Lipids and lipid-like compounds for the delivery of therapeutic lipid nanoparticles (LNPs)

Abstract

This disclosure relates to lipid-based compositions and methods for administering related therapeutic agents. In particular, this disclosure relates to lipid-like substituted aryl and / or heteroaryl compounds, substituted piperazines, and / or other aryl and / or heteroaryl lipid compounds as LNP delivery materials that can be incorporated into lipid-based compositions to increase the delivery efficiency of therapeutic agents to target tissues. In one embodiment, this disclosure provides lipid particles comprising any of the above compounds.

Description

[Technical Field]

[0001] Cross-references to related applications This application is a U.S. Provisional Patent Application No. 63 / 494,871, filed on April 7, 2023, under Section 119(e) of the U.S. Patent Act, for the invention titled "LIPID-LIKE SUBSTITUTED ARYL AND HETEROARYL COMPOUNDS AS LIPID NANOPARTICLE (LNP) DELIVERY MATERIALS AND THERAPEUTIC USES THEREOF", a U.S. Provisional Patent Application No. 63 / 494,872, filed on April 7, 2023, for the invention titled "SUBSTITUTED PIPERAZINE IONIZABLE LIPIDS USEFUL FOR THERAPEUTIC LIPID NANOPARTICLE (LNP) DELIVERY", and a U.S. Provisional Patent Application No. 63 / 494,872, filed on January 24, 2024, for the invention titled "SUBSTITUTED PIPERAZINE IONIZABLE LIPIDS USEFUL FOR THERAPEUTIC LIPID NANOPARTICLE We claim priority to U.S. Provisional Patent Application No. 63 / 624,550 for “(LNP) DELIVERY”, U.S. Provisional Patent Application No. 63 / 501,566, filed on 11 May 2023, for the invention “ARYL AND HETEROARYL LIPID COMPOUNDS, LIPID NANOPARTICLE (LNP) FORMULATIONS, AND THERAPEUTIC USES THEREOF”, and U.S. Provisional Patent Application No. 63 / 624,073, filed on 23 January 2024, for the invention “ARYL AND HETEROARYL LIPID COMPOUNDS, LIPID NANOPARTICLE (LNP) FORMULATIONS, AND THERAPEUTIC USES THEREOF”. The entirety of the aforementioned patent applications is incorporated herein by reference.

[0002] field This disclosure relates to lipid-based compositions. In particular, this disclosure relates to (1) ionizable lipid-like substituted aryl and heteroaryl compounds as lipid nanoparticle (LNP) delivery materials, (2) substituted piperazines as ionizable lipids, and (3) other aryl and heteroaryl lipid compounds that can be incorporated into lipid-based compositions to increase the delivery efficiency of therapeutic agents to one or more tissues of interest. [Background technology]

[0003] background Nucleic acid therapy holds great potential for treating diseases at the individual target gene level. However, a safe and effective delivery system is essential to fully realize the potential of nucleic acid therapeutics. Non-specific delivery of nucleic acid therapeutics to all organs and tissues often leads to off-site (untargeted and / or non-targeted) effects and toxicity. Preferred delivery of nucleic acid therapeutics to target organs or tissues where specific effects are desired is an ongoing goal of drug delivery in general, particularly the delivery of nucleic acid-based drugs. Unfortunately, there are virtually no nanoparticle delivery system options that can target specific tissues without introducing ligand-based targeting strategies (i.e., active targeting). Therefore, in the art, there is an unmet need for delivery modes that can achieve tissue-specific delivery of nucleic acid cargoes based solely on the structural components of such delivery modes (e.g., via non-active targeting modes). [Overview of the project] [Means for solving the problem]

[0004] Brief Overview This disclosure provides a variety of novel lipids and lipid-like compounds.

[0005] Certain aspects of this disclosure provide novel ionizable lipid-like chemicals (e.g., lipid-like substituted aryl and heteroaryl compounds represented by formula I) designed and synthesized according to the techniques disclosed herein. These novel ionizable lipid-like substituted aryl and heteroaryl compounds have been shown to be formulated into lipid nanoparticles (LNPs) that provide stable and efficient LNP formulations comparable to or better than conventional benchmark lipids.

[0006] Some aspects of this disclosure provide novel ionizable lipid-like chemicals (e.g., ionizable substituted piperazine lipids represented by formula VII) designed and synthesized according to the techniques disclosed herein. These novel ionizable lipid-like chemicals have also been shown to be formulated into lipid nanoparticles (LNPs) to provide stable and efficient LNP formulations that are comparable to or better than conventional benchmark lipids.

[0007] Further aspects of this disclosure provide novel ionizable lipid-like chemicals (e.g., aryl and heteroaryl lipid compounds represented by formula IX) designed and synthesized according to the techniques disclosed herein. These novel aryl and heteroaryl lipid compounds have also been shown to be formulated into lipid nanoparticles (LNPs) to provide stable and efficient LNP formulations that are comparable to or better than benchmark lipids of the prior art.

[0008] This disclosure is at least in part based on the discovery that lipid-like substituted aryl and heteroaryl compounds can be used to form novel ionizable lipids having properties advantageous when used in lipid particles for in vivo delivery of therapeutic agents. In particular, the techniques herein provide lipid-based nanoparticle compositions and formulations that can specifically target cargo moieties (e.g., nucleic acid cargoes) to specific tissues of interest without requiring ligand-based targeting strategies. The lipid-like substituted aryl and heteroaryl compounds disclosed herein may include the following general structures.

[0009] In one embodiment, the present disclosure relates to a compound of formula I: [ka] or its salt or isomer [in the formula, X is either CH or N. a and b are independently between 2 and 5. m1, m2, m3, and m4 are independently between 4 and 10. E1, E2, E3, and E4 are independently -O(CO)O-, -(CO)O-, or -O(CO)-. T1, T2, T3, and T4 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R1 and R2 are independently H, or C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. R3 is independently H, or C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, [ka] And, G is O, -(CO)NR3-, -NR3(CO)-, -(CO)O-, or -CH2(CO)NR3-, L is a C1-C4 alkyl group that is bonded or substituted as needed. Z is CH or N, R4 and R5 are independently H, C1-C3 alkyl, C2-C3 alkenyl or C2-C3 alkynyl, or [ka] Is it, Alternatively, R4 or R5 may form a 3-7 membered ring containing 0-2 heteroatoms as needed, or R4 and R5 may form a substituted 3-7 membered ring together with L.

[0010] In some embodiments, one or more sets of R1 and R2, a and b, m1 to m4, E1 to E4, and T1 to T4 are the same.

[0011] In some embodiments, R1 and R2, a and b, m1 to m4, E1 to E4, and one or more of T1 to T4 are different.

[0012] In some embodiments, R1 and R2, a and b, m1 to m4, E1 to E4, and T1 to T4 are all the same.

[0013] In some embodiments, R1 and R2 are H.

[0014] In some embodiments, a and b are independently 3, 4, or 5.

[0015] In some embodiments, a and b are 3.

[0016] In some embodiments, m1, m2, m3, and m4 are independently 6, 7, 8, or 9.

[0017] In some embodiments, m1, m2, m3, and m4 are 8.

[0018] In some embodiments, E1, E2, E3, and E4 are -(CO)O-.

[0019] In some embodiments, E1, E2, E3, and E4 are -O(CO)- or -O(CO)O-.

[0020] In some embodiments, T1, T2, T3, or T4 are replaced as needed by C5~C 18 Alkyl, C5~C 18 Alkenyl, and C5~C 18Independently selected from the group consisting of alkynyl, and optionally, T1, T2, T3, and T4 are each C5-C 18 alkyl, C5-C 18 alkenyl, and C5-C 18 independently selected from the group consisting of alkynyl.

[0021] In some embodiments, T1, T2, T3, or T4 is independently selected from the group consisting of C5-C 12 alkyl, C5-C 12 alkenyl, and C5-C 12 independently selected from the group consisting of alkynyl, and optionally, T1, T2, T3, and T4 are each C5-C 12 alkyl, C5-C 12 alkenyl, and C5-C 12 independently selected from the group consisting of alkynyl.

[0022] In some embodiments, T1, T2, T3, or T4 is independently selected from the group consisting of C5-C 10 alkyl, C5-C 10 alkenyl, and C5-C 10 independently selected from the group consisting of alkynyl, and optionally, T1, T2, T3, and T4 are each C5-C 10 alkyl, C5-C 10 alkenyl, and C5-C 10 independently selected from the group consisting of alkynyl.

[0023] In some embodiments, T1, T2, T3, or T4 is independently selected from the group consisting of C5-C8 alkyl, C5-C8 alkenyl, and C5-C8 alkynyl, and optionally, T1, T2, T3, and T4 are each independently selected from the group consisting of C5-C8 alkyl, C5-C8 alkenyl, and C5-C8 alkynyl.

[0024] In some embodiments, T1, T2, T3, or T4 are independently selected from the group consisting of C6-C8 alkyl, C6-C8 alkenyl, and C6-C8 alkynyl, each of which is optionally substituted.

[0025] In some embodiments, T1, T2, T3, or T4 are independently selected from the group consisting of C7 or C8 alkyl, C7 or C8 alkenyl, and C7 or C8 alkynyl, each of which is optionally substituted.

[0026] In some embodiments, G is O, -(CO)NR 3 -, and -NR 3 It is (CO)-.

[0027] In some embodiments, R3 is H or methyl.

[0028] In some embodiments, R3 is [ka] That is the case.

[0029] In some embodiments, L is a bond.

[0030] In some embodiments, L is a C1 alkyl group.

[0031] In some embodiments, L is a C2 alkyl group.

[0032] In some embodiments, L is a C3 alkyl group.

[0033] In some embodiments, L is a C4 alkyl group.

[0034] In some embodiments, R4, R5, or R4 and R5 are either absent or H.

[0035] In some embodiments, R4, R5, or R4 and R5 are [ka] That is the case.

[0036] In some embodiments, R 4 and R 5 These are independently C1, C2, or C3 alkyl groups.

[0037] In some embodiments, R 4 and R 5 It forms a six-membered ring which may be substituted as needed and may contain one or two heteroatoms.

[0038] In some embodiments, at least one of the one or two heteroatoms is nitrogen (N).

[0039] In some embodiments, at least one hydrogen atom of the six-membered ring is substituted with a methyl group.

[0040] In some embodiments, at least one atom is a nitrogen atom.

[0041] In some embodiments, the 6-membered ring contains 1, 2, or 3 double bonds, and optionally the 6-membered ring contains 3 double bonds.

[0042] In some embodiments, R4 and R5 are methyl groups.

[0043] In some embodiments, R4 and R5 form a five-membered ring which is optionally substituted and contains one or two heteroatoms and one or two double bonds.

[0044] In some embodiments, at least one of the one or two heteroatoms is nitrogen (N).

[0045] In some embodiments, T1, T2, T3, and T4 are alkyl groups selected independently from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as needed.

[0046] In some embodiments, T1, T2, T3, and T4 are octane or tridecane.

[0047] In some embodiments, T1, T2, T3, and T4 are independently substituted as needed with buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, de The alkenyl is selected from the group consisting of ca-2-ene, deca-3-ene, deca-4-ene, deca-5-ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and alkenyl groups containing two or more double bonds.

[0048] In some embodiments, T1, T2, T3, and T4 are independently substituted as needed with buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, de The alkynyl is selected from the group consisting of k-2-ine, deca-3-ine, deca-4-ine, deca-5-ine, deca-6-ine, undeca-1-ine, undeca-2-ine, undeca-3-ine, undeca-4-ine, undeca-5-ine, undeca-6-ine, undeca-7-ine, dodeca-1-ine, dodeca-2-ine, dodeca-3-ine, dodeca-4-ine, dodeca-5-ine, dodeca-6-ine, dodeca-8-ine, and alkynyl groups containing two or more triple bonds.

[0049] In some embodiments, X is N.

[0050] In one embodiment, the present disclosure relates to a compound of formula II: [ka] or its salt or isomer [in the formula, X is either CH or N. L1, L2, and L3 are independently -O- and -(CO)NR x -, -NR x (CO)-, -(CO)O-, -CH2(CO)NR x - and if A is CH, then all of L1, L2, and L3 are -(CO)NR x -That is not the case, R x These are H, C1-C6 alkyl, or C3-C6 cycloalkyl, G1 and G2 are given by Equation III: [ka] or a salt or isomer thereof, where, n1 is 3, 4, 5, 6, 7, 8, 9, or 10. L4 is -(CO)O- or -O(CO)-, R6 is branched from C5 to C 20 It is alkyl, G3 is equation (III), equation (IV), equation (V), or equation (VI): [ka] And, R7 and R8 are independently functionalized C1-C5 alkyl groups as needed. n2, n3, and n4 are independently 0, 1, 2, or 3. X1 is C, N, or O. R9 and R 10 [These independently provide H or, optionally, a functionalized C1-C5 alkyl group.]

[0051] In one embodiment, the present disclosure provides compounds selected from the group consisting of the following: [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka] [ka]

[0052] In one embodiment, the present disclosure relates to a lipid of formula I: [ka] or its salt or isomer [in the formula, X is either CH or N. a and b are independently between 2 and 5. m1, m2, m3, and m4 are independently between 4 and 10. E1, E2, E3, and E4 are independently -O(CO)O-, -(CO)O-, or -O(CO)-. T1, T2, T3, and T4 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R1 and R2 are independently H, or C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. R3 is independently H, or C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, [ka] And, G is O, -(CO)NR3-, -NR3(CO)-, -(CO)O-, or -CH2(CO)NR3-, L is a C1-C4 alkyl group that is bonded or substituted as needed. Z is CH or N, R4 and R5 are independently H, C1-C3 alkyl, C2-C3 alkenyl or C2-C3 alkynyl, or [ka] Is it, Alternatively, R4 or R5 may form a 3-7 membered ring containing 0-2 heteroatoms as needed, or R4 and R5 may form a substituted 3-7 membered ring together with L. The present invention provides a pharmaceutical composition containing the following:

[0053] In some embodiments, one or more sets of R1 and R2, a and b, m1 to m4, E1 to E4, and T1 to T4 are the same.

[0054] In some embodiments, R1 and R2, a and b, m1 to m4, E1 to E4, and one or more of T1 to T4 are different.

[0055] In some embodiments, R1 and R2, a and b, m1 to m4, E1 to E4, and T1 to T4 are all the same.

[0056] In some embodiments, R1 and R2 are H.

[0057] In some embodiments, a and b are independently 1, 2, or 3.

[0058] In some embodiments, a and b are 1.

[0059] In some embodiments, m1, m2, m3, and m4 are independently 4, 5, 6, or 7.

[0060] In some embodiments, m1, m2, m3, and m4 are 6.

[0061] In some embodiments, E1, E2, E3, and E4 are -(CO)O-.

[0062] In some embodiments, E1, E2, E3, and E4 are -O(CO)- or -O(CO)O-.

[0063] In some embodiments, T1, T2, T3, or T4 are replaced as needed by C5~C 18 Alkyl, C5~C 18 Alkenyl, and C5~C 18 C5~C, independently selected from the group consisting of alkynyls, with T1, T2, T3, and T4 substituted as needed. 18 Alkyl, C5~C 18 Alkenyl, and C5~C 18 It is independently selected from the group consisting of alkynnyls.

[0064] In some embodiments, T1, T2, T3, or T4 are replaced as needed by C5~C 12 Alkyl, C5~C 12 Alkenyl, and C5~C 12 C5~C, independently selected from the group consisting of alkynyls, with T1, T2, T3, and T4 substituted as needed. 12 Alkyl, C5~C 12 Alkenyl, and C5~C 12 It is independently selected from the group consisting of alkynnyls.

[0065] In some embodiments, T1, T2, T3, or T4 are replaced as needed by C5~C 10 Alkyl, C5~C 10 Alkenyl, and C5~C 10 C5~C, independently selected from the group consisting of alkynyls, with T1, T2, T3, and T4 substituted as needed. 10 Alkyl, C5~C 10 Alkenyl, and C5~C 10 It is independently selected from the group consisting of alkynnyls.

[0066] In some embodiments, T1, T2, T3, or T4 are independently selected from the group consisting of C5-C8 alkyl, C5-C8 alkenyl, and C5-C8 alkynyl, each optionally substituted as needed.

[0067] In some embodiments, T1, T2, T3, or T4 are independently selected from the group consisting of C6-C8 alkyl, C6-C8 alkenyl, and C6-C8 alkynyl, each of which is optionally substituted.

[0068] In some embodiments, T1, T2, T3, or T4 are independently selected from the group consisting of C7 or C8 alkyl, C7 or C8 alkenyl, and C7 or C8 alkynyl, each of which is optionally substituted.

[0069] In some embodiments, G is O, -(CO)NR 3 -, and -NR 3 It is (CO)-.

[0070] In some embodiments, R3 is H or methyl.

[0071] In some embodiments, R3 is [ka] That is the case.

[0072] In some embodiments, L is a bond.

[0073] In some embodiments, L is a C1 alkyl group.

[0074] In some embodiments, L is a C2 alkyl group.

[0075] In some embodiments, L is a C3 alkyl group.

[0076] In some embodiments, L is a C4 alkyl group.

[0077] In some embodiments, R4, R5, or R4 and R5 are either absent or H.

[0078] In some embodiments, R4, R5, or R4 and R5 are [ka] That is the case.

[0079] In some embodiments, R 4 and R 5 These are independently C1, C2, or C3 alkyl groups.

[0080] In some embodiments, R 4 and R 5 It forms a six-membered ring which may be substituted as needed and may contain one or two heteroatoms.

[0081] In some embodiments, at least one of the one or two heteroatoms is nitrogen (N).

[0082] In some embodiments, at least one hydrogen atom of the six-membered ring is substituted with a methyl group.

[0083] In some embodiments, at least one atom is a nitrogen atom.

[0084] In some embodiments, the 6-membered ring contains 1, 2, or 3 double bonds, and optionally the 6-membered ring contains 3 double bonds.

[0085] In some embodiments, R4 and R5 are methyl groups.

[0086] In some embodiments, R4 and R5 form a five-membered ring which is optionally substituted and contains one or two heteroatoms and one or two double bonds.

[0087] In some embodiments, at least one of the one or two heteroatoms is nitrogen (N).

[0088] In some embodiments, T1, T2, T3, and T4 are alkyl groups selected independently from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as needed.

[0089] In some embodiments, T1, T2, T3, and T4 are octane or tridecane.

[0090] In some embodiments, T1, T2, T3, and T4 are independently substituted as needed with buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, de The alkenyl is selected from the group consisting of ca-2-ene, deca-3-ene, deca-4-ene, deca-5-ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and alkenyl groups containing two or more double bonds.

[0091] In some embodiments, T1, T2, T3, and T4 are independently substituted as needed with buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, de The alkynyl is selected from the group consisting of k-2-ine, deca-3-ine, deca-4-ine, deca-5-ine, deca-6-ine, undeca-1-ine, undeca-2-ine, undeca-3-ine, undeca-4-ine, undeca-5-ine, undeca-6-ine, undeca-7-ine, dodeca-1-ine, dodeca-2-ine, dodeca-3-ine, dodeca-4-ine, dodeca-5-ine, dodeca-6-ine, dodeca-8-ine, and alkynyl groups containing two or more triple bonds.

[0092] In some embodiments, X is N.

