Cationic lipids and methods for their preparation

Abstract

The present invention relates to a cationic lipid and a method for preparing the same, and more particularly, to a cationic lipid which forms (or is capable of forming) a complex with an anionic drug due to its special structure, thereby being useful for drug delivery, and a method for preparing the same.

Description

Technical Field

[0001] This invention relates to a cationic lipid and its preparation method. More specifically, this invention relates to a cationic lipid that, due to its special structure, readily forms a complex with anionic drugs, thereby enabling it to be used for drug delivery, and its preparation method. Background Technology

[0002] In therapies using anionic drugs, including nucleic acids, safe and efficient drug delivery technologies have long been researched, and various vectors and delivery methods have been developed. Vectors are mainly divided into viral vectors utilizing adenoviruses, retroviruses, etc., and non-viral vectors utilizing cationic lipids, cationic polymers, etc. It is known that viral vectors pose risks such as exposure to nonspecific immune responses, and their commercial application faces many challenges due to complex manufacturing processes. Therefore, in recent years, research has shifted towards using non-viral vectors to overcome these drawbacks. Compared to viral vectors, non-viral vectors have the advantage of fewer side effects in terms of in vivo safety and lower production costs in terms of cost-effectiveness.

[0003] Representative non-viral vectors for delivering nucleic acid substances are complexes of cationic lipids and nucleic acids (lipoplexes) and complexes of polycationic polymers and nucleic acids (polyplexes). These cationic lipids or polycationic polymers stabilize anionic drugs by forming complexes through electrostatic interactions with them, thereby improving intracellular delivery efficiency. For these reasons, they have been extensively studied (De Paula D, Bentley MV, Mahato RI, Hydrophobization and bioconjugation for enhanced siRNA delivery and targeting, RNA 13 (2007) 431-56; Gary DJ, Puri N, Won YY, Polymer-based siRNA delivery: Perspectives on the fundamental and phenomenological distinctions from polymer-based DNA delivery, J Controlrelease 121 (2007) 64-73).

[0004] However, polycationic polymers exhibit cytotoxicity due to their multivalent cationic charges, posing challenges for practical applications; while nucleic acid-cationic lipid complexes suffer from poor stability in blood, making them unsuitable for in vivo therapeutic use. Furthermore, ionic liposomes containing cationic lipids, neutral lipids, and fusion lipids also suffer from drawbacks such as complex synthesis methods for the cationic lipids used, cytotoxicity, and low intracellular nucleic acid delivery efficiency. Summary of the Invention

[0005] Technical issues The purpose of this invention is to provide a cationic lipid with a specific structure and a method for preparing the same, wherein the cationic lipid can readily form a complex with anionic drugs, thereby enabling its use in drug delivery.

[0006] Technical solution A first aspect of the present invention provides a cationic lipid having the structure shown in Formula 1: [Formula 1] , In Equation 1 above, R1 is a substituted or unsubstituted alkylene group. R2, R3, and R4 are each independently a substituted or unsubstituted alkylene group. R5, R6, and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group, or each independently a -R group. 10 -(L4) n -R 11 , R 10 Each is independently a substituted or unsubstituted alkylene group. R 11 Each can be an independently substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. L1, L2, L3, and L4 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atoms, alkyl groups, and alkenyl groups. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion.

[0007] More specifically, in Equation 1 above, R1 represents C with or without substitution. 1-6 Alkylene R2, R3, and R4 are each independently substituted or unsubstituted C. 3-12 Alkylene R5, R6, and R7 are each independently monovalent carbon, either substituted or unsubstituted, saturated or unsaturated. 3-20 hydrocarbon group, R8 and R9 are independently substituted or unsubstituted C. 1-6 Alkyl or C 3-6 Carbon cyclic groups, or individually -R 10 -(L4) n -R 11 , R 10 Each is independently either substituted or unsubstituted C 3-12 Alkylene R 11 Each is an independent monovalent C, whether substituted or unsubstituted, saturated or unsaturated. 3-20 hydrocarbon group, L1, L2, L3, and L4 are each independently selected from -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, C 2-6 imidene group, C 6-20 Aromatic and C 3-20 The group composed of heteroarylene groups, where L' is a direct bond and C' is a ligand. 1-13 Alkylene or C 2-13 The subalkenyl group, and R' is independently chosen from hydrogen atoms and C. 1-18 Alkyl and C 2-18 Group composed of alkenyl groups. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion of inorganic or organic acids.

[0008] Even more specifically, in Equation 1 above, R1 is C with or without substitution. 3-4 Alkylene R2, R3, and R4 are each independently substituted or unsubstituted C. 6-8 Alkylene R5, R6, and R7 are each independently monovalent carbon, either substituted or unsubstituted, saturated or unsaturated.5-15 hydrocarbon group, R8 and R9 are independently substituted or unsubstituted C. 1-2 Alkyl groups, or individually -R 10 -(L4) n -R 11 , R 10 Each is independently either substituted or unsubstituted C 6-8 Alkylene R 11 Each is an independent monovalent C, whether substituted or unsubstituted, saturated or unsaturated. 5-15 hydrocarbon group, L1, L2, L3, and L4 are each independently selected from -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -P(O)(OR')O-, -SS-, and C 2-5 The group consisting of alkenyl groups, where R' is independently chosen from hydrogen and C atoms. 1-6 The group consisting of alkyl groups, n is 0 or 1, and X - For F - Cl - ,Br - I - Nitrate anion, benzoate anion, methanesulfonate anion, acetate anion (CH3COO) - (i.e., AcO) - ) or trihaloacetate anion (CF3COO) - ).

[0009] More specifically, cationic lipids can be cationic lipids having structures selected from the following formulas A to Q:

[0010] A second aspect of the present invention provides a method for preparing cationic lipids having the structure shown in Formula 1-1, the method comprising the step of reacting a compound of Formula (a) with a compound of Formula (b): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Equation 1-1] , in, R1 is a substituted or unsubstituted alkylene group. R3 is an substituted or unsubstituted alkylene group, R6 can be an independent monovalent hydrocarbon group, whether substituted or unsubstituted, saturated or unsaturated. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group. L3 is independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atoms, alkyl groups, and alkenyl groups. X - It is a pharmaceutically acceptable monovalent anion.

[0011] A third aspect of the present invention provides a method for preparing cationic lipids having the structures shown in Formula 1-2 or Formula 1-3, 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 a compound of Formula (c) with a compound of Formula (d): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Equation (c)] (R6-L3-R3-)NH-R1-N + (-R8)(-R9)(-R3-L3-R6), [Equation (d)] R7-L2-R2-X, [Equation 1-2] , [Equation 1-3] , in, R1 is a substituted or unsubstituted alkylene group. R2 and R3 are each independently substituted or unsubstituted alkylene groups. R6 and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group. L2 and L3 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atom, alkyl, and alkenyl. X - It is a pharmaceutically acceptable monovalent anion.

