Cationic lipid and preparation method thereof

Abstract

The present invention relates to a cationic lipid and a method for preparing the same, and more particularly, to a cationic lipid which promotes the formation of a complex with an anionic drug material so as to be useful for drug delivery, and a method for preparing the cationic lipid.

Description

Technical Field

[0001] The present invention relates to cationic lipids and methods for preparing the same, and more particularly, to a cationic lipid that is easily complexed with anionic drugs and can be used for drug delivery, and a method for manufacturing the same. Background Art

[0002] In therapies using anionic drugs including nucleic acids, safe and effective drug delivery technologies have been studied for a long time, and various carriers and delivery technologies have been developed. Carriers are mainly classified into viral carriers using adenoviruses, retroviruses, etc. and non-viral carriers using cationic lipids, cationic polymers, etc. It is known that viral carriers have risks such as non-specific immune responses, and there are many problems in commercialization due to the complexity of the production process. Therefore, recent research is moving in the direction of improving these drawbacks by using non-viral carriers. Compared with viral carriers, non-viral carriers have the advantages of fewer side effects in terms of in vivo safety and lower production costs in terms of economy.

[0003] Representative non-viral carriers for delivering nucleic acid substances are complexes of cationic lipids and nucleic acids (lipoplexes) and complexes of polycationic polymers and nucleic acids (polyplexes). Such cationic lipids or polycationic polymers form complexes by electrostatic interaction with anionic drugs to stabilize the anionic drugs and increase intracellular delivery, and for these reasons, various studies have been conducted on them (De Paula D, Bentley MV, Mahato RI, Hydrophobization and bioconjugation for enhanced siRNA delivery and targeting, RNA 13 (2007) 431-56; Gary DJ, Puri N, Won YY, Polymer-based siRNA delivery: Perspectives on the fundamental and phenomenological distinctions from polymer-based DNA delivery, J Control release 121 (2007) 64-73).

[0004] However, polycationic polymers have cytotoxicity due to their multivalent cationic charges, which poses problems in their practical applications. Moreover, conventional cationic lipids used in ion lipid nanoparticles (including cationic lipids, neutral lipids, and fusogenic lipids) have the disadvantages of complex synthesis methods, cytotoxicity, and low intracellular nucleic acid delivery efficiency. Summary of the Invention

[0005] Technical Problem

[0006] An object of the present invention is to provide a cationic lipid having a specific structure and a preparation method thereof. The cationic lipid can easily form a complex with anionic drugs and can thus be used for drug delivery.

[0007] Technical Solution

[0008] A first aspect of the present invention provides a lipid having the structure shown in the following formula (1):

[0009] [Formula 1]

[0010]

[0011] Wherein in the above formula (1),

[0012] M 1 and M 2 are each independently a divalent linker group,

[0013] R 1 and R 2 are each independently a substituted or unsubstituted carbocyclic group or heterocyclic group,

[0014] R 3 is a hydrogen atom or an optionally substituted or unsubstituted organic group containing one or more heteroatoms,

[0015] R 4 to R 7 are each independently a hydrogen atom, or a substituted or unsubstituted saturated or unsaturated hydrocarbon group, and

[0016] a and b are each independently an integer from 1 to 20.

[0017] According to an embodiment of the present invention, in the above formula (1), M 1 and M 2may each independently be selected from the group consisting of: -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -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), wherein M' may be a direct bond, C 1-13 alkylene or C 2-13 alkenylene, and R' may each independently be selected from the group consisting of a hydrogen atom, C 1-18 alkyl, and C 2-18 alkenyl.

[0018] According to one embodiment of the present invention, in the above formula (1), R 1 and R 2 may each independently be selected from the group consisting of C 3-20 cycloalkyl, C 3-20 cycloalkenyl, C 6-20 aryl, C 3-20 heterocycloalkyl, C 3-20 heterocycloalkenyl, and C 3-20 heteroaryl, wherein they may each independently be unsubstituted or substituted with C 1-18 alkyl or C 2-18 alkenyl, and wherein the heterocycloalkyl, heterocycloalkenyl, or heteroaryl may each independently have one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S.

[0019] According to one embodiment of the present invention, in the above formula (1), R 3 may be selected from the group consisting of a hydrogen atom, substituted or unsubstituted C 1-6 alkyl, substituted or unsubstituted C 3-6 carbocyclic group, -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR, and -CQ(R) 2 wherein R may each independently be selected from the group consisting of a hydrogen atom, C 1-3 alkyl, and C 2-3 alkenyl; Q may be selected from the group consisting of a carbocyclic group, a heterocyclic group, -OR, -O(CH 2 ) n N(R) 2, -C(O)OR, -OC(O)R, -CX 3 , -CX 2 H, -CXH 2 , -CN, -N(R) 2 , -C(O)N(R) 2 , -N(R)C(O)R, -N(R)S(O) 2 R, -N(R)C(O)N(R) 2 , -N(R)C(S)N(R) 2 , -N(R)R 12 , N(R)S(O)2R 12 , -O(CH 2 ) n OR, -N(R)C(=NR 13 )N(R) 2 , -N(R)C(=CHR 13 )N(R) 2 , -OC(O)N(R) 2 , -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O) 2 R, -N(OR)C(O)OR, -N(OR)C(O)N(R) 2 , -N(OR)C(S)N(R) 2 , -N(OR)C(=NR 13 )N(R) 2 , -N(OR)C(=CHR 13 )N(R) 2 , -C(=NR 13 )N(R) 2 , -C(=NR 13 )R, -C(O)N(R)OR and -C(R)N(R) 2 C(O)OR; where n is independently an integer from 1 to 5; R 12 is selected from the group consisting of C 3-6 carbocyclic groups and heterocyclic groups; R 13 is selected from the group consisting of H, CN, NO 2 , C 1-6 alkyl, -OR, -S(O) 2 R, -S(O) 2 N(R) 2、 C 2-6 alkenyl, C 3-6 carbocyclic groups and heterocyclic groups; each R is independently selected from the group consisting of a hydrogen atom, C 1-3 alkyl and C 2-3 alkenyl; each X is independently selected from the group consisting of F, Cl, Br, and I, where when R3 is -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR or -CQ(R) 2 when, (i) when n is 1, 2, 3, 4 or 5, Q is not -N(R) 2 , or (ii) when n is 1 or 2, Q is not a 5-, 6- or 7-membered heterocycloalkyl group.

[0020] According to an embodiment of the present invention, in the above formula (1), R 4 to R 7 can each independently be selected from the group consisting of a hydrogen atom, C 1-3 alkyl and C 2-3 alkenyl.

[0021] According to an embodiment of the present invention, in the above formula (1), a and b can each independently be an integer from 1 to 15.

[0022] More specifically, in the above formula (1), M 1 and M 2 can each independently be selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)- and -C(O)- (where M' and R' are the same as those defined above).

[0023] More specifically, in the above formula (1), R 1 and R 2 can each independently be selected from the group consisting of C 3-20 cycloalkyl and C 3-20 heterocycloalkyl, each of which can independently be unsubstituted or substituted by C 1-18 alkyl or C 2-18 alkenyl, and the heterocycloalkyl can have one or more (e.g., 1 to 3) heteroatoms selected from N, O and S.

[0024] More specifically, in the above formula (1), R 3 can be selected from the group consisting of a hydrogen atom, substituted or unsubstituted C 1-6 alkyl and substituted or unsubstituted C 3-6 carbocyclic group (e.g., C 3-6 cycloalkyl).

[0025] More specifically, in the above formula (1), R 4 to R 7 can each independently be a hydrogen atom or C 1-3 alkyl.

[0026] More specifically, in the above formula (1), a and b can each independently be an integer from 3 to 13.

[0027] Even more specifically, in the above formula (1), M 1 and M 2 can each independently be selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R')-, and -N(R')C(O) (where R' is the same as defined above).

[0028] Even more specifically, in the above formula (1), R 1 and R 2 can each independently be a substituted or unsubstituted C 3-15 cycloalkyl.

[0029] Even more specifically, in the above formula (1), R 3 can be a hydrogen atom, or a substituted or unsubstituted C 1-3 alkyl.

[0030] Even more specifically, in the above formula (1), R 4 to R 7 can be a hydrogen atom.

[0031] Even more specifically, in the above formula (1), a and b can each independently be an integer from 5 to 11, and even more specifically an integer from 5 to 9.

[0032] Even more specifically, the lipid can be a lipid having a structure selected from Structures A to R of the following formulas:

[0033]

[0034] The second aspect of the present invention provides a method for preparing a lipid having the structure shown in formula (1'), the method comprising the following steps: (1) reacting a compound of formula (a) with a compound of formula (b) to obtain a compound of formula (c); and (2) reacting the compound of formula (c) with a compound of formula (d):

[0035] [Formula a]

[0036] R 1 -OH

[0037] [Formula b]

[0038]

[0039] [Formula c]

[0040]

[0041] [Formula d]

[0042] R 3 -NH 2

[0043] [Formula 1']

[0044]

[0045] wherein,

[0046] M 1 , R 1 , R 3 , R 4 , R 5 and a are the same as those defined in the above formula (1), and

[0047] X is independently selected from the group consisting of F, Cl, Br, and I.

[0048] The third aspect of the present invention provides a method for preparing a lipid having the structure shown in formula (1), the method 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 the compound of formula (c') with a compound of formula (d) to obtain a compound of formula (e); and (3) reacting the compound of formula (e) with the compound of formula (c) obtained from the second aspect of the present invention:

[0049] [Formula a']

[0050] HO-R 2

[0051] [Formula b']

[0052]

[0053] [Formula c']

[0054]

[0055] [Formula d]

[0056] R 3 -NH 2

[0057] [Formula e]

[0058]

[0059] [Formula c]

[0060]

[0061] [Formula 1]

[0062]

[0063] Among them,

[0064] M 1 、M 2 、R 1 to R 7 、a and b are the same as those defined in the above formula (1), and

[0065] X is independently selected from the group consisting of F, Cl, Br, and I.

[0066] The fourth aspect of the present invention provides a method for preparing a lipid having the structure shown in formula (1'), the method comprising the following steps: (1) reacting a compound of formula (i) with a compound of formula (ii) to obtain a compound of formula (iii); and (2) reacting the compound of formula (iii) with a compound of formula (iv):

[0067] [Formula i]

[0068] R 1 -M 1 -H

[0069] [Formula ii]

[0070]

[0071] [Formula iii]

[0072]

[0073] [Formula iv]

[0074] R 3 -NH 2

[0075] [Formula 1']

[0076]

[0077] Among them,

[0078] M 1 、R 1 、R 3 、R 4 、R 5 、and a are the same as those defined in the above formula (1), and

[0079] X is independently selected from the group consisting of F, Cl, Br, and I.

