOROFORM-d): δ 0.05 - 0.16 (m, 2 H) 0.16 - 0.34 (m, 2 H) 0.57 - 0.67 (m, 2 H) 0.67 -0.80 (m, 1 H) 1.02 (br s, 9 H) 1.05 (br s, 2 H) 1.07 - 1.15 (m, 6 H) 1.16 - 1.29 (m, 2 H) 1.34 - 1.44 (m, 4 H) 1.48 - 1.66(m, 2 H) 2.05 (t, 2 H) 3.06 - 3.33 (m, 2 H) 3.97 - 4.07 (m, 1H)

[0103] (3) Synthesis of compound (E) In a 100 mL three-necked RBF, 1-cyclopropylnonyl 8-bromooctanoate (4.78 g, 12.3 mmol, 2.50 eq), 2-aminoethanol (MEA) (0.30 g, 4.91 mmol, 1.00 eq), and Na2CO3 (521 mg, 4.91 mmol, 1.00 eq) were added to EtOH (5 mL), and the mixture was purged three times with nitrogen. The mixture was stirred at 95 °C for 16 hours. The mixture in the reactor was vacuum concentrated to obtain the residue. The obtained residue was purified using a silica column with DCM:MeOH = 100:1 → 10:1 to obtain the compound of formula (E) (1.00 g, 1.47 mmol, 30.0% yield) as a yellow oily substance. 1 H NMR (400 MHz, CHLOROFORM-d): δ = 4.27 (td, 2H), 3.53 (t, 2H), 2.57 (t, 2H), 2.47 - 2.40 (m, 4H), 2.30 (t, 4H), 1.70 - 1.57 (m, 10H), 1.48 - 1.39 (m, 4H), 1.36 - 1.26 (m, 33H), 0.95 (dt, 2H), 0.89 (t, 6H), 0.60 - 0.42 (m, 4H), 0.41 - 0.22 (m, 4H)

[0104] Example 6 The compound of formula (F) below was prepared according to the synthesis scheme shown in Figure 6.

[0105] [ka]

[0106] (1) Synthesis of 2-cyclopropyldecanoic acid 25.0 g, 250 mmol, 1.00 eq of 2-cyclopropylacetic acid was added to a 2000 mL three-necked RBF vessel, followed by the addition of THF (250 mL) and cooling with nitrogen. Then, sodium hydride (NaH) (11.0 g, 275 mmol, 60% purity, 1.10 eq) was added and the mixture was stirred at 0°C for 30 minutes. Next, at the same temperature and under the same conditions, lithium diisopropylamide (LDA) (2 M, 137 mL, 1.10 eq) was added and the mixture was stirred for 30 minutes. Then, 1-iodooctane (60.0 g, 250 mmol, 1.00 eq) was added at 25°C and the mixture was stirred at 45°C for 12 hours under a nitrogen atmosphere. The reactor was neutralized with 100 mL of water and 1 M HCl (600 mL, pH=4), and extracted with ethyl acetate (300 mL x 3 times). The extracted organic layer was dried with anhydrous Na2SO4 and concentrated under vacuum. The concentrated residue was purified using a silica column with a petroleum ether:siRNA ratio of 20:1 → 5:1 to obtain 2-cyclopropyldecanoic acid (42.6 g, 201 mmol, yield 80.4%) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d): δ 11.88 (s, 1H), 1.84 - 1.70 (m, 1H), 1.70 - 1.54 (m, 2H), 1.42 - 1.19 (m, 12H), 1.00 - 0.81 (m, 4H), 0.63 - 0.45 (m, 2H), 0.32 (qd, 1H), 0.23 - 0.10 (m, 1H)