[0012] A fourth aspect of the present invention provides a method for preparing cationic lipids having the structures shown in Formulas 1-4, 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 (f): [Equation (a)] R6-L3-R3-X, [Formula (b')] H2N-R1-NH2, [Formula (c')] (R6-L3-R3-)NH-R1-NH(-R3-L3-R6), [Equation (d)] R7-L2-R2-X, [Equation (e)] (R6-L3-R3-)(R7-L2-R2-)N-R1-N(-R2-L2-R7)(-R3-L3-R6), [Equation (f)] R8-X, [Equations 1-4] , in, R1 is a substituted or unsubstituted alkylene group. R2 and R3 are each independently substituted or unsubstituted alkylene groups. R6 and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 groups are each independently a substituted or unsubstituted alkyl or carbocyclic group. L2 and L3 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atom, alkyl, and alkenyl. X - It is a pharmaceutically acceptable monovalent anion.

[0013] A fifth aspect of the present invention provides a method for preparing cationic lipids having the structures shown in formulas 1-5, 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 a compound of formula (c”) with a compound of formula (d): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Formula (c”)] (R6-L3-R3-)NH-R1-N(-R8)(-R9), [Equation (d)] R7-L2-R2-X, [Equations 1-5] , in, R1 is a substituted or unsubstituted alkylene group. R2 and R3 are each independently substituted or unsubstituted alkylene groups. R6 and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group. L2 and L3 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atom, alkyl, and alkenyl. X - It is a pharmaceutically acceptable monovalent anion.

[0014] A sixth aspect of the invention provides a composition for drug delivery, the composition comprising a cationic lipid according to the invention.

[0015] Beneficial effects The cationic lipids according to the invention, due to their special structure (possessing both quaternary ammonium and tertiary amine), can readily form complexes with anionic drugs, and the drugs can be effectively delivered to target living tissues by utilizing these complexes. Attached Figure Description

[0016] Figure 1 This is the reaction scheme for the synthesis process of compound A in Example 1.

[0017] Figure 2 This is the reaction scheme for the synthesis process of compound B in Example 2.

[0018] Figure 3 This is the reaction scheme for the synthesis process of compound C in Example 3.

[0019] Figure 4 This is the reaction scheme for the synthesis process of compounds of formula D and formula E carried out in Example 4.

[0020] Figure 5 This is the reaction scheme for the synthesis process of compound F in Example 5.

[0021] Figure 6 This is the reaction scheme for the synthesis process of compound G in Example 6.

[0022] Figure 7 This is the reaction scheme for the synthesis process of compound H in Example 7. Detailed Implementation

[0023] The present invention will now be described in detail.

[0024] The cationic lipid provided in the first aspect of the present invention has the structure shown in Formula 1: [Formula 1] , In Equation 1 above, R1 is a substituted or unsubstituted alkylene group. R2, R3, and R4 are each independently a substituted or unsubstituted alkylene group. R5, R6, and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group, or each independently a -R group. 10 -(L4) n -R 11 , R 10 Each is independently a substituted or unsubstituted alkylene group. R 11 Each can be an independently substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. L1, L2, L3, and L4 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atoms, alkyl groups, and alkenyl groups. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion.

[0025] As used herein, for any group, the expression "substituted or unsubstituted" means, unless otherwise stated, that the group is either unsubstituted or substituted by one or more substituents selected from: -OH, halogen atoms, C 1-6 Alkyl, C 1-6 Alkoxy, C 1-6 Haloalkyl, C 1-6 Halogenated alkoxy groups, C 3-20 cycloalkyl, C 3-20 Heterocyclic alkyl, C 6-20 Aryl or C 3-20 Mixed aromatic compounds.

[0026] As used herein, for any group (e.g., heteroaryl, heterocycloalkyl, etc.), the expression "hetero-" means that, unless otherwise stated, the group has one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0027] As used in this article, a "monovalent hydrocarbon group" can be branched or unbranched, cyclic or acyclic, or aromatic.

[0028] According to one embodiment of the present invention, in formula 1 above, R1 can be substituted or unsubstituted C. 1-6 Alkylene R2, R3, and R4 can each independently be substituted or unsubstituted C. 3-12 Alkylene R5, R6, and R7 can each independently be a monovalent carbon, either substituted or unsubstituted, saturated or unsaturated. 3-20 hydrocarbon group, R8 and R9 can be independently substituted or unsubstituted C. 1-6 Alkyl or C 3-6 The carbonyl group, or each of them can be independently -R 10 -(L4) n -R 11 , R 10 Each can be independently substituted or unsubstituted C. 3-12 Alkylene R 11 Each can be an independently substituted or unsubstituted, saturated or unsaturated monovalent carbon. 3-20 hydrocarbon group, L1, L2, L3, and L4 can be independently selected from -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, C 2-6 imidene group, C 6-20 Aromatic and C 3-20 The group consisting of heteroarylene groups, where L' can be a direct bond, C 1-13 Alkylene or C 2-13 The subalkenyl group, and R' can be independently chosen from hydrogen atoms and C atoms. 1-18 Alkyl and C 2-18 Group composed of alkenyl groups. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion of inorganic or organic acids.

[0029] Even more specifically, in Equation 1 above, R1 can be substituted or unsubstituted C. 3-4 Alkylene R2, R3, and R4 can each independently be substituted or unsubstituted C. 6-8 Alkylene R5, R6, and R7 can each independently be a monovalent carbon, either substituted or unsubstituted, saturated or unsaturated. 5-15 hydrocarbon group, R8 and R9 can be independently substituted or unsubstituted C. 1-2 Alkyl groups, or each can be independently -R 10 -(L4) n -R 11 , R 10 Each can be independently substituted or unsubstituted C. 6-8 Alkylene R 11 Each can be an independently substituted or unsubstituted, saturated or unsaturated monovalent carbon. 5-15 hydrocarbon group, L1, L2, L3, and L4 can each be independently selected from -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -P(O)(OR')O-, -SS-, and C. 2-5 The group consisting of alkenyl groups, where R' can be independently chosen from hydrogen and C atoms. 1-6 The group consisting of alkyl groups, n is 0 or 1, and X - It can be F - Cl - ,Br - I - nitrate anion (NO3) - ), benzoate anion (C6H5COO) - ), mesylate anion, acetate anion (CH3COO) - (i.e., AcO) - ) or trihaloacetate anion (CF3COO) - ).

[0030] More specifically, cationic lipids can be cationic lipids having structures selected from the following formulas A to Q:

[0031] A second aspect of the present invention provides a method for preparing cationic lipids having the structure shown in Formula 1-1, the method comprising the step of reacting a compound of Formula (a) with a compound of Formula (b): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Equation 1-1] , in, R1, R3, R6, R8, R9, L3 and X - Same as defined above.

[0032] According to a second aspect of the invention, in an embodiment of a method for preparing cationic lipids, the reaction between the compound of formula (a) and the compound of formula (b) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN) etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, N,N-diisopropylethylamine (DIEA) etc.) at an elevated temperature (e.g., 80°C to 100°C), but is not limited thereto.

[0033] A third aspect of the present invention provides a method for preparing cationic lipids having the structures shown in Formula 1-2 or Formula 1-3, 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 a compound of Formula (c) with a compound of Formula (d): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Equation (c)] (R6-L3-R3-)NH-R1-N + (-R8)(-R9)(-R3-L3-R6), [Equation (d)] R7-L2-R2-X, [Equation 1-2] , [Equation 1-3] , in, R1 to R3, R6 to R9, L2, L3 and X- Same as defined above.