[0080] The fifth aspect of the present invention provides a method for preparing a lipid having the structure shown in formula (1), the method 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 the compound of formula (iii') with a compound of formula (iv) to obtain a compound of formula (v); and (3) reacting the compound of formula (v) with the compound of formula (iii) obtained from the fourth aspect of the present invention:

[0081] [Formula i']

[0082] H-M 2 -R 2

[0083] [Formula ii']

[0084]

[0085] [Formula iii']

[0086]

[0087] [Formula iv]

[0088] R 3 -NH 2

[0089] [Formula v]

[0090]

[0091] [Formula iii]

[0092]

[0093] [Formula 1]

[0094]

[0095] Wherein,

[0096] M 1 、M 2 、R 1 to R 7 、a and b are the same as those defined in formula (1) above, and

[0097] X are each independently selected from the group consisting of F, Cl, Br, and I.

[0098] The sixth aspect of the present invention provides a composition for drug delivery, which comprises the lipid according to the present invention.

[0099] Advantages of the Invention

[0100] Lipids with a specific structure according to the present invention can easily form complexes with anionic drugs, and by utilizing the complexes, the drugs can be effectively delivered to target living tissues. Description of the Drawings

[0101] Figure 1 is the reaction scheme of the lipid synthesis process carried out in Example 1.

[0102] Figure 2 is the reaction scheme of the lipid synthesis process carried out in Example 2.

[0103] Figure 3 is the reaction scheme of the lipid synthesis process carried out in Example 3.

[0104] Figure 4 is the reaction scheme of the lipid synthesis process carried out in Example 4.

[0105] Figure 5 is the reaction scheme of the lipid synthesis process carried out in Example 5.

[0106] Figure 6 is the reaction scheme of the lipid synthesis process carried out in Example 6.

[0107] Figure 7 is the reaction scheme of the lipid synthesis process carried out in Example 7.

[0108] Figure 8 is the reaction scheme of the lipid synthesis process carried out in Example 8.

[0109] Figure 9 is the reaction scheme of the lipid synthesis process carried out in Example 9.

[0110] Figure 10 is the reaction scheme of the lipid synthesis process carried out in Example 10.

[0111] Figure 11 is the reaction scheme of the lipid synthesis process carried out in Example 11.

[0112] Figure 12 is the reaction scheme of the lipid synthesis process carried out in Example 12.

[0113] Figure 13 is the reaction scheme of the lipid synthesis process carried out in Example 13.

[0114] Figure 14 is the reaction scheme of the lipid synthesis process carried out in Example 14.

[0115] Figure 15 is the reaction scheme of the lipid synthesis process carried out in Example 15.

[0116] Figure 16 is the reaction scheme of the lipid synthesis process carried out in Example 16.

[0117] Figure 17 is the reaction scheme of the lipid synthesis process carried out in Example 17.

[0118] Figure 18 is the reaction scheme of the lipid synthesis process carried out in Example 18. Detailed implementation mode

[0119] The present invention will be explained in detail below.

[0120] The lipid provided by the first aspect of the present invention has the structure shown in the following formula (1):

[0121] [Formula 1]

[0122]

[0123] Wherein in the above formula (1),

[0124] M 1 and M 2 are each independently a divalent linking group,

[0125] R 1 and R 2 are each independently a substituted or unsubstituted carbocyclic group or heterocyclic group,

[0126] R 3 is a hydrogen atom or an optionally substituted or unsubstituted organic group containing one or more heteroatoms,

[0127] R 4 to R 7 are each independently a hydrogen atom, or a substituted or unsubstituted saturated or unsaturated hydrocarbon group, and

[0128] a and b are each independently an integer from 1 to 20.

[0129] As used herein, unless otherwise specified, the expression "substituted or unsubstituted" for any group means that the group is unsubstituted or substituted by one or more substituents selected from -OH, halogen atoms, C 1-8 alkyl (more specifically C 3-7 alkyl) or C 1-8 haloalkyl (more specifically C 3-7 haloalkyl).

[0130] According to an embodiment of the present invention, in the above formula (1), M 1 and M 2may each independently be selected from the group consisting of: -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -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, still more specifically C 6-10 arylene) and heteroarylene (more specifically C having one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S 3-20 heteroarylene, still more specifically C 3-10 heteroarylene), wherein M' may be a direct bond, C 1-13 alkylene (more specifically C 1-6 alkylene) or C 2-13 alkenylene (more specifically C 2-6 alkenylene), and R' may each independently be selected from the group consisting of a hydrogen atom, C 1-18 alkyl (more specifically C 1-10 alkyl, still more specifically C 1-6 alkyl) and C 2-18 alkenyl (more specifically C 2-10 alkenyl, still more specifically C 2-6 alkenyl).

[0131] According to one embodiment of the present invention, in the above formula (1), R 1 and R 2 may each independently be selected from the group consisting of substituted or unsubstituted C 3-20 cycloalkyl (more specifically C 3-15 cycloalkyl, still more specifically C 6-15 cycloalkyl), substituted or unsubstituted C 3-20 cycloalkenyl (more specifically C 3-15 cycloalkenyl, still more specifically C 6-15 cycloalkenyl), substituted or unsubstituted C 6-20 aryl (more specifically C 6-14 aryl), substituted or unsubstituted C 3-20 heterocycloalkyl (more specifically C 3-15 heterocycloalkyl), substituted or unsubstituted C 3-20 heterocycloalkenyl (more specifically C 3-15 heterocycloalkenyl) and substituted or unsubstituted C 3-20 heteroaryl (more specifically C 3-15 heteroaryl), wherein the heterocycloalkyl, heterocycloalkenyl, and heteroaryl may each independently have one or more (e.g., 1 to 3) heteroatoms selected from N, O, and S.

[0132] According to one embodiment of the present invention, in the above formula (1), R 3 may be selected from the group consisting of a hydrogen atom, a substituted or unsubstituted C 1-6 alkyl group, a substituted or unsubstituted C 3-6 carbocyclic group, -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR, and -CQ(R) 2 wherein each R may be independently selected from the group consisting of a hydrogen atom, a C 1-3 alkyl group, and a C 2-3 alkenyl group; Q may be selected from the group consisting of a carbocyclic group, a heterocyclic group, -OR, -O(CH 2 ) n N(R) 2 , -C(O)OR, -OC(O)R, -CX 3 , -CX 2 H, -CXH 2 , -CN, -N(R) 2 , -C(O)N(R) 2 , -N(R)C(O)R, -N(R)S(O) 2 R, -N(R)C(O)N(R) 2 , -N(R)C(S)N(R) 2 , -N(R)R 12 , N(R)S(O)2R 12 , -O(CH 2 ) n OR, -N(R)C(=NR 13 )N(R) 2 , -N(R)C(=CHR 13 )N(R) 2 , -OC(O)N(R) 2 , -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O) 2 R, -N(OR)C(O)OR, -N(OR)C(O)N(R) 2 , -N(OR)C(S)N(R) 2 , -N(OR)C(=NR 13 )N(R) 2 , -N(OR)C(=CHR 13 )N(R) 2 , -C(=NR 13 )N(R) 2 , -C(=NR 13)R, -C(O)N(R)OR, and -C(R)N(R) 2 C(O)OR, where n is independently an integer from 1 to 5; R 12 is selected from the group consisting of C 3-6 carbocyclic groups and heterocyclic groups; R 13 is selected from the group consisting of H, CN, NO 2 , C 1-6 alkyl, -OR, -S(O) 2 R, -S(O) 2 N(R) 2、 C 2-6 alkenyl, C 3-6 carbocyclic groups and heterocyclic groups; each R is independently selected from the group consisting of a hydrogen atom, C 1-3 alkyl, and C 2-3 alkenyl; each X is independently selected from the group consisting of F, Cl, Br, and I, provided that when R 3 is -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR, or -CQ(R) 2 , (i) when n is 1, 2, 3, 4, or 5, Q is not -N(R) 2 , or (ii) when n is 1 or 2, Q is not a 5-, 6-, or 7-membered heterocycloalkyl.

[0133] According to one embodiment of the present invention, in the above formula (1), R 4 to R 7 can each independently be selected from the group consisting of a hydrogen atom, C 1-3 alkyl, and C 2-3 alkenyl.

[0134] According to one embodiment of the present invention, in the above formula (1), a and b can each independently be an integer from 1 to 15, and more specifically an integer from 3 to 13.

[0135] Even more specifically, in the above formula (1), M 1 and M 2 can each independently be selected from the group consisting of -C(O)O-, -OC(O)-, -C(O)N(R’)-, and -N(R’)C(O)-, where R’ is the same as defined above.

[0136] Even more specifically, in the above formula (1), R 1 and R 2 can each independently be a substituted or unsubstituted C 3-15 cycloalkyl.

[0137] Still more specifically, in the above formula (1), R 3 can be a hydrogen atom, or a substituted or unsubstituted C 1-3 alkyl group.

[0138] Still more specifically, in the above formula (1), R 4 to R 7 can be a hydrogen atom.

[0139] Still more specifically, in the above formula (1), a and b can each independently be an integer from 5 to 11, and even more specifically an integer from 5 to 9.

[0140] Even more specifically, the lipid can be a lipid having a structure selected from Structures A to R represented by the following formulas:

[0141]

[0142] The second aspect of the present invention provides a method for preparing a lipid having the structure represented by formula (1’) included in the above formula (1), the method comprising the following steps: (1) reacting a compound of formula (a) with a compound of formula (b) to obtain a compound of formula (c); and (2) reacting the compound of formula (c) with a compound of formula (d):

[0143] [Formula a]

[0144] R 1 -OH

[0145] [Formula b]

[0146]

[0147] [Formula c]

[0148]

[0149] [Formula d]

[0150] R 3 -NH 2

[0151] [Formula 1’]

[0152]

[0153] Wherein,

[0154] M 1 、R 1 、R 3 、R 4 、R 5 and a are the same as those defined above, and

[0155] X is independently selected from the group consisting of F, Cl, Br, and I.

[0156] In one embodiment of the method for preparing a lipid according to the second aspect of the present invention, the reaction of step (1) can be carried out in a solvent (such as dichloromethane (DCM) or toluene) in the presence of a catalyst (such as triethylamine (TEA) or sulfuric acid) at room temperature (such as 20 °C to 30 °C) or elevated temperature (such as from 40 °C to 150 °C); the reaction of step (2) can be carried out in a solvent (such as ethanol (EtOH) or dioxane) at elevated temperature (such as from 40 °C to 150 °C) optionally in the presence of Na 2 CO 3 or a catalyst (such as N,N-diisopropylethylamine (DIEA)), but not limited thereto.

[0157] According to one embodiment, before the reaction of the compound of formula (a) and the compound of formula (b) in step (1), the compound of formula (b) can be pretreated, and the pretreatment can be carried out in a solvent (such as dichloromethane (DCM)) in the presence of dimethylformamide (DMF) using a chloride (such as oxalyl chloride) at low temperature (such as -10 °C to 10 °C) or room temperature (such as 20 °C to 30 °C), but not limited thereto.