[0107] (2) Synthesis of 7-bromoheptyl 2-cyclopropyldecanoate In a 2000 mL three-necked RBF, 2-cyclopropyldecanoic acid (10.0 g, 47.1 mmol, 1.00 eq), 7-bromoheptan-1-ol (11.0 g, 56.5 mmol, 1.20 eq), EDCI (11.7 g, 61.2 mmol, 1.30 eq), and DMAP (5.75 g, 47.1 mmol, 1.00 eq) were added together with DCM (100 mL), and the mixture was purged three times with nitrogen. The mixture was stirred under a nitrogen atmosphere at 25°C for 16 hours, and the reaction mixture was heated to 25°C and poured into water (100 mL) to separate the organic and aqueous layers. The aqueous layer was extracted with DCM (100 mL each, three times). The organic layer was collected, concentrated under vacuum, dried over anhydrous Na2SO4, and filtered. The residue after filtration was purified using a silica column with a ratio of petroleum ether:siRNA = 20:1 → 5:1 to obtain 7-bromoheptyl 2-cyclopropyldecanoate (9.40 g, 24.1 mmol, 51.2%) as a pale yellow oily substance. 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.10 (t, J = 6.6 Hz, 2H), 3.42 (t, 2H), 1.89 - 1.84 (m, 1H), 1.81 - 1.51 (m, 6H), 1.50 - 1.35 (m, 6H), 1.27 (br s, 12H), 0.89 (t, 4H), 0.61 - 0.37 (m, 2H), 0.24 (s, 1H), 0.13 (qd, 1H)

[0108] (3) Synthesis of 7-(methylamino)heptyl 2-cyclopropyldecanoate 7-bromoheptyl 2-cyclopropyldecanoate (4.00 g, 10.3 mmol, 1.00 eq) and methylamine (2 M in THF, 185 mL, 36.0 eq) were added to a 100 mL three-necked RBF container, and the mixture was stirred under a nitrogen atmosphere at 50°C for 16 hours. The stirred reaction mixture was vacuum concentrated and extracted with DCM (30 mL x 3 times), and the organic layer was stored. The stored organic layer was vacuum concentrated and purified using a silica column with DCM:methanol = 20:1 → 10:1 to obtain 7-(methylamino)heptyl 2-cyclopropyldecanoate (0.45 g, 1.33 mmol, 12.9%) as a yellow oil. 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.19 - 3.97 (m, 2H), 2.59 (t, 2H), 2.45 (s, 3H), 1.79 - 1.69 (m, 1H), 1.67 - 1.46 (m, 6H), 1.43 - 1.19 (m, 18H), 0.88 (t, 4H), 0.60 - 0.40 (m, 2H), 0.30 - 0.06 (m, 2H)

[0109] (4) Synthesis of the compound of formula (F) 7-(methylamino)heptyl 2-cyclopropyldecanoate (0.40 g, 1.18 mmol, 1.00 eq) was added to a 50 mL three-necked RBF, and 7-bromoheptyl 2-cyclopropyldecanoate (0.60 g, 1.53 mmol, 1.30 eq) and Na2CO3 (0.25 g, 2.36 mmol, 2.00 eq) in dioxane (2 mL) were added. The mixture was stirred at 100 °C for 16 hours, then cooled and poured into water (5 mL) to separate the organic and aqueous layers. The aqueous layer was extracted with siRNA (5 mL x 3 times), the organic layer was collected, concentrated under vacuum, washed with saturated Na2CO3 aqueous solution (5 mL), dried over anhydrous Na2SO4, and filtered. The concentrated residue was purified using a silica column with a DCM:methanol ratio of 20:1 → 10:1 to obtain the compound of formula (F) (0.36 g, 556 μmol, 47.2%) as a pale yellow oil. 1H NMR: (400 MHz, CHLOROFORM-d) δ 4.09 (dt, 4H), 2.72 - 2.07 (m, 7H), 1.73 (br dd, 2H), 1.67 - 1.50 (m, 12H), 1.40 - 1.23 (m, 36H), 0.88 (br t, 8H), 0.59 - 0.50 (m, 2H), 0.45 (s, 2H), 0.24 (s, 2H), 0.13 (br d, 2H)

[0110] Example 7 The compound of formula (G) below was prepared according to the synthesis scheme shown in Figure 7.