[0034] According to a third aspect of the invention, in an embodiment of a method for preparing cationic lipids, the reaction between the compound of formula (a) and the compound of formula (b) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, N,N-diisopropylethylamine (DIEA), etc.) at an elevated temperature (e.g., 80°C to 100°C), but is not limited thereto. Furthermore, the reaction between the compound of formula (c) and the compound of formula (d) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, N,N-diisopropylethylamine (DIEA), etc.) at an elevated temperature (e.g., 80°C to 100°C), but is not limited thereto.

[0035] A fourth aspect of the present invention provides a method for preparing cationic lipids having the structures shown in Formulas 1-4, 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 (f): [Equation (a)] R6-L3-R3-X, [Formula (b')] H2N-R1-NH2, [Formula (c')] (R6-L3-R3-)NH-R1-NH(-R3-L3-R6), [Equation (d)] R7-L2-R2-X, [Equation (e)] (R6-L3-R3-)(R7-L2-R2-)N-R1-N(-R2-L2-R7)(-R3-L3-R6), [Equation (f)] R8-X, [Equations 1-4] , in, R1 to R3, R6 to R8, L2, L3 and X - Same as defined above.

[0036] According to a fourth aspect of the invention, in an embodiment of a method for preparing cationic lipids, the reaction between the compound of formula (a) and the compound of formula (b') can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, etc.) at an elevated temperature (e.g., 40°C to 60°C), but is not limited thereto. Furthermore, the reaction between the compound of formula (c') and the compound of formula (d) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, etc.) at an elevated temperature (e.g., 40°C to 60°C), but is not limited thereto. Furthermore, the reaction between the compound of formula (e) and the compound of formula (f) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, etc.) at a temperature, for example, 20°C to 40°C, but is not limited thereto.

[0037] A fifth aspect of the present invention provides a method for preparing cationic lipids having the structures shown in formulas 1-5, 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 a compound of formula (c”) with a compound of formula (d): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Formula (c”)] (R6-L3-R3-)NH-R1-N(-R8)(-R9), [Equation (d)] R7-L2-R2-X, [Equations 1-5] , in, R1 to R3, R6 to R9, L2, L3 and X - Same as defined above.

[0038] According to a fifth aspect of the invention, in an embodiment of a method for preparing cationic lipids, the reaction between the compound of formula (a) and the compound of formula (b) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, N,N-diisopropylethylamine (DIEA), etc.) at an elevated temperature (e.g., 80°C to 100°C), but is not limited thereto. Furthermore, the reaction between the compound of formula (c”) and the compound of formula (d) can be carried out in a solvent (e.g., ethanol or acetonitrile (ACN), etc.) in the presence of a catalyst (e.g., Na2CO3, K2CO3, N,N-diisopropylethylamine (DIEA), etc.) at an elevated temperature (e.g., 80°C to 100°C), but is not limited thereto.

[0039] The cationic lipids described in this invention contain both quaternary ammonium and tertiary amines, and therefore can readily form complexes with anionic drugs, thus enabling their use in drug delivery.

[0040] Therefore, a sixth aspect of the present invention provides a composition for drug delivery, the composition comprising a cationic lipid according to the present invention.

[0041] In one implementation, the drug may be selected from nucleic acids, peptides, viruses, or combinations thereof.

[0042] "Nucleic acid" can be, for example, DNA, RNA, siRNA, shRNA, miRNA, mRNA, aptamers, antisense oligonucleotides, or combinations thereof, but is not limited to these.

[0043] "Polypeptide" can refer to a protein that is active in the body, such as an antibody or fragment thereof, a cytokine, a hormone or its analogue, or a protein that can be recognized as an antigen through a series of processes in the body, including polypeptide sequences of antigens, analogues or their precursors.

[0044] In one embodiment, the lipids of the present invention can form a complex with a drug, and the complex is encapsulated in a nanoparticle structure formed of an amphiphilic block copolymer.

[0045] In one embodiment, the amphiphilic block copolymer may be an AB-type block copolymer comprising a hydrophilic A block and a hydrophobic B block. In an aqueous environment, the AB-type block copolymer forms core-shell polymer nanoparticles, wherein the hydrophobic B block forms the core (inner wall) and the hydrophilic A block forms the shell (outer wall).

[0046] In one embodiment, the hydrophilic A block may be one or more selected from the group consisting of polyalkylene glycols, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide and its derivatives.

[0047] More specifically, the hydrophilic A block can be one or more selected from the group consisting of monomethoxy polyethylene glycol (mPEG), monoacetoxy polyethylene glycol, polyethylene glycol, copolymers of polyethylene and propylene glycol, and polyvinylpyrrolidone.

[0048] Furthermore, if necessary, the ends of the hydrophilic A-block can be chemically bound to functional groups or ligands capable of reaching specific tissues or cells, or functional groups capable of promoting intracellular delivery, to control the in vivo distribution of the polymer nanoparticle carrier or improve the delivery efficiency of the nanoparticle carrier into cells. In one embodiment, the functional group or ligand may be selected from one or more of the group consisting of monosaccharides, polysaccharides, vitamins, peptides, proteins, and cell surface receptor antibodies. More specifically, the functional group or ligand may be selected from one or more of the group consisting of anisamide, vitamin B9 (folic acid), vitamin B12, vitamin A, galactose, lactose, mannose, hyaluronic acid, RGD peptide, NGR peptide, transferrin, transferrin receptor antibody, etc.

[0049] The hydrophobic B-block is a biocompatible, biodegradable polymer, and in one embodiment, it can be selected from one or more of the group consisting of polyesters, polyanhydrides, polyamino acids, polyorthoesters, and polyphosphazenes.

[0050] More specifically, the hydrophobic B-block may be selected from one or more of the group consisting of polylactide (PLA), polyglycolic acid, polycaprolactone, polydioxanone, copolymers of polylactide and glycolic acid, copolymers of polylactide and polydioxanone, copolymers of polylactide and polycaprolactone, and copolymers of polyglycolic acid and polycaprolactone.

[0051] Furthermore, 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 can be modified by chemically binding the hydroxyl groups at the end of the hydrophobic B-block with tocopherol, cholesterol, or fatty acids having 10 to 24 carbon atoms.

[0052] The present invention will now be described in more detail with reference to the following embodiments. However, these embodiments are for illustrative purposes only, and the scope of the invention is not limited thereto.

[0053] Example Example 1 according to Figure 1 The synthetic scheme shown prepared a compound of formula A.