[0158] The third aspect of the present invention provides a method for preparing a lipid having the structure shown in the above formula (1), the method 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 the compound of formula (c') with a compound of formula (d) to obtain a compound of formula (e); and (3) reacting the compound of formula (e) with the compound of formula (c) obtained from the second aspect of the present invention:

[0159] [Formula a']

[0160] HO-R 2

[0161] [Formula b']

[0162]

[0163] [Formula c']

[0164]

[0165] [Formula d]

[0166] R 3 -NH 2

[0167] [Formula e]

[0168]

[0169] [Formula c]

[0170]

[0171] [Formula 1]

[0172]

[0173] wherein,

[0174] M 1 、M 2 、R 1 to R 7 、a and b are the same as defined above, and

[0175] X is independently selected from the group consisting of F, Cl, Br, and I.

[0176] In one embodiment of the method for preparing a lipid according to the third aspect of the present invention, the reaction of step (1) can be carried out in a solvent (such as dichloromethane (DCM) or toluene) in the presence of a catalyst (such as triethylamine (TEA) or sulfuric acid) at room temperature (such as 20°C to 30°C) or elevated temperature (such as from 40°C to 150°C); the reactions of step (2) and step (3) can each independently be carried out in a solvent (such as ethanol (EtOH)) at elevated temperature (such as from 40°C to 150°C) optionally in the presence of Na 2 CO 3 or a catalyst (such as N,N-diisopropylethylamine (DIEA)), but not limited thereto.

[0177] According to one embodiment, before the reaction of the compound of formula (a') and the compound of formula (b') in step (1), the compound of formula (b') can be pretreated, and the pretreatment can be carried out in a solvent (such as dichloromethane (DCM)) in the presence of dimethylformamide (DMF) using a chloride (such as oxalyl chloride) at low temperature (such as -10°C to 10°C) or room temperature (such as 20°C to 30°C), but not limited thereto.

[0178] The fourth aspect of the present invention provides a method for preparing a lipid having the structure shown in the above formula (1'), the method comprising the following steps: (1) reacting a compound of formula (i) with a compound of formula (ii) to obtain a compound of formula (iii); and (2) reacting the compound of formula (iii) with a compound of formula (iv):

[0179] [Formula i]

[0180] R 1 -M1 -H

[0181] [Formula ii]

[0182]

[0183] [Formula iii]

[0184]

[0185] [Formula iv]

[0186] R 3 -NH 2

[0187] [Formula 1']

[0188]

[0189] wherein,

[0190] M 1 、R 1 、R 3 、R 4 、R 5 and a are the same as those defined above, and

[0191] X is independently selected from the group consisting of F, Cl, Br, and I.

[0192] In one embodiment of the method for preparing a lipid according to the fourth aspect of the present invention, the reaction of step (1) can be carried out in a solvent (such as dichloromethane (DCM) or toluene) in the presence of a catalyst (such as triethylamine (TEA) or sulfuric acid) at room temperature (such as 20 °C to 30 °C) or elevated temperature (such as from 40 °C to 150 °C); the reaction of step (2) can be carried out in a solvent (such as ethanol (EtOH) or dioxane) at elevated temperature (such as from 40 °C to 150 °C) optionally in the presence of Na 2 CO 3 or a catalyst (such as N,N-diisopropylethylamine (DIEA)), but not limited thereto.

[0193] The fifth aspect of the present invention provides a method for preparing a lipid having the structure shown in the above formula (1), the method 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 the compound of formula (iii') with a compound of formula (iv) to obtain a compound of formula (v); and (3) reacting the compound of formula (v) with the compound of formula (iii) obtained from the fourth aspect of the present invention:

[0194] [Formula i']

[0195] H-M 2 -R 2

[0196] [Formula ii']

[0197]

[0198] [Formula iii']

[0199]

[0200] [Formula iv]

[0201] R 3 -NH 2

[0202] [Formula v]

[0203]

[0204] [Formula iii]

[0205]

[0206] [Formula 1]

[0207]

[0208] Wherein,

[0209] M 1 、M 2 、R 1 to R 7 、a and b are the same as defined above, and

[0210] X is independently selected from the group consisting of F, Cl, Br, and I.

[0211] In one embodiment of the method for preparing lipids according to the fifth aspect of the present invention, the reaction of step (1) can be carried out in a solvent (such as dichloromethane (DCM) or toluene) in the presence of a catalyst (such as triethylamine (TEA) or sulfuric acid) at room temperature (such as 20 °C to 30 °C) or elevated temperature (such as from 40 °C to 150 °C); the reactions of steps (2) and (3) can each independently be carried out in a solvent (such as ethanol (EtOH)) at elevated temperature (such as from 40 °C to 150 °C) optionally in the presence of Na 2 CO 3 or a catalyst (such as N,N-diisopropylethylamine (DIEA)), but not limited thereto.

[0212] The lipids having the structure shown in Formula 1 of the present invention can easily form complexes with anionic drugs and thus can be used for drug delivery.

[0213] Accordingly, a sixth aspect of the present invention provides a composition for drug delivery, which comprises a lipid having the structure shown in Formula 1 of the present invention.

[0214] In one embodiment, the drug can be selected from nucleic acids, polypeptides, viruses, or combinations thereof.

[0215] The "nucleic acid" can be, for example, DNA, RNA, siRNA, shRNA, miRNA, mRNA, aptamer, antisense oligonucleotide, or combinations thereof, but is not limited thereto.

[0216] The "polypeptide" can refer to a protein having activity in vivo, such as an antibody or a fragment thereof, a cytokine, a hormone or an analogue thereof, or a protein that can be recognized as an antigen through a series of processes in vivo, including a polypeptide sequence of an antigen, an analogue or a precursor thereof.

[0217] In one embodiment, the lipid of the present invention can form a complex with a drug, and the complex is encapsulated within a nanoparticle structure formed by an amphiphilic block copolymer.

[0218] In one embodiment, the amphiphilic block copolymer can be an A-B type block copolymer, which comprises a hydrophilic A block and a hydrophobic B block. In an aqueous environment, the A-B type block copolymer forms a core-shell type polymer nanoparticle, wherein the hydrophobic B block forms the core (inner wall) and the hydrophilic A block forms the shell (outer wall).

[0219] In one embodiment, the hydrophilic A block can be one or more selected from the group consisting of polyalkylene glycols, polyvinyl alcohol, polyvinylpyrrolidone, polyacrylamide, and derivatives thereof.

[0220] 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, a copolymer of polyethylene and propylene glycol, and polyvinylpyrrolidone.

[0221] In addition, if needed, the terminal of the hydrophilic A block can be chemically conjugated with a functional group or ligand capable of reaching a specific tissue or cell, or with a functional group capable of facilitating intracellular delivery, to control the in vivo distribution of the polymeric nanoparticle carrier formed by the amphiphilic block copolymer and polylactate, or to increase the efficiency of delivering the nanoparticle carrier to cells. In one embodiment, the functional group or ligand can 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 can be one or more selected from the group consisting of anisamide, vitamin B9 (folic acid), vitamin B12, vitamin A, galactose, lactose, mannose, hyaluronic acid, RGD peptide, NGR peptide, transferrin, antibodies against transferrin receptor, and the like.

[0222] The hydrophobic B block is a biocompatible and biodegradable polymer, which can be, in one embodiment, one or more selected from the group consisting of polyesters, polyanhydrides, polyamino acids, polyorthoesters, and polyphosphazines.

[0223] More specifically, the hydrophobic B block can be one or more selected from the group consisting of poly(lactic acid) (PLA), poly(glycolic acid), polycaprolactone, poly(dioxanone-2-one), copolymers of poly(lactic acid) and poly(glycolic acid), copolymers of poly(lactic acid) and poly(dioxanone-2-one), copolymers of poly(lactic acid) and polycaprolactone, and copolymers of poly(glycolic acid) and polycaprolactone.

[0224] In addition, in one embodiment, to increase the hydrophobicity of the hydrophobic B block and thus improve the stability of the nanoparticles, the hydrophobic B block can be modified by chemically conjugating the hydroxyl group at the terminal of the hydrophobic B block with tocopherol, cholesterol, or a fatty acid having 10 to 24 carbons.

[0225] The present invention will be explained in more detail with reference to the following examples. However, the examples are only for illustrating the present invention, and the scope of the present invention is not limited in any way.

[0226] Examples

[0227] Example 1

[0228] 1-1. Prepare Compound A of the following formula according to the Figure 1 synthesis scheme shown.

[0229] [Formula A]

[0230]

[0231] Synthesis of 4-pentylcyclohexyl 8-bromooctanoate

[0232] In a 250 mL three-necked round-bottom flask (RBF), 8-bromooctanoic acid (2.00 g, 8.96 mmol, 1.00 equivalent (eq)), dichloromethane (DCM) (40 mL), and dimethylformamide (DMF) (0.5 mL) were added together, and oxalyl chloride (2.28 g, 17.9 mmol, 2.00 equivalents) was added thereto at 0 °C under a nitrogen atmosphere. The resulting mixture was stirred at 25 °C for 4 hours under a nitrogen atmosphere, then 4-pentylcyclohexan-1-ol (2.29 g, 13.5 mmol, 1.50 equivalents) and triethylamine (TEA) (1.36 g, 13.5 mmol, 1.50 equivalents) were added, and the mixture was continuously stirred at 25 °C for 12 hours under a nitrogen atmosphere. The reaction mixture was concentrated in vacuo, and the concentrated residue was purified using a silica gel column with petroleum ether:ethyl acetate (EtOAc) = 1:0 → 50:1 to obtain 4-pentylcyclohexyl 8-bromooctanoate (2.46 g, 6.55 mmol, 73.1% yield) as a pale yellow oil.

[0233] 1 H NMR (400 MHz, chloroform-d): δ 0.76 - 0.86 (m, 3 H), 0.88 - 1.94 (m, 28H), 2.21 (dt, 2 H), 3.33 (td, 2 H), 4.52 - 4.66 (m, 1 H), 4.87 - 4.95 (m, 1H)

[0234] Synthesis of 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate

[0235] In a 100 mL three-necked RBF, 4-pentylcyclohexyl 8-bromooctanoate (2.46 g, 6.55 mmol, 1.00 equiv), 2-aminoethan-1-ol (2.00 g, 32.8 mmol, 5.00 equiv), and ethanol (EtOH) (50 mL) were added, and the mixture was stirred at 80 °C for 16 h under a nitrogen atmosphere. The reaction mixture was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography with petroleum ether:EtOAc = 1:0 → 50:1 to obtain 4-pentylcyclohexyl 8-((2-hydroxyethyl)amino)octanoate (2.00 g, 5.62 mmol, 85.8% yield) as a yellow solid.