[0093] In one embodiment, the present disclosure relates to a lipid of formula II: [ka] or its salt or isomer [in the formula, X is either CH or N. L1, L2, and L3 are independently -O- and -(CO)NR x -, -NR x (CO)-, -(CO)O-, -CH2(CO)NR x - and if A is CH, then all of L1, L2, and L3 are -(CO)NR x -That is not the case, R x These are H, C1-C6 alkyl, or C3-C6 cycloalkyl, G1 and G2 are given by Equation III: [ka] or a salt or isomer thereof, where, n1 is 3, 4, 5, 6, 7, 8, 9, or 10. L4 is -(CO)O- or -O(CO)-, R6 is branched from C5 to C 20 It is alkyl, G3 is equation (III), equation (IV), equation (V), or equation (VI): [ka] And, R7 and R8 are independently functionalized C1-C5 alkyl groups as needed. n2, n3, and n4 are independently 0, 1, 2, or 3. X1 is C, N, or O. R9 and R 10 [These are independently H or, if necessary, functionalized C1-C5 alkyl groups.] The present invention provides a pharmaceutical composition containing the following:

[0094] In one embodiment, the present disclosure provides lipid particles comprising any of the above compounds.

[0095] In some embodiments, the disclosure further provides a therapeutic agent.

[0096] In some embodiments, the therapeutic agent is nucleic acid.

[0097] In one embodiment, the present disclosure provides a pharmaceutical composition comprising the lipid particles and a pharmaceutically acceptable additive, carrier, or diluent.

[0098] Other aspects of this disclosure are at least in part based on the discovery that substituted piperazines can be used to form novel ionizable lipids having properties advantageous when used in lipid particles for therapeutic agent delivery. In particular, the techniques of this specification provide lipid-based nanoparticle compositions and formulations that can specifically target cargo moieties (e.g., nucleic acid cargoes) to specific tissues of interest without requiring ligand-based targeting strategies. The ionizable substituted piperazine lipids disclosed herein include a head group having the following structure:

[0099] [ka] In the formula, the protonable piperazine (e.g., pH titrable) head group is connected via a linker having the following structure, C5~C 20 The hydrocarbon chains, for example, alkyl or alkenyl chains, are bonded to each hydrocarbon chain, which independently has 0 to 5 (e.g., 0, 1, 2, 3, 4, or 5) double bonds. [ka] In the formula, L5 is -O(CO)-, -CH2-O(CO)-, -CH2(CO)O-, or -O(CO)-NH-, n5 is 2, 3, 4, 5, 6, 7, or 8, and G4 is a bond, -(CO)O-, or -O(CO)-. C5~C 20The hydrocarbon chain may be bonded to the G4 atom of the linker. The ionizable substituted piperazine lipids disclosed herein provide stable and efficient lipid nanoparticle (LNP) formulations for delivering therapeutic oligonucleotides to specific target tissues of interest.

[0100] One aspect of this disclosure relates to a chemical compound of formula VII: [ka] or its salt or isomer [in the formula, L6 is -O(CO)-, -CH2-O(CO)-, -CH2(CO)O-, or -O(CO)-NH-, n6 and n7 are independently 1, 2, 3, 4, 5, 6, 7, or 8. G5 and G6 are independently a bond, -(CO)O- or -O(CO)-, R 11 and R 12 These are C5~C, which are substituted independently as needed. 20 Alkyl or C5-C 20 [Provides alkenil]

[0101] In some embodiments, R 11 and R 12 They are the same.

[0102] In some embodiments, R 11 and R 12 They are different.

[0103] In some embodiments, R 11 or R 12 C8~C are substituted as needed. 20 Alkyl and C8-C 20 Independently selected from the group consisting of alkenyls, and R as needed. 11 and R 12 C8~C are substituted as needed. 20 Alkyl and C8-C 20 It is independently selected from the group consisting of alkenils.

[0104] In some embodiments, n6 and n7 are independently 4, 5, 6, 7, or 8.

[0105] In some embodiments, R 11 or R 12 contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0106] In some embodiments, R 11 or R 12 is independently selected from the group consisting of optionally substituted C8 - C 17 alkyl and C8 - C 17 alkenyl, and optionally R 11 and R 12 are independently selected from the group consisting of optionally substituted C8 - C 17 alkyl and C8 - C 17 alkenyl.

[0107] In some embodiments, n6 and n7 are independently 5, 6, or 7.

[0108] In some embodiments, R 11 or R 12 contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0109] In some embodiments, R 11 or R 12 is independently selected from the group consisting of optionally substituted C 10 - C 17 alkyl and C 10 - C 17 alkenyl, and optionally R 11 and R 12 are independently selected from the group consisting of optionally substituted C 10 - C 17 alkyl and C 10 - C 17 alkenyl.

[0110] In some embodiments, n6 and n7 are independently 5, 6, or 7.

[0111] In some embodiments, R 11 or R 12 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0112] In some embodiments, R 11 or R 12 C is replaced as needed. 12 ~C 17 Alkyl and C 12 ~C 17 Independently selected from the group consisting of alkenyls, and R as needed. 11 and R 12 C is replaced as needed. 12 ~C 17 Alkyl and C 12 ~C 17 It is independently selected from the group consisting of alkenils.

[0113] In some embodiments, n6 and n7 are independently 5, 6, or 7.

[0114] In some embodiments, R 11 or R 12 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0115] In some embodiments, R 11 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Selected from the group consisting of alkenyls, R 12 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Selected from the group consisting of alkenyls.

[0116] In some embodiments, n6 and n7 are 5.

[0117] In some embodiments, G5 and G6 are independently bonded or -(CO)O-.

[0118] In some embodiments, R 11 or R 12 C8, C is substituted as needed. 12 , C 15 or C 17 Alkyl, and C8, C 12 , C 15 or C 17 Independently selected from the group consisting of alkenyls, and R as needed. 11 and R 12 C8, C is substituted as needed. 12 , C 15 or C 17 Alkyl, and C8, C 12 , C 15 or C 17 It is independently selected from the group consisting of alkenils.

[0119] In some embodiments, R 11 and R 12 These are alkyl groups independently selected from the group consisting of octane, nonane, decane, undecane, and dodecane, each of which is substituted as needed.

[0120] In some embodiments, R 11 and R 12 This is an alkyl group independently selected from the group consisting of octane, nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane, and eicosane.

[0121] In some embodiments, R 11 and R 12These are independently Octa-1-en, Octa-2-en, Octa-3-en, Octa-4-en, Nona-1-en, Nona-2-en, Nona-3-en, Nona-4-en, Nona-5-en, Deca-1-en, Deca-2-en, Deca-3-en, Deca-4-en, Deca-5-en, Deca-6-en, Undeka-1-en, Undeka-2-en, Undeka-3-en, Undeka-4-en, Undeka-5-en, Undeka-6-en, Undeka-7-en, Dodeka-1-en, Dodeka-2-en, Dodeka-3-en, Dodeka-4-en, Dodeka-5-en, Dodeka-6-en, Trideka-1-en, Trideka-2-en, Trideka-3-en, Trideka-4-en, Trideka-5-en, Trideka-6-en, Trideka-7-en, Tetradeca-1-en, Tetradeca-2-en, Tetradeca-3-en, Tetradeca-4-en, Tetradeca-5-en, Tetradeca-6-en, Tetradeca-7-en, Pentadeca-1-en, Pentadeca-2-en, Pentadeca-3-en, Pentadeca-4-en, Pentadeca-5-en, Pentadeca-6-en, Pentadeca- 7-yen, hexadeca-1-yen, hexadeca-2-yen, hexadeca-3-yen, hexadeca-4-yen, hexadeca-5-yen, hexadeca-6-yen, hexadeca-7-yen, hexadeca-8-yen, heptadeca-1-yen, heptadeca-2-yen, heptadeca-3-yen, heptadeca-4-yen, heptadeca-5-yen, heptadeca-6-yen, heptadeca-7-yen, heptadeca-8-yen, octadeca-1-yen, octadeca-2-yen, octadeca-3-yen, octadeca-4-yen, octadeca-5-yen, O The alkenil is selected from the group consisting of tatadeca-6-en, octadeca-7-en, octadeca-8-en, octadeca-9-en, nonadeca-1-en, nonadeca-2-en, nonadeca-3-en, nonadeca-4-en, nonadeca-5-en, nonadeca-6-en, nonadeca-7-en, nonadeca-8-en, nonadeca-9-en, icosa-1-en, icosa-2-en, icosa-3-en, icosa-4-en, icosa-5-en, icosa-6-en, icosa-7-en, icosa-8-en, and icosa-9-en.

[0122] In some embodiments, R11 and R 12 It independently includes one or more additional double bonds, and optionally R 11 and R 12 It contains one double bond independently.

[0123] One aspect of this disclosure relates to a chemical compound of formula VIII: [ka] or its salt or isomer [in the formula, m5 is 4, 5, 6, 7, or 8. G7 is a bond, -(CO)O- or -O(CO)-, R 13 and R 14 These are C8~C, which are substituted independently as needed. 20 Alkyl or C8-C 20 [Provides alkenil]

[0124] In some embodiments, R 13 and R 14 They are the same.

[0125] In some embodiments, R 13 and R 14 They are different.

[0126] In some embodiments, R 13 or R 14 C8~C are substituted as needed. 19 Alkyl and C8-C 19 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 19 Alkyl and C8-C 19 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 18 Alkyl and C8-C 18Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 18 Alkyl and C8-C 18 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 16 Alkyl and C8-C 16 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 16 Alkyl and C8-C 16 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 15 Alkyl and C8-C 15 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 15 Alkyl and C8-C 15 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C9~C are substituted as needed. 12 Alkyl and C9-C 12 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14C9~C are substituted as needed. 12 Alkyl and C9-C 12 It is independently selected from the group consisting of alkenils.

[0127] In some embodiments, m5 is 4, 5, 6, 7, or 8.

[0128] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0129] In some embodiments, m5 is 5, 6, or 7.

[0130] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0131] In some embodiments, R 13 or R 14 C is replaced as needed. 10 ~C 17 Alkyl and C 10 ~C 17 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C is replaced as needed. 10 ~C 17 Alkyl and C 10 ~C 17 It is independently selected from the group consisting of alkenils.

[0132] In some embodiments, m5 is 5, 6, or 7.

[0133] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0134] In some embodiments, R13 or R 14 C is replaced as needed. 12 ~C 17 Alkyl and C 12 ~C 17 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C is replaced as needed. 12 ~C 17 Alkyl and C 12 ~C 17 It is independently selected from the group consisting of alkenils.

[0135] In some embodiments, m5 is 5, 6, or 7.

[0136] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0137] In some embodiments, R 13 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Selected from the group consisting of alkenyls, R 14 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Selected from the group consisting of alkenyls.

[0138] In some embodiments, m5 is 5.

[0139] In some embodiments, G7 is a bond or -(CO)O-.

[0140] In some embodiments, R 13 or R 14 C8, C is substituted as needed. 12 , C 15 or C 17 Alkyl, and C8, C 12 , C 15 or C 17C8, C 12 , C 15 or C 17 Alkyl, and C8, C 12 , C 15 or C 17 It is independently selected from the group consisting of alkenils.

[0141] In some embodiments, R 13 and R 14 These are alkyl groups independently selected from the group consisting of octane, nonane, decane, undecane, and dodecane, each of which is substituted as needed.

[0142] In some embodiments, R 13 and R 14 This is an alkyl group independently selected from the group consisting of octane, nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane, and eicosane.

[0143] In some embodiments, R 13 and R 14These are independently Octa-1-en, Octa-2-en, Octa-3-en, Octa-4-en, Nona-1-en, Nona-2-en, Nona-3-en, Nona-4-en, Nona-5-en, Deca-1-en, Deca-2-en, Deca-3-en, Deca-4-en, Deca-5-en, Deca-6-en, Undeka-1-en, Undeka-2-en, Undeka-3-en, Undeka-4-en, Undeka-5-en, Undeka-6-en, Undeka-7-en, Dodeka-1-en, Dodeka-2-en, Dodeka-3-en, Dodeka-4-en, Dodeka-5-en, Dodeka-6-en, Trideka-1-en, Trideka-2-en, Trideka-3-en, Trideka-4-en, Trideka-5-en, Trideka-6-en, Trideka-7-en, Tetradeca-1-en, Tetradeca-2-en, Tetradeca-3-en, Tetradeca-4-en, Tetradeca-5-en, Tetradeca-6-en, Tetradeca-7-en, Pentadeca-1-en, Pentadeca-2-en, Pentadeca-3-en, Pentadeca-4-en, Pentadeca-5-en, Pentadeca-6-en, Pentadeca- 7-yen, hexadeca-1-yen, hexadeca-2-yen, hexadeca-3-yen, hexadeca-4-yen, hexadeca-5-yen, hexadeca-6-yen, hexadeca-7-yen, hexadeca-8-yen, heptadeca-1-yen, heptadeca-2-yen, heptadeca-3-yen, heptadeca-4-yen, heptadeca-5-yen, heptadeca-6-yen, heptadeca-7-yen, heptadeca-8-yen, octadeca-1-yen, octadeca-2-yen, octadeca-3-yen, octadeca-4-yen, octadeca-5-yen, O The alkenil is selected from the group consisting of tatadeca-6-en, octadeca-7-en, octadeca-8-en, octadeca-9-en, nonadeca-1-en, nonadeca-2-en, nonadeca-3-en, nonadeca-4-en, nonadeca-5-en, nonadeca-6-en, nonadeca-7-en, nonadeca-8-en, nonadeca-9-en, icosa-1-en, icosa-2-en, icosa-3-en, icosa-4-en, icosa-5-en, icosa-6-en, icosa-7-en, icosa-8-en, and icosa-9-en.

[0144] In some embodiments, R13 and R 14 It independently includes one or more additional double bonds, and optionally R 13 and R 14 It contains one double bond independently.

[0145] In one embodiment, the present disclosure relates to a chemical compound selected from the group consisting of: [ka] (9,9'-di(heptadecan-9-yl)=O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate), SM-048); [ka] ((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=dioleate, SM-074); [ka] (9,9'-Bis(2-butyloctyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate), SM-076); [ka] (1-(heptadecan-9-yl)=9-(4-(2-(2-(1-(2-(4-(2-(2-(2-(4-((oleoyloxy)phenyl)acetoxy)ethyl)piperidine-1-yl)acetyl)piperazine-1-yl)-2-oxoethyl)piperidine-4-yl)ethoxy)-2-oxoethyl)phenyl)=nonanedioate, SM-077); [ka] ((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=bis(9-((2-heptylnonanoyl)oxy)nonanoate), SM-079); [ka] (9,9'-Bis(2-ethylhexyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate), SM-083); [ka] (Di(heptadecan-9-yl)9,9'-(((((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(oxy))bis(carbonyl))bis(azandiyl))dinonanoate;SM-085); [ka] (9,9'-di(heptadecan-9-yl)O'1,O1-(((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(methylene))=di(nonanedioate);SM-088); [ka] (7,7'-Dinonyl O'1,O1-((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(heptanedioate);SM-100); [ka] (Di(heptadecan-9-yl)9,9'-((2,2'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(acetyl))bis(oxy))dinonanoete;SM-109); [ka] (Di(heptadecan-9-yl)O,O'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=diglutarate;SM-113); [ka] (Bis(9-(heptadecan-9-yloxy)-9-oxononyl)O,O'-(((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))=diterephthalate;SM-125), as well as their salts and isomers are provided.

[0146] In one embodiment, the present disclosure provides lipid particles comprising any of the compounds disclosed herein.

[0147] In some embodiments, the lipid particles further comprise a therapeutic agent.

[0148] In some embodiments, the therapeutic agent is nucleic acid.

[0149] In one embodiment, the present disclosure provides a pharmaceutical composition comprising one of the above-mentioned lipid particles and optionally comprising pharmaceutically acceptable additives, carriers, or diluents.

[0150] In one embodiment, the disclosure provides nucleic acid-lipid particles for delivering nucleic acid cargo to a target, wherein the nucleic acid-lipid particles are compounds selected from the group consisting of: [ka] (9,9'-di(heptadecan-9-yl)=O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate), SM-048); [ka] ((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=dioleate, SM-074); [ka] (9,9'-Bis(2-butyloctyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate), SM-076); [ka] (1-(heptadecan-9-yl)=9-(4-(2-(2-(1-(2-(4-(2-(2-(2-(4-((oleoyloxy)phenyl)acetoxy)ethyl)piperidine-1-yl)acetyl)piperazine-1-yl)-2-oxoethyl)piperidine-4-yl)ethoxy)-2-oxoethyl)phenyl)=nonanedioate, SM-077); [ka] ((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=bis(9-((2-heptylnonanoyl)oxy)nonanoate), SM-079); [ka] (9,9'-Bis(2-ethylhexyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate), SM-083); [ka] (Di(heptadecan-9-yl)9,9'-(((((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(oxy))bis(carbonyl)bis(azandiyl))dinonanoate;SM-085); [ka] (9,9'-di(heptadecan-9-yl)O'1,O1-(((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(methylene))=di(nonanedioate);SM-088); [ka] (7,7'-Dinonyl O'1,O1-((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(heptanedioate);SM-100); [ka] (Di(heptadecan-9-yl)9,9'-((2,2'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(acetyl))bis(oxy))dinonanoete;SM-109); [ka] (Di(heptadecan-9-yl)O,O'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=diglutarate;SM-113); [ka] (Bis(9-(heptadecan-9-yloxy)-9-oxononyl)O,O'-(((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))=diterephthalate; SM-125) The compounds further constitute approximately 30-70 mol% or approximately 40-60 mol% or approximately 50 mol% of the total lipids present in the nucleic acid-lipid particles, and optionally constitute 45 mol%, 46 mol%, 47 mol%, 48 mol%, 49 mol%, 50 mol%, 51 mol%, 52 mol%, 53 mol%, 54 mol%, or 55 mol% of the total lipids present in the nucleic acid-lipid particles.

[0151] In some embodiments, the nucleic acid-lipid particles include a conjugated lipid that inhibits particle aggregation, comprising 0.01-2% of the total lipids present, optionally comprising a polyethylene glycol (PEG)-lipid complex, optionally having an average molecular weight of 550-5000 daltons for the PEG-lipid complex, optionally being a PEG5000-lipid complex, optionally being a PEG2000-lipid complex, optionally comprising one or more of 1,2-dimiristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2k) and 1,2-distearoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DSG-PEG2k), optionally comprising 1,2-dimiristoyl-rac-glycero- The PEG2000-lipid complex is 3-methoxypolyethylene glycol-2000 (DMG-PEG2k), and optionally the PEG2000-lipid complex is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)-polyethylene glycol methoxy (DOPE-mPEG2k), and optionally the PEG2000-lipid complex is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-mPEG2k) complexed with methoxylpoly(ethylene glycol), and optionally the nucleic acid-lipid particles contain PEG-lipid complexes at concentrations selected from the group consisting of approximately 0.5 mol% of the total lipids present in the nucleic acid-lipid particles, approximately 1.0 mol% of the total lipids present in the nucleic acid-lipid particles, and approximately 0.5 to 3.0 mol% of the total lipids present in the nucleic acid-lipid particles.

[0152] In some embodiments, the PEG-lipid complex is DMG-PEG2k, which constitutes approximately 1.5 mol% of the total lipids present in the nucleic acid-lipid particles.