[0054] [Formula A] , (1) Synthesis of 6-bromohexyl 2-hexyldecanoate In a 2000 mL three-necked round-bottom flask (RBF), 2-hexyldecanoic acid (100 g, 390 mmol, 1.00 equivalent), 6-bromohex-1-ol (91.8 g, 507 mmol, 1.30 equivalent), and toluene (1000 mL) were added, followed by the addition of H₂SO₄ (7.65 g, 78.0 mmol, 0.20 equivalent). The mixture was degassed and purged with N₂ gas, then stirred at 120 °C for 16 hours. After cooling to room temperature, the mixture was concentrated under reduced pressure to remove the solvent. The residue was purified by silica gel column chromatography (petroleum ether:ethyl acetate (EtOAc) = 10:1 → 1:100) to obtain 6-bromohexyl 2-hexyldecanoate (135 g, 322 mmol, yield 82.5%) as a pale yellow oil.

[0055] (2) Synthesis of compound A In 500 mL of three-necked RBF, add 15.0 g (35.8 mmol, 3.50 equivalents) of 6-bromohexyl 2-hexyldecanoate and N. 1 N 1 Dimethylpropane-1,3-diamine and ethanol (EtOH) (150 mL) were mixed, and then Na₂CO₃ (3.25 g, 30.7 mmol, 3.00 equivalent) was added to the mixture. After purging three times with N₂ gas, the mixture was stirred at 90 °C for 48 hours. After cooling to room temperature, the solvent was removed under reduced pressure. The residue was extracted with a saturated aqueous solution of Na₂CO₃ and dichloromethane (DCM), and the DCM layer was collected. The solvent was removed under reduced pressure to obtain residue A. Residue A was dissolved in EtOH (60 mL), and H₂O (60 mL) and excess Na₂CO₃ were added. The mixture was stirred at room temperature for 16 hours. After filtering the mixture, EtOH was removed from the filtrate under reduced pressure. The residue solution was extracted with DCM, and the DCM layer was collected. The solvent was removed under reduced pressure to obtain residue B. Residue B was purified by reversed-phase HPLC (column: C1 250×80mm, 10μm; mobile phase: [HCl aqueous solution-ACN]; gradient: 55%-85%, 20 min) to obtain compound A (12.0 g, 10.7 mmol, yield 35.5%) as a yellow solid.

[0056] 1 H NMR (400 MHz, deuterated chloroform): δ4.07-4.04 (m, 4H), 3.45 (td, 2H), 3.28 (s,6H), 3.08 (br, 4H), 2.72 (br, H), 2.33-2.27 (m, 3H), 1.84 (br, 6H), 1.68-1.45 (m,20H), 1.45-1.42 (m, 18H), 1.32-1.25 (m, 62H), 0.89 (t, 18H) Example 2 according to Figure 2 The synthetic scheme shown prepared a compound of formula B.

[0057] [Formula B] , (1) Synthesis of heptadecano-9-yl 8-bromooctanoate In 1000 mL of three-necked RBF, heptadecano-9-ol (43.00 g, 168.00 mmol, 1.00 equivalent), 8-bromooctanoic acid (44.90 g, 201.00 mmol, 1.20 equivalent), 4-dimethylaminopyridine (DMAP) (20.50 g, 168.00 mmol, 1.00 equivalent), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (48.20 g, 252.00 mmol, 1.50 equivalent), and triethylamine (TEA) (17.00 g, 168.00 mmol, 23.30 mL, 1.00 equivalent) were added, and DCM (430 mL) was added. After purging three times with N2, the mixture was stirred at 25 °C under N2 for 16 hours. Add 500 mL of water to the reaction mixture at 20 °C, quench, and then use 900 mL of DCM (300 mL) 3) Extraction of the mixture. The collected organic layer was concentrated under reduced pressure, and the residue was then purified by silica gel column chromatography (petroleum ether:EtOAc=100:1→1:1) to obtain heptadecano-9-yl 8-bromooctanoate (36.00 g, 78.00 mmol, yield 46.5%) as a pale yellow oil.

[0058] (2) Synthesis of compound B In 100 mL of three-necked RBF, add heptadecano-9-yl 8-bromooctanoate (3.16 g, 6.85 mmol, 3.50 equivalents), N 1 N 11,3-Dimethylpropane-1,3-diamine (0.20 g, 1.96 mmol, 1.00 equivalent) and N,N-diisopropylethylamine (DIEA) (1.26 g, 9.79 mmol, 1.70 mL, 5.00 equivalent) were mixed, and EtOH (5 mL) was added. After purging three times with N2, the mixture was stirred at 95 °C for 16 hours under N2 conditions. The reaction mixture was filtered and concentrated under reduced pressure to remove the solvent. The mixture was extracted with saturated Na2CO3 solution and 300 mL (100 mL x 3) of DCM, and the collected organic layer was concentrated under reduced pressure to obtain residue A. Residue A was dissolved in EtOH (50 mL), and then 60 mL of water and excess Na2CO3 were added to the mixture, and the mixture was stirred at 25 °C for 16 hours. The mixture was filtered and concentrated under reduced pressure to remove EtOH, and then the aqueous layer was extracted with 300 mL (100 mL x 3) of DCM. The collected organic layer was concentrated under reduced pressure to obtain residue B. Residue B was purified by preparative HPLC (column: C1 250×80 mm, 10 μm; mobile phase: [HCl aqueous solution-ACN]; gradient: 55%-85%, 20 min), and concentrated under reduced pressure to remove solvent, thereby obtaining compound of formula B (1.52 g, 1.19 mmol, yield 60.6%), which was a pale yellow viscous substance.

[0059] 1 H NMR (400 MHz, deuterated chloroform): δ 4.86 (t, J =6.2 Hz, 3H), 4.15 (br d, J= 1.3 Hz, 2H), 3.44 - 3.32 (m, 4H), 3.28 (s, 6H), 3.14 - 2.94 (m, 4H), 2.72 - 2.58 (m, 2H), 2.28 (dt, J =2.1, 7.4 Hz, 6H), 1.96 - 1.74 (m, 12H), 1.65 - 1.58 (m, 6H), 1.51 (brd, J =5.8 Hz, 12H), 1.41 - 1.25 (m, 84H), 0.94 - 0.83 (m, 18H) Example 3 according to Figure 3 The synthetic scheme shown prepared a compound of formula C.

[0060] [Formula C] , (1) Synthesis of 1-cyclopropyl octyl 5-bromopentanoic acid In 500 mL of three-necked RBF, 1-cyclopropyloctyl-1-ol (15.00 g, 88.90 mmol, 1.00 equivalent), DMAP (2.15 g, 17.60 mmol, 0.20 equivalent), and TEA (17.80 g, 176.00 mmol, 24.50 mL, 2.00 equivalent) were added, along with 150 mL of DCM. After purging the mixture three times with N2, 5-bromopentanoyl chloride (26.40 g, 132.00 mmol, 17.70 mL, 1.50 equivalent) was added to the solution at 0 °C. After stirring the solution at 25 °C for 16 hours, 100 mL of H2O was added to the reaction mixture at 25 °C for quenching. The solution was then quenched with 1500 mL of DCM (500 mL...). 3) The extract mixture was dried with Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10:1) to obtain 1-cyclopropyl octyl 5-bromopentanoate (20.0 g, 60.0 mmol, yield 68.1%), as a colorless oil.