[0236] 1 H NMR (400 MHz, chloroform-d): δ 0.76 - 0.86 (m, 3 H), 0.88 - 1.94 (m, 27H), 2.32 (t, 2 H), 2.51 (t, 2 H), 2.71 (t, 2H), 3.54 (d, 2 H), 3.91 - 4.00(m, 1 H), 4.11 - 4.31 (m, 1 H)

[0237] 1-4. Synthesis of cyclopentadecyl 8-bromooctanoate

[0238] In a 250 mL three-necked RBF, cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 equiv), 8-bromooctanoic acid (4.93 g, 22.1 mmol, 1 equiv), sulfuric acid (H 2 SO 4 )(217 mg, 2.21 mmol, 0.10 equiv), and toluene (100 mL) were added, and the mixture was stirred at 120 °C for 16 h under a nitrogen atmosphere. After evaporation of the solvent, the residue was purified by silica gel column chromatography with petroleum ether:EtOAc = 1:0 → 50:1 to obtain cyclopentadecyl 8-bromooctanoate (2.60 g, 6.03 mmol, 27.3% yield) as a colorless oil.

[0239] 1 H NMR (400 MHz, chloroform-d): δ 4.89 (quin, 1H), 3.41 (dt, 2H), 2.28(t, 2H), 1.85 (quin, 2H), 1.73 - 1.16 (m, 36H)

[0240] 1-5. Synthesis of Compound A

[0241] In a 100 mL three-necked RBF, 8-bromooctanoic acid cyclopentadecyl ester (1.60 g, 3.71 mmol, 1.00 equiv), 8-((2-hydroxyethyl)amino)octanoic acid 4-pentylcyclohexyl ester (1.32 g, 3.71 mmol, 1.00 equiv), N,N-diisopropylethylamine (DIEA) (527 mg, 4.08 mmol, 1.10 equiv) and ethanol (30 mL) were added, and the mixture was stirred at 80 ° C for 48 hours. Subsequently, after evaporation of the solvent, the residue was purified using a silica gel column with petroleum ether: EtOAc = 10: 1 → 1: 1, and purified again by preparative HPLC (folic acid conditions) with NaHCO 3 The organic layer was washed with aqueous solution (300 mL), and then concentrated and extracted with DCM (200 mL x 2). The organic layer was stirred for 2 h under anhydrous Na 2 SO 4 It was dried over 400 °C and filtered, and the filtrate was concentrated to obtain the compound of formula A (0.240 g, 340 μmol, 9.16% yield) as a yellow oil.

[0242] 1 H NMR (400 MHz, deuterated chloroform): δ 4.91 (br s, 1H), 4.82 (quin, 1H), 3.46 (br t, 2H), 2.51 (br d, 2H), 2.37 (br t, 4H), 2.21 (td, 4H), 1.58 - 1.42 (m,14H), 1.33 - 1.17 (m, 52H), 0.83 - 0.80 (m, 3H)

[0243] Example 2

[0244] 2-1. Based on Figure 2 The synthetic scheme shown below prepares the compound of formula B.

[0245] [Formula B]

[0246]

[0247] 2-2. Synthesis of cyclopentadecyl 6-bromohexanoate

[0248] In a 100 mL three-necked RBF, add cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 equiv), 6-bromohexanoic acid (6.46 g, 33.1 mmol, 1.50 equiv), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) (5.08 g, 26.5 mmol, 1.20 equiv), 4-dimethylaminopyridine (DMAP) (540 mg, 4.42 mmol, 0.20 equiv), TEA (4.47 g, 44.2 mmol, 6.15 mL, 2.00 equiv) and DCM (50 mL), and purge the mixture with nitrogen three times. Stir the resulting mixture at 25 °C under a nitrogen atmosphere for 16 h. After terminating the reaction by adding 200 mL of water at 20 °C, extract the mixture with 600 mL of DCM (200 mL x 3). Collect the organic layer, dry it over Na 2 SO 4 , filter, and concentrate the filtrate in vacuo. Purify the concentrated residue using a silica gel column with petroleum ether:EtOAc = 100:1 → 1:1 to obtain cyclopentadecyl 6-bromohexanoate (2.20 g, 5.45 mmol, 24.7% yield) as a yellow oil.

[0249] 1 1H NMR (400 MHz, chloroform-d‌): δ 4.83 (quin, 1H), 3.50 - 3.30 (m, 2H), 2.23 (t, 2H), 1.87 - 1.68 (m, 2H), 1.63 - 1.37 (m, 10H), 1.26 (br s, 22H)

[0250] 2 - 3. Synthesis of 4-propylcyclohexyl 6-bromohexanoate

[0251] In a 250 mL three-necked RBF, 4-propylcyclohexan-1-ol (10.0 g, 70.3 mmol, 1.00 equiv) in DCM (100 mL), 6-bromohexanoic acid (16.5 g, 84.4 mmol, 1.20 equiv), EDCI (20.2 g, 105 mmol, 1.50 equiv), DMAP (8.59 g, 70.3 mmol, 1.00 equiv) and TEA (7.11 g, 70.3 mmol, 9.79 mL, 1.00 equiv) were added. The mixture was purged with nitrogen three times and stirred at 25 °C for 16 h in a nitrogen atmosphere. The reaction mixture was neutralized by adding 100 mL of water at 20 °C, extracted with 600 mL of DCM (200 mL x 3), and then the organic layer was collected and concentrated in vacuo. The concentrated residue was purified using a silica column with petroleum ether:EtOAc = 100:1 → 10:1 to obtain 4-propylcyclohexyl 6-bromohexanoate (6.80 g, 21.3 mmol, 30.3% yield) as a yellow oil.

[0252] 1 H NMR (400 MHz, chloroform-d): δ 0.76 - 0.86 (m, 3 H), 0.88 - 1.94 (m, 19 H), 2.21 (dt, 2 H), 3.33 (td, 2 H), 4.52 - 4.66 (m, 1 H)

[0253] 2-4. Synthesis of 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate

[0254] In a 250 mL three-necked RBF, 4-propylcyclohexyl 6-bromohexanoate (3.00 g, 9.40 mmol, 1.00 equiv), 2-aminoethan-1-ol (2.87 g, 47.0 mmol, 2.84 mL, 5.00 equiv) and EtOH (60 mL) were added. The mixture was purged with nitrogen three times and stirred at 85 °C for 16 h in a nitrogen atmosphere. The reaction mixture was cooled to room temperature and concentrated to remove the solvent. The residue was purified by column chromatography (SiO 2 , DCM:MeOH = 100:1 → 1:1) to obtain 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate (0.70 g, 2.34 mmol, 24.9% yield) as a yellow oil.

[0255] 11H NMR (400 MHz, chloroform-d): δ 5.04 - 4.60 (m, 1H), 3.70 - 3.61 (m, 2H), 2.79 (t, 2H), 2.64 (dt, 2H), 2.35 - 2.24 (m, 2H), 2.01 - 1.94 (m, 2H), 1.87 - 1.74 (m, 2H), 1.65 (qd, 2H), 1.56 - 1.48 (m, 4H), 1.41 - 1.16 (m, 9H), 1.07 - 0.94 (m, 1H), 0.89 (dt, 3H)

[0256] 2-5. Synthesis of Compound B

[0257] In a 50 mL three-necked RBF, 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate (371 mg, 1.24 mmol, 1.00 equiv), 4-propylcyclohexyl 6-bromohexanoate (0.50 g, 1.24 mmol, 1.00 equiv), N,N-diisopropylethylamine (DIEA) (800 mg, 6.20 mmol, 1.08 mL, 5 equiv) were added together with 1,4-dioxane (10 mL). The mixture was purged with nitrogen three times and stirred at 95 °C for 16 h under a nitrogen atmosphere. The reaction mixture was cooled to room temperature and concentrated under reduced pressure to remove the solvent. The residue was purified by column chromatography (SiO 2 , DCM:MeOH = 100:1 → 10:1) to obtain Compound B (0.21 g, 337.64 μmol, 27.2% yield) as a yellow oil.

[0258] 1 1H NMR (400 MHz, chloroform-d): δ 4.94 - 4.52 (m, 2H), 3.79 (br s, 2H), 3.06 - 2.63 (m, 6H), 2.29 - 2.17 (m, 4H), 1.93 - 1.81 (m, 1H), 1.79 - 1.37 (m, 17H), 1.36 - 1.09 (m, 34H), 0.99 - 0.86 (m, 1H), 0.85 - 0.76 (m, 3H)

[0259] Example 3

[0260] 3-1. Prepare the following Compound C according to the synthetic scheme shown Figure 3 below.

[0261] [Formula C]

[0262]

[0263] Synthesis of cyclopentadecyl 10 - bromodecanoate

[0264] In a 100 mL three - necked RBF, cyclopentadecanol (5.00 g, 22.1 mmol, 1.00 equiv), 10 - bromodecanoic acid (8.32 g, 33.1 mmol, 1.50 equiv), DMAP (540 mg, 4.42 mmol, 0.20 equiv), EDCI (5.08 g, 26.5 mmol, 1.20 equiv) and TEA (4.47 g, 44.2 mmol, 2.00 equiv) were added together with DCM (50 mL). The mixture was purged with nitrogen three times and stirred at 50 °C for 16 h in a nitrogen atmosphere. The reaction mixture was cooled to 20 °C, water was added to terminate the reaction, and extraction was carried out with 600 mL of DCM (200 mL x 3), then the organic layer was collected, dried over Na 2 SO 4 dried, filtered and the filtrate was concentrated. The obtained residue was purified by column chromatography (SiO 2 , petroleum ether:EtOAc = 100:1 → 1:1) to give cyclopentadecyl 10 - bromodecanoate (2.60 g, 5.66 mmol, 25.6% yield) as a yellow oil.

[0265] 1 1H NMR (400 MHz, chloroform - d): δ 4.90 (quin, 1H), 3.58 - 3.36 (m, 2H), 2.27 (t, 2H), 1.90 - 1.72 (m, 2H), 1.64 - 1.53 (m, 6H), 1.43 - 1.28 (m, 34H)

[0266] Synthesis of 4 - heptylcyclohexan - 1 - ol

[0267] In a 500 mL three - necked RBF, LiAlH 4(A solution of 2.50 M in THF, 61.1 mL, 1.20 eq) was added together with THF (250 mL). The mixture was cooled to 0 °C, and a solution of 4-heptylcyclohexan-1-one (25.0 g, 127 mmol, 1.00 eq) in THF (250 mL) was slowly added thereto over 20 minutes under a nitrogen atmosphere. Then the temperature of the mixed solution was raised to 25 °C, and the mixture was stirred for 3 hours under a nitrogen atmosphere. The reaction mixture was cooled to 0 °C, and then water (120 mL) was added in a nitrogen atmosphere, taking care not to generate a large amount of foam. Then, a 15% aqueous NaOH solution (12 mL) was slowly added to the reaction mixture at 0 °C. After 5 minutes, water (36 mL) was added at the same temperature, and the temperature was slowly raised to 25 °C, and the mixture was stirred for 15 minutes. The resulting mixture was filtered, and the filter cake was concentrated in vacuo to obtain 4-heptylcyclohexan-1-ol (50.0 g, 252 mmol, 98.9% yield) as a colorless powder.