[0111] [ka]

[0112] (1) Synthesis of 1-cyclopropylheptan-1-ol Cyclopropanecarboxyl (27.0 g, 385 mmol, 1.00 eq) and THF (500 mL) were added to a 2000 mL three-necked RBF. Magnesium hexyl bromide (1 M in THF, 500 mL, 1.30 eq) was slowly added to this mixture at 0°C. The mixture was stirred at 25°C for 4 hours. The reaction mixture was poured into a saturated aqueous solution of NH4Cl, and the organic layer and aqueous layer were separated. The aqueous layer was extracted with HCl (500 mL x 3 times), the organic layer was collected, and the mixture was concentrated under vacuum. The residue after concentration was purified using a silica column with petroleum ether:HCl = 50:1 → 10:1 to obtain 1-cyclopropylheptan-1-ol (46 g, 294.37 mmol, yield 76.4%) as a colorless oil. 1 H NMR (400 MHz, CHLOROFORM-d): δ 2.86 (td, 1H), 1.67 - 1.38 (m, 6H), 1.36 - 1.27 (m, 6H), 0.91 - 0.87 (m, 3H), 0.59 - 0.42 (m, 2H), 0.32 - 0.17 (m, 2H)

[0113] (2) Synthesis of 1-cyclopropylheptyl 8-bromooctanoate 1-Cyclopropylheptan-1-ol (10.0 g, 63.9 mmol, 1.00 eq) and 8-bromooctanoic acid (12.5 g, 63.9 mmol, 1.00 eq) in 100 mL of DCM were added to a 2000 mL three-necked RBF. Next, DMAP (10.2 g, 83.19 mmol, 1.30 eq) and EDCI (15.9 g, 83.2 mmol, 1.30 eq) were added to the mixture. The mixture was stirred at 25°C for 16 hours. The reaction mixture was poured into H2O (100 mL), extracted, and then re-extracted with DCM (100 mL x 3 times). The organic layer was stored and concentrated under vacuum. The concentrated residue was purified using a silica column with a ratio of petroleum ether:ethyl = 50:1 → 10:1 to obtain 1-cyclopropylheptyl 8-bromooctanoate (7.50 g, 22.5 mmol, 35.1% yield) as a colorless oil. 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.28 (td, 1H), 3.59 - 3.36 (m, 2H), 2.35 - 2.26 (m, 2H), 1.92 - 1.72 (m, 2H), 1.69 - 1.60 (m, 4H), 1.50 - 1.41 (m, 2H), 1.37 - 1.24 (m, 12H), 1.01 - 0.84 (m, 4H), 0.62 - 0.50 (m, 1H), 0.50 - 0.41 (m, 1H), 0.37 (td, 1H), 0.26 (qd, 1H)

[0114] (3) Synthesis of the compound of formula (G) MeNH2 (2M in THF, 200.76 mL, 48.4 eq) was added to a 500 mL three-necked RBF flask, and 1-cyclopropylheptyl 8-bromooctanoate (3.00 g, 8.30 mmol, 1.00 eq) was added to the flask under a nitrogen atmosphere. The mixture was stirred at 80°C for 16 hours, and then the solvent was evaporated. The residue after evaporation was purified using a silica column with petroleum ether:siRNA = 50:1 → 10:1 to obtain the compound of formula (G) (0.50 g, 844 μmol, yield 10.2%) as a yellow oily substance. 1 H NMR (400 MHz, CHLOROFORM-d): δ 0.26 (dq, 2 H) 0.34 - 0.40 (m, 2 H) 0.42 - 0.49 (m, 2 H) 0.50 - 0.58 (m, 2 H) 0.89 (br t, 6 H) 0.93 - 0.98 (m, 2 H) 1.26 - 1.34 (m, 26 H) 1.47 (br s, 4 H) 1.57 - 1.70 (m, 10 H) 2.13 - 2.41 (m, 11 H) 4.27 (dt, 2 H)

[0115] Example 8 The compound of formula (H) below was prepared according to the synthesis scheme shown in Figure 8. The synthesis method was the same as in Example 5, but "cyclopropanecarbaldehyde" was replaced with "cyclopentanecarbaldehyde" and the same molar equivalent was used.