[0061] (2) Synthesis of 1-cyclopropyl octyl 5-iodovalanoate In 500 mL of three-necked RBF, 1-cyclopropyl octyl 5-bromopentanoate (20.00 g, 60.00 mmol, 1.00 equivalent) and KI (19.90 g, 120.00 mmol, 2.00 equivalent) were added, followed by the addition of acetonitrile (ACN) (200 mL). The mixture was purged three times with N2 and stirred at 90 °C for 16 hours under N2 conditions. The mixture was filtered and concentrated under reduced pressure to obtain 1-cyclopropyl octyl 5-iodopentanoate (18.00 g, 47.30 mmol, yield 78.9%) as a yellow oil.

[0062] (3) Synthesis of compound C Add N to 50 mL of three-necked RBF. 1 N 11,3-Dimethylpropane-1,3-diamine (0.20 g, 1.96 mmol, 0.33 equivalents), 1-cyclopropyl octyl 5-iodopentanoate (2.23 g, 5.87 mmol, 1.00 equivalents), and K₂CO₃ (812.00 mg, 5.87 mmol, 3.00 equivalents) were added, along with EtOH (2 mL) and ACN (2 mL). The mixture was purged three times with N₂ and stirred at 80 °C for 12 hours under N₂ conditions. The mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: C1 250 × 80 mm, 10 μm; mobile phase: [HCl aqueous solution-ACN]; gradient: 55%-85%, 20 min) to obtain compound of formula C (0.19 g, 221.00 μmol, yield 11.3%) as a yellow viscous substance.

[0063] 1 H NMR (400 MHz, deuterated chloroform): δ 0.26 (dt, J =9.57, 4.85 Hz, 3 H), 0.34 (dq, J =9.54, 4.87 Hz, 3 H), 0.40 - 0.49 (m, 3 H), 0.51 - 0.60 (m, 3 H), 0.83 - 0.92 (m, 9 H), 0.92 - 1.01 (m, 3 H), 1.27 (br s, 30 H), 1.39 - 1.47 (m, 4 H), 1.55 - 1.69 (m, 10 H), 1.70 - 1.78 (m, 2 H), 1.78 - 1.92 (m, 4 H), 2.32 (t, J =7.00 Hz, 4 H), 2.36 - 2.46 (m, 6 H), 2.50 (br t, J =5.88 Hz, 2 H), 3.38 (s, 6 H), 3.48 - 3.60 (m, 2 H), 3.60 - 3.74 (m, 2 H), 4.14 - 4.33 (m, 3 H) Example 4 according to Figure 4 The synthetic scheme shown prepared compounds of formula D and formula E.

[0064] [Form D] , [Formula E] , (1) Synthesis of undecane-3-yl 8-bromooctanoate In 500 mL of three-necked RBF, undecane-3-ol (25.00 g, 145.00 mmol, 1.00 equivalent) dissolved in toluene (250 mL) was added, followed by the addition of 8-bromooctanoic acid (48.60 g, 218.00 mmol, 1.50 equivalent). Then, H₂SO₄ (2.13 g, 21.8 mmol, 0.15 equivalent) was added. The mixture was purged three times with N₂ and stirred at 120 °C for 16 hours. The mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc = 10:1 → 1:100) to obtain undecane-3-yl 8-bromooctanoate (40.00 g, 106.00 mmol, yield 73.1%) as a yellow oil.

[0065] (2) Synthesis of heptadecano-9-yl 8-bromooctanoate In 5000 mL of three-necked RBF, heptadecan-9-ol (235.00 g, 916.00 mmol, 1.00 equivalent) dissolved in toluene (2.35 L) was added, followed by the addition of 8-bromooctanoic acid (245.00 g, 1.10 mol, 1.20 equivalent). Then, H₂SO₄ (18.00 g, 183.00 mmol, 0.20 equivalent) was added. The mixture was purged three times with N₂ and stirred at 120 °C for 16 hours. The mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc = 10:1 → 1:100) to obtain heptadecan-9-yl 8-bromooctanoate (470.00 g, 1.02 mol, yield 55.6%) as a yellow oil.

[0066] (3) Chloride 8-(heptadecane-9-yloxy)-N-(3-((8-(heptadecane-9-yloxy)-8-oxooctyl)amino Synthesis of (N,N-dimethyl-8-oxooctane-1-ammonium) In 250 mL of three-necked RBF, add heptadecano-9-yl 8-bromooctanoate (10.00 g, 21.70 mmol, 2.20 equivalents), N 1 N 1Dimethylpropane-1,3-diamine (1.01 g, 9.85 mmol, 1.00 equivalent), Na₂CO₃ (2.09 g, 19.70 mmol, 2.00 equivalent), and DIEA dissolved in EtOH (100 mL) (3.82 g, 29.50 mmol, 3.00 equivalent) were added. The mixture was stirred at 90 °C for 16 hours, and the reaction mixture was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (DCM:MeOH = 10:1 → 1:100), and 6 g of the purified product was further purified by reversed-phase HPLC to obtain 8-(heptadecane-9-yloxy)-N-(3-((8-(heptadecane-9-yloxy)-8-oxooctyl)amino)propyl)-N,N-dimethyl-8-oxooctane-1-ammonium chloride (2.00 g, 2.31 mmol, yield 23.5%) as a yellow solid.

[0067] (4) Synthesis of compounds of formula D and formula E In 50 mL of three-necked RBF, undecane-3-yl 8-bromooctanoate (314.00 mg, 833.00 μmol, 1.20 equivalents), 8-(heptadecano-9-yloxy)-N-(3-((8-(heptadecano-9-yloxy)-8-oxooctyl)amino)propyl)-N,N-dimethyl-8-oxooctane-1-ammonium chloride (0.60 g, 694.00 μmol, 1.00 equivalents), and K₂CO₃ dissolved in acetonitrile (6 mL) (115.00 mg, 833.00 μmol, 1.20 equivalents) were added. The mixture was stirred at 90 °C for 24 hours, and then the reaction mixture was concentrated under reduced pressure. The residue was purified by reversed-phase HPLC (column: C1 250×80 mm, 10 μm; mobile phase: [HCl aqueous solution-ACN]; gradient: 45%-75%, 25 min) to obtain compound D (0.20 g, 167.00 μmol, yield 24.1%) (yellow oil) and compound E (0.30 g, 258.00 μmol, yield 37.2%) (white solid).

[0068] [Compounds of type D] 1 H NMR (400 MHz, deuterated chloroform): δ 11.60 (br s, 1H), 4.92 - 4.75 (m, 3H), 4.16 (br s, 2H), 3.48 - 3.33 (m, 4H), 3.29 (s, 6H), 3.14 - 2.96 (m, 4H), 2.66 (br s, 2H), 2.28 (tt, J=2.3, 7.3 Hz, 6H), 2.21 (br s, 2H), 1.82 (br s, 6H), 1.65 - 1.58 (m, 6H), 1.57 - 1.46 (m, 12H), 1.42 - 1.34 (m, 16H), 1.26 (br s, 54H), 0.88 (t, J =6.6 Hz, 18H) [Compounds of Formula E] 1 H NMR (400 MHz, deuterated chloroform): δ 11.60 (br s, 1H), 4.90 - 4.76 (m, 3H), 4.14 (br d, J =7.1 Hz, 2H), 3.48 - 3.32 (m, 4H), 3.29 (s, 6H), 3.14 - 2.97 (m, 4H), 2.65 (br s, 2H), 2.32 - 2.25 (m, 8H), 1.82 (br s, 6H), 1.65 - 1.58 (m, 6H), 1.57 -1.46 (m, 12H), 1.43 - 1.34 (m, 16H), 1.32 - 1.24 (m, 59H), 0.93 - 0.82 (m, 18H) Example 5 according to Figure 5 The synthetic scheme shown prepared a compound of the following formula F.