[0153] In some embodiments, the nucleic acid-lipid particles contain one or more noncationic lipids that constitute 20 mol% to 80 mol% of the total lipids present in the lipid-nucleic acid particles, and optionally one or more noncationic lipids contain cholesterol or a derivative thereof.

[0154] In some embodiments, the nucleic acid-lipid particles contain cholesterol or a derivative in a concentration range selected from the group consisting of 35 mol% to 45 mol% of the total lipids present in the nucleic acid-lipid particles, 45 mol% to 55 mol% of the total lipids present in the nucleic acid-lipid particles, and 55 mol% to 65 mol% of the total lipids present in the nucleic acid-lipid particles, wherein the cholesterol or derivative may optionally constitute about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, or about 45% of the total lipids present in the nucleic acid-lipid particles, and optionally the cholesterol or derivative may constitute about 40% of the total lipids present in the nucleic acid-lipid particles.

[0155] In some embodiments, the nucleic acid-lipid particles contain one or more noncationic lipids other than cholesterol or its derivatives, and optionally, one or more noncationic lipids other than cholesterol or its derivatives constitute 5 mol% to 20 mol% of the total lipids present in the lipid-nucleic acid particles, and optionally, one or more noncationic lipids other than cholesterol or its derivatives constitute about 10 mol% of the total lipids present in the nucleic acid-lipid particles.

[0156] In some embodiments, one or more noncationic lipids other than cholesterol or its derivatives include noncationic lipids selected from the group consisting of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), and β-sitosterol, and optionally one or more noncationic lipids other than cholesterol or its derivatives are DOPE.

[0157] In some embodiments, the nucleic acid cargo comprises synthetic or naturally occurring RNA or DNA, or derivatives thereof, and optionally the nucleic acid cargo is modified RNA, which optionally is selected from the group consisting of modified mRNA, modified antisense oligonucleotides, and modified siRNA, and optionally the modified mRNA encodes a nucleic acid regulatory factor.

[0158] In some embodiments, the nucleic acid cargo includes 2'-O-methyl modified nucleotides, nucleotides containing a 5'-phosphorothioate group, terminal nucleotides linked to cholesteryl derivatives, 2'-deoxy-2'-fluoro modified nucleotides, 5'-methoxy modified nucleotides (e.g., 5'-methoxyuridine), 2'-deoxy modified nucleotides, locked nucleotides, debased nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides, phosphoramidates, nucleotides containing unnatural bases; phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotryesters, aminoalkylphosphotryesters The modifications include one or more modifications selected from the group consisting of internucleoside links or skeletons, including tel, methyl and other alkylphosphonates including 3'-alkylene phosphonates and chiral phosphonates, phosphinates, phosphoramidates including 3'-aminophosphoramidates and aminoalkylphosphoramidates, thionophosphoramidates, thionoalkylphosphonates, thionoalkylphosphotryesters, and boranophosphates having the usual 3'-5' linkage, their 2'-5' linked analogs, and internucleoside links or skeletons having inverted polarity, including those in which pairs of adjacent nucleoside units are linked from 3'-5' to 5'-3' or 2'-5' to 5'-2'.

[0159] In some embodiments, the nucleic acid-lipid particles contain approximately 50 mol% of the total lipids present in the nucleic acid-lipid particles of SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, or SM-125, approximately 38.5 mol% of the total lipids present in the nucleic acid-lipid particles of cholesterol, approximately 10 mol% of the total lipids present in the nucleic acid-lipid particles of DOPE, and approximately 1.5 mol% of the total lipids present in the nucleic acid-lipid particles of DMG-PEG2k.

[0160] In one embodiment, the present disclosure provides a compound having the following structure. [ka]

[0161] Further aspects of this disclosure are at least in part based on the discovery that other aryl and heteroaryl lipid compounds can be used to form novel ionizable lipids having properties advantageous when used in lipid particles for in vivo delivery of therapeutic agents. In particular, the techniques of this specification provide lipid-based nanoparticle compositions and formulations that can specifically target cargo moieties (e.g., nucleic acid cargoes) to specific tissues of interest without requiring ligand-based targeting strategies. Certain aryl and heteroaryl lipid compounds disclosed herein may include the following general structures:

[0162] In one embodiment, the present disclosure relates to a compound of formula IX: [ka] or its salt or isomer [in the formula, X2, X3, and X4 are independently CH or N. G8, G9, and G 10 These are independently O, -(CO)O-, -CH2O(CO)-, and -(CH2)2(CO)NR 15 -,-(CH2)O(CO)NR 15 -, CH2(CO)NR 15 -, -(CO)NR 15 -, -NR 15 (CO)-, or NR 15 (CO)O-, where X2, X3, and X4 are all CH, then G8, G9, and G 10 Not all of them are -(CO)NR1-, a1, b1, and c1 are independently 0, 1, 2, 3, or 4. m6, m7, and m8 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. E5, E6, and E7 are independently -(CO)O- or -O(CO)-, T5, T6, and T7 are independently branched or unbranched C5~C22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R 15 [These are H, or optionally functionalized C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl].

[0163] In some embodiments, two or more sets of m6-m8, E5-E7, and T5-T7 are the same.

[0164] In some embodiments, one or more of m6-m8, E5-E7, and T5-T7 are different.

[0165] In some embodiments, two of the following are the same: m6-m8, E5-E7, and T5-T7.

[0166] In some embodiments, a1, b1, or c1 is 2, 3, or 4.

[0167] In some embodiments, a1, b1, and c1 are 2, 3, and 4.

[0168] In some embodiments, a1, b1, or c1 is 3 or 4.

[0169] In some embodiments, a1 and b1 are 3, and c1 is 4.

[0170] In some embodiments, a1, b1, and c1 are 3.

[0171] In some embodiments, m6, m7, and m8 are 1 to 8.

[0172] In some embodiments, m6, m7, or m8 are 5-8, and m6, m7, and m8 are 5-8 as needed.

[0173] In some embodiments, m6, m7, or m8 are 5-6, and m6, m7, and m8 are 5-6 as needed.

[0174] In some embodiments, m6, m7, or m8 is 6, and m6, m7, and m8 are 6 as needed.

[0175] In some embodiments, one of X2, X3, or X4 is N, and optionally two of X2, X3, or X4 are N.

[0176] In some embodiments, X2, X3, and X4 are CH.

[0177] In some embodiments, G8, G9, or G 10 Two of them are the same, and G8, G9, and G as needed. 10 All of them are the same.

[0178] In some embodiments, G8, G9, or G 10 One or more of them are different.

[0179] In some embodiments, one of X2, X3, or X4 is N, and G8, G9, and G 10 ha-(CO)NR 15 - is

[0180] In some embodiments, G8, G9, and G 10 It is -(CO)O-.

[0181] In some embodiments, G8, G9, and G 10 Two of them are -(CO)NR 15 - is

[0182] In some embodiments, G8, G9, and G 10 One of them is -NR 15 It is (CO)-.

[0183] In some embodiments, G8, G9, and G10 All of -NR 15 It is (CO)-.

[0184] In some embodiments, G8, G9, and G 10 One of them is -(CO)NR 15 - and G8, G9, and G 10 Two of them are -NR 15 It is (CO)-.

[0185] In some embodiments, G8, G9, and G 10 One of them is -(CO)NR 15 - and G8, G9, and G 10 Two of them are O.

[0186] In some embodiments, T5, T6, or T7 are replaced as needed by C5-C 18 Alkyl, C5~C 18 Alkenyls, and C5~C 18 C5-C are independently selected from the group consisting of alkynyls, and T5, T6, and T7 are substituted as needed. 18 Alkyl, C5~C 18 Alkenyls, and C5~C 18 It is independently selected from the group consisting of alkynnyls.

[0187] In some embodiments, T5, T6, or T7 are replaced as needed by C5-C 12 Alkyl, C5~C 12 Alkenyls, and C5~C 12 C5-C are independently selected from the group consisting of alkynyls, and T5, T6, and T7 are substituted as needed. 12 Alkyl, C5~C 12 Alkenyls, and C5~C 12 It is independently selected from the group consisting of alkynnyls.

[0188] In some embodiments, T5, T6, or T7 are replaced as needed by C5-C 10 Alkyl, C5~C 10 Alkenyls, and C5~C 10 C5-C are independently selected from the group consisting of alkynyls, and T5, T6, and T7 are substituted as needed. 10 Alkyl, C5~C 10 Alkenyls, and C5~C 10 It is independently selected from the group consisting of alkynnyls.

[0189] In some embodiments, T5, T6, or T7 are independently selected from the group consisting of C5-C8 alkyl, C5-C8 alkenyl, and C5-C8 alkynyl, each optionally substituted as needed.

[0190] In some embodiments, T5, T6, or T7 are independently selected from the group consisting of C6-C8 alkyl, C6-C8 alkenyl, and C6-C8 alkynyl, each of which is optionally substituted.

[0191] In some embodiments, T5, T6, or T7 are independently selected from the group consisting of C7 or C8 alkyl, C7 or C8 alkenyl, and C7 or C8 alkynyl, each of which is optionally substituted.

[0192] In some embodiments, T5, T6, or T7 are independently a C8 alkyl, C8 alkenyl, or C8 alkynyl, each optionally substituted, and T5, T6, and T7 are optionally a C8 alkyl, C8 alkenyl, or C8 alkynyl, each optionally substituted.

[0193] In some embodiments, E5, E6, or E7 is -(CO)O-, and E5, E6, and E7 are -(CO)O- as needed.

[0194] In some embodiments, m6, m7, or m8 is 8, and m6, m7, and m8 are 8 as needed.

[0195] In some embodiments, T5, T6, and T7 are alkyl groups selected independently from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as needed.

[0196] In some embodiments, T5, T6, and T7 are octane or tridecane.

[0197] In some embodiments, T5, T6, and T7 are independently substituted as needed with buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca The alkenyl is selected from the group consisting of -2-ene, deca-3-ene, deca-4-ene, deca-5-ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and alkenyl groups containing two or more double bonds.

[0198] In some embodiments, T5, T6, and T7 are independently substituted as needed with buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, deca The alkynyl is selected from the group consisting of -2-yne, deca-3-yne, deca-4-yne, deca-5-yne, deca-6-yne, undeca-1-yne, undeca-2-yne, undeca-3-yne, undeca-4-yne, undeca-5-yne, undeca-6-yne, undeca-7-yne, dodeca-1-yne, dodeca-2-yne, dodeca-3-yne, dodeca-4-yne, dodeca-5-yne, dodeca-6-yne, dodeca-8-yne, and alkynyl groups containing two or more triple bonds.

[0199] In some embodiments, R 15 H is H.

[0200] In one embodiment, the present disclosure provides compounds selected from the group consisting of: [ka] (SM-016; 1-Ethylhexyl 9-[3-[[3,5-Bis[3-[Bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate) [ka] (SM-062; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-((((pyridine-2,4,6-tricarbonyl)tris(azandiyl))tris(propane-3,1-diyl))tris(azantriyl))hexanonanoate), [ka] (SM-065; Tris(3-(bis(9-(octan-3-yloxy)-9-oxononyl)amino)propyl)=benzene-1,3,5-tricarboxylate), [ka] (SM-067; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(4-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)butanamide)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-068; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-(((benzene-1,3,5-triyltris(azandiyl))tris(4-oxobutane-4,1-diyl))tris(azantriyl))hexanonaate), [ka] (SM-070; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)carbamoyl)-1,3-phenylene)bis(azandiyl))bis(4-oxobutan-4,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-072; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)carbamoyl)-1,3-phenylene)bis(oxy))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-073; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(2-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-2-oxoethyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-107; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(3-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-3-oxopropyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-111; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(((5-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)pentanoyl)oxy)methyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), and [ka] (SM-112; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(19-ethyl-8-(9-(octane-3-yloxy)-9-oxononyl)-3,17-dioxo-2,18-dioxa-4,8-diazatetracosyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate).

[0201] In one embodiment, the present disclosure relates to lipids of formula IX: [ka] or its salt or isomer [in the formula, X2, X3, and X4 are independently CH or N. G8, G9, and G 10 These are independently O, -(CO)O-, -CH2O(CO)-, and -(CH2)2(CO)NR 15 -,-(CH2)O(CO)NR 15 -, CH2(CO)NR 15 -, -(CO)NR 15 -, -NR 15 (CO)-, or NR 15 (CO)O-, where X2, X3, and X4 are all CH, then G8, G9, and G 10 All of the following are -(CO)NR 15 -That is not the case, a1, b1, and c1 are independently 0, 1, 2, 3, or 4. m6, m7, and m8 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. E5, E6, and E7 are independently -(CO)O- or -O(CO)-, T5, T6, and T7 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R1 is H, or optionally a functionalized C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. The present invention provides a pharmaceutical composition containing the following:

[0202] In some embodiments, two or more sets of m6-m8, E5-E7, and T5-T7 are the same.

[0203] In some embodiments, one or more of m6-m8, E5-E7, and T5-T7 are different.

[0204] In some embodiments, two of the following are the same: m6-m8, E5-E7, and T5-T7.

[0205] In some embodiments, a1, b1, or c1 is 2, 3, or 4.

[0206] In some embodiments, a1, b1, and c1 are 2, 3, or 4.

[0207] In some embodiments, a1, b1, or c1 is 3 or 4.

[0208] In some embodiments, a1 and b1 are 3, and c1 is 4.

[0209] In some embodiments, a1, b1, and c1 are 3.

[0210] In some embodiments, m6, m7, or m8 are 1 to 8.

[0211] In some embodiments, m6, m7, or m8 are 5-8, and m6, m7, and m8 are 5-8 as needed.

[0212] In some embodiments, m6, m7, or m8 are 5-6, and m6, m7, and m8 are 5-6 as needed.

[0213] In some embodiments, m6, m7, or m8 is 6, and m6, m7, and m8 are 6 as needed.

[0214] In some embodiments, one of X2, X3, or X4 is N, and optionally two of X2, X3, or X4 are N.

[0215] In some embodiments, X2, X3, and X4 are CH.

[0216] In some embodiments, G8, G9, or G 10 Two of them are the same, and G8, G9, and G as needed. 10 All of them are the same.

[0217] In some embodiments, G8, G9, or G 10 One or more of them are different.

[0218] In some embodiments, one of X2, X3, or X4 is N, and G8, G9, and G 10 ha-(CO)NR 15 - is

[0219] In some embodiments, G8, G9, and G 10 It is -(CO)O-.

[0220] In some embodiments, G8, G9, and G 10 Two of them are -(CO)NR 15 - is

[0221] In some embodiments, G8, G9, and G 10One of them is -NR 15 It is (CO)-.

[0222] In some embodiments, G8, G9, and G 10 All of -NR 15 It is (CO)-.

[0223] In some embodiments, G8, G9, and G 10 One of them is -(CO)NR 15 - and G8, G9, and G 10 Two of them are -NR 15 It is (CO)-.

[0224] In some embodiments, G8, G9, and G 10 One of them is -(CO)NR 15 - and G8, G9, and G 10 Two of them are O.

[0225] In some embodiments, T5, T6, or T7 are replaced as needed by C5-C 18 Alkyl, C5~C 18 Alkenyls, and C5~C 18 C5-C are independently selected from the group consisting of alkynyls, and T5, T6, and T7 are substituted as needed. 18 Alkyl, C5~C 18 Alkenyls, and C5~C 18 It is independently selected from the group consisting of alkynnyls.

[0226] In some embodiments, T5, T6, or T7 are replaced as needed by C5-C 12 Alkyl, C5~C 12 Alkenyls, and C5~C 12 C5-C are independently selected from the group consisting of alkynyls, and T5, T6, and T7 are substituted as needed. 12 Alkyl, C5~C 12 Alkenyls, and C5~C 12It is independently selected from the group consisting of alkynnyls.

[0227] In some embodiments, T5, T6, or T7 are replaced as needed by C5-C 10 Alkyl, C5~C 10 Alkenyls, and C5~C 10 C5-C are independently selected from the group consisting of alkynyls, and T5, T6, and T7 are substituted as needed. 10 Alkyl, C5~C 10 Alkenyls, and C5~C 10 It is independently selected from the group consisting of alkynnyls.

[0228] In some embodiments, T5, T6, or T7 are independently selected from the group consisting of C5-C8 alkyl, C5-C8 alkenyl, and C5-C8 alkynyl, each optionally substituted as needed.

[0229] In some embodiments, T5, T6, or T7 are independently selected from the group consisting of C6-C8 alkyl, C6-C8 alkenyl, and C6-C8 alkynyl, each of which is optionally substituted.

[0230] In some embodiments, T5, T6, or T7 are independently selected from the group consisting of C7 or C8 alkyl, C7 or C8 alkenyl, and C7 or C8 alkynyl, each of which is optionally substituted.

[0231] In some embodiments, T5, T6, or T7 are independently a C8 alkyl, C8 alkenyl, or C8 alkynyl, each optionally substituted, and T5, T6, and T7 are optionally a C8 alkyl, C8 alkenyl, or C8 alkynyl, each optionally substituted.

[0232] In some embodiments, E5, E6, or E7 is -(CO)O-, and E5, E6, and E7 are -(CO)O- as needed.

[0233] In some embodiments, m6, m7, or m8 is 8, and m6, m7, and m8 are 8 as needed.

[0234] In some embodiments, T5, T6, and T7 are alkyl groups selected independently from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as needed.

[0235] In some embodiments, T5, T6, and T7 are octane or tridecane.

[0236] In some embodiments, T5, T6, and T7 are independently substituted as needed with buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca The alkenyl is selected from the group consisting of -2-ene, deca-3-ene, deca-4-ene, deca-5-ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and alkenyl groups containing two or more double bonds.

[0237] In some embodiments, T5, T6, and T7 are independently substituted as needed with buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, deca The alkynyl is selected from the group consisting of -2-yne, deca-3-yne, deca-4-yne, deca-5-yne, deca-6-yne, undeca-1-yne, undeca-2-yne, undeca-3-yne, undeca-4-yne, undeca-5-yne, undeca-6-yne, undeca-7-yne, dodeca-1-yne, dodeca-2-yne, dodeca-3-yne, dodeca-4-yne, dodeca-5-yne, dodeca-6-yne, dodeca-8-yne, and alkynyl groups containing two or more triple bonds.

[0238] In some embodiments, R 15 H is H.

[0239] In some embodiments, the lipids are selected from the group consisting of: [ka] (SM-016; 1-Ethylhexyl 9-[3-[[3,5-Bis[3-[Bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate) [ka] (SM-062; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-((((pyridine-2,4,6-tricarbonyl)tris(azandiyl))tris(propane-3,1-diyl))tris(azantriyl))hexanonanoate), [ka] (SM-065; Tris(3-(bis(9-(octan-3-yloxy)-9-oxononyl)amino)propyl)=benzene-1,3,5-tricarboxylate), [ka] (SM-067; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(4-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)butanamide)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-068; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-(((benzene-1,3,5-triyltris(azandiyl))tris(4-oxobutane-4,1-diyl))tris(azantriyl))hexanonaate), [ka] (SM-070; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)carbamoyl)-1,3-phenylene)bis(azandiyl))bis(4-oxobutan-4,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-072; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)carbamoyl)-1,3-phenylene)bis(oxy))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-073; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(2-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-2-oxoethyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-107; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(3-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-3-oxopropyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), [ka] (SM-111; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(((5-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)pentanoyl)oxy)methyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), and [ka] (SM-112; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(19-ethyl-8-(9-(octane-3-yloxy)-9-oxononyl)-3,17-dioxo-2,18-dioxa-4,8-diazatetracosyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate).