[0116] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 4.26 (td, 2H), 3.54 (t, 2H), 2.55 (t, 2H), 2.50 - 2.41 (m, 4H), 2.28 (t, 4H), 1.70 - 1.56 (m, 10H), 1.49 - 1.39 (m, 4H), 1.35 - 1.21 (m, 49H), 0.95 (dt, 2H), 0.89 (t, 6H)

[0117] Example 9 The compound of formula (I) below was prepared according to the synthesis scheme shown in Figure 9. The synthesis method was the same as in Example 5, but "cyclopropanecarbaldehyde" was replaced with "cyclopentanecarbaldehyde" and "octylmagnesium bromide" was replaced with "7-methyloctylmagnesium bromide," and the same molar equivalents were used.

[0118] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 4.28 (td, 2H), 3.50 (t, 2H), 2.58 (t, 2H), 2.49 - 2.40 (m, 4H), 2.30 (t, 4H), 1.71 - 1.60 (m, 10H), 1.50 - 1.41 (m, 4H), 1.35 - 1.20 (m, 49H), 0.97 (dt, 2H), 0.92 (t, 6H)

[0119] Example 10 The compound of formula (J) below was prepared according to the synthesis scheme shown in Figure 10. The synthesis method was the same as in Example 7, but "magnesium hexyl bromide" was replaced with "(3,7-dimethyloxyl)magnesium bromide" and the same molar equivalent was used.

[0120] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): 4.19 (dt, 2 H), 2.41 - 2.15 (m, 11 H), 1.71 - 1.51 (m, 10 H), 1.47 (br s, 4 H), 1.33 - 1.25 (m, 22 H), 0.98 - 0.93 (m, 2 H), 0.91 - 0.88 (m, 18H), 0.58 - 0.50 (m, 2 H), 0.49 - 0.42 (m, 2 H), 0.40 - 0.34 (m, 2 H), δ 0.26 (dq, 2 H)

[0121] Example 11 The compound of formula (K) below was prepared according to the synthesis scheme shown in Figure 11. The synthesis method was the same as in Example 5, but "2-aminoethanol" was replaced with "2-methoxyethane-1-amine" and the same molar equivalent was used.

[0122] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 4.25 (td, 2H), 3.69 (t, 2H), 3.35 (s, 3H), 2.60 (t, 2H), 2.45 - 2.39 (m, 4H), 2.38 (t, 4H), 1.72 - 1.57 (m, 10H), 1.43 - 1.34 (m, 4H), 1.30 - 1.25 (m, 33H), 0.95 (dt, 2H), 0.88 (t, 6H), 0.60 - 0.42 (m, 4H), 0.41 - 0.22 (m, 4H)

[0123] Example 12 The compound of formula (L) below was prepared according to the synthesis scheme shown in Figure 12. The synthesis method was the same as in Example 3 above. "1-Cyclopropylundecyl 8-bromooctanoate" and "1-Cyclopropylnonyl 10-bromodecanoate" were prepared and reacted with "methylamine" to produce the compound of formula (L).

[0124] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.28 (td, 2H), 2.41 (br t, 8H), 2.22 (s, 3H), 1.80 - 1.66 (m, 8H), 1.50 - 1.39 (m, 4H), 1.39 - 1.23 (m, 48H), 1.02 (m, 2H), 0.87 (m, 6H), 0.61 - 0.22 (m, 8H)

[0125] Example 13 The compound of formula (M) below was prepared according to the synthesis scheme shown in Figure 13. The synthesis method was the same as in Example 5 above. "1-Cyclopropylundecyl 8-bromooctanoate" and "1-Cyclohexylnonyl 8-bromooctanoate" were prepared and reacted with "2-aminoethanol" to produce the compound of formula (M).

[0126] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 4.30-4.27 (m, 2H), 3.52 (t, 2H), 2.57 (t, 2H), 2.47 - 2.41 (m, 4H), 2.31-2.29 (m, 4H), 1.70 - 1.57 (m, 8H), 1.48 - 1.41 (m, 4H), 1.36 - 1.26 (m, 57H), 0.94 (dt, 1H), 0.88 (t, 6H), 0.60 - 0.42 (m, 2H), 0.41 - 0.22 (m, 2H)

[0127] Example 14 The compound of formula (N) below was prepared according to the synthesis scheme shown in Figure 14. The synthesis method was the same as in Example 1, but "cyclopropanecarbaldehyde" was replaced with "bicyclo[2.2.1]heptane-2-carbaldehyde" and the same molar equivalent was used.