[0069] [Formula F] , (1) Synthesis of 8,8'-(propane-1,3-diylbis(azadiyl))dioctanoic acid di(heptadecane-9-yl) ester In 100 mL of three-necked RBF, heptadecano-9-yl 8-bromooctanoate (2.00 g, 4.33 mmol, 2.00 equivalent), propane-1,3-diamine (160.00 mg, 2.17 mmol, 1.00 equivalent), and K₂CO₃ (598.00 mg, 4.33 mmol, 2.00 equivalent) were added, and ACN (20 mL) was added. After purging the mixture three times with N₂, the solution was stirred at 45 °C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (DCM:MeOH = 10:1) to obtain 8,8'-(propane-1,3-diylbis(azadiyl))dioctanoate di(heptadecano-9-yl) ester (0.80 g, 0.96 mmol, yield 11.0%) as a white solid.

[0070] (2)8,8'-(9,33-diethyl-11,31-dioxo-10,32-dioxa-19,23-diazatane-) Synthesis of 19,23-diyl)dioctanoic acid di(heptadecyl-9-yl) ester In 50 mL of three-necked RBF, 0.80 g (0.96 mmol, 1.00 equivalent) of 8,8'-(propane-1,3-diylbis(azadiyl))dioctanoic acid di(heptadecano-9-yl) ester (8.80 g, 0.96 mmol, 1.00 equivalent), 722.00 mg (1.92 mmol, 1.00 equivalent) of undecanoic acid 3-yl ester (8.00 mg, 1.92 mmol, 1.00 equivalent) and 264.00 mg (1.92 mmol, 2.00 equivalent) of K₂CO₃ were added, and 8 mL of ACN was added. The solution was stirred at 55 °C for 16 hours. The reaction mixture was filtered and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (water (HCl)-ACN) and concentrated under reduced pressure to remove ACN. 10 mL of NaHCO₃ aqueous solution was added to the ACN-removed aqueous solution, and the mixture was neutralized overnight. Then, 5 mL of DCM was added. 2) The mixture was extracted, dried with Na2SO4, filtered, and then subjected to reduced pressure to obtain 8,8'-(9,33-diethyl-11,31-dioxo-10,32-dioxa-19,23-diazattracarbamo-19,23-diyl)dioctanoic acid di(heptadecane-9-yl) ester (0.40 g, 0.28 mmol, yield 29.2%), which was a yellow oil.

[0071] (3) Synthesis of compounds of formula F In 50 mL of three-necked RBF, 0.40 g (0.28 mmol, 1.00 equivalent) of 8,8'-(9,33-diethyl-11,31-dioxo-10,32-dioxa-19,23-diazattracarbamo-19,23-diyl)dioctanoic acid di(heptadecan-9-yl) ester (42.50 mg, 0.31 mmol, 1.10 equivalent) and K₂CO₃ (42.50 mg, 0.31 mmol, 1.10 equivalent) were dissolved in THF (4 mL), and CH₃I (39.70 mg, 0.28 mmol, 1.00 equivalent) was added. The solution was stirred at 25 °C for 16 hours. 2 M HCl (10 mL) was added to the reaction mixture for quenching, and distilled water (DW) (5 mL) was added for dilution, followed by EtOAc (5 mL). 2) Extraction was performed. The collected organic layer was concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: C1 250×80mm, 10μm; mobile phase: [HCl aqueous solution-ACN]; gradient: 45%-75%, 25 min) and concentrated under reduced pressure to remove ACN. NaHCO3 aqueous solution (10 mL) was added to the ACN-removed aqueous solution, and the mixture was neutralized overnight. Subsequently, it was extracted with DCM (5 mL). 2) The mixture was extracted, dried with Na2SO4, filtered, and then subjected to reduced pressure to obtain compound F (0.40 g, 0.28 mmol, yield 29.2%), which was a yellow oil.

[0072] 1 HNMR: (400 MHz, deuterated chloroform) δ 0.74 - 1.00 (m, 24 H) 1.18 - 1.44 (m, 98 H)1.47 - 1.56 (m, 14 H) 1.57 (br s, 8 H) 1.64 - 1.88 (m, 10 H) 2.18 - 2.38 (m,9 H) 2.65 (ddd, J= 10.98, 5.16, 1.88 Hz, 1 H) 2.90 - 3.15 (m, 3 H) 3.20 - 3.48 (m, 8 H) 3.92 - 4.12 (m, 2 H) 4.84 (dt, J =19.35, 6.14 Hz, 4 H) Example 6 according to Figure 6 The synthetic scheme shown prepared compounds of the following formula G.

[0073] [Form G] , (1) Synthesis of 2-octyldecanoic acid In 1000 mL of three-necked RBF, nonanoic acid (60.00 g, 348.00 mmol, 1.00 equivalent) and THF (100 mL) were placed under N2 and 0 °C. Under N2 and 0 °C, lithium diisopropylaminolithium (LDA) (2 M, 383.00 mL, 2.20 equivalent) was added to the mixture, and the mixture was stirred at 0 °C for 30 min. Subsequently, 1-iodooctane (92.00 g, 383.00 mmol, 1.00 equivalent) was added at room temperature, and the mixture was stirred at 45 °C for 16 h. The reaction mixture was quenched by adding 1 N HCl (1 L), extracted with EtOAc (2 L), dried over Na2SO4, filtered, and concentrated to obtain the residue. After purging three times with N2, the solution was stirred at 45 °C for 16 h. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc=100:1→10:1) to obtain 2-octyldecanoic acid (67.00 g, 236.00 mmol, yield 67.6%), which was a yellow oil.

[0074] (2) Synthesis of 7-bromoheptyl 2-octyldecanoate In 250 mL of a three-necked RBF, 7-bromoheptane-1-ol (5.14 g, 26.40 mmol, 1.50 equivalent), octyldecanoic acid (5.00 g, 17.60 mmol, 1.00 equivalent), and H₂SO₄ (259 mg, 2.64 mmol, 0.15 equivalent) dissolved in toluene (50 mL) were added, and the mixture was purged three times with N₂. The mixture was stirred at 120 °C for 16 hours and concentrated under reduced pressure to obtain the residue. The residue was purified by silica gel column chromatography (petroleum ether:EtOAc = 10:1 → 1:100) to obtain 7-bromoheptyl 2-octyldecanoate (5.50 g, 11.90 mmol, yield 67.8%) as a yellow oil.