[0268] 1 H NMR (400 MHz, chloroform-d): δ 3.47 (tt, H), 1.95 - 1.84 (m, 2H), 1.75 - 1.65 (m, 2H), 1.42 (s, 1H), 1.24 - 1.14 (m, 14H), 1.11 - 1.08 (m, 2H), 0.83 - 0.80 (m, 3H)

[0269] 3 - 4. Synthesis of 4-heptylcyclohexyl 10-bromodecanoate

[0270] In a 250 mL three-necked RBF, 4-heptylcyclohexan-1-ol (10.0 g, 50.4 mmol, 1.00 eq), 10-bromodecanoic acid (15.2 g, 60.5 mmol, 1.20 eq), EDCI (14.5 g, 75.6 mmol, 1.50 eq), DMAP (6.16 g, 50.4 mmol, 1.00 eq), TEA (5.10 g, 50.4 mmol, 1.00 eq) and DCM (100 mL) were added. The mixture was purged with nitrogen three times and stirred at 25 °C for 16 hours under a nitrogen atmosphere. The reaction mixture was neutralized by adding 200 mL of water at 20 °C, extracted with 600 mL of DCM (200 mL x 3), and then the organic layer was collected and concentrated in vacuo. By column chromatography (SiO 2, the concentrated residue was purified with petroleum ether:EtOAc = 100:1→1:1) to obtain 4-heptylcyclohexyl 10-bromodecanoate (6.60 g, 15.3 mmol, 30.3% yield), which is a white solid.

[0271] 1 H NMR (400 MHz, chloroform-d): δ 4.59 (tt, 1H), 3.46 (t, 1H), 3.33 (t,1H), 2.19 (t, 2H), 1.87 (br dd, 2H), 1.82 - 1.67 (m, 4H), 1.58 - 1.50 (m,2H), 1.38 - 1.32 (m, 2H), 1.21 (br d, 23H), 0.97 - 0.86 (m, 2H), 0.81 (t, 3H)

[0272] 3 - 5. Synthesis of 4-heptylcyclohexyl 10-((2-hydroxyethyl)amino)decanoate

[0273] In a 250 mL three-necked RBF, 4-heptylcyclohexyl 10-bromodecanoate (2.30 g, 5.33 mmol, 1.00 equiv) and 2-aminoethan-1-ol (1.63 g, 26.6 mmol, 5 equiv) were added together with EtOH (60 mL). The mixture was purged with nitrogen three times and stirred at 95 °C for 16 h in a nitrogen atmosphere. The reaction mixture was concentrated in vacuo and the concentrated residue was purified by column chromatography (SiO 2 , DCM:MeOH = 100:1→1:1) to obtain 4-heptylcyclohexyl 10-((2-hydroxyethyl)amino)decanoate (1.30 g, 3.16 mmol, 59.2% yield), which is a white solid.

[0274] 1 H NMR (400 MHz, chloroform-d): δ4.66 (tt, 1H), 3.83 - 3.73 (m, 2H), 2.96 - 2.88 (m, 2H), 2.81 - 2.73 (m, 2H), 2.26 (t, 2H), 1.95 (br dd, 2H), 1.78 (brd, 2H), 1.62 (td, 4H), 1.28 (br d, 24H), 1.18 (br d, 2H), 1.04 - 0.96 (m,2H), 0.89 (t, 3H)

[0275] 3 - 6. Synthesis of Compound C

[0276] In a 50 mL three - necked RBF, 10 - ((2 - hydroxyethyl)amino)decanoic acid 4 - heptylcyclohexyl ester (0.80 g, 1.94 mmol, 1.00 equiv), 10 - bromodecanoic acid cyclopentadecyl ester (893 mg, 1.94 mmol, 1.00 equiv) and DIEA (1.26 g, 9.72 mmol, 1.69 mL, 5.00 equiv) were added together with EtOH (3 mL). The mixture was purged with nitrogen three times and stirred at 95 °C for 16 h in a nitrogen atmosphere. The reaction mixture was cooled to room temperature, concentrated in vacuo, and the concentrated residue was purified by chromatography (SiO 2 , DCM:MeOH = 100:1 → 10:1) to afford Compound C (0.35 g, 447 μmol, 22.9% yield) as a white solid.

[0277] 1 1H NMR (400 MHz, chloroform - d): δ 4.96 - 4.61 (m, 2H), 3.77 - 3.52 (m,2H), 2.84 - 2.37 (m, 6H), 2.27 (dt, 4H), 2.01 - 1.92 (m, 2H), 1.79 (br d,2H), 1.68 - 1.46 (m, 18H), 1.37 - 1.25 (m, 56H), 1.00 (br d, 1H), 0.89 (br t,3H)

[0278] Example 4

[0279] 4 - 1. According to Figure 4 the synthetic scheme shown, prepare the following Compound D.

[0280] [Formula D]

[0281]

[0282] 4-2. Synthesis of (E)-2-(hept-1-en-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane

[0283] In a 500 mL three-necked RBF, hept-1-yne (64.4 g, 670 mmol, 1.00 equiv), TEA (Et 3 N, 6.78g, 670mmol, 0.10 equivalents), 4,4,5,5-tetramethyl-1,3,2-dioxaborolane (90.0g, 703mmol, 1.05 equivalents), zirconium chloride and cyclopentane (17.9g, 670mmol, 0.10 equivalents), the mixture was purged with nitrogen three times, and then stirred at 60°C for 16 hours in a nitrogen environment. The reaction mixture was cooled to room temperature and filtered, and the filtrate was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10: 1) to obtain (E)-2-(hept-1-ene-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborolane (102g, 455mmol, 67.9% yield) as a yellow oil.

[0284] 1 H NMR: (400 MHz, deuterated chloroform): δ 6.64 (td, 1H), 5.43 (td, 1H), 2.19 -2.12 (m, 2H), 1.44 - 1.39 (m, 2H), 1.30 (br d, 4H), 1.27 (s, 12H), 0.90 -0.87 (m, 3H)

[0285] 4-3. Synthesis of 4,4,5,5-tetramethyl-2-((1R,2R)-2-pentylcyclopropyl)-1,3,2-dioxaborolane

[0286] In a 1000mL three-necked RBF, diethylzinc (1M, 178.5mL, 2.00 equivalents) was added together with distilled DCM (80mL) purified under a nitrogen atmosphere, and a solution of trifluoroacetic acid (TFA) (20.4g, 178mmol, 2.00 equivalents) dissolved in DCM (40mL) was gradually added at 0°C. In addition, a solution of diiodomethane (47.8g, 178mmol, 2.00 equivalents) dissolved in DCM (40mL) was added while stirring at 0°C for 30 minutes. The reaction mixture was stirred for 30 minutes, and then a solution of (E)-2-(hept-1-ene-1-yl)-4,4,5,5-tetramethyl-1,3,2-dioxaborane (20.0g, 89.2mmol, 1.00 equivalents) in DCM (40mL) was added at 0°C, and the mixture was stirred for 2 hours at 25°C under a nitrogen atmosphere. The reaction mixture was neutralized with water, extracted with DCM (1000 mL x 3), the organic layer was collected, and 2 SO 4 The product was dried over 400 °C and filtered, the filtrate was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10: 1 → 1: 100) to give 4,4,5,5-tetramethyl-2-((1R,2R)-2-pentylcyclopropyl)-1,3,2-dioxaborolane (14.0 g, 58.8 mmol, 65.9% yield) as a yellow oil.

[0287] 1 H NMR (400 MHz, deuterated chloroform): δ 1.42 - 1.37 (m, 2H), 1.32 - 1.26 (m,6H), 1.26 - 1.24 (m, 2H), 1.22 (s, 10H), 0.89 (br t, 4H), 0.67 (dt, 1H), 0.43- 0.33 (m, 1H), -0.42 (td, 1H)

[0288] 4-4. Synthesis of (1R,2R)-2-pentylcyclopropan-1-ol

[0289] In a 500 mL three-necked RBF, 4,4,5,5-tetramethyl-2-((1R,2R)-2-pentylcyclopropyl)-1,3,2-dioxaborolane (8.00 g, 33.6 mmol, 1.00 equiv) and THF (160 mL) were added, and NaOH (2.69 g, 67.2 mmol, 2.00 equiv) was slowly added to the mixture at 0°C, and H 2 O 2(7.87 g, 69.4 mmol, 30% purity, 2.07 equiv.), and the mixture was stirred at 25 °C for 16 hours. 2 SO 3 (2 equivalents) of ice water was poured into the reaction mixture to terminate the reaction, followed by extraction with DCM (30 mL x 3), and the organic layer was collected and stirred at room temperature for 2 hours. 2 SO 4 The filtrate was concentrated in vacuo and the concentrated residue was purified by silica gel column chromatography (petroleum ether:EtOAc=10:1→1:100) to obtain (1R,2R)-2-pentylcyclopropan-1-ol (2.50 g, 19.5 mmol, 58.1% yield) as a yellow oil.

[0290] 1 H NMR (400 MHz, deuterated chloroform): δ 3.20 (td, 1H), 1.84 - 1.67 (m, 1H), 1.41- 1.35 (m, 2H), 1.33 - 1.26 (m, 4H), 1.23 - 1.06 (m, 2H), 0.94 - 0.87 (m,4H), 0.68 (ddd, 1H), 0.31 (q, 1H)

[0291] 4-5. Synthesis of (1R,2R)-2-pentylcyclopropyl 8-bromooctanoate

[0292] In a 100 mL three-necked RBF, (1R,2R)-2-pentylcyclopropan-1-ol (2.50 g, 19.5 mmol, 1.00 equiv) was added with DCM (25 mL), and 8-bromooctanoic acid (5.22 g, 23.4 mmol, 1.20 equiv), EDCI (4.49 g, 23.4 mmol, 1.20 equiv), DMAP (476 mg, 3.90 mmol, 0.20 equiv) and Et 3 N (3.95g, 39.00mmol, 2.00 equivalents). The mixture was purged with nitrogen three times and stirred at 25 ° C for 16 hours in a nitrogen environment. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10: 1 → 1: 100) to obtain (1R, 2R) -2-pentylcyclopropyl 8-bromooctanoate (3.50g, 10.5mmol, 53.9% yield) as a yellow oil.