[0240] In one embodiment, the present disclosure provides lipid particles comprising any of the above compounds.

[0241] In some embodiments, the lipid particles further comprise a therapeutic agent.

[0242] In some embodiments, the therapeutic agent is nucleic acid.

[0243] In one embodiment, the present disclosure provides a pharmaceutical composition comprising the lipid particles and a pharmaceutically acceptable additive, carrier, or diluent.

[0244] definition Unless otherwise specifically stated or as is evident from the context, the term “about” as used herein is understood to mean within the normal acceptable range in the art, for example, within two standard deviations of the mean. “About” can be understood to mean within 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.5%, 0.1%, 0.05%, or 0.01% of the stated value.

[0245] In certain embodiments, the terms “approximately” or “about” mean, unless otherwise stated or evident from the context, a range of values ​​that are 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) the stated reference value (except where such a number can exceed 100% of the possible value).

[0246] Unless otherwise evident from the context, all figures provided herein are modified by the term “approximately”.

[0247] As used herein, “fully closed RNA” and “circular RNA” refer to closed-loop oligoribonucleotide molecules in which the free 3' and 5' ends found in linear RNA morphology join together to form a closed loop, which is thought to stabilize the RNA morphology and make it long-lasting. Not to be bound by theory, this is thought to be because the absence of free ends makes such fully closed / circular RNA resistant to digestion by exonucleases. Certain such closed-loop RNAs have recently been designed to contain a translatable sequence (e.g., mRNA) in a form commercially called “Endless RNA®” or “eRNA” (see, e.g., U.S. Publication Nos. 2022 / 0257794 and 2022 / 0143062, and U.S. Patent No. 10,953,033). Therefore, fully closed or circular RNA refers to mRNA that is circular and reads sequentially. While not strictly adhering to theory, circular RNA is described as a versatile synthetic RNA platform that directs cells to express desired therapeutic proteins, and its innate stability allows for long-lasting protein expression (in contrast to the transient existence of translatable linear RNA). In addition, circular RNA is described as non-immunogenic, making repeated re-administration possible, and its inherent stability allows for multiple administration routes, including intravenous, subcutaneous, topical, and intratracheal administration.

[0248] The term "lipids" refers to a group of organic compounds that include, but are not limited to, fatty acid esters, and are characterized by being insoluble in water but soluble in many organic solvents. Lipids are usually divided into at least three classes: (1) "simple lipids" which include fats, oils, and waxes; (2) "compound lipids" which include phospholipids and glycolipids; and (3) "derived lipids" such as steroids.

[0249] As used herein, the term “cationic lipid” refers to any lipid species that carries a net positive charge at a selected pH, such as physiological pH. Cationic lipids may have a head group that is always positively charged in aqueous solution (“obligate cationic lipid”). For example, an obligate cationic lipid may have a quaternary amine as its head group, but is not limited to this.

[0250] Alternatively, cationic lipids may have a proton-accepting head group in solution such that they exist primarily as cations when the lipid is less than pKa and primarily as a neutral portion when the lipid is greater than pKa, for example, a pH titrable amino head group, or substituted aryl and heteroaryl compounds (e.g., in the case of "ionizable substituted aryl and heteroaryl complex lipids" as defined below, and / or in the case of "ionizable substituted piperazine lipids" as defined below). For example, but not limited to, ionizable lipids may have primary, secondary, or tertiary amines (e.g., alkylamino or dialkylamino head groups) as their head group. In some embodiments, the ionizable lipid may have a protonable tertiary amine (e.g., pH titrable) head group;C 18The compounds include hydrocarbon chains, for example, alkyl, alkenyl, or alkynyl chains (where each hydrocarbon chain independently has 0 to 3 (e.g., 0, 1, 2, or 3) double bonds), and ether, ester, or ketal links between the head group and the hydrocarbon chain. In some embodiments, ionizable substituted aryl and heteroaryl compounds may have a general structure according to formula I disclosed herein, or substituted aryl or heteroaryl compounds may have a general structure according to formula IX disclosed herein.

[0251] In some embodiments, ionizable substituted piperazine lipids may include piperazines such as ((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl)bis(2-phenylacetate) having the following structure.

[0252] [ka]

[0253] Examples of obligate cationic lipids include, but are not limited to, dimethyldioctadecylammonium bromide (DDAB), N-(4-carboxybenzyl)-N,N-dimethyl-2,3-bis(oleoyloxy)propane-1-aminium (DOBAQ), 1,2-dioleoyl-3-trimethylammonium-propane or 18:1 TAP, double-chain or gemini-type cationic lipids (DOTAP), 1,2-di-O-octadecenyl-3-trimethylammoniumpropane chloride (DOTMA), ethylphosphatidylcholine (EPC), and trimethylsphingosine.

[0254] Various forms of the obligate cationic lipid EPC, and related forms of obligate cationic phosphatidylcholine, are commercially available. Ethyl phosphatidylcholine, 18:1 EPC (Cl salt), also known as 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (chloride salt), has the following structure. [ka]

[0255] 18:0 EPC (Cl salt) is also known as 1,2-distearoyl-sn-glycero-3-ethylphosphocholine (chloride salt) and has the following structure. [ka]

[0256] 16:0 EPC (Cl salt) is also known as 1,2-dipalmitoyl-sn-glycero-3-ethylphosphocholine (chloride salt) and has the following structure. [ka]

[0257] 14:0 EPC (Cl salt) is also known as 1,2-dimyristoyl-sn-glycero-3-ethylphosphocholine (chloride salt) and has the following structure. [ka]

[0258] 12:0 EPC (Cl salt) is also known as 1,2-dilauroyl-sn-glycero-3-ethylphosphocholine (chloride salt) and has the following structure. [ka]

[0259] 14:1 EPC (Tf salt) is also known as 1,2-dimyristreoyl-sn-glycero-3-ethylphosphocholine (Tf salt) and has the following structure. [ka]

[0260] 16:0~18:1 EPC (Cl salt) is also known as 1-palmitoyl-2-oleoyl-sn-glycero-3-ethylphosphocholine (chloride salt) and has the following structure. [ka]

[0261] 18:1 EPC (Cl salt) is also known as 1,2-dioleoyl-sn-glycero-3-ethylphosphocholine (chloride salt) and has the following structure. [ka]

[0262] 16:0~18:0 PC is also known as 1-palmitoyl-2-stearoyl-sn-glycero-3-phosphocholine and has the following structure. [ka]

[0263] 16:0 / 16:1 (9Z)-PC is also known as 1-(1-enyl-palmitoyl)-2-palmitreoyl-sn-glycero-3-phosphocholine and has the following structure. [ka]

[0264] 16:0~18:2 PC is also known as 1-palmitoyl-2-linoleoyl-sn-glycero-3-phosphocholine and has the following structure. [ka]

[0265] 18:0~18:1(9Z)-PC is also known as 1-stearoyl-2-oleoyl-sn-glycero-3-phosphocholine and has the following structure. [ka]

[0266] 18:0~18:2(9Z,12Z)-PC is also known as 1-octadecanyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine and has the following structure. [ka]

[0267] 18:1~18:2(9Z,12Z)-PC is also known as 1-(9Z,12Z-octadecadienoyl)-2-(9Z-octadecenoyl)-glycero-3-phosphocholine and has the following structure. [ka]

[0268] As used herein, the terms “ionizable lipid” or “ionizable cationic lipid” refer to lipids that become cationic (protonated) when the pH drops below the pKa of the lipid’s ionizable group, but gradually approach neutrality as the pH value increases. When lipids are components of lipid-nucleic acid particles, they can associate with negatively charged polynucleic acids at pH values ​​below the pKa. Certain examples of such ionizable lipids include lipids and salts thereof having one, two, three or more fatty acid or fatty hydrocarbon chains, and pH titrable amino head groups (e.g., alkylamino or dialkylamino head groups).

[0269] In some embodiments, the ionizable substituted piperazine lipid comprises a protonable piperazine (e.g., pH titrable) head group, one or more hydrocarbon chains, e.g., alkyl or alkenyl chains, where each of the one or more hydrocarbon chains may or may not contain one or more double bonds, and the head group is bonded to the hydrocarbon chain via a linker having the following structure. [ka] In the formula, L5 is -O(CO)-, -CH2-O(CO)-, -CH2(CO)O-, or -O(CO)-NH-, n5 is 2, 3, 4, 5, 6, 7, or 8, and G4 is a bond, -(CO)O-, or -O(CO)-. In some embodiments, the ionizable substituted piperazine lipids may include substituted piperazines having a head group having the following five ring structures. [ka] In the formula, the wavy lines indicate the linkage points of the linkers described above.

[0270] Examples of ionizable lipids, but not limited to, include 1,2-dioleoyl-3-dimethylammonium-propane (DODAP), 9-heptadecanyl 8-{(2-hydroxyethyl)[6-oxo-6-(undecyloxy)hexyl]amino}octanoate (SM-102), and disulfanediylbis(ethane-2,1-diyl)bis(piperidine-1,4-diyl)bis(ethane-2,1-diyl)bis(oxy)bis(2-oxoethane-2,1-diyl)bis(4,1-phenylene)=dioleate. (SS-OP), Dimethylsphingosine, 3-(N-(N',N'-dimethylaminoethane)-carbamoyl)cholesterol (DC-cholesterol), C12-200; N4-Cholesteryl-Spermine HCl salt (GL67); N1-[2-((1S)-1-[(3-aminopropyl)amino]-4-[di(3-amino-propyl)amino]butylcarboxamide)ethyl]-3,4-di[oleyloxy]-benzamide (MVL5); 1,2-Distearyloxy-N,N-dimethyl-3-aminopropane (DSDM A); 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane (DODMA); 1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane (DLinDMA); 1,2-dilinolelenyloxy-N,N-dimethyl-3-aminopropane (DLenDMA); 1,2-di-γ-linolelenyloxy-N,N-dimethylaminopropane (γ-DLenDMA); 1,2-dilinoleyloxy-keto-N,N-dimethyl-3-aminopropane (DLinK-DMA); 1,2-dilinoleyl-4-(2-dimethyl (Aminoethyl)-[1,3]-dioxolane (DLinKC2-DMA) (also known as DLin-C2K-DMA, XTC2, and C2K); 2,2-dilinoleyl-4-(3-dimethylaminopropyl)[1,3]-dioxolane (DLin-K-C3-DMA); 2,2-dilinoleyl-4-(4-dimethylaminobutyl)[1,3]-dioxolane (DLin-K-C4-DMA); 1,2-dilinolenyloxy-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (γ-DLen-C2K-DMA);1,2-di-γ-linolenyloxy-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (γ-DLen-C2K-DMA); dilinoleylmethyl-3-dimethylaminopropionate (DLin-M-C2-DMA) (also known as MC2); (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate (DLin-M-C3-DMA) (also known as MC3); 3-(dilinoleylmethoxy)-N,N-dimethylpropane-1- Amine (DLin-MP-DMA) (also known as 1-B11); 2-({8-[(3β)-cholesta-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1-amine (octyl-CLinDMA); (2R)2-({8-[(3β)-cholesta-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1-amine (R-octyl-CLinDMA); (2S)2-({8-[(3β)-cholesta-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-1-amine(S-octyl-CLinDMA); (2S)-1-{7-[(3β)-cholesta-5-en-3-yloxy]heptyloxy}-3-[(4Z)-deca-4-en-1-yloxy]-N,N-dimethylpropan-2-amine; (2R)-1-{4-[(3β)-cholesta-5-en-3-yloxy] [sta-5-en-3-yloxy]butoxy}-3-[(4Z)-deca-4-en-1-yloxy]-N,N-dimethylpropan-2-amine; 1-[(2R)-1-{4-[(3β)-cholesta-5-en-3-yloxy]butoxy}-3-(octyloxy)propan-2-yl]guanidine; 1-[(2R)-1-{7-[(3β)-cholesta-5-en-3-yloxy]heptyloxy}-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-dien-1-yloxy]propan-2-amine;1-[(2R)-1-{4-[(3β)-cholesta-5-en-3-yloxy]butoxy}-N,N-dimethyl-3-[(9Z,12Z)-octadeca-9,12-diene-1-yloxy]propan-2-amine;(2S)-1-({6-[(3β))-cholesta-5-en-3-yloxy]hexyl}oxy)-N,N-dimethyl-3-[(9Z)-octadeca-9-en-1-yloxy]propan-2-amine;(3β)-3-[6-{[(2S)-3-[(9Z)-octadeca-9-en-1-yloxyl] -2-(pyrrolidine-1-yl)propyl]oxy}hexyl)oxy]cholest-5-ene; (2R)-1-{4-[(3β)-cholest-5-ene-3-yloxy]butoxy}3-(octyloxy)propan-2-amine; (2R)-1-({8-[(3β)-cholest-5-ene-3-yloxy]octyl}oxy)-N,N-dimethyl-3-(pentyloxy)propan-2-amine; (2R)-1-({8-[(3β)-cholest-5-ene-3-yloxy]octyl}oxy)-3-(heptyloxy )-N,N-dimethylpropane-2-amine;(2R)-1-({8-[(3β)-cholesta-5-en-3-yloxy]octyl}oxy)-N,N-dimethyl-3-[(2Z)-penta-2-en-1-yloxy]propane-2-amine;(2S)-1-butoxy-3-({8-[(3β)-cholesta-5-en-3-yloxy]octyl}oxy)-N,N-dimethylpropane-2-amine;(2S-1-({8-[(3β)-cholesta-5-en-3-yloxy]octyl}oxy)-3-[2,2,3,3 ,4,4,5,5,6,6,7,7,8,8,9,9-Hexadecafluorononyl)oxy]-N,N-dimethylpropane-2-amine;2-amino-2-{[(9Z,12Z)-Octadeca-9,12-dien-1-yloxy]methyl}propane-1,3-diol;2-amino-3-({9-[(3β,8ξ,9ξ,14ξ,17ξ,20ξ)-Cholesta-5-en-3-yloxy]nonyl}oxy)-2-{[(9Z,12Z)-Octadeca-9,12-dien-1-yloxy]methyl}propane-1-ol;2-amino-3-({6-[(3β,8ξ,9ξ,14ξ,17ξ,20ξ)-cholest-5-en-3-yloxy]hexyl}oxy)-2-{[(9Z)-octadec-9-en-1-yloxy]methyl}propan-1-ol);(20Z,23Z)-N,N-dimethylnonacosa -20,23-diene-10-amine; (17Z,20Z)-N,N-dimethylhexacosa-17,20-diene-9-amine; (16Z,19Z)-N,N-dimethylpentacosa-16,19-diene-8-amine; (13Z,16Z)-N,N-dimethyldocosa-13,16-diene-5-amine; (12Z,15Z)-N,N-dimethylhenicosa-12,15-diene-4-amine; (14Z,17Z)-N,N-dimethyltricosa-14,17-diene-6-amine; (15Z,18Z)-N,N-dimethyltetracosa-15,18-diene- 7-amine; (18Z,21Z)-N,N-dimethylheptacosa-18,21-diene-10-amine; (15Z,18Z)-N,N-dimethyltetracosa-15,18-diene-5-amine; (14Z,17Z)-N,N-dimethyltricosa-14,17-diene-4-amine; (19Z,22Z)-N,N-dimethyloctacosa-19,22-diene-9-amine; (18Z,21Z)-N,N-dimethylheptacosa-18,21-diene-8-amine; (17Z,20Z)-N,N-dimethylhexacosa-17,20-diene-7-amine; (16 (Z,19Z)-N,N-dimethylpentacosa-16,19-diene-6-amine; (22Z,25Z)-N,N-dimethylhentriaconta-22,25-diene-10-amine; (21Z,24Z)-N,N-dimethyltriaconta-21,24-diene-9-amine; (18Z)-N,N-dimethylheptacosa-18-en-10-amine; (17Z)-N,N-dimethylhexacosa-17-en-9-amine; (19Z,22Z)-N,N-dimethyloctacosa-19,22-diene-7-amine; N,N-dimethylheptacosa-10-amine;(20Z,23Z)-N-ethyl-N-methylnonacosa-20,23-dien-10-amine;1-[(11Z,14Z)-1-nonylicosa-11,14-dien-1-yl]pyrrolidine;(20Z)-N,N-dimethylheptacosa-20-en-10-amine;(15Z)-N,N-dimethylheptacosa-15-en-10-amine;(14Z)-N,N-dimethylnonacosa-14-en-10-amine;(17Z)-N,N-dimethylnonacosa-17-en-10-amine;(24Z)-N,N-dimethyltritriaconta-24-en-10 -amine; (20Z)-N,N-dimethylnonacosa-20-en-10-amine; (22Z)-N,N-dimethylhenthriaconta-22-en-10-amine; (16Z)-N,N-dimethylpentacosa-16-en-8-amine; (12Z,15Z)-N,N-dimethyl-2-nonylhenicosa-12,15-dien-1-amine; (13Z,16Z)-N,N-dimethyl-3-nonyldocosa-13,16-dien-1-amine; N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]heptadecane-8-amine; 1-[(1S [(1S,2R)-2-hexylcyclopropyl]-N,N-dimethylnonadecane-10-amine; N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]nonadecane-10-amine; N,N-dimethyl-21-[(1S,2R)-2-octylcyclopropyl]henicosane-10-amine; N,N-dimethyl-1-[(1S,2S)-2-{[(1R,2R)-2-pentylcyclopropyl]methyl}cyclopropyl]nonadecane-10-amine; N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]hexadecane -8-amine; N,N-dimethyl-1-[(1R,2S)-2-undecylcyclopropyl]tetradecane-5-amine; N,N-dimethyl-3-{7-[(1S,2R)-2-octylcyclopropyl]heptyl}dodecane-1-amine; 1-[(1R,2S)-2-heptylcyclopropyl]-N,N-dimethyloctadecane-9-amine; 1-[(1S,2R)-2-decylcyclopropyl]-N,N-dimethylpentadecane-6-amine; N,N-dimethyl-1-[(1S,2R)-2-octylcyclopropyl]pentadecane-8-amine;(11E,20Z,23Z)-N,N-dimethylnonacosa-11,20,23-triene-10-amine; 2,2-dilinoleyl-5-dimethylaminomethyl-[1,3]-dioxane (DLin-K6-DMA), 2,2-dilinoleyl-4-N-methylpepiazino-[1,3]-dioxolane (DL (in-K-MPZ), 2,2-dioleoyl-4-dimethylaminomethyl-[1,3]-dioxolane (DO-K-DMA), 2,2-distearoyl-4-dimethylaminomethyl-[1,3]-dioxolane (DS-K-DMA), 2,2-dilinoleyl-4-N-morpholino-[1,3]-dioxolane (DLin-K-MA), 2,2-dilinoleyl-4-trimethylamino-[1,3]-dioxolancholine; (DLin-K-TMA.Cl), 2,2-dilinoleyl-4,5-bis(dimethylaminomethyl)-[1,3]-dioxolane (DLin-K2-DMA), 2,2-dilinoleyl-4-methylpiperzine-[1,3]-dioxolane (D-Lin-KN-methylpiperzine), DLen- C2K-DMA, γ-DLen-C2K-DMA, DPan-C2K-DMA, DPan-C3K-DMA, DLen-C2K-DMA, γ-DLen-C2K-DMA, DPan-C2K-DMA, TLinDMA, C2-TLinDMA, C3-TLinDMA, 1,2-di-γ-linolenyloxy-N,N-dimethylaminopropane (γ-DLenDMA), 1,2-dilinoleyloxy-(N,N- Dimethyl)-butyl-4-amine (C2-DLinDMA), 1,2-Dilinoleoyloxy-(N,N-dimethyl)-butyl-4-amine (C2-DLinDAP), CP-LenMC3, CP-γ-LenMC3, CP-MC3, CP-DLen-C2K-DMA, CP-γDLen-C2K-DMA, CP-C2K-DMA, CP-DODMA, CP-DPetroDMA, CP-DLinDMA, CP-DLenDMA, CP-γDLenDMA, 1,2-Dioeylcarbamoyloxy-3-dimethylaminopropane (DO-C-DAP), 1,2-Dimyristreoyl-3-dimethylaminopropane (DMDAP), 1,2-Dioleoyl-3-trimethylaminopropane chloride (DOTAP).Cl), 1,2-dilinoleylcarbamoyloxy-3-dimethylaminopropane (DLin-C-DAP), 1,2-dilinoleoxy-3-(dimethylamino)acetoxypropane (DLin-DAC), 1,2-dilinoleoxy-3-morpholinopropane (DLin-MA), 1,2-dilinoleyl-3-dimethylaminopropane (DLinDAP), 1,2-dilinoleylthio-3-dimethylamino Propane (1,2-dilinoleylthio-3-dimethylarninopropane) (DLin-S-DMA), 1-linoleoyl-2-linoleyloxy-3-dimethylaminopropane (DLin-2-DMAP), 1,2-dilinoleoyloxy-3-trimethylaminopropane chloride (DLin-TMA.Cl), 1,2-dilinoleoyl-3-trimethylaminopropane (1,2-dilinoleoyl-3-trimethylaminopropane) chloride (DLin-TAP.Cl), 1,2-dilinoleyloxy-3-(N-methylpiperazino)propane (DLin-MPZ), 3-(N,N-dilinoleylamino)-1,2-propanediol (DLinAP), 3-(N,N-dioleylamino)-1,2-propanediol (3-(N,N-dioleylamino)-1,2-propanedio) (DOAP), 1,2-dilinoleyloxo-3-(2-N,N-dimethylamino)ethoxypropane (DLin-EG-DMA), 3-dimethylamino-2-(cholesta-5-ene-3-beta-oxybutane-4-oxy)-1-(cis,cis-9,12-octadecadienoxy)propane (CLinDMA), 2-[5'-(cholesta-5-ene-3-beta-oxy)-3'-oxapentoxy)-3 Examples include -dimethyl(dimethy)-1-(cis,cis-9',1-2'-octadecadienoxy)propane (2-[5'-(cholest-5-en-3-beta-oxy)-3'-oxapentoxy)-3-dimethy-1-(cis,cis-9',1-2'-octadecadienoxy)propane) (CpLinDMA), N,N-dimethyl-3,4-dioleyloxybenzylamine (DMOBA), 1,2-N,N'-dioleylcarbamyl-3-dimethylaminopropane (DOcarbDAP), and 1,2-N,N'-dilinoleylcarbamyl-3-dimethylaminopropane (DLincarbDAP), as well as their pharmaceutically acceptable salts and any stereoisomers thereof.