[0075] (3) Synthesis of 7-iodoheptyl 2-octyldecanoate In 250 mL of three-necked RBF, 7-bromoheptyl 2-octyldecanoate (5.50 g, 11.90 mmol, 1.00 equivalent) was added, followed by the addition of 55 mL of ACN. Then, KI (3.96 g, 23.80 mmol, 2.00 equivalent) was added, and the mixture was purged three times with N2. The mixture was stirred at 90 °C for 24 hours, filtered, and concentrated under reduced pressure to obtain 7-iodoheptyl 2-octyldecanoate (5.00 g, 9.83 mmol, yield 82.5%) as a yellow oil.

[0076] Synthesis of compound G (4) In 250 mL of three-necked RBF, 7-iodoheptyl 2-octyldecanoate (5.00 g, 9.83 mmol, 4.00 equivalents), 2,2'-((3-aminopropyl)azadiyl)bis(ethane-1-ol) (399.00 mg, 2.46 mmol, 1.00 equivalents), and K₂CO₃ (42.50 mg, 0.31 mmol, 1.10 equivalents) dissolved in ACN (50 mL) were added, and the mixture was purged three times with N₂. The solution was stirred at 80 °C for 16 hours, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by preparative HPLC (column: C1 250×80mm, 10μm; mobile phase: [HCl aqueous solution-ACN]; gradient: 60%-90%, 25 min) to obtain compound of formula G (0.25 g, 0.19 mmol, yield 99.2%), which was a gray oil.

[0077] 1 H NMR (400 MHz, deuterated chloroform): δ 5.04 - 4.62 (m, 2H), 4.06 (t, J =6.8 Hz, 6H), 3.95 - 3.86 (m, 2H), 3.70 - 3.61 (m, 4H), 3.35 - 3.21 (m, 6H), 2.70 (br s, 2H), 2.61 (br s, 4H), 2.31 (tt, J =5.4, 8.8 Hz, 3H), 1.85 - 1.69 (m, 10H), 1.66 - 1.53 (m, 12H), 1.46 - 1.36 (m, 24H), 1.26 (s, 65H), 0.93 - 0.84 (m, 18H) Example 7 according to Figure 7 The synthetic scheme shown prepared a compound with the following formula H.

[0078] [Formula H] , (1) Synthesis of 6-((3-(dimethylamino)propyl)amino)hexyl ester of 2-hexyldecanoic acid In a 100 mL three-necked round-bottom flask (RBF), add N 1 N 11,3-Dimethylpropane-1,3-diamine (1.0 g, 9.79 mmol, 1.22 mL, 1 equivalent), 6-bromohexyl 2-hexyldecanoate (4.11 g, 9.79 mmol, 1 equivalent), and ethanol (20 mL) were added. N,N-Diisopropylethylamine (6.32 g, 48.93 mmol, 8.52 mL, 5.0 equivalent) was added to the mixture. The mixture was stirred at 90 °C for 3 days. After cooling to room temperature, silica (1 g) was added to the mixture, and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography with petroleum ether:EtOAc = 1:1 → 2:1 to obtain 6-((3-(dimethylamino)propyl)amino)hexyl 2-hexyldecanoate (1.08 g, yield 25.0%) as a pale yellow oil.

[0079] (2) Synthesis of compounds of formula H In 500 mL of three-necked RBF, 6-((3-(dimethylamino)propyl)amino)hexyl 2-hexyldecanoate (1.08 g, 2.45 mmol, 1 equivalent), 6-bromohexyl 2-hexyldecanoate (1.03 g, 2.45 mmol, 1 equivalent), and ethanol (10 mL) were added. N,N-diisopropylethylamine (633.40 mg, 4.90 mmol, 853.63 μL, 2 equivalents) was added to the mixture, and the mixture was stirred at 90 °C for 3 days. After cooling to room temperature, silica (1 g) was added to the mixture, and the solvent was removed under vacuum. The residue was purified by silica gel column chromatography with petroleum ether:EtOAc = 3:1 → 1:1 to obtain compound of formula H (61.94 mg, yield 2.3%) as a pale yellow oil.

[0080] 1 H NMR (400 MHz, deuterated chloroform): δ 4.07-4.04 (m, 4H), 3.45 (td, 2H), 3.28 (s,6H), 3.08 (br, 4H), 2.72 (br, H), 2.33-2.27 (m, 3H), 1.84 (br, 6H), 1.68-1.45 (m,20H), 1.45-1.42 (m, 18H), 1.32-1.25 (m, 62H), 0.89 (t, 18H) [Example of preparation of a composition for drug delivery] 1. Raw material preparation According to the table below, dissolve the materials required for formulation preparation in the respective diluents to achieve the desired concentration. During dissolution, place the materials at room temperature, then add the solvent to dissolve them.

[0081] 2. Raw material mixing Take the required amounts of raw materials and mix them to achieve an N / P ratio (amino group of lipids / phosphate group of mRNA) of 6, and for each compound of formula A through H: DOPE:cholesterol:DMG-PEG = 50:10:38.5:1.5. Add ethanol to the ethanol layer to bring the total molecular weight of all raw materials below 12.5 mM, and mix the aqueous and ethanol phases at a volume ratio of 3:1. After mixing, to reduce the total ethanol content, change the buffer as follows: concentrate the mixture by centrifuging at 4000 rpm using an Amicon-Ultra tube filter (Merck Millipore, UFC505096 or UFC805024, pore size: 50K, volume: 0.5 mL, 4 mL, or 15 mL), then dilute with PBS and centrifuge to concentrate again. Repeat this process to change the buffer.

[0082] The more specific process is as follows: 1) Prepare two autoclaved tubes (tubes (A) and (B)).

[0083] 2) In tube (A), add the molar amounts of each compound of formula A to H, DOPE, cholesterol and DMG-PEG calculated according to the experimental conditions in sequence, and mix by vortexing.

[0084] 3) In the ethanol phase, add ethanol if necessary to ensure that the total molecular weight of all raw materials is within 12.5 mM.

[0085] 4) In tube (B), mix the mRNA and 20 mM sodium acetate buffer (pH 4.6). At this point, calculate the ratio and add the buffer proportionally so that the total volume of the aqueous phase is three times the total volume of the ethanol phase.

[0086] 5) Use a microfluidic device (Ignite, Precision Nanosystems) to mix tubes (A) and (B). The microfluidic operating conditions are: C:R flow rate ratio (FRR) = 3:1, and total flow rate (TRR) is 12 mL / min.

[0087] 6) Centrifuge the mixture obtained from step 5) at 4000 rpm using an Amicon-Ultra tubular filter to concentrate it. Repeat the process of dilution with PBS and centrifugation to remove excess ethanol, and then concentrate to a final concentration of x mg / ml (theoretical concentration).

[0088] 7) Once the formulation has been concentrated to the required concentration, it is sterilized using a filter membrane with a pore size of 0.22 μm.

[0089] 3. Evaluation of formulation performance 1) For the prepared formulation, particle characteristics (i.e., ζ-mean particle size (Z-mean), polydispersity index (PDI) and ζ-potential) were confirmed using a particle size analyzer (dynamic light scattering, DLS), and the results are shown in Table 1 below.

[0090] 2) The encapsulation efficiency of the prepared formulation was confirmed by Ribo-green assay, and the results are shown in Table 1 below.