[0128] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ 4.35 (td, 2H), 2.35-2.29 (m, 8H), 2.20 (s, 3H), 2.18 - 2.15 (m, 2H), 1.75 - 1.66 (m, 8H), 1.54 - 1.23 (m, 60H), 0.89 (t, 6H)

[0129] Example 15 The compound of formula (O) below was prepared according to the synthesis scheme shown in Figure 15. The synthesis method was the same as in Example 5, but "cyclopropanecarbaldehyde" was replaced with "bicyclo[3.1.1]heptane-3-carbaldehyde" and the same molar equivalent was used.

[0130] [ka] 1 H NMR (400 MHz, CHLOROFORM-d): δ = 4.40 (td, 2H), 3.54 (t, 2H), 2.56 (t, 2H), 2.47 - 2.40 (m, 4H), 2.30 (t, 4H), 2.22 (m, 2H), 1.71 - 1.58 (m, 10H), 1.48 - 1.26 (m, 54H), 0.99 - 0.79 (m, 10H)

[0131] Manufacturing examples of drug delivery compositions: Manufacturing of formulations using each compound of formula (A-G) 1. Preparation of raw materials According to Table 1 below, the raw materials necessary for the manufacture of the formulation were dissolved in each dilution solvent and prepared to the required concentration. During dissolution, the raw materials were kept at room temperature and dissolved by adding the solvent.

[0132] [Table 1]

[0133] 2. Mixing of raw materials The required amounts of raw materials were mixed so that the N / P ratio (amine group of lipids: phosphate group of mRNA) was 6, and the ratio of each compound in formula (A-G):DOPE:cholesterol:DMG-PEG was 50:10:38.5:1.5. Ethanol was added to the ethanol layer so that the total molecular weight 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. After mixing, buffer exchange was performed as follows to reduce the total ethanol content: The mixed solution was concentrated by centrifugation at 4,000 rpm using an Amicon-Ultra tube filter (Merk Millipore, UFC505096 or UFC805024, pore size: 50K, volume: 15 mL), then diluted with PBS and concentrated by centrifugation. This process was repeated to perform buffer exchange.

[0134] The more specific steps are as follows: 1) Two autoclaved tubes were prepared (tubes (A) and (B)). 2) Each compound of formula (A-G), along with DOPE, cholesterol, and DMG-PEG, were sequentially added to tube (A) in molar amounts calculated according to the experimental conditions, and mixed by vortexing. 3) Ethanol was added to the ethanol phase as needed, so that the total molecular weight of all raw materials was 12.5 mM or less. 4) In tube (B), mRNA was mixed with 20 mM sodium acetate buffer (pH 4.6). The ratio was calculated so that the total volume of the aqueous phase was three times that of the ethanol phase. 5) Mixing of tube (A) and tube (B) was performed using a microfluidic (Ignite, Precision Nanosystems). The operating conditions for the microfluidic were FRR (flow rate ratio) C:R = 3:1 and 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), and the process of concentration and dilution was repeated to remove excess ethanol, and finally concentrated to x mg / mL (theoretical concentration).

[0135] 3. Evaluation of the physical properties of the formulation 1) The particle characteristics of the manufactured formulations were confirmed using a particle size analyzer (dynamic light scattering method, DLS), and the results are shown in Table 2 below. 2) The mRNA inclusion efficiency was confirmed by the ribogreen assay, and the results are shown in Table 2 below.