[0293] 1H NMR (400 MHz, deuterated chloroform): δ 3.85 - 3.79 (m, 1H), 3.41 (t, 2H),2.26 (t, 2H), 1.86 (quin, 2H), 1.65 - 1.58 (m, 2H), 1.46 - 1.38 (m, 4H), 1.36- 1.27 (m, 10H), 1.00 (ddd, 1H), 0.89 (br t, 3H), 0.78 (ddd, 1H), 0.52 (q,1H)

[0294] 4-6. Synthesis of cyclohexyl 8-bromooctanoate

[0295] In a 1000 mL three-necked RBF, 8-bromooctanoic acid (15.0 g, 67.2 mmol, 1.00 equiv) was placed with DCM (150 mL), trifluoroacetic anhydride (TFAA) (14.1 g, 67.2 mmol, 1.00 equiv) was added under nitrogen, and the mixture was stirred at 0 to 25 ° C for 2.5 hours under nitrogen. Cyclohexanol (33.7 g, 336 mmol, 35.1 mL, 5.00 equiv) was added to the mixture at 0 ° C, the mixture was stirred at 25 ° C for 16 hours under nitrogen, the reaction mixture was neutralized with 100 mL of water at 20 ° C, and extracted with 600 mL of DCM (200 mL x 3). The organic layer was collected and precipitated with Na 2 SO 4 The mixture was dried and filtered, and the filtrate was concentrated in vacuo and purified by silica gel column chromatography (SiO 2 , petroleum ether:EtOAc=100:1→10:1) to give cyclohexyl 8-bromooctanoate (7.10 g, 23.3 mmol, 34.6% yield) as a colorless oil.

[0296] 1 H NMR (400 MHz, deuterated chloroform): δ 4.77 (br d, 1H), 3.42 (t, 2H), 2.30 (t,2H), 1.93 - 1.82 (m, 4H), 1.74 (br dd, 2H), 1.67 - 1.55 (m, 4H), 1.47 - 1.31(m, 10H)

[0297] 4-7. Synthesis of cyclohexyl 8-((2-hydroxyethyl)amino)octanoate

[0298] In a 250 mL three-necked RBF, 8-bromooctanoic acid cyclohexyl ester (3.00 g, 9.83 mmol, 1.00 equiv), 2-aminoethan-1-ol (3.00 g, 49.1 mmol, 5.00 equiv), Na 2 CO 3 (1.04 g, 9.83 mmol, 1.00 equiv) and 1,4-dioxane (60 mL), the mixture was purged with nitrogen three times, and stirred at 80 ° C. for 16 hours under a nitrogen environment. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (DCM: MeOH = 100: 1 → 10: 1) to give 8-((2-hydroxyethyl) amino) cyclohexyl caprylate (2.80 g, 9.81 mmol, 99.8% yield) as a yellow oil.

[0299] 1 H NMR (400 MHz, deuterated chloroform): δ 4.77 (td, 1H), 3.75 - 3.67 (m, 2H), 2.91- 2.80 (m, 5H), 2.68 (t, 2H), 2.29 (t, 2H), 1.88 - 1.82 (m, 2H), 1.77 - 1.70(m, 2H), 1.66 - 1.60 (m, 2H), 1.55(br d, 2H), 1.46 - 1.28 (m, 12H)

[0300] 4-8. Synthesis of Compound D

[0301] In a 50mL three-necked RBF, 8-((2-hydroxyethyl)amino)octanoic acid cyclohexyl ester (942mg, 3.30mmol, 1.10 equivalents) and EtOH (10mL) were added, to which (1R, 2R)-2-pentylcyclopropyl 8-bromooctanoate (1.00g, 3.00mmol, 1.00 equivalents) and DIEA (1.94g, 15.00mmol, 5.00 equivalents) were added, and the mixture was stirred at 90°C for 16 hours under nitrogen. The reaction mixture was filtered, the filtrate was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (DCM:MeOH=10:1→1:100) to obtain the compound of formula D (0.15g, 279μmol, 9.30% yield) as a yellow oil.

[0302] 1 H NMR (400 MHz, deuterated chloroform): δ5.23 - 4.94 (m, 2H), 4.63 - 4.56 (m,1H), 4.12 (br s, 2H), 3.68 - 3.57 (m, 2H), 3.56 - 3.37 (m, 4H), 2.40 - 2.27(m, 2H), 2.04 (br d, 6H), 1.91 - 1.76 (m, 2H), 1.74 - 1.55 (m, 17H), 1.54 -1.46 (m, 2H), 1.43 - 1.25 (m, 5H), 1.21 (s, 2H), 0.90 (d, 6H)

[0303] Example 5

[0304] 5-1. Based on Figure 5 The synthetic scheme shown below prepares the compound of formula E.

[0305] [Formula E]

[0306]

[0307] 5-2. Synthesis of cyclopentadecanecarbonitrile

[0308] In a 2000 mL three-necked RBF, cyclopentadecanone (25.0 g, 111 mmol, 1.00 equiv), potassium 2-methylpropan-2-olate (25.0 g, 223 mmol, 2.00 equiv) and 2-methylpropan-2-ol (250 mL) were added together with THF (500 mL), the mixture was cooled to 0°C, and then 1-(isocyanomethylsulfonyl)-4-methyl-benzene (32.6 g, 167 mmol, 1.50 equiv) was slowly added thereto at 0°C for 1 hour, and the mixture was stirred at 20°C for 11 hours. 2 The reaction mixture was diluted with HO (400 mL), extracted with EtOAc (400 ml), and washed with H 2 O (400 mL), and the organic layer was washed with Na 2 SO 4 The filtrate was concentrated in vacuo and the concentrated residue was purified by silica gel column chromatography (petroleum ether:EtOAc=1:0→5:1) to give cyclopentadecanecarbonitrile (20 g, 84.96 mmol, 76.25% yield) as a colorless oil.

[0309] 1 H NMR (400 MHz, deuterated chloroform): δ 1.33 (br s, 20 H), 1.44 - 1.53 (m, 4 H), 1.67 (q, 4 H), 2.59 (quin, 1 H)

[0310] 5-3. Synthesis of cyclopentadecane carboxylic acid

[0311] In a 250mL three-necked RBF, cyclopentadecane carbonitrile (14.0g, 59.5mmol, 1.00 equivalent), KOH (6.00M, 69.4mL, 7.00 equivalent) and EtOH (70mL) were added, the mixture was purged with nitrogen three times, and stirred at 100°C for 16 hours under a nitrogen atmosphere. The reaction mixture was concentrated in vacuo, the concentrated residue was acidified to pH 4 with 4M aqueous hydrochloric acid solution (50mL), and extracted with EtOAc (50mLx3). The organic layer was collected and heated to 40 ℃ for 12 hours. 2 SO 4 The product was dried over 400 °C and filtered, and the filtrate was concentrated in vacuo. The concentrated residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10: 1 → 1: 1) to give cyclopentadecanecarboxylic acid (1.20 g, 4.72 mmol, 7.93% yield) as a white solid.

[0312] 1 H NMR (400 MHz, deuterated chloroform): δ ppm 1.27 - 1.46 (m, 24 H), 1.55 - 1.72 (m, 4 H), 2.44 (quin, 1 H), 10.15 - 11.75 (m, 1 H)

[0313] 5-4. Synthesis of 7-bromoheptylcyclopentadecanecarboxylate

[0314] In a 50 mL three-necked RBF, cyclopentadecanecarboxylic acid (1.20 g, 4.72 mmol, 1.10 equiv) and toluene (12 mL) were added, to which 7-bromoheptan-1-ol (837 mg, 4.29 mmol, 1.00 equiv) was added, followed by H 2 SO 4The mixture was purged with nitrogen three times and stirred for 16 hours at 120 ° C in a nitrogen environment. The reaction mixture was concentrated in vacuo and the concentrated residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10: 1) to obtain 7-bromoheptylcyclopentadecanate (0.90 g, 2.09 mmol, 48.6% yield) as a colorless oil.

[0315] 1 H NMR (400 MHz, deuterated chloroform): δ 1.29 - 1.40 (m, 30 H), 1.56 - 1.64 (m, 6H), 1.87 (quin, 2 H), 2.34 - 2.46 (m, 1 H), 3.41 (t, 2 H), 4.07 (t, 2 H)

[0316] 5-5. Synthesis of 7-bromoheptyl 4-pentylcyclohexane-1-carboxylate

[0317] In a 250 mL three-necked RBF, 4-pentylcyclohexane-1-carboxylic acid (4.47 g, 22.9 mmol, 1.00 eq) and toluene (50 mL) were added, to which 7-bromoheptan-1-ol (5.00 g, 25.2 mmol, 1.00 eq) was added, followed by H 2 SO 4 The mixture was purged with nitrogen three times and stirred for 16 hours at 120 ° C. under a nitrogen atmosphere. The reaction mixture was concentrated in vacuo and the concentrated residue was purified by silica gel column chromatography (petroleum ether: EtOAc = 10: 1) to give 7-bromoheptyl 4-pentylcyclohexane-1-carboxylate (7.00 g, 18.7 mmol, 81.4% yield) as a colorless oil.

[0318] 1 H NMR (400 MHz, deuterated chloroform): δ0.86 - 0.90 (m, 3 H), 1.18 - 1.32 (m, 10H), 1.32 - 1.40 (m, 5 H), 1.41 - 1.59 (m, 6 H), 1.59 - 1.68 (m, 2 H), 1.81 -1.90 (m, 2 H), 1.91 - 2.02 (m, 2 H), 2.46 - 2.54 (m, 1 H), 3.41 (t, 2 H),4.02 - 4.12 (m, 2 H)

[0319] 5-6. Synthesis of 7-((2-hydroxyethyl)amino)heptyl 4-pentylcyclohexane-1-carboxylate

[0320] In a 250mL three-necked RBF, 7-bromoheptyl 4-pentylcyclohexane-1-carboxylate (3.00g, 7.99mmol, 1.00 equivalent), 2-aminoethan-1-ol (2.44g, 40.0mmol, 2.41mL, 5.00 equivalent) and 1,4-dioxane (90mL) were added, the mixture was purged with nitrogen three times, and stirred at 100°C for 16 hours under a nitrogen environment. The reaction mixture was concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (DCM: MeOH = 10: 1) to give 7-((2-hydroxyethyl)amino)heptyl 4-pentylcyclohexane-1-carboxylate (1.70g, 4.78mmol, 59.8% yield) as a yellow oil.