[0091] [Table 1]

Claims

1. A cationic lipid having the structure shown in Formula 1: [Formula 1] ， in, In Equation 1 above, R1 is a substituted or unsubstituted alkylene group. R2, R3, and R4 are each independently a substituted or unsubstituted alkylene group. R5, R6, and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group, or each independently a -R group. 10 -(L4) n -R 11 , R 10 Each is independently a substituted or unsubstituted alkylene group. R 11 Each can be an independently substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. L1, L2, L3, and L4 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atoms, alkyl groups, and alkenyl groups. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion.

2. The cationic lipid according to claim 1, wherein... R1 is C with or without substitution. 1-6 Alkylene R2, R3, and R4 are each independently substituted or unsubstituted C. 3-12 Alkylene R5, R6, and R7 are each independently monovalent carbon, either substituted or unsubstituted, saturated or unsaturated. 3-20 hydrocarbon group, R8 and R9 are independently substituted or unsubstituted C. 1-6 Alkyl or C 3-6 Carbon cyclic groups, or individually -R 10 -(L4) n -R 11 , R 10 Each is independently either substituted or unsubstituted C 3-12 Alkylene R 11 Each is an independent monovalent C, whether substituted or unsubstituted, saturated or unsaturated. 3-20 hydrocarbon group, L1, L2, L3, and L4 are each independently selected from -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, C 2-6 imidene group, C 6-20 Aromatic and C 3-20 The group composed of heteroarylene groups, where L' is a direct bond and C' is a ligand. 1-13 Alkylene or C 2-13 The subalkenyl group, and R' is independently chosen from hydrogen atoms and C. 1-18 Alkyl and C 2-18 Group composed of alkenyl groups. n is 0 or 1, and X - It is a pharmaceutically acceptable monovalent anion of inorganic or organic acids.

3. The cationic lipid according to claim 1, wherein... R1 is C with or without substitution. 3-4 Alkylene R2, R3, and R4 are each independently substituted or unsubstituted C. 6-8 Alkylene R5, R6, and R7 are each independently monovalent carbon, either substituted or unsubstituted, saturated or unsaturated. 5-15 hydrocarbon group, R8 and R9 are independently substituted or unsubstituted C. 1-2 Alkyl groups, or individually -R 10 -(L4) n -R 11 , R 10 Each is independently either substituted or unsubstituted C 6-8 Alkylene R 11 Each is an independent monovalent C, whether substituted or unsubstituted, saturated or unsaturated. 5-15 hydrocarbon group, L1, L2, L3, and L4 are each independently selected from -C(O)O-, -OC(O)-, -C(O)N(R')-, -N(R')C(O)-, -P(O)(OR')O-, -SS-, and C 2-5 The group consisting of alkenyl groups, where R' is independently chosen from hydrogen and C atoms. 1-6 The group consisting of alkyl groups, n is 0 or 1, and X - For F - Cl - ,Br - I - Nitrate anion, benzoate anion, methanesulfonate anion, acetate anion (CH3COO) - ) or trihaloacetate anion (CF3COO) - ).

4. The cationic lipid according to claim 1, wherein the cationic lipid is a cationic lipid having a structure selected from the following formulas A to Q:

5. A method for preparing cationic lipids having the structure shown in Formula 1-1, the method comprising the step of reacting a compound of Formula (a) with a compound of Formula (b): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Equation 1-1] ， in, R1 is a substituted or unsubstituted alkylene group. R3 is an substituted or unsubstituted alkylene group, R6 can be an independent monovalent hydrocarbon group, whether substituted or unsubstituted, saturated or unsaturated. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group. L3 is independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atoms, alkyl groups, and alkenyl groups. X - It is a pharmaceutically acceptable monovalent anion.

6. A method for preparing cationic lipids, said cationic lipids having the structure shown in Formula 1-2 or Formula 1-3, said method comprising the following steps: (1) Reacting the compound of formula (a) with the compound of formula (b) to obtain the compound of formula (c); and (2) Reacting the compound of formula (c) with the compound of formula (d): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Equation (c)] (R6-L3-R3-)NH-R1-N + (-R8)(-R9)(-R3-L3-R6), [Equation (d)] R7-L2-R2-X, [Equation 1-2] ， [Equation 1-3] ， in, R1 is a substituted or unsubstituted alkylene group. R2 and R3 are each independently substituted or unsubstituted alkylene groups. R6 and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group. L2 and L3 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atom, alkyl, and alkenyl. X - It is a pharmaceutically acceptable monovalent anion.

7. A method for preparing cationic lipids having the structures shown in Formulas 1-4, the method comprising the following steps: (1) Reacting the compound of formula (a) with the compound of formula (b') to obtain the compound of formula (c'); (2) Reacting the compound of formula (c') with the compound of formula (d) to obtain the compound of formula (e); and (3) Reacting the compound of formula (e) with the compound of formula (f): [Equation (a)] R6-L3-R3-X, [Formula (b')] H2N-R1-NH2, [Formula (c')] (R6-L3-R3-)NH-R1-NH(-R3-L3-R6), [Equation (d)] R7-L2-R2-X, [Equation (e)] (R6-L3-R3-)(R7-L2-R2-)N-R1-N(-R2-L2-R7)(-R3-L3-R6), [Equation (f)] R8-X, [Equations 1-4] ， in, R1 is a substituted or unsubstituted alkylene group. R2 and R3 are each independently substituted or unsubstituted alkylene groups. R6 and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 groups are each independently a substituted or unsubstituted alkyl or carbocyclic group. L2 and L3 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atom, alkyl, and alkenyl. X - It is a pharmaceutically acceptable monovalent anion.

8. A method for preparing cationic lipids, the cationic lipids having the structures shown in Formulas 1-5, the method comprising the following steps: (1) Reacting the compound of formula (a) with the compound of formula (b) to obtain the compound of formula (c”); and (2) Reacting the compound of formula (c”) with the compound of formula (d): [Equation (a)] R6-L3-R3-X, [Equation (b)] H2N-R1-N(-R8)(-R9), [Formula (c”)] (R6-L3-R3-)NH-R1-N(-R8)(-R9), [Equation (d)] R7-L2-R2-X, [Equations 1-5] ， in, R1 is a substituted or unsubstituted alkylene group. R2 and R3 are each independently substituted or unsubstituted alkylene groups. R6 and R7 are each independently a substituted or unsubstituted, saturated or unsaturated monovalent hydrocarbon group. R8 and R9 are each independently a substituted or unsubstituted alkyl or carbocyclic group. L2 and L3 are each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-L'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -C(O)-, -C(S)-, -C(S)S-, -SC(S)-, -CH(OH)-, -P(O)(OR')O-, -S(O)2-, -SS-, alkenyl, arylene, and heteroarylene, where L' is a direct bond, alkylene, or alkenyl, and R' is independently selected from the group consisting of hydrogen atom, alkyl, and alkenyl. X - It is a pharmaceutically acceptable monovalent anion.

9. A composition for drug delivery, said composition comprising a cationic lipid according to any one of claims 1 to 4.

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