[0136] [Table 2]

Claims

1. A lipid having the structure shown in the following formula (1). 【Chemistry 1】 (In the formula, M 1 and M 2 Each of these is 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)-, where M' is a direct bond, C1-13 alkylene, or C2-13 alkenylene, and R' is independently selected from the group consisting of a hydrogen atom, C1-18 alkyl, and C2-18 alkenyl. R 1 and R 2 Each of these is independently selected from the group consisting of unsubstituted or C3-20 cycloalkyl groups substituted with a hydroxyl group or a C1-6 alkyl group, and C3-20 heterocycloalkyl groups substituted with a hydroxyl group or a C1-6 alkyl group. R 3 R is selected from the group consisting of a hydrogen atom, an unsubstituted or hydroxyl-substituted C1-6 alkyl group, an unsubstituted or hydroxyl-substituted or C1-6 alkyl group C3-6 carbocyclic group, and -(CH2)n OR, where R is selected from the group consisting of C1-3 alkyl and C2-3 alkenyl, and n is an integer from 1 to 5. R 4 ~R 11 Each of these is independently a hydrogen atom or a C1-3 alkyl group. Me is a methyl group, a, b, c, and d are each independent integers between 3 and 11.

2. M 1 and M 2 are each independently —C(O)O— or —OC(O)—, R 1 and R 2 Each of these is independently an unsubstituted or C3-6 cycloalkyl group substituted with a hydroxyl group or a C1-6 alkyl group. R 3 R is a hydrogen atom, an unsubstituted C1-3 alkyl group, or -(CH2)n OR, where R is a C1-3 alkyl group and n is an integer from 1 to 5. R 4 ~R 11 It is a hydrogen atom, The lipid according to claim 1, wherein a, b, c, and d are each independently integers from 5 to 9.

3. The lipid according to claim 1, wherein the lipid has a structure selected from the following formulas (A to O). 【Chemistry 2】 【Transformation 3】

4. A method for producing a lipid having the structure shown in formula (1'), comprising the following steps: (1) A step of reacting the compound of formula (a) with the compound of formula (b) to obtain the compound of formula (c); (2) A step of reacting the compound of formula (c) with the compound of formula (d) to obtain the compound of formula (e); and (3) A step of reacting the compound of formula (e) with the compound of formula (f). 【Chemistry 4】 (In the formula, M 1 , R 1 , R 3 , R 4 , R 5 , R 8 , R 9 Me, a, and b are synonymous with those defined in claim 1, and each X is independently selected from the group consisting of F, CI, BR, and I.

5. A method for producing a lipid having the structure shown in formula (1), comprising the following steps: (1) A step of reacting the compound of formula (a') with the compound of formula (b') to obtain the compound of formula (c'); (2) A step of reacting the compound of formula (c') with the compound of formula (d') to obtain the compound of formula (e'); and (3) A step of reacting the compound of formula (e') and the compound of formula (e) obtained in claim 4 with the compound of formula (f). 【Transformation 5】 (In the formula, M 1 M 2 , R 1 ~R 11 Me, a, b, c, and d are synonymous with those defined in claim 1, and each X is independently selected from the group consisting of F, CI, BR, and I.

6. A method for producing a lipid having the structure shown in formula (1'), comprising the following steps: (1) A step of reacting the compound of formula (i) with the compound of formula (ii) to obtain the compound of formula (iii); (2) A step of reacting the compound of formula (iii) with the compound of formula (iv) to obtain the compound of formula (v); (3) A step of reacting the compound of formula (v) with the compound of formula (vi) to obtain the compound of formula (vii); and (4) A step of reacting the compound of formula (v) with the compound of formula (vii). 【Transformation 6】 (In the formula, M 1 , R 1 , R 3 , R 4 , R 5 , R 8 , R 9 Me, a, and b are synonymous with those 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 the structure shown in formula (1), comprising the following steps: (1) A step of reacting the compound of formula (i') with the compound of formula (ii') to obtain the compound of formula (iii'); (2) A step of reacting the compound of formula (iii') with the compound of formula (iv') to obtain the compound of formula (v'); (3) A step of reacting the compound of formula (v') with the compound of formula (vi) to obtain the compound of formula (vii'); and (4) A step of reacting the compound of formula (vii') with the compound of formula (v) obtained in claim 6. 【Transformation 7】 (In the formula, M 1 M 2 , R 1 ~R 11 Me, a, b, c, and d are the same as those defined in claim 1, and each X is independently selected from the group consisting of F, CI, BR, and I.

8. A drug delivery composition comprising the lipid described in any one of claims 1 to 3.

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