[0321] 1 H NMR (400 MHz, deuterated chloroform): δ 0.88 (t, 3 H), 1.19 - 1.28 (m, 8 H), 1.34 (br s, 6 H), 1.41 - 1.71 (m, 8 H), 1.84 - 2.01 (m, 6 H), 2.42 - 2.54 (m,1 H), 2.63 (t, 2 H), 2.74 - 2.86 (m, 2 H), 3.50 - 3.79 (m, 2 H), 3.99 - 4.13 (m, 2 H)

[0322] 5-7. Synthesis of Compound E

[0323] In a 50mL three-necked RBF, 7-((2-hydroxyethyl)amino)heptyl 4-pentylcyclohexane-1-carboxylate (90mg, 209μmol, 1.00 equivalent) and 1,4-dioxane (5mL) were added, 7-bromoheptylcyclopentadecanecarboxylate (74.2mg, 209μmol, 1.00 equivalent) and DIEA (135mg, 1.04mmol, 5.00 equivalent) were added thereto, the mixture was purged with nitrogen three times, and stirred at 105°C for 16 hours in a nitrogen environment. The reaction mixture was cooled to 25°C, water (10mL) was added to quench the reaction, and the mixture was extracted with 30mL of EtOAc (10mLx3). The organic layer was collected and precipitated under Na 2 SO 4 It was dried over 400 °C and filtered, and the filtrate was concentrated in vacuo. The concentrated residue was purified by silica gel column chromatography (DCM: MeOH = 10: 1) to obtain the compound of formula E (70 mg, 99.1 μmol, 47.5% yield) as a yellow oil.

[0324] 1 H NMR (400 MHz, deuterated chloroform): δ 0.88 (t, 3 H), 1.23 - 1.40 (m, 46 H), 1.45 - 1.50 (m, 4 H), 1.52 - 1.66 (m, 12 H), 1.72 - 2.05 (m, 4 H), 2.37 -2.55 (m, 6 H), 2.63 (br t, 2 H), 3.57 (br t, 2 H), 4.02 - 4.09 (m, 4 H)

[0325] Example 6

[0326] 6-1. According to Figure 6 The synthetic scheme shown prepares the compound of formula F below.

[0327] [Formula F]

[0328]

[0329] 6-2. Synthesis of 10-cyclopentadecanyl bromodecanate

[0330] The synthesis was carried out by the method described in Example 3-2.

[0331] 6-3. Synthesis of 4-propylcyclohexyl 6-bromohexanoate

[0332] The synthesis was carried out by the method described in Examples 2-3.

[0333] 6-4. Synthesis of 4-propylcyclohexyl 6-((2-hydroxyethyl)amino)hexanoate

[0334] The synthesis was carried out by the method described in Examples 2-4.

[0335] 6-5. Synthesis of Compound F

[0336] In 50mL three-necked RBF, 6-((2-hydroxyethyl)amino)hexanoic acid 4-propylcyclohexyl ester (0.10g, 1.00 equivalent), 10-bromodecane acid cyclopentadecane ester (153mg, 1.00 equivalent) and DIEA (44mg, 1.00 equivalent) were added together with EtOH (5mL), the mixture was purged with nitrogen three times, and stirred at 95°C for 16 hours in a nitrogen environment. The reaction mixture was cooled to room temperature and concentrated in vacuo, and the concentrated residue was purified by silica gel column chromatography (DCM:MeOH=100:1→10:1), to obtain formula F compound (70mg, 30.9% yield), as a yellow oil.

[0337] 1 H NMR (400 MHz, deuterated chloroform): δ 5.03 - 4.62 (m, 2H), 4.01 - 3.72 (m,2H), 3.10 - 2.75 (m, 6H), 2.35 - 2.25 (m, 4H), 1.95 (br d, 1H), 1.59 (br s,18H), 1.40 - 1.17 (m, 42H), 1.05 - 0.96 (m, 1H), 0.92 - 0.87 (m, 3H)

[0338] Example 7

[0339] According to the invention, except that 3-pentylcyclopentane-1-ol is used instead of 4-pentylcyclohexane-1-ol. Figure 7 The compound of formula G was prepared in the same manner as in Example 1.

[0340] [Formula G]

[0341]

[0342] 1 H NMR (400 MHz, deuterated chloroform): δ5.03 - 4.77 (m, 2H), 4.09 - 3.88 (m,2H), 3.04 - 2.88 (br, 6H), 2.41 - 2.31 (m, 4H), 1.89 - 1.56 (br, 14H), 1.51 -0.96 (br, 53H), 0.88 (t, 3H)

[0343] Example 8

[0344] According to the invention, except that 6-bromohexanoic acid was used instead of 8-bromooctanoic acid and 3-pentylcyclopentane-1-ol was used instead of 4-pentylcyclohexane-1-ol. Figure 8 The compound of formula H was prepared in the same manner as in Example 1.

[0345] [Formula H]

[0346]

[0347] 1 H NMR (400 MHz, deuterated chloroform): δ 4.68 (m, 1H), 4.48 (m, 1H), 3.91 (m,2H), 3.02 - 2.89 (br, 6H), 2.35 - 2.29 (m, 4H), 1.92 - 1.61 (m, 14H), 1.58 -1.01 (m, 45H), 0.90 (t, 3H)

[0348] Example 9

[0349] According to the invention, except that 10-bromodecanoic acid was used instead of 8-bromooctanoic acid and 3-pentylcyclopentane-1-ol was used instead of 4-pentylcyclohexane-1-ol. Figure 9 The following compound of formula I was prepared in the same manner as in Example 1.

[0350] [Formula I]

[0351]

[0352] 1 H NMR (400 MHz, deuterated chloroform): δ4.70 (m, 1H), 4.47 (m, 1H), 3.90 (m,2H), 2.99 - 2.79 (br, 6H), 2.31 - 2.26 (m, 4H), 1.92 - 1.61 (m, 14H), 1.58 -1.01 (m, 61H), 0.85 (t, 3H)

[0353] Example 10

[0354] According to the invention, cyclopentadecanol was replaced with cyclohexanol and 4-pentylcyclohexane-1-ol was replaced with 3-pentylcyclopentane-1-ol. Figure 10 The compound of formula J was prepared in the same manner as in Example 1.

[0355] [Formula J]

[0356]

[0357] 1 H NMR (400 MHz, deuterated chloroform): δ 4.68 - 4.61 (m, 2H), 3.94 (m, 2H), 3.12- 2.94 (br, 6H), 2.44 (m, 4H), 1.91 - 1.10 (m 45H), 0.88 (t, 3H)

[0358] Embodiment 11

[0359] Except that aminocyclopentadecane is used instead of cyclopentadecanol, according to Figure 11 The compound of formula K was prepared in the same manner as in Example 1.

[0360] [Formula K]

[0361]

[0362] 1 H NMR (400 MHz, deuterated chloroform): δ 8.10 (br, 1H), 4.67 - 4.63 (m, 1H), 3.95- 3.88 (m, 2H), 3.64 - 3.59 (m, 1H), 3.11 - 2.89 (br, 6H), 2.31 - 2.19 (m,4H), 1.95 - 1.25 (m, 65H), 0.88 (t, 3H)

[0363] Example 12

[0364] According to the invention, except that 1-amino-4-pentylcyclohexane is used instead of 4-pentylcyclohexane-1-ol. Figure 12 The compound of formula L was prepared in the same manner as in Example 1.

[0365] [Formula L]

[0366]

[0367] 1 H NMR (400 MHz, deuterated chloroform): δ 8.12 (br, 1H), 4.50 - 4.43 (m, 1H), 3.88- 3.76 (m, 2H), 3.49 - 3.40 (m, 2H), 3.36 - 3.33 (m, 1H), 3.03 - 2.90 (br,6H), 2.31 - 2.90 (m, 4H), 1.92 - 1.20 (m, 65H), 0.90 (t, 3H)

[0368] Embodiment 13

[0369] According to the invention, except that aminocyclopentadecane is used instead of cyclopentadecanol and 1-amino-4-pentylcyclohexane is used instead of 4-pentylcyclohexane-1-ol. Figure 13 The compound of formula M was prepared in the same manner as in Example 1.

[0370] [Formula M]

[0371]

[0372] 1 H NMR (400 MHz, deuterated chloroform): δ 8.10 (br, 2H), 3.84 - 3.78 (m, 2H), 3.69- 3.61 (m, 2H), 3.11 - 2.98 (br, 6H), 2.33 - 2.19 (m, 4H), 1.90 - 1.22 (m,65H), 0.91 (t, 3H)

[0373] Embodiment 14

[0374] Except that (1R,2R)-2-pentylcyclopropan-1-ol prepared in Example 4-4 was used instead of 4-pentylcyclohexane-1-ol, Figure 14 The compound of formula N was prepared in the same manner as in Example 1.

[0375] [Formula N]

[0376]

[0377] 1 H NMR (400 MHz, deuterated chloroform): δ 4.56 - 4.45 (m, 1H), 3.56 - 3.52 (m,2H), 3.42 (br, 1H), 3.11 - 3.05 (br, 4H), 2.77 - 2.70 (br, 2H), 2.44 - 2.38(m, 4H), 1.91 - 1.21 (m, 57H), 0.9 (t, 3H), 0.62 - 0.55 (m, 2H)

[0378] Embodiment 15

[0379] In addition to using methylamine (CH 3 -NH 2 ) instead of 2-aminoethane-1-ol, according to Figure 15 The compound of formula O was prepared in the same manner as in Example 1.

[0380] [Formula O]

[0381]

[0382] 1 H NMR (400 MHz, deuterated chloroform): δ 4.41 - 4.38 (m, 1H), 4.22 - 4.18 (m,1H), 3.00 - 2.82 (m, 4H), 2.75 (s, 3H), 2.42 - 2.39 (m, 4H), 1.92 - 1.20 (m,65H), 0.90 (t, 3H)

[0383] Example 16

[0384] In addition to using methylamine (CH 3 -NH 2 ) instead of 2-aminoethan-1-ol, 10-bromodecanoic acid instead of 8-bromooctanoic acid, and 4-heptylcyclohexane-1-ol instead of 4-pentylcyclohexane-1-ol, according to Figure 16 The compound of formula P was prepared in the same manner as in Example 1.

[0385] [Formula P]

[0386]

[0387] 1 H NMR (400 MHz, deuterated chloroform): δ 4.43 - 4.40 (m, 1H), 4.21 - 4.18 (m,1H), 3.12 - 2.92 (m, 4H), 2.80 (s, 3H), 2.42 - 2.39 (m, 4H), 1.93 - 1.19 (m,73H), 0.90 (t, 3H)

[0388] Embodiment 17

[0389] Except that cyclodecyl alcohol is used instead of cyclopentadecanol, according to Figure 17 The compound of the following formula Q was prepared in the same manner as in Example 1.

[0390] [Formula Q]

[0391]

[0392] 1 H NMR (400 MHz, deuterated chloroform): δ 4.48 - 4.41 (m, 1H), 4.19 - 4.17 (m,1H), 3.54 - 3.49 (m, 2H), 3.02 - 2.92 (br, 4H), 2.78 - 2.71 (br, 2H), 2.35 -2.31 (m, 4H), 1.91 - 1.19 (m, 55H), 0.88 (t, 3H)

[0393] Embodiment 18

[0394] In addition to using cyclodecyl alcohol instead of cyclopentadecanol and using methylamine (CH 3 -NH 2 ) instead of 2-aminoethane-1-ol, according to Figure 18 The compound of formula R was prepared in the same manner as in Example 1.