[0271] As used herein, the term “noncationic lipid” refers to any uncharged, anionic, or amphoteric lipid. At physiological pH, such lipids include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebroside, diacylglycerol, phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamine, N-succinylphosphatidylethanolamine, N-glutarylphosphatidylethanolamine, lysylphosphatidylglycerol, palmitoyloleyolphosphatidylglycerol (POPG), and other anionic modifying groups attached to neutral lipids. In some embodiments, the noncationic lipids used in this disclosure are 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), and / or 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE). In some embodiments, the noncationic lipids are cholesterol (CHE) and / or β-sitosterol.

[0272] In some embodiments, the noncationic lipids present in the lipid particles include, or consist of, a mixture of one or more phospholipids and cholesterol or its derivatives.

[0273] In certain embodiments, the lipid compositions of the present disclosure may include lipids such as “neutral lipids,” “helper lipids,” and / or “stealth lipids.”

[0274] "Neutral lipids" suitable for use in the lipid compositions of this disclosure include, for example, a variety of neutral, uncharged, or amphoteric lipids. In some embodiments, the neutral lipids disclosed herein may include, for example, diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebroside, and diacylglycerol. Other examples of neutral phospholipids suitable for use in this disclosure include, but are not limited to, distearoyl phosphatidylcholine (DSPC), phosphocholine (DOPC), dimyristoyl phosphatidylcholine (DMPC), phosphatidylethanolamine (PE), egg phosphatidylcholine (EPC), palmitoyl oleoyl phosphatidylcholine (POPC), dioleoyl phosphatidylethanolamine (DOPE), dilinoleoyl phosphatidylcholine, distearoyl phosphatidylethanolamine (DSPE), dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), palmitoyl oleoyl phosphatidylethanolamine (POPE), lysophosphatidylethanolamine, and combinations thereof. In one embodiment, the neutral phospholipid may be selected from the group consisting of dioleoyl phosphatidylethanolamine (DOPE). In another embodiment, the neutral phospholipid may be distearoylphosphatidylcholine (DSPC). While not strictly theoretical, it has been explained that the neutral lipid functions to stabilize LNPs and improve their processing.

[0275] "Helper lipids" are lipids that enhance transfection (e.g., transfection of nanoparticles containing bioactive drugs). While not strictly theoretical, the mechanism by which helper lipids enhance transfection includes enhancing particle stability. In certain embodiments, helper lipids enhance fusogenicity. Helper lipids include the above-mentioned "neutral lipids," which include, but are not limited to, distearoyl phosphatidylcholine (DSPC), phosphocholine (DOPC), dimyristoyl phosphatidylcholine (DMPC), phosphatidylethanolamine (PE), egg phosphatidylcholine (EPC), palmitoyl oleoyl phosphatidylcholine (POPC), dioleoyl phosphatidylethanolamine (DOPE), dilinoleoyl phosphatidylcholine, distearoyl phosphatidylethanolamine (DSPE), dimyristoyl phosphatidylethanolamine (DMPE), dipalmitoyl phosphatidylethanolamine (DPPE), palmitoyl oleoyl phosphatidylethanolamine (POPE), lysophosphatidylethanolamine and combinations thereof, as well as steroids and sterols. Helper lipids suitable for use in this disclosure include, but are not limited to, neutral lipids, cholesterol, and PEG-cholesterol. In one embodiment, the helper lipid may be cholesterol. In one embodiment, the helper lipid may be PEG-cholesterol.

[0276] "Stealth lipids" are lipids that alter the length of time nanoparticles can exist in vivo (e.g., in the blood). While not strictly theoretical, stealth lipids can assist in formulation processes, for example, by reducing particle aggregation and controlling particle size. The stealth lipids used herein can modulate the pharmacokinetic properties of LNPs. Suitable stealth lipids for use in the lipid compositions of this disclosure include, but are not limited to, stealth lipids having hydrophilic head groups linked to the lipid moiety. Information on suitable stealth lipids for use in the lipid compositions of this disclosure, and the biochemistry of such lipids, can be found in Romberg et al, Pharmaceutical Research, Vol. 25, No. 1, 2008, pg. 55-71 and Hoekstra et al, Biochimica et Biophysica Acta 1660 (2004) 41-52. In one embodiment, the hydrophilic head group of the stealth lipid includes a polymer moiety selected from polymers based on PEG (sometimes called poly(ethylene oxide)), poly(oxazoline), poly(vinyl alcohol), poly(glycerol), poly(N-vinylpyrrolidone), polyamino acids, and polyN-(2-hydroxypropyl)methacrylamide. The stealth lipid may include a lipid moiety. In some embodiments, the lipid moiety of the stealth lipid can be derived from diacylglycerol or diacylglycamide, which includes about C4 to about C 40The lipid moiety comprises a dialkylglycerol or dialkylglycamide group having an alkyl chain length independently containing saturated or unsaturated carbon atoms, where the chain may contain one or more functional groups such as amides or esters. The dialkylglycerol or dialkylglycamide group may further contain one or more substituted alkyl groups. In some embodiments, the stealth lipid may include α-methoxy-ω-(3-oxopropoxy), polyoxyethylene (methoxyPEG, aldehyde), PEG2k-DMG, PEG2k-DSG, PEG2k-DSPE, PEG2K-DOPE, PEG5k-DOPE, methoxyPEG aldehyde 20k, PEG2K-cholesterol, and analogues.

[0277] The term “lipid nanoparticles (LNPs),” as used herein, refers to compositions of different types of nanoscale particles in which lipid-containing particles function as carriers for crossing cell membranes and biological barriers, delivering compounds to target cells and tissues of humans and other organisms. As used herein, “lipid nanoparticles” in this disclosure may further include additional lipids and other components. Other lipids may be included for various purposes, such as preventing lipid oxidation or binding ligands to the surface of the lipid nanoparticles. Lipid nanoparticles in this disclosure may include any of a number of lipids, including amphiphilic, neutral, cationic, and anionic lipids. Such lipids may be used alone or in combination and may also include bilayer stabilizing components such as polyamide oligomers (see, e.g., U.S. Patent No. 6,320,017), peptides, proteins, surfactants (detergents), lipid derivatives, and PEG compounded with phosphatidylethanolamine and PEG compounded with ceramide (see, e.g., U.S. Patent No. 5,885,613).

[0278] As used herein, the “PEG” complex lipids that inhibit particle aggregation refer to one or more polyethylene glycol (PEG)-lipid complexes, polyamide (ATTA)-lipid complexes, and mixtures thereof. In one embodiment, the PEG-lipid complex is one or more PEG-dialkyloxypropyl (DAA), PEG-diacylglycerol (DAG), PEG-phospholipid, PEG-ceramide, and mixtures thereof. In one embodiment, the PEG-DAG complex is PEG-dilauroylglycerol (C) 12 ), PEG-dimyristoylglycerol (C 14 ), PEG-Dipalmitoylglycerol (C 16 ), and PEG-distearoylglycerol (C 18 ) is one or more of the following. In one embodiment, the PEG-DAA complex is PEG-dilauryloxypropyl (C 12 ), PEG-Dimyristyloxypropyl (C 14 ), PEG-Dipalmityloxypropyl(C 16 ), and PEG-distearyloxypropyl (C 18 ) is one or more of the above. In some embodiments, PEG is 2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG-DMG or PEG2k-DMG) and / or 1,2-distearoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG-DSG).

[0279] As used herein, the term "N / P ratio" refers to the molar ratio of nitrogen (N) to phosphorus (P) between the cationic aminolipid and the negatively charged phosphate group of the nucleic acid.

[0280] The "polydispersion index" or "PDI," as used herein, is a measure of heterogeneity based on sample size. Polydispersion can arise from the size distribution within a sample, or from the aggregation or clumping of samples during isolation or analysis.

[0281] When used herein, "zeta potential" or "surface charge" refers to the degree of electrostatic repulsion between adjacent, similarly charged particles in a dispersion. For sufficiently small molecules and particles, a high zeta potential imparts stability, i.e., the solution or dispersion resists aggregation.

[0282] As used herein, the term nucleic acid “cargo” refers to nucleic acids intended to be delivered to cells or tissues (in some embodiments, therapeutic nucleic acids for delivery to cells or tissues).

[0283] As used herein, the term “nucleic acid-lipid nanoparticles” refers to the lipid nanoparticles described above that associate with or encapsulate one or more nucleic acids to deliver one or more nucleic acid cargoes to a tissue.

[0284] As used herein, “encapsulated” can refer to nucleic acid-lipid nanoparticle formulations that provide complete encapsulation, partial encapsulation, association by ionic or van der Waals forces, or all of the aforementioned. In one embodiment, the nucleic acid is completely encapsulated in nucleic acid-lipid nanoparticles.

[0285] As used herein, “nucleic acid” refers to synthetic or naturally occurring RNA or DNA, or derivatives thereof. In one embodiment, the cargo and / or agent of the Disclosure is a nucleic acid, such as double-stranded RNA (dsRNA). In one embodiment, the nucleic acid or nucleic acid cargo is single-stranded DNA or RNA, double-stranded DNA or RNA, or a DNA-RNA hybrid. For example, double-stranded DNA may be a structural gene, a gene containing regulatory and terminal regions, or a self-replicating system such as a virus or plasmid DNA. Double-stranded RNA may be, for example, dsRNA or another RNA interference reagent. Single-stranded nucleic acid may be, for example, mRNA, antisense oligonucleotide, ribozyme, microRNA, or triple-helix-forming oligonucleotide. In certain embodiments, the nucleic acid or nucleic acid cargo may include modified RNA, where modified RNA is one or more modified mRNA, modified antisense oligonucleotide, and modified siRNA. In some embodiments, the nucleic acid cargo of the Disclosure includes, or is, modified mRNA encoding a nucleic acid regulatory factor.

[0286] As used herein, the term “modified nucleic acid” includes 2'-O-methyl modified nucleotides, nucleotides containing a 5'-phosphorothioate group, terminal nucleotides linked to cholesteryl derivatives, 2'-deoxy-2'-fluoro modified nucleotides, 5'-methoxy modified nucleotides (e.g., 5'-methoxyuridine), 2'-deoxy modified nucleotides, locked nucleotides, debased nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides, phosphoramidates, nucleotides containing unnatural bases; phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, and aminoalkyl phosphotriesters. This refers to any non-natural nucleic acids, selected from the group including, but not limited to, methyl and other alkyl phosphonates, phosphinates, 3'-aminophosphoamides and aminoalkylphosphoamides, thionophosphoamides, thionoalkylphosphonates, thionoalkylphosphotryesters, and boranophosphates having the usual 3'-5' linkage, their 2'-5' linked analogs, and internucleoside linkages or skeletons having inverted polarity, including those in which pairs of adjacent nucleoside units are linked from 3'-5' to 5'-3' or 2'-5' to 5'-2'.

[0287] As used herein, the term “nucleic acid modulating controller” refers to mRNA encoding a protein regulatory component, but a reference to “nucleic acid modulating controller” may also refer to the protein regulatory component itself expressed by the mRNA. In certain embodiments, the protein regulatory component encoded by the mRNA includes a zinc finger protein (ZFP) or other form of DNA or RNA-binding domain (DBD or RBD) associated with (or tethered to, as necessary) one or more epigenetic regulators or nucleases (the epigenetic regulators or nucleases are generally referred to as effectors, effector domains, or effector moieties). While not strictly theoretical, the advantage of the nucleic acid regulatory factors described herein is that they provide sustained gene programming only at the confluence point where (1) mRNA encoding the nucleic acid regulatory factor is expressed, (2) the nucleic acid binds to a ZFP or other nucleic acid binding domain, and (3) the relevant effector domain can exert activity (i.e., the effector domain can alter the epigenomic state (for example, in the case of an epigenomic regulator)).

[0288] As used herein, the terms “effector moiety” or “effector domain” refer to a domain that, when localized to an appropriate site within a cell, such as the cell nucleus, can alter the expression of a target gene. In some embodiments, the effector moiety recruits components of a transcription mechanism. In some embodiments, the effector moiety inhibits the recruitment of components of a transcription factor or repressor. In some embodiments, the effector moiety includes an epigenetic modification moiety (e.g., an epigenetic modification of a target DNA sequence). Specific examples of effector moieties, but not limited to, include, in particular, effectors that can bind to a Kruppel-associated box (KRAB) domain (KRAB is a domain of approximately 75 amino acids found at the N-terminus of about one-third of eukaryotic Kruppel-type C2H2 zinc finger proteins (ZFPs)), and genetically engineered prokaryotic DNA methyltransferase MQ1.

[0289] As used herein, “epigenetic modification moiety” refers to a domain that, when appropriately localized to a nucleic acid (e.g., by a targeted moiety), alters i) the structure of chromatin, e.g., two-dimensional structure, and / or ii) one or more epigenetic markers (e.g., DNA methylation, histone methylation, histone acetylation, histone SUMOylation, histone phosphorylation, and RNA-associated silencing). In some embodiments, the epigenetic modification moiety includes an enzyme, or a functional fragment or variant thereof, that affects one or more epigenetic markers (e.g., by increasing or decreasing their levels). In some embodiments, the epigenetic modification moiety includes a DNA methyltransferase, a histone methyltransferase, a CREB-binding protein (CBP), or a functional fragment of any of these.

[0290] As used herein, the term “expression regulatory sequence” refers to a nucleic acid sequence that increases or decreases the transcription of a gene, and includes, but is not limited to, promoters and enhancers. “Enhancing sequence” refers to a subtype of expression regulatory sequence that increases the potential for gene transcription. “Silencing or repressing sequence” refers to a subtype of expression regulatory sequence that decreases the potential for gene transcription.

[0291] As used herein, the term “repressor” refers to one or more functional agents or entities that reduce the expression of a target gene within a cell and specifically bind to a DNA sequence (e.g., a DNA sequence associated with the target gene, or a transcriptional regulatory element operably linked to the target gene). In certain embodiments, the repressor comprises at least one targeting moiety and optionally one effector moiety.

[0292] As used herein, the term “targeting region” means an active agent or entity that specifically targets, for example, a regulatory or anchoring sequence, promoter, enhancer, or CTCF site of a genomic sequence (e.g., an expression regulatory sequence or anchoring sequence, promoter, enhancer, or CTCF site). In some embodiments, the genomic sequence element is adjacent to and / or operably linked to a target gene (e.g., MYC).

[0293] As used herein, “localization” refers to the location of the lipids, peptides, or other components of the lipid particles of this disclosure within a living organism and / or tissue. In some embodiments, localization can be detected in individual cells. In some embodiments, labels, such as fluorescent labels, optionally fluorescently labeled lipids, or optionally Cy7, can be used to detect localization. In some embodiments, the label of the lipid nanoparticles may be quantum dots or lipids detectable by stimulated Raman scattering. In other embodiments, the label is any fluorophore known in the art, i.e., a fluorophore that exhibits excitation and emission in the ultraviolet, visible, or infrared spectrum. In some embodiments, localization is detected or further demonstrated by immunohistochemistry or immunofluorescence.

[0294] As used herein, the term “activity” refers to any detectable effect mediated by the components or compositions of the present disclosure. In some embodiments, “activity” may refer, as used herein, to a measurable effect (whether direct or surrogate) of, for example, the lipid particle cargo of the present disclosure. Examples of activity, but not limited to, include the intracellular expression of nucleic acid cargo (e.g., mRNA, CRISPR / Cas systems, RNAi agents, nucleic acid regulatory factors, etc.) and the resulting effects, which may be measured as needed at the cellular, tissue, organ, and / or biological levels.

[0295] As used herein, “multiple doses” refers to two or more doses of a lipid nanoparticle formulation given to a subject as part of a treatment regimen.

[0296] As used herein, the term “subjects” includes humans and mammals (e.g., mice, rats, pigs, cats, dogs, and horses). In many embodiments, subjects are mammals, particularly primates, and especially humans. In some embodiments, subjects are livestock such as cattle, sheep, goats, cows, pigs, and similar animals; poultry such as chickens, ducks, geese, turkeys, and similar animals; and domesticated animals, particularly pets, such as dogs and cats. In some embodiments (e.g., particularly in the context of research), subject mammals are, for example, rodents (e.g., mice, rats, hamsters), rabbits, primates, or pigs and similar animals such as inbred pigs.

[0297] As used herein, “administration” to a subject may include, as necessary, intravenous injection, inhalation, intravenous, intra-arterial, intratracheal, topical, parenteral administration, or direct injection into tissue.

[0298] The term "treatment" includes the administration of compositions for preventing or delaying the onset of symptoms, complications, or biochemical signs of a disease (e.g., cancer, including tumor formation, growth, and / or metastasis), reducing symptoms, or inhibiting or suppressing further development of the disease, condition, or disorder. Treatment may be prophylactic (preventing or delaying the onset of the disease, or preventing the manifestation of its clinical or subclinical symptoms) or therapeutic suppression or mitigation of symptoms after the onset of the disease.

[0299] As used herein, “pharmaceutical composition” comprises a pharmacologically effective amount of lipid particles, optionally nucleic acid lipid nanoparticles (NLNPs), and a pharmaceutically acceptable carrier. As used herein, “pharmacologically effective amount,” “therapeutic effective amount,” or simply “effective amount” refers to the amount of nucleic acid effective to produce an intended pharmacological, therapeutic, or preventive effect. For example, if a given clinical treatment is considered effective when a measurable parameter associated with a disease or disorder is reduced by at least 25%, then the therapeutic effective amount of a drug for treating that disease or disorder is the amount required to induce at least a 25% reduction in that parameter.

[0300] The term "pharmacologically acceptable carrier" refers to a carrier for administering a therapeutic agent. Such carriers include, but are not limited to, saline, buffered saline, glucose, water, glycerol, ethanol, and combinations thereof.

[0301] As used herein, “lung tissue” can refer to any cells within the lung organ, including but not limited to the group including epithelium, endothelium, interstitial connective tissue, blood vessels, hematopoietic tissue, lymphoid tissue, and pleura. In preferred embodiments, nucleic acid-lipid nanoparticles target lung tissue. In some other embodiments, nucleic acid-lipid nanoparticles can target other cells or tissues, including but not limited to cells or tissues of the brain, nerves, skin, eyes, pharynx, larynx, heart, blood vessels, hematopoietic system (e.g., white blood cells or red blood cells), breasts, liver, pancreas, spleen, esophagus, gallbladder, stomach, intestines, colon, kidneys, bladder, ovaries, uterus, cervix, prostate, muscle, bone, thyroid gland, parathyroid gland, adrenal gland, and pituitary gland.

[0302] As used herein, “localization” refers to the location of the lipids, peptides, or other components of the lipid particles of this disclosure within a living organism and / or tissue. In some embodiments, localization can be detected in individual cells. In some embodiments, labels, such as fluorescent labels, optionally fluorescently labeled lipids, or optionally Cy7, can be used to detect localization. In some embodiments, the label of the lipid nanoparticles may be quantum dots or lipids detectable by stimulated Raman scattering. In other embodiments, the label is any fluorophore known in the art, i.e., a fluorophore that exhibits excitation and emission in the ultraviolet, visible, or infrared spectrum. In some embodiments, localization is detected or further demonstrated by immunohistochemistry or immunofluorescence.

[0303] As used herein, the term “lung disease or disorder” may include, but is not limited to, diseases or disorders selected from lung cancer, pneumonia, pulmonary fibrosis, COPD, asthma, bronchiectasis, sarcoidosis, pulmonary hypertension, emphysema, alpha-1 antitrypsin deficiency, aspergillosis, bronchiolitis, bronchitis, pneumoconiosis, coronavirus, Middle East respiratory syndrome, severe acute respiratory syndrome, cystic fibrosis, Legionnaires' disease, influenza, pertussis, pulmonary embolism, and tuberculosis.

[0304] As used herein, “Joint disease or disorder” may include, but is not limited to, diseases or disorders selected from rheumatoid arthritis, psoriatic arthritis, gout, tendinitis, bursitis, carpal tunnel syndrome, and osteoarthritis.

[0305] As used herein, “inflammatory disease or disorder” includes, but is not limited to, inflammatory bowel disease, peritonitis, osteomyelitis, cachexia, pancreatitis, trauma-induced shock, bronchial asthma, allergic rhinitis, cystic fibrosis, acute bronchitis, acute severe bronchitis, osteoarthritis, rheumatoid arthritis, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, arthritis, osteoarthritis, gout, spondyloarthritis, ankylosing spondylitis, arthritis associated with vasculitis syndrome, neurogenic polyarteritis nodosa, hypersensitivity vasculitis, rugenic granulomatosis, and rheumatic polyposis. This may include diseases or disorders selected from among myalgia, inflammatory cell arteritis, calcium polycystic arthropathy, corrosive gout, non-arthritis, bursitis, hay fever, purulent inflammation (e.g., tennis elbow), neuropathic joint disease, intra-articular hemorrhage, Henoch-Schlein purpura, hypertrophic osteoarthritis, multiple hemorrhoids, scoliosis, hemochromatosis, hyperlipoproteinemia, hypogammaglobulinemia, COPD, acute respiratory distress syndrome, acute lung injury, bronchopulmonary dysplasia, and systemic lupus erythematosus (SLE).

[0306] As used herein, “epidermal disease or disorder” may include, but is not limited to, diseases or disorders selected from psoriasis, atopic dermatitis, scleroderma, eczema, rosacea, seborrheic dermatitis, melanoma, actinic keratosis, ichthyosis, Glover's disease, verruca vulgaris, keratocanthoma, and seborrheic keratosis.

[0307] As used herein, the term "or" is understood to be inclusive unless otherwise specifically stated or evident from the context. As used herein, the terms "a," "an," and "the" are understood to be singular or plural unless otherwise specifically stated or evident from the context.

[0308] In this specification, a range can be expressed as from a certain value with "approximately" attached and / or to another specific value with "approximately" attached. When such a range is expressed, another aspect includes from a certain value and / or to another specific value. Similarly, when a value is expressed as an approximation by using the antecedent "approximately", it is understood that a particular value forms another aspect. Furthermore, it is understood that each endpoint of a range is important, both in relation to the other endpoint and independently of the other endpoint. Also, it is understood that numerous values ​​are disclosed in this specification, and each value is disclosed in this specification not only as the value itself but also as a specific value with "approximately" attached. Furthermore, throughout this application, data is provided in numerous different forms, and it is understood that this data represents endpoints and starting points, as well as ranges, for any combination of data points. For example, when a specific data point "10" and a specific data point "15" are disclosed, it is understood that values ​​greater than 10 and 15, equal to or greater than 10 and 15, less than 10 and 15, equal to or less than 10 and 15, and between 10 and 15 are disclosed. It is also understood that each unit between two specific units is disclosed. For example, if 10 and 15 are disclosed, then 11, 12, 13, and 14 are also disclosed.

[0309] The ranges provided herein are understood to be abbreviations for all values ​​within that range. For example, the range 1 to 50 is understood to include any number, combination of numbers, or subranges of the group consisting of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, or 50, and all decimal values ​​that lie between the aforementioned integers, such as 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, and 1.9. With respect to subranges, "nested subranges" extending from any endpoint of the range are specifically intended. For example, nested subranges of the exemplary range 1-50 may include 1-10, 1-20, 1-30, and 1-40 in one direction, or 50-40, 50-30, 50-20, and 50-10 in the other direction.

[0310] The transitional term “comprising,” which is synonymous with “including,” “contains,” or “characterized by,” is comprehensive or open-ended and does not exclude additional, unlisted elements or method steps. In contrast, the transitional phrase “consisting of” excludes any elements, steps, or components not specified in the claims. The transitional phrase “essentially consisting of” limits the claims to the specified materials or steps “and not substantially affect the basic and novel features of the disclosure.”

[0311] As used herein, the term "alkyl" refers to a linear or branched saturated hydrocarbon group having 1 to 22 carbon atoms ("C"). 1~22 "alkyl" refers to an alkyl group consisting of 3 to 22 carbon atoms. In some embodiments, an alkyl group consists of 3 to 22 carbon atoms ("C"). 3~22 Alkyl) and / or 7 to 22 carbon atoms ("C") 7~22 The alkyl group may have 7 to 18 carbon atoms ("C"). In some embodiments, the alkyl group may have 7 to 18 carbon atoms ("C").7~18 Alkyl) and / or 7 to 12 carbon atoms ("C") 7~12 Alkyl("C") may have 7 to 8 carbon atoms. In some embodiments, the alkyl group may have 7 to 8 carbon atoms. 7~8 ("alkyl"). In some embodiments, the alkyl group has 7 to 9 carbon atoms ("C"). 7~9 ("alkyl"). In some embodiments, the alkyl group may have 7 to 10 carbon atoms ("C"). 7~10 ("alkyl"). In some embodiments, the alkyl group has 7 to 11 carbon atoms ("C"). 7~11 ("alkyl"). In some embodiments, the alkyl group may have 8 to 12 carbon atoms ("C"). 8~12 ("alkyl"). In some embodiments, the alkyl group has 9 to 12 carbon atoms ("C"). 9~12 ("alkyl"). In some embodiments, the alkyl group has 10 to 12 carbon atoms ("C"). 10~12 (alkyl). In some embodiments, the alkyl group has 11 to 12 carbon atoms ("C"). 11~12 Alkyl). Further examples of alkyl groups include n-heptyl (C7), n-octyl (C8), n-nonyl (C9), and n-decyl (C 10 ), n-undecyl(C 11 ), n-dodecyl(C 12 Examples include ), and similar items.

[0312] The "alkyl" group, as used herein, may be unsubstituted or optionally substituted. Unless otherwise specified, each alkyl group may independently be optionally substituted, i.e., unsubstituted ("unsubstituted alkyl") or substituted with one or more substituents ("substituted alkyl"). Preferred substituents include hydroxyl, nitro, amino (e.g., -NH2 or dialkylamino), imino, cyano, halo (e.g., F, Cl, Br, I, and analogous atoms), haloalkyl (e.g., -CCl3, -CF3, and analogous groups), thio, sulfonyl, thioamide, amidino, imidino, oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamide, carboxyl, formyl, alkyl, alkoxy, alkoxy-alkyl, alkylcarbonyl, alkylcarbonyloxy (e.g., -OCOR), aminocarbonyl, arylcarbonyl, aralkylcarbonyl, carbonylamino, heteroarylcarbonyl This may include, but is not limited to, groups such as heteroaralkyl-carbonyl, alkylthio, aminoalkyl, cyanoalkyl, carbamoyl (e.g., -NHCOOR- or -OCONHR-), urea (e.g., -NHCONHR-), cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, (=O), thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amide, N-amide, C-carboxy, O-carboxy, nitro, amino, heterocyclic, -CN, and similar groups. When used herein, "alkyl" can form one or more substituted alkyl groups and / or functionalized alkyl groups together with other groups, e.g., those provided above.

[0313] The “alkyl” group as defined above may further contain one or more (e.g., 1, 2, 3, 4, etc.) heteroatoms in the parent chain (e.g., “heteroalkyl,” e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus, and analogous atoms), where one or more heteroatoms are inserted between adjacent carbon atoms in the parent carbon chain, and / or one or more heteroatoms are inserted between carbon atoms and the parent molecule, i.e., between bonding points. In certain embodiments, the heteroalkyl group is a saturated group having 1 to 22 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("heteroC"). 1~22 "Alkyl" refers to a saturated group having 3 to 22 carbon atoms and / or 7 to 22 carbon atoms, as well as 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 3~22 "Alkyl" and / or "hetero-C" 7~22 This refers to an alkyl group. In some embodiments, a heteroalkyl group may have 7 to 18 carbon atoms and / or 7 to 12 carbon atoms, as well as 1, 2, 3, 4, or other heteroatoms (hetero C 7~18 "Alkyl" and / or "hetero-C" 7~12 ("Alkyl"). In some embodiments, heteroalkyl groups may have 7 to 8 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~8 ("Alkyl"). In some embodiments, heteroalkyl groups may have 7 to 9 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~9 ("Alkyl"). In some embodiments, a heteroalkyl group has 7 to 10 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 7~10 ("Alkyl"). In some embodiments, a heteroalkyl group has 7 to 11 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~11 ("Alkyl"). In some embodiments, a heteroalkyl group has 8 to 12 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 8~12("Alkyl"). In some embodiments, a heteroalkyl group has 9 to 12 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 9~12 ("Alkyl"). In some embodiments, a heteroalkyl group has 10 to 12 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 10~12 ("Alkyl"). In some embodiments, a heteroalkyl group has 11 to 12 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 11~12 Alkyl).

[0314] As used herein, the term “alkenyl” means a branched carbon chain having 2 to 22 carbon atoms and containing at least one double bond (e.g., 1, 2, 3, 4 carbon-carbon double bonds, etc.). 2~22 The alkenyl group contains 3 to 22 carbon atoms ("C"). In some embodiments, the alkenyl group contains 3 to 22 carbon atoms ("C"). 3~22 "Alkenyl") and / or 7 to 22 carbon atoms ("C") 7~22 It may have an alkenyl group. In some embodiments, the alkenyl group has 7 to 18 carbon atoms ("C"). 7~18 "Alkenyl") and / or 7 to 12 carbon atoms ("C") 7~12 It may have an alkenyl group. In some embodiments, the alkenyl group has 7 to 8 carbon atoms ("C"). 7~8 ("Alkenyl"). In some embodiments, the alkenyl group has 7 to 9 carbon atoms ("C"). 7~9 ("Alkenyl"). In some embodiments, the alkenyl group may have 7 to 10 carbon atoms ("C"). 7~10 ("Alkenyl"). In some embodiments, the alkenyl group has 7 to 11 carbon atoms ("C"). 7~11 ("Alkenyl"). In some embodiments, the alkenyl group may have 8 to 12 carbon atoms ("C"). 8~12 ("Alkenyl"). In some embodiments, the alkenyl group has 9 to 12 carbon atoms ("C"). 9~12 ("Alkenyl"). In some embodiments, the alkenyl group has 10 to 12 carbon atoms ("C").10~12 ("Alkenyl"). In some embodiments, the alkenyl group has 11 to 12 carbon atoms ("C"). 11~12 Alkenyl). Further examples of alkenyl groups include n-heptyl (C7), n-octyl (C8), n-nonyl (C9), and n-decyl (C 10 ), n-undecyl(C 11 ), n-dodecyl(C 12 Examples include ), and analogues. One or more carbon-carbon double bonds may be internal (e.g., 2-butenyl) or terminal (e.g., 1-butenyl). C 2~4 Examples of alkenyl groups include ethenyl (C2), 1-propenyl (C3), 2-propenyl (C3), 1-butenyl (C4), 2-butenyl (C4), butadienyl (C4), and analogues. 2~6 As an example of an alkenyl group, the aforementioned C 2~4 Examples of alkenyl groups include pentenyl (C5), pentadienyl (C5), hexenyl (C6), and analogues. Further examples of alkenyls include heptenyl (C7), octenyl (C8), octatrienyl (C8), and analogues.

[0315] Unless otherwise specified, each case of the alkenyl group is independently substituted as necessary, i.e., unsubstituted ("unsubstituted alkenyl") or substituted with one or more substituents, e.g., 1 to 5 substituents, 1 to 3 substituents, or 1 substituent ("substituted alkenyl"). In certain embodiments, the alkenyl group is unsubstituted C 3~22 It is an alkenyl. In certain embodiments, the alkenyl group is a substituted C 3~22 It is an alkenyl. Exemplary substituents are listed above for "alkyl" and can be similarly used for "alkenyl" here.

[0316] The term "heteroalkenyl," as used herein, refers to the alkenyl group as defined above, further comprising one or more (e.g., 1, 2, 3, 4, etc.) heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus, and analogous atoms), where one or more heteroatoms are inserted between adjacent carbon atoms in the parent carbon chain, and / or one or more heteroatoms are inserted between carbon atoms and the parent molecule, i.e., between bonding points. In some embodiments, the heteroalkenyl group is an unsaturated group having 2 to 22 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C 2~22 "Alkenyl" refers to an unsaturated group having 7 to 18 carbon atoms and / or 7 to 12 carbon atoms, as well as 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 7~18 "Alkenyl" or "HeteroC" 7~12 This refers to an "alkenyl" group. In some embodiments, a heteroalkenyl group can have 7 to 8 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~8 ("Alkenyl"). In some embodiments, the heteroalkenyl group may have 7 to 9 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~9 ("Alkenyl"). In some embodiments, the heteroalkenyl group has 7 to 10 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 7~10 ("Alkenyl"). In some embodiments, the heteroalkenyl group has 7 to 11 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 7~11 ("Alkenyl"). In some embodiments, the heteroalkenyl group has 8 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 8~12 ("Alkenyl"). In some embodiments, the heteroalkenyl group has 9 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 9~12 ("Alkenyl"). In some embodiments, the heteroalkenyl group has 10 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 10~12("Alkenyl"). In some embodiments, the heteroalkenyl group has 11 to 12 carbon atoms and 1, 2, 3, 4 heteroatoms, etc. ("hetero C"). 11~12 Alkenyl). Further examples of alkenyl groups include n-heptyl (C7), n-octyl (C8), n-nonyl (C9), and n-decyl (C 10 ), n-undecyl(C 11 ), n-dodecyl(C 12 Examples of alkenyls include heptenyl (C7), octenyl (C8), octatrienyl (C8), and analogues. One or more carbon-carbon double bonds may be internal (e.g., 2-butenyl) or terminal (e.g., 1-butenyl). Examples of alkenyls include heptenyl (C7), octenyl (C8), octatrienyl (C8), and analogues.

[0317] As used herein, the term "alkynyl" means branched as necessary and containing 2 to 22 carbon atoms ("C"). 2~22 The alkynyl group comprises a carbon chain containing at least one carbon-carbon triple bond (i.e., C≡C). In some embodiments, the alkynyl group comprises 3 to 22 carbon atoms ("C"). 3~22 "Alkynyl") and / or 7 to 22 carbon atoms ("C") 7~22 It may have an alkynyl group. In some embodiments, the alkynyl group has 7 to 18 carbon atoms ("C"). 7~18 Alkynyl) and / or 7 to 12 carbon atoms ("C") 7~12 It may have an alkynyl group. In some embodiments, the alkynyl group has 7 to 8 carbon atoms ("C"). 7~8 ("Alkynyl"). In some embodiments, the alkynyl group has 7 to 9 carbon atoms ("C"). 7~9 ("Alkynyl"). In some embodiments, the alkynyl group may have 7 to 10 carbon atoms ("C"). 7~10 ("Alkynyl"). In some embodiments, the alkynyl group has 7 to 11 carbon atoms ("C"). 7~11 ("Alkynyl"). In some embodiments, the alkynyl group may have 8 to 12 carbon atoms ("C"). 8~12("Alkynyl"). In some embodiments, the alkynyl group has 9 to 12 carbon atoms ("C"). 9~12 In some embodiments, the alkynyl group has 10 to 12 carbon atoms ("C"). 10~12 ("Alkynyl"). In some embodiments, the alkynyl group has 11 to 12 carbon atoms ("C"). 11~12 Alkinil).