[0395] [Formula R]

[0396]

[0397] 1 H NMR (400 MHz, deuterated chloroform): δ4.48 - 4.41 (m, 1H), 4.19 - 4.17 (m,1H), 3.01 - 2.93 (br, 4H), 2.75 (br, 3H), 2.34 - 2.30 (m, 4H), 1.90 - 1.21(m, 55H), 0.90 (t, 3H)

[0398] [Preparation Example of Composition for Drug Delivery]

[0399] 1. Preparation of Raw Materials

[0400] According to the following table, dissolve the substances required for preparing the preparation in various dilution solvents and prepare them to the required concentrations. When dissolving, keep the substances at room temperature and then add the solvent to dissolve them.

[0401]

[0402] 2. Mixing of Raw Materials

[0403] Mix the required amounts of raw materials to satisfy an N / P ratio (amine group of lipid / phosphate group of mRNA) of 6, with various compounds of Formula A to Formula F: DOPE:cholesterol:DMG-PEG = 50:10:38.5:1.5. Add ethanol to the ethanol layer to keep the total molecular amount of all raw materials within 12.5 mM, and keep the volume ratio of the aqueous phase and the ethanol phase at 3:1 and mix. After mixing, to reduce the total ethanol content, perform buffer exchange as follows: Concentrate the mixed solution to 1 / 3 level by centrifuging it at 4000 rpm using an Amicon-Ultra tube filter (Merk Millipore, UFC505096 or UFC805024, pore size: 50K, volume: 15 mL), then dilute it with three times the volume of PBS and centrifuge to concentrate. Repeat this process six times to exchange the buffer.

[0404] The more specific procedure is as follows:

[0405] 1) Prepare two autoclaved tubes (tube (A) and tube (B)).

[0406] 2) In tube (A), sequentially add the molar amounts of various compounds of Formula A to Formula F, DOPE, cholesterol, and DMG-PEG calculated according to the experimental conditions, and mix by vortexing.

[0407] 3) Add ethanol to the ethanol phase if necessary to keep the total molecular amount of all raw materials within 12.5 mM.

[0408] 4) In tube (B), mix the mRNA and 20 mM sodium acetate buffer (pH 4.6) (= prepared by diluting to 20 mM with 3 M sodium acetate buffer and titrating to pH 4.6 using 1 M HCl). At the same time, calculate the ratio and add so that the total volume of the aqueous phase is three times that of the ethanol phase.

[0409] 5) Use a microfluidics (Ignite, Precision Nanosystem) device to mix tube (A) and tube (B). The microfluidic operating conditions are a flow rate ratio (FRR) of C:R = 3:1 and a total flow rate (TRR) of 12 mL / min.

[0410] 6) Concentrate the mixture obtained in step 5) to 1 / 3 level by centrifugation at 4000 rpm using an Amicon-Ultra tube filter (50K), then dilute 3-fold with PBS and concentrate by centrifugation. Repeat this process 6 times to concentrate to the desired volume.

[0411] 3. Evaluation of formulation performance

[0412] 1) For the prepared formulation, use a particle size analyzer (dynamic light scattering, DLS) to confirm the particle characteristics (i.e., ζ average particle size (Z-average), polydispersity index (PDI), and ζ potential), and the results are shown in Table 1 below.

[0413] 2) For the prepared formulation, confirm the mRNA encapsulation efficiency by the Ribo-green assay, and the results are shown in Table 1 below.

[0414] [Table 1]

[0415]

Claims

1. A lipid having the structure shown in the following formula (1): [Formula 1] Wherein in the above formula (1), M 1 and M 2 each independently represents a divalent linking group, R 1 and R 2 each independently represents a substituted or unsubstituted carbocyclic group or heterocyclic group, R 3 is a hydrogen atom or a substituted or unsubstituted organic group optionally containing one or more heteroatoms, R 4 to R 7 each independently represents a hydrogen atom, or a substituted or unsubstituted saturated or unsaturated hydrocarbon group, and a and b are each independently an integer from 1 to 20.

2. The lipid according to claim 1, wherein M 1 and M 2 each independently selected from the group consisting of: -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, -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), wherein M' is a direct bond, C 1-13 alkylene or C 2-13 alkenylene, and each R' is independently selected from the group consisting of a hydrogen atom, C 1-18 alkyl, and C 2-18 alkenyl, R 1 and R 2 each independently selected from the group consisting of: C 3-20 cycloalkyl, C 3-20 cycloalkenyl, C 6-20 aryl, C 3-20 heterocycloalkyl, C 3-20 heterocycloalkenyl and C 3-20 heteroaryl, each of said groups being independently unsubstituted or substituted by C 1-18 alkyl or C 2-18 alkenyl, R 3 selected from the group consisting of a hydrogen atom, a substituted or unsubstituted C 1-6 alkyl group, a substituted or unsubstituted C 3-6 carbocyclic group, -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR and -CQ(R) 2 wherein each R is independently selected from the group consisting of a hydrogen atom, a C 1-3 alkyl group and a C 2-3 alkenyl group; Q is selected from the group consisting of a carbocyclic group, a heterocyclic group, -OR, -O(CH 2 ) n N(R) 2 , -C(O)OR, -OC(O)R, -CX 3 , -CX 2 H, -CXH 2 , -CN, -N(R) 2 , -C(O)N(R) 2 , -N(R)C(O)R, -N(R)S(O) 2 R, -N(R)C(O)N(R) 2 , -N(R)C(S)N(R) 2 , -N(R)R 12 , N(R)S(O)2R 12 , -O(CH 2 ) n OR, -N(R)C(=NR 13 )N(R) 2 , -N(R)C(=CHR 13 )N(R) 2 , -OC(O)N(R) 2 , -N(R)C(O)OR, -N(OR)C(O)R, -N(OR)S(O) 2 R, -N(OR)C(O)OR, -N(OR)C(O)N(R) 2 , -N(OR)C(S)N(R) 2 , -N(OR)C(=NR 13 )N(R) 2 , -N(OR)C(=CHR 13 )N(R) 2 , -C(=NR 13 )N(R) 2 , -C(=NR 13 )R, -C(O)N(R)OR and -C(R)N(R) 2 a group consisting of C(O)OR; wherein n is each independently an integer from 1 to 5; R 12 is selected from the group consisting of C 3-6 carbocyclic groups and heterocyclic groups; R 13 is selected from the group consisting of H, CN, NO 2 , C 1-6 alkyl, -OR, -S(O) 2 R, -S(O) 2 N(R) 2、 C 2-6 alkenyl, C 3-6 carbocyclic groups and heterocyclic groups; each R is independently selected from the group consisting of a hydrogen atom, C 1-3 alkyl and C 2-3 alkenyl; each X is independently selected from the group consisting of F, Cl, Br, and I, provided that when R 3 is -(CH 2 ) n Q, -(CH 2 ) n CHQR, -CHQR or -CQ(R) 2 then (i) when n is 1, 2, 3, 4, or 5, then Q is not -N(R) 2 , or (ii) when n is 1 or 2, then Q is not a 5-, 6-, or 7-membered heterocycloalkyl, R 4 to R 7 each independently selected from the group consisting of a hydrogen atom, C 1-3 alkyl, and C 2-3 alkenyl, and a and b are each independently an integer from 1 to 15.

3. The lipid according to claim 2, wherein M 1 and M 2 each independently selected from the group consisting of -C(O)O-, -OC(O)-, -OC(O)-M'-C(O)O-, -C(O)N(R')-, -N(R')C(O)-, and -C(O)-, where M' and R' are the same as those defined in claim 2 R 1 and R 2 each independently selected from the group consisting of C 3-20 cycloalkyl and C 3-20 heterocycloalkyl, wherein C 3-20 cycloalkyl and C 3-20 heterocycloalkyl are each independently unsubstituted or substituted by C 1-18 alkyl or C 2-18 alkenyl, R 3 selected from the group consisting of a hydrogen atom, a substituted or unsubstituted C 1-6 alkyl group, and a substituted or unsubstituted C 3-6 carbocyclic group, R 4 to R 7 each independently represents a hydrogen atom or a C 1-3 alkyl group, and a and b are each independently an integer from 3 to 13.

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

5. The lipid according to claim 4, wherein the lipid is a lipid having a structure selected from the group consisting of the structures of the following formulas A to R:

6. A method for preparing a lipid having the structure shown in formula (1'), the method comprising the following steps: (1) Reacting a compound of formula (a) with a compound of formula (b) to obtain a compound of formula (c); and (2) Reacting the compound of formula (c) with a compound of formula (d): [Formula a] R 1 -OH [Formula b] [Formula c] [Formula d] R 3 -NH 2 [Formula 1'] Wherein, M 1 、R 1 、R 3 、R 4 、R 5 and a are the same as those defined in claim 1, and X is each independently selected from the group consisting of F, Cl, Br, and I.

7. A method for preparing a lipid having the structure shown in formula (1), the method 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 the compound of formula (c') with a compound of formula (d) to obtain a compound of formula (e); and (3) Reacting the compound of formula (e) with the compound of formula (c) obtained in claim 6: [Formula a'] HO-R 2 [Formula b'] [Formula c'] [Formula d] R 3 -NH 2 [Formula e] [Formula c] [Formula 1] Wherein, M 1 、M 2 、R 1 to R 7 、a and b are the same as those defined in claim 1, and X is each independently selected from the group consisting of F, Cl, Br, and I.

8. A method for preparing a lipid having the structure shown in formula (1'), the method comprising the following steps: (1) Reacting a compound of formula (i) with a compound of formula (ii) to obtain a compound of formula (iii); and (2) Reacting the compound of formula (iii) with a compound of formula (iv): [Formula i] R 1 -M 1 -H [Formula ii] [Formula iii] [Formula iv] R 3 -NH 2 [Formula 1'] Wherein, M 1 、R 1 、R 3 、R 4 、R 5 and a are the same as those defined in claim 1, and X is each independently selected from the group consisting of F, Cl, Br, and I.

9. A method for preparing a lipid having the structure shown in formula (1), the method 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 the compound of formula (iii') with a compound of formula (iv) to obtain a compound of formula (v); and (3) Reacting the compound of formula (v) with the compound of formula (iii) obtained in claim 8: [Formula i'] H-M 2 -R 2 [Formula ii'] [Formula iii'] [Formula iv] R 3 -NH 2 [Formula v] [Formula iii] [Formula 1] Wherein, M 1 、M 2 、R 1 to R 7 、a and b are the same as those defined in claim 1, and X is each independently selected from the group consisting of F, Cl, Br, and I.

10. A composition for drug delivery, which comprises the lipid according to any one of claims 1 to 5.

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