[0318] The alkynyl may be unsubstituted or substituted with respect to "alkyl" as described above or in the various embodiments provided herein. Examples of alkynyl groups include, but are not limited to, ethynyl, 1-propynyl, 2-propynyl, 1-, 2-, or 3-butynyl, and analogues.

[0319] The term "heteroalkynyl," as used herein, refers to the alkynyl group as defined above, further comprising one or more (e.g., 1, 2, 3, 4, etc.) heteroatoms (e.g., oxygen, sulfur, nitrogen, boron, silicon, phosphorus, and analogous atoms), where one or more heteroatoms are inserted between adjacent carbon atoms in the parent carbon chain, and / or one or more heteroatoms are inserted between carbon atoms and the parent molecule, i.e., between bonding points. In some embodiments, the heteroalkynyl group is an unsaturated group having 2 to 22 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero-C"). 2~22 The term "heteroalkynyl" refers to an unsaturated group having 7 to 18 carbon atoms and / or 7 to 12 carbon atoms, as well as 1, 2, 3, 4, or other heteroatoms ("heteroC"). 7~18 "Alkinyl" or "HeteroC" 7~12 This refers to an "alkynyl" group. In some embodiments, a heteroalkynyl group can have 7 to 8 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~8 ("Alkynyl"). In some embodiments, the heteroalkynyl group may have 7 to 9 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C").7~9 In some embodiments, the heteroalkynyl group has 7 to 10 carbon atoms and 1, 2, 3, 4, or other heteroatoms ("hetero C"). 7~10 ("Alkynyl"). In some embodiments, the heteroalkynyl group has 7 to 11 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("heteroC"). 7~11 ("Alkynyl"). In some embodiments, the heteroalkynyl group has 8 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("heteroC"). 8~12 ("Alkynyl"). In some embodiments, the heteroalkynyl group has 9 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("heteroC"). 9~12 ("Alkynyl"). In some embodiments, the heteroalkynyl group has 10 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("heteroC"). 10~12 In some embodiments, the heteroalkynyl group has 11 to 12 carbon atoms and 1, 2, 3, 4, etc. heteroatoms ("hetero C"). 11~12 Alkinil).

[0320] As used herein, "carbocykrill" or "carbocyclic formula" has 3 to 8 ring carbon atoms ("C"). 3~8 "Carbocyclyl" refers to a radical of a non-aromatic cyclic hydrocarbon group having zero heteroatoms in the non-aromatic ring system. In some embodiments, the carbocyclyl group has 3 to 7 ring carbon atoms ("C"). 3~7 Carbocyclyl). In some embodiments, the carbocyclyl group has 3 to 6 ring carbon atoms ("C"). 3~6 Carbocyclyl). In some embodiments, the carbocyclyl group has 4 to 6 ring carbon atoms ("C"). 4~6 Carbocyclyl). In some embodiments, the carbocyclyl group has 5 to 6 ring carbon atoms ("C"). 5~6 Carbocyclyl). In some embodiments, the carbocyclyl group has 5 to 8 ring carbon atoms ("C"). 5~8 Carbocyclyl). Exemplary C 3~6Examples of carbocyclyl groups include, but are not limited to, cyclopropyl (C3), cyclopropenyl (C3), cyclobutyl (C4), cyclobutenyl (C4), cyclopentyl (C5), cyclopentenyl (C5), cyclohexyl (C6), cyclohexenyl (C6), cyclohexadienyl (C6), and analogues. 3~8 The carbocyryl group is not limited to the aforementioned C 3~6 Examples include the carbocyclyl group, as well as cycloheptyl (C7), cycloheptenyl (C7), cycloheptadienyl (C7), cycloheptatrielinyl (C7), cyclooctyl (C8), cyclooctenyl (C8), bicyclo[2.2.1]heptanyl (C7), bicyclo[2.2.2]octanyl (C8), and analogues. As the above examples show, in certain embodiments, the carbocyclyl group may be monocyclic ("monocyclic carbocyclyl") or polycyclic (e.g., containing condensed, cross-linked, or spirocyclic systems such as bicyclic ("bicyclic carbocyclyl") or tricyclic ("tricyclic carbocyclyl")), and may be saturated or contain one or more carbon-carbon double or triple bonds. "Carbocyclyl" also includes cyclic systems in which the carbocykyl ring as defined above is fused with one or more aryl or heteroaryl groups on the carbocykyl ring, in which case the carbon number still indicates the carbon number in the carbocyclic system. Unless otherwise specified, each case of a carbocykyl group is independently either unsubstituted ("unsubstituted carbocykyl") or substituted with one or more substituents ("substituted carbocykyl"). In certain embodiments, the carbocykyl group is an unsubstituted C 3~10 It is a carbocyclyl. In certain embodiments, the carbocyclyl group is a substituted C 3~10 It is carbocyclyl.

[0321] In some embodiments, "carbocyrill" or "carbocyclic" refers to a "cycloalkyl" group, i.e., a monocyclic saturated carbocyclyl group having 3 to 8 ring carbon atoms ("C"). 3~8It is called a "cycloalkyl" group. In some embodiments, the cycloalkyl group has 3 to 6 ring carbon atoms ("C"). 3~6 ("Cycloalkyl"). In some embodiments, the cycloalkyl group has 4 to 6 ring carbon atoms ("C"). 4~6 ("Cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 6 ring carbon atoms ("C"). 5~6 ("Cycloalkyl"). In some embodiments, the cycloalkyl group has 5 to 8 ring carbon atoms ("C"). 5~8 Cycloalkyl). C 5~6 Examples of cycloalkyl groups include cyclopentyl (C5) and cyclohexyl (C5). 3~6 As an example of a cycloalkyl group, the aforementioned C 5~6 Examples include cycloalkyl groups, as well as cyclopropyl (C3) and cyclobutyl (C4). 3~8 As an example of a cycloalkyl group, the aforementioned C 3~6 Examples include cycloalkyl groups, as well as cycloheptyl (C7) and cyclooctyl (C8). Unless otherwise specified, each case of a cycloalkyl group is independently either unsubstituted ("unsubstituted cycloalkyl") or substituted with one or more substituents ("substituted cycloalkyl"). In certain embodiments, the cycloalkyl group is unsubstituted C 3~8 It is a cycloalkyl group. In certain embodiments, the cycloalkyl group is a substituted C 3~8 It is a cycloalkyl group.

[0322] The term "heterocyclic" refers to a saturated or unsaturated aromatic or non-aromatic group having 1 to 8 cyclic carbon atoms and 1 to 4 cyclic heteroatoms such as nitrogen, oxygen, sulfur, boron, phosphorus, silicon, and analogous atoms, where the nitrogen and sulfur atoms are oxidized as necessary, and the nitrogen atom is quaternized as necessary. Heterocyclic groups can have a monocyclic or multiple fused rings. Heterocyclics containing more than one ring can be fused, spiro, or bridging rings, or any combination thereof. In fused ring systems, one or more fused rings can be aryl or heteroaryl.Examples of heterocyclic groups include dihydropyranyl, thiazolinyl, thiazolidinyl, tetrahydrothiophenyl, 2,3-dihydrobenzo[b]thiophen-2-yl, 4-amino-2-oxopyrimidine-1(2H)-yl, benzimidazolyl, benzofuranyl, benzofuranyl, benzopyrazolyl, benzotriazolyl, benzothiophenyl, benzoxazolyl, carbazolyl, carborinyl, sinnolinyl, furanyl, imidazolyl, indolinyl, indolyl, i Ndazinyl, indazolyl, isobenzofuranil, isoindolyl, isoquinolyl, isothiazolyl, isoxazolyl, naphtopyridinyl, oxadiazolyl, oxazolyl, oxazoline, isoxazolline, oxetanyl, pyranyl, pyrazinyl, pyrazolyl, pyridadinyl, pyridopyridinyl, pyridadinyl, pyridyl, pyrimidyl, pyrrolyl, quinazolinyl, quinolyl, quinoxalinyl, tetrahydropyranyl, tetrazolyl, tetrazolopyridyl, thiadi Azolyl, thiazolyl, thienyl, triazolyl, azetidinyl, 1,4-dioxanyl, hexahydroazepinyl, piperazinyl, piperidinyl, pyrrolidinyl, morpholinyl, thiomorpholinyl, dihydrobenzimidazolyl, dihydrobenzofuranyl, dihydrobenzothiophenyl, dihydrobenzoxazolyl, dihydrofuranyl, dihydroimidazolyl, dihydroindolyl, dihydroisoxazolyl, dihydroisothiazolyl, dihydrooxadiazolyl Examples include, but are not limited to, dihydrooxazolyl, dihydropyrazine, dihydropyrizolyl, dihydropyrimidinyl, dihydropyrrolyl, dihydroquinolinyl, dihydrotetrazolyl, dihydrothiadiazolyl, dihydrothiazolyl, dihydrothienyl, dihydrotriazolyl, dihydroazetidinyl, methylenedioxybenzoyl, tetrahydrofuranil, and tetrahydrothienyl, their N-oxides, and analogues."Heterocycle" as disclosed herein means, for example, hydroxyl, nitro, amino (e.g., -NH2 or dialkylamino), imino, cyano, halo (e.g., F, Cl, Br, I, and analogous atoms), haloalkyl (e.g., -CCl3, -CF3, and analogous atoms), thio, sulfonyl, thioamide, amidino, imidino, oxo, oxamidino, methoxamidino, imidino, guanidino, sulfonamide, carboxyl, formyl, alkyl, alkoxy, alkoxy-alkyl, alkylcarbonyl, alkylcarbonyloxy (e.g., -OCOR), aminocarbonyl, arylcarbonyl, aralkylcarbonyl, carbonylamino, heteroarylcarbonyl, The compounds may be substituted as needed with one or more substituents, including but not limited to heteroaralkyl-carbonyl, alkylthio, aminoalkyl, cyanoalkyl, carbamoyl (e.g., -NHCOOR- or -OCONHR-), urea (e.g., -NHCONHR-), cycloalkyl, aryl, heteroaryl, heteroalicyclic, hydroxy, alkoxy, aryloxy, mercapto, alkylthio, arylthio, cyano, halo, carbonyl, (=O), thiocarbonyl, O-carbamyl, N-carbamyl, O-thiocarbamyl, N-thiocarbamyl, C-amide, N-amide, C-carboxy, O-carboxy, nitro, amino, heterocyclic, -CN, and analogues. For example, but not limited to, additional substituents as needed include fluorine, chlorine, bromine, and iodine atoms, as well as CF3, CN, OH, =O, SH, =S, NH2, =NH, N3, and NO2 groups. Substituents as needed include C1-C. 10 Alkyl, C1-C 10 Heteroalkyl, C2~C 10 Alkenyl, C2~C 10 Heteroalkenyl, C2~C 10 Alkinyl, C2~C 10 This also includes heteroalkynyls and their analogues. Exemplary substituents include F, Cl, Br, OH, SH, =O, NH2, amino, and C. 1~4 Alkyl (e.g., methyl, ethyl, t-butyl), C 1~4These are heteroalkyl cyclopropyl, SF5, NO, NO2, NMe2, CONH2, CH2NMe2, NHSO2Me, C(CH3)2CN, COMe, OMe, SMe, COOMe, COOEt, CH2COOH, OCH2COOH, COOH, SOMe, SO2Me, cyclopropyl, SO2NH2, SO2NHMe, SO2CH2CH2OH, NHCH2CH2OH, CH2CH2OCH3, SF5, SO2NMe2, NO, NO2, OCF3, SO2CF3, CN, or CF3.

[0323] In a heterocyclic group containing one or more nitrogen atoms, the bond site may be a carbon or nitrogen atom, as long as the valence allows. The heterocyclic group may be a monocyclic ("monocyclic heterocycle") or a condensed, bridged, or spirocyclic system ("bicyclic heterocycle"), and may be saturated or partially unsaturated. A heterocyclic or bicyclic system may contain one or more heteroatoms in one or both rings. A "heterocycle" includes a ring system in which the heterocyclic ring as defined above is fused with one or more carbocyrill groups and the bond site is on either a carbocyrill or heterocyclyl ring, or a ring system in which the heterocyclic ring as defined above is fused with one or more aryl or heteroaryl groups and the bond site is on a heterocyclyl ring, in which case the number of ring members still refers to the number of ring members in the heterocyclic system. Unless otherwise specified, each case of heterocyclyl is independently substituted as necessary, i.e., unsubstituted ("unsubstituted heterocyclyl") or substituted with one or more substituents ("substituted heterocycle"). In certain embodiments, the heterocyclic group is an unsubstituted 3- to 8-membered heterocycle. In certain embodiments, the heterocyclic group is a substituted 3- to 8-membered heterocycle.

[0324] In some embodiments, the heterocyclic group is a 3-8 membered non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, where each heteroatom is independently selected from nitrogen, oxygen, sulfur, boron, phosphorus, and silicon ("5-10 membered heterocycle"). In some embodiments, the heterocyclic group is a 5-8 membered non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-8 membered heterocycle"). In some embodiments, the heterocyclic group is a 5-6 membered non-aromatic ring system having a ring carbon atom and 1-4 ring heteroatoms, where each heteroatom is independently selected from nitrogen, oxygen, and sulfur ("5-6 membered heterocycle"). In some embodiments, the 5-6 membered heterocycle has 1-3 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocycle has 1-2 ring heteroatoms selected from nitrogen, oxygen, and sulfur. In some embodiments, the 5-6 membered heterocycle has one ring heteroatom selected from nitrogen, oxygen, and sulfur.

[0325] The term "non-aromatic heterocycle" refers to monocyclic or polycyclic compounds that are not aromatic and contain one or more heteroatoms in their ring structure. Such heteroatoms are preferably selected from the group consisting of S, N, and O. Examples of non-aromatic heterocycles include, but are not limited to, pyrrolidine, piperidine, piperazine, morpholine, and thiomorpholine.

[0326] As used herein, the expression "substituted as necessary" means that at least one hydrogen atom present on a group (e.g., a carbon or nitrogen atom) is replaced by an acceptable substituent, such as a substituent that, when substituted, results in a stable compound, such as one that does not spontaneously undergo transformation by rearrangement, cyclization, elimination, or other reactions. Heteroatoms such as nitrogen may have substituents such as any preferred substituents described herein that satisfy the valence of the heteroatom and form a stable moiety.

[0327] The embodiments described below and in the claims can be understood in consideration of the above definitions.

[0328] Other features and advantages of this disclosure will become apparent from the following description of preferred embodiments and the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as those commonly understood by those skilled in the art to which this disclosure belongs. Methods and materials similar or equivalent to those described herein may be used in the implementation or testing of this disclosure, but preferred methods and materials are described below. All published foreign patents and patent applications cited herein are incorporated herein by reference. All other published references, documents, manuscripts and scientific documents cited herein are incorporated herein by reference. In the event of any conflict, this specification, including definitions, shall prevail. In addition, materials, methods and examples are illustrative and not intended to be limiting. [Modes for carrying out the invention]

[0329] Detailed explanation In certain embodiments, this disclosure provides novel ionizable substituted aryl and heteroaryllipid-like compounds (e.g., as represented by Formula I) designed and synthesized according to the techniques disclosed herein. These novel ionizable substituted aryl and heteroaryllipid-like compounds have been shown to be formulated into lipid nanoparticles (LNPs) that provide stable and efficient LNP formulations comparable to or better than conventional benchmark lipids.

[0330] This disclosure also provides novel ionizable lipid-like chemicals (e.g., ionizable substituted piperazine lipids represented by formula VII) designed and synthesized according to the techniques disclosed herein. These novel ionizable lipid-like chemicals have also been formulated into lipid nanoparticles (LNPs) and have been shown to provide stable and efficient LNP formulations that are comparable to or better than benchmark lipids of the prior art.

[0331] This disclosure further provides other novel aryl and heteroaryl lipid compounds (e.g., as represented by formula IX) designed and synthesized according to the techniques disclosed herein. These other aryl and heteroaryl lipid compounds have also been formulated into lipid nanoparticles (LNPs) and have been shown to provide stable and efficient LNP formulations that are comparable to or better than benchmark lipids of the prior art.

[0332] This disclosure is at least in part based on the discovery that novel ionizable lipids having advantageous properties when used in lipid particles for therapeutic agent delivery can be formed using ionizable substituted aryl and heteroaryl lipid-like compounds. In particular, the techniques herein provide lipid-based nanoparticle compositions and formulations that can specifically target cargo moieties (e.g., nucleic acid cargoes) to specific tissues of interest without requiring ligand-based targeting strategies. The lipid-like substituted aryl and heteroaryl compounds disclosed herein have the following general structures: [ka] or its salt or isomer [in the formula, X is either CH or N. a and b are independently between 2 and 5. m1, m2, m3, and m4 are independently between 4 and 10. E1, E2, E3, and E4 are independently -O(CO)O-, -(CO)O-, or -O(CO)-. T1, T2, T3, and T4 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R1 and R2 are independently H, or C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. R3 is independently H, or C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, [ka] And, G is O, -(CO)NR3-, -NR3(CO)-, -(CO)O-, or -CH2(CO)NR3-, L is a C1-C4 alkyl group that is bonded or substituted as needed. Z is CH or N, R4 and R5 are independently H, C1-C3 alkyl, C2-C3 alkenyl or C2-C3 alkynyl, or [ka] Is it, Alternatively, R4 or R5 may form a 3-7 membered ring containing 0-2 heteroatoms as needed, or R4 and R5 may form a substituted 3-7 membered ring together with L. It may include.

[0333] The lipid-like substituted aryl and heteroaryl compounds disclosed herein have the following general structure: [ka] or its salt or isomer [in the formula, X is either CH or N. L1, L2, and L3 are independently -O- and -(CO)NR x -, -NR x (CO)-, -(CO)O-, -CH2(CO)NR x - and if A is CH, then all of L1, L2, and L3 are -(CO)NR x -That is not the case, R x These are H, C1-C6 alkyl, or C3-C6 cycloalkyl, G1 and G2 are given by Equation III: [ka] or a salt or isomer thereof, where, n1 is 3, 4, 5, 6, 7, 8, 9, or 10. L4 is -(CO)O- or -O(CO)-, R6 is branched from C5 to C 20 It is alkyl, G3 is equation (III), equation (IV), equation (V), or equation (VI): [ka] And, R7 and R8 are independently functionalized C1-C5 alkyl groups as needed. n2, n3, and n4 are independently 0, 1, 2, or 3. X1 is C, N, or O. R9 and R 10 [These are independently H or, if necessary, functionalized C1-C5 alkyl groups.] It may also include that.

[0334] This disclosure is also at least in part based on the discovery that substituted piperazines can be used to form novel ionizable lipids having properties advantageous when used in lipid particles for therapeutic agent delivery. In particular, the techniques herein provide lipid-based nanoparticle compositions and formulations that can specifically target cargo portions (e.g., nucleic acid cargoes) to specific tissues of interest without requiring ligand-based targeting strategies. The ionizable substituted piperazine lipids disclosed herein include a head group having the following structure: [ka] In the formula, the protonable piperazine (e.g., pH titrable) head group is connected via a linker having the following structure, C5~C 20The hydrocarbon chains, for example, alkyl or alkenyl chains, are bonded to each hydrocarbon chain, which independently has 0 to 5 (e.g., 0, 1, 2, 3, 4, or 5) double bonds. [ka] In the formula, L5 is -O(CO)-, -CH2-O(CO)-, -CH2(CO)O-, or -O(CO)-NH-, n5 is 2, 3, 4, 5, 6, 7, or 8, and G4 is a bond, -(CO)O-, or -O(CO)-. C5~C 20 The hydrocarbon chain may be bonded to the G atom of the linker.

[0335] This disclosure is at least in part based on the discovery that other aryl and heteroaryl lipid compounds can be used to form novel ionizable lipids having properties advantageous when used in lipid particles for therapeutic agent delivery. In particular, the techniques herein provide lipid-based nanoparticle compositions and formulations that can specifically target cargo moieties (e.g., nucleic acid cargoes) to specific tissues of interest without requiring ligand-based targeting strategies. Other aryl and heteroaryl compounds disclosed herein have the following general structure represented by formula IX: [ka] or its salt or isomer [in the formula, X2, X3, and X4 are independently CH or N. G8, G9, and G 10 These are independently O, -(CO)O-, -CH2O(CO)-, and -(CH2)2(CO)NR 15 -,-(CH2)O(CO)NR 15 -, CH2(CO)NR 15 -, -(CO)NR 15 -, -NR 15 (CO)-, or NR 15 (CO)O-, where X2, X3, and X4 are all CH, then G8, G9, and G 10All of the following are -(CO)NR 15 -That is not the case, a1, b1, and c1 are independently 0, 1, 2, 3, or 4. m6, m7, and m8 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. E5, E6, and E7 are independently -(CO)O- or -O(CO)-, T5, T6, and T7 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R 15 [This is H, or optionally a functionalized C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl.] It may include.

[0336] The substituted aryl and heteroaryl lipid-like compounds, ionizable substituted piperazine lipids, and / or other ionizable aryl and / or heteroaryl lipid compounds disclosed herein provide stable and efficient lipid nanoparticle (LNP) formulations for delivering therapeutic oligonucleotides to specific target tissues of interest.

[0337] Conventional LNPs consist of four main components: an ionizable or cationic lipid for encapsulating mRNA, an amphiphilic helper phospholipid for increased efficacy, cholesterol for structural stability, and polyethylene glycol (PEG)-lipid for steric stability. This first generation of LNPs can be thought of as "only one ionizable lipid-LNP" or "single LNP." Traditionally, effective intracellular delivery materials have relied on an optimal balance of ionizable amines (pKa between 6.0 and 6.5) that bind to RNA and release it, and hydrophobicity to stabilize the nanoparticles. Thus, there has been a thorough focus on developing ionizable lipids, which have proven to be highly effective delivery platforms for the liver and hepatocytes. However, modifying the chemical structure of ionizable / cationic lipids to achieve different pKa values ​​and creating libraries is a validated but time-consuming and costly labor-intensive task. Certain aspects of this disclosure provide ionizable substituted aryl and heteroaryl lipid-like compounds that have the remarkable ability to provide stable and efficient LNP formulations for delivering therapeutic oligonucleotides to specific target tissues of interest. While not strictly adhering to theory, certain ionizable substituted aryl and heteroaryllipid-like compounds disclosed herein (e.g., SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, S M-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, and SM-173) appear to be able to shift the directivity of the LNP vectors disclosed herein to a specific tissue of interest without requiring any further active targeting components in the LNPs of this disclosure.

[0338] [Table 1-1] [Table 1-2] [Table 1-3] [Table 1-4] [Table 1-5] [Table 1-6]

[0339] The novel ionizable substituted aryl and heteroaryllipid-like compounds disclosed herein have the general structure described in Formula I or Formula II herein, and include their (R) and / or (S) enantiomers.

[0340] This disclosure also provides ionizable substituted piperazine lipids that have a remarkable ability to provide stable and efficient LNP formulations for delivering therapeutic oligonucleotides to specific target tissues of interest. While not bound by theory, certain ionizable substituted piperazine lipids disclosed herein (e.g., SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, and SM-125) appear capable of shifting the directivity of the LNP vectors disclosed herein to specific tissues of interest without requiring additional active targeting components in the LNPs of this disclosure.

[0341] [Table 2-1] [Table 2-2] [Table 2-3]

[0342] The novel ionizable substituted piperazine lipids disclosed herein have the general structure described in Formula VII or Formula VIII herein, and include their (R) and / or (S) enantiomers.

[0343] This disclosure further provides ionizable substituted piperazine lipids that have the remarkable ability to provide stable and efficient LNP formulations for delivering therapeutic oligonucleotides to specific target tissues of interest. Without being bound by theory, certain ionizable substituted piperazine lipids disclosed herein (e.g., SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, and SM-125) appear capable of shifting the directivity of the LNP vectors disclosed herein to specific tissues of interest without requiring additional active targeting components in the LNPs of this disclosure.

[0344] This disclosure also provides other aryl and heteroaryl lipid compounds that have a remarkable ability to provide stable and efficient LNP formulations for delivering therapeutic oligonucleotides to specific target tissues of interest. Without being bound by theory, certain aryl and heteroaryl lipid compounds disclosed herein (e.g., SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and SM-112) appear capable of shifting the directivity of the LNP vectors disclosed herein to specific tissues of interest without requiring additional active targeting components in the LNPs of this disclosure.

[0345] Some of the additional aryl and heteroaryllipid compounds disclosed herein have the general structure described in Formula IX herein and include their (R) and / or (S) enantiomers.

[0346] In some embodiments, the techniques of this specification provide improved lipid-based compositions for delivering therapeutic agents, particularly nucleic acid therapeutic agents. As disclosed herein, these lipid-based compositions are effective in increasing cargo release efficiency from lipid-based compositions such as LNPs. Furthermore, this disclosure demonstrates that the activity of these improved lipid-based compositions depends on the presence of certain novel ionizable substituted aryl and heteroaryl lipid-like compounds, certain novel ionizable substituted piperazine lipids, and / or other ionizable aryl and / or heteroaryl lipid compounds disclosed herein.

[0347] Within the scope of this disclosure, lipid-based compositions comprising ionizable substituted aryl and heteroaryl lipid-like compounds, ionizable substituted piperazine lipids, and / or other ionizable aryl and / or heteroaryl lipid compounds disclosed herein are intended to be used for a variety of purposes, such as the delivery of encapsulated therapeutic agents to cells in vitro and / or in vivo. In this regard, this disclosure provides a method for treating a disease or disorder in a subject requiring such treatment by contacting the subject with a suitable therapeutic agent, such as nucleic acids (e.g., siRNA, ASO, tRNA, miRNA, mRNA, DNA, and analogues), proteins, peptides, and other macromolecules, in combination with the lipid-based compositions disclosed herein.

[0348] Nucleic acid therapy possesses a well-known and astonishing potential to treat diseases at the genetic level. However, a safe and effective delivery system is essential for nucleic acid therapy. Non-specific delivery to organs and tissues often results in effects and toxicity outside the target site. The delivery of therapeutic agents to specific organs of interest is a well-recognized need in lipid-nanoparticle development and in drug development in general. The concept of targeting only the cause of disease without harming other parts of the body was described by Ehrlich 120 years ago. However, existing methods do not provide a defined or well-known methodology for developing nanoparticles that target specific tissues without introducing additional ligand-based targeting strategies. Therefore, organ-specific targeting of lipid nanoparticles based on the structural affinity of lipids to tissues, as disclosed herein, satisfies a well-established need in reducing effects and toxicity outside the target site.

[0349] The ability to deliver nucleic acid therapeutics specifically to certain tissues or organs while avoiding activation in other tissues or organs is essential for effectively treating many tissue or organ-specific diseases. This disclosure includes, for example, substituted aryl and heteroaryl lipid-like compounds such as SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, and / or SM-173; for example, SM-04 8. By incorporating ionizable substituted piperazine lipids such as SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, and / or SM-125; and / or SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112 into the LNP, we demonstrate that the vector's directivity can be shifted to a specific tissue of interest without requiring an active target component in the LNP.

[0350] In some embodiments, the lipid-based compositions disclosed herein are particularly useful for the delivery of nucleic acid therapeutics (e.g., siRNA, ASO, tRNA, miRNA, mRNA, DNA, and analogues). The lipid-based compositions disclosed herein can be used to modulate the expression of target genes and proteins both in vitro and in vivo by contacting tissues / cells with lipid-based compositions containing the lipids disclosed herein that carry a cargo such as a therapeutic nucleic acid (e.g., siRNA) capable of reducing the expression of a desired target gene.

[0351] The technologies described herein provide ionizable substituted aryl and heteroaryl lipid-like compounds that enable the formulation of pharmaceutical compositions for delivering therapeutic agents, such as nucleic acids (e.g., siRNA, ASO, tRNA, miRNA, mRNA, DNA, and analogues), proteins, peptides, and other macromolecules, in vitro or in vivo.

[0352] Exemplary embodiments of ionizable substituted aryl and heteroaryl lipid-like compounds of the present disclosure, as well as lipid-based compositions containing them, and their synthesis and LNP formulations are described in further detail below.

[0353] Lipids In certain embodiments, the disclosure provides novel ionizable substituted aryl and heteroaryllipid-like compounds having the general structure of formula I. [ka]

[0354] Ionizable substituted aryl and heteroaryllipid-like compounds feature a design that includes a central six-membered heterocyclic ring skeleton with three side chain groups at the 2, 4, and 6 positions of the heterocyclic ring.

[0355] This disclosure also provides novel ionizable substituted piperazine lipids of the general structure of formula VII. The ionizable substituted piperazine lipids disclosed herein feature a design comprising a piperazine head group having the following structure: [ka] In the formula, the protonable piperazine (e.g., pH titrable) head group is connected via a linker having the following structure, C5~C 20 The hydrocarbon chains, for example, alkyl or alkenyl chains, are bonded to each hydrocarbon chain, which independently has 0 to 5 (e.g., 0, 1, 2, 3, 4, or 5) double bonds. [ka] In the formula, L5 is -O(CO)-, -CH2-O(CO)-, -CH2(CO)O-, or -O(CO)-NH-, n5 is 2, 3, 4, 5, 6, 7, or 8, and G4 is a bond, -(CO)O-, or -O(CO)-. C5~C 20 The hydrocarbon chain may be bonded to the G atom of the linker. The ionizable substituted piperazine lipids disclosed herein provide stable and efficient lipid nanoparticle (LNP) formulations for delivering therapeutic oligonucleotides to specific target tissues of interest.

[0356] This disclosure also provides other ionizable substituted aryl and heteroaryllipid-like compounds of the general structure of formula IX. Certain ionizable aryl and heteroaryllipid compounds feature a design comprising a six-membered heterocyclic ring skeleton with three side chain groups at the 2, 4, and 6 positions of the heterocyclic ring, where each of the atoms at positions 1, 3, and 5 can be either CH or N. Exemplary ionizable aryl and heteroaryllipid compounds are disclosed herein.

[0357] [Table 3-1] [Table 3-2] [Table 3-3] [Table 3-4]

[0358] Certain aspects of the present disclosure provide novel ionizable lipids that can be advantageously used in lipid-based compositions of the present disclosure for delivering therapeutic agents to tissues / cells in vivo.

[0359] Within the scope of this disclosure, ionizable lipids (e.g., ionizable substituted aryl and / or heteroaryl lipid-like compounds, ionizable substituted piperazine lipids, and / or other ionizable aryl and / or heteroaryl lipid compounds) are intended to include racemic mixtures or mixtures of one or more diastereomers. In some embodiments, the ionizable lipids are enriched with one enantiomer such that they contain at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% enantiomer excess. In some embodiments, the ionizable lipids are enriched with one diastereomer such that the cationic lipid contains at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95% diastereomer excess. In some embodiments, the ionizable lipids are chiralally pure (e.g., including a single optical isomer). In some embodiments, the ionizable lipids are enriched with one optical isomer (e.g., an optically active isomer) to contain an isomer excess of at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, or 95%. This disclosure provides the synthesis of ionizable lipids of formulas I, VII, and / or IX, either as racemic mixtures or in optically pure forms.

[0360] As used herein, the term “salt” includes any anionic and cationic complexes, such as complexes formed between ionizable substituted aryl or heteroaryllipid-like compounds, ionizable substituted piperazine lipids, and / or other ionizable aryl and / or heteroaryllipid compounds disclosed herein and one or more anions. Examples of anions include, for example, hydrides, fluorides, chlorides, bromides, iodides, oxalates (e.g., hemioxalates), phosphates, phosphonates, hydrogen phosphates, dihydrogen phosphates, oxides, carbonates, bicarbonates, nitrates, nitrites, nitrides, bisulfites, sulfides, sulfites, bisulfates, sulfates, thiosulfates, bisulfates, borates, formates, acetates, benzoates, citrates, tartrates, lactates, acrylates, polyacrylates, fumarates, maleates, itaconates, and glycols. Examples of inorganic and organic anions include, but are not limited to, salts of acid salts, gluconates, malates, mandelates, tiglic acid, ascorbic acid, salicylates, polymethacrylates, perchlorates, chlorates, chlorites, hypochlorites, bromates, hypobromites, iodates, alkylsulfonates, arylsulfonates, arsenates, arsenites, chromates, dichromates, cyanides, cyanates, thiocyanates, hydroxides, peroxides, permanganates, and mixtures thereof. In certain embodiments, the salts of substituted aryl and heteroaryllipid-like compounds disclosed herein are crystalline salts.

[0361] As used herein, the term "alkyl" includes linear or branched, acyclic or cyclic saturated aliphatic hydrocarbons containing 1 to 24 carbon atoms. Representative saturated linear alkyls include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, and analogues, while saturated branched alkyls include, but are not limited to, isopropyl, sec-butyl, isobutyl, tert-butyl, isopentyl, and analogues. Representative saturated cyclic alkyls include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, and analogues, while unsaturated cyclic alkyls include, but are not limited to, cyclopentenyl, cyclohexenyl, and analogues.

[0362] As used herein, the term “alkenyl” includes alkyl groups as defined above, which contain at least one double bond between adjacent carbon atoms. Alkenyls include both cis and trans isomers. Representative linear and branched alkenyls include, but are not limited to, ethirenyl, propyrenyl, 1-butenyl, 2-butenyl, isobutyrenyl, 1-pentenyl, 2-pentenyl, 3-methyl-1-butenyl, 2-methyl-2-butenyl, 2,3-dimethyl-2-butenyl, and analogues. Cyclic alkenyls are also intended for lipids in this disclosure.

[0363] As used herein, the term “alkynyl” includes any alkyl or alkenyl as defined above, further containing at least one triple bond between adjacent carbon atoms. Representative linear and branched alkynyls include, but are not limited to, acetylenyl, propynyl, 1-butynyl, 2-butynyl, 1-pentynyl, 2-pentynyl, 3-methyl-1-butynyl, and analogues.

[0364] As used herein, the term “acyl” includes any alkyl, alkenyl, or alkynyl in which the carbon at the bonding site is substituted with an oxo group as defined below. -C(=O)alkyl, -C(=O)alkenyl, and -C(=O)alkynyl are non-limiting examples of acyl groups.

[0365] As used herein, the term “heterocyclic” includes monocyclic (e.g., 5, 6, 7-membered, and similar numbers), bicyclic (e.g., 7, 8, 9, 10-membered, and similar numbers), or saturated, unsaturated, or aromatic heterocyclic rings containing one or two heteroatoms independently selected from nitrogen, oxygen, and sulfur, wherein the nitrogen and sulfur heteroatoms may be oxidized as necessary, and the nitrogen heteroatom may be quaternized as necessary, and also includes bicyclic rings in which any of the above heterocyclic rings are fused to a benzene ring. Heterocyclic rings may be bonded via any heteroatom or carbon atom. Heterocycles include, but are not limited to, heteroaryls as defined below, as well as morpholinyl, pyrrolidinonyl, piperidinyl, piperidinyl, hydantoinyl, valerolactamyl, oxyranyl, oxetanyl, tetrahydrofuranyl, tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl, tetrahydrothiophenyl, tetrahydrothiopyranyl, and analogues.

[0366] As used herein, the terms “optionally substituted alkyl,” “optionally substituted alkenyl,” “optionally substituted alkynyl,” “optionally substituted acyl,” and “optionally substituted heterocycle” mean that, if substituted, at least one hydrogen atom is replaced by the substituent. In the case of an oxo substituent (=O), two hydrogen atoms are replaced. In this regard, substituents include, but are not limited to, oxo, halogen, heterocycle, -CN, -NRxRy, -NRxC(=O)Ry, -NRxSO2Ry, -C(=O)Rx, -C(=O)ORx, -C(=O)NRxRy, -SOnRx, and -SOnNRxRy, where n is 0, 1, or 2, and Rx and Ry are the same or different, independently of hydrogen, alkyl , or heterocyclic, and each alkyl and heterocyclic substituent may be further substituted with one or more of the following: oxo, halogen, -OH, -CN, alkyl, -ORx, heterocyclic, -NRxRy, -NRxC(=O)Ry, -NRxSO2Ry, -C(=O)ORx, -C(=O)ORx, -C(=O)NRxRy, -C(O-R1)(O-R2), -SOnRx, and -SOnNRxRy. The term "substituted as necessary" when used before a list of substituents means that each substituent in the list may be substituted as necessary as described herein.

[0367] As used herein, the term "halogen" includes fluorine, chlorine, bromine, and iodine.

[0368] In some embodiments, the present disclosure relates to substituted aryl and heteroaryllipid-like compounds of formula I having the following structure: [ka] or its salt or isomer [in the formula, X is either CH or N, a and b are independently between 2 and 5. m1, m2, m3, and m4 are independently between 4 and 10. E1, E2, E3, and E4 are independently -O(CO)O-, -(CO)O-, or -O(CO)-. T1, T2, T3, and T4 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R1 and R2 are independently H, or C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl. R3 is independently H, or C1-C6 alkyl, C2-C6 alkenyl or C2-C6 alkynyl, [ka] And, G is O, -(CO)NR3-, -NR3(CO)-, -(CO)O-, or -CH2(CO)NR3-, L is a C1-C4 alkyl group that is bonded or substituted as needed. Z is CH or N, R4 and R5 are independently H, C1-C3 alkyl, C2-C3 alkenyl or C2-C3 alkynyl, or [ka] Is it, Alternatively, R4 or R5 may form a 3-7 membered ring containing 0-2 heteroatoms as needed, or R4 and R5 may form a substituted 3-7 membered ring together with L.

[0369] In some embodiments, the present disclosure relates to substituted aryl and heteroaryllipid-like compounds having the following structures: [ka] or its salt or isomer [in the formula, X is either CH or N, L1, L2, and L3 are independently -O- and -(CO)NR x -, -NR x (CO)-, -(CO)O-, -CH2(CO)NR x - and if A is CH, then all of L1, L2, and L3 are -(CO)NR x -That is not the case, R x These are H, C1-C6 alkyl, or C3-C6 cycloalkyl, G1 and G2 are given by Equation III: [ka] or a salt or isomer thereof, where, n1 is 3, 4, 5, 6, 7, 8, 9, or 10. L4 is -(CO)O- or -O(CO)-, R6 is branched from C5 to C 20 It is alkyl, G3 is equation (III), equation (IV), equation (V), or equation (VI): [ka] And, R7 and R8 are independently functionalized C1-C5 alkyl groups as needed. n2, n3, and n4 are independently 0, 1, 2, or 3. X1 is C, N, or O. R9 and R 10 [These independently provide H or, optionally, a functionalized C1-C5 alkyl group.]

[0370] In certain embodiments, the present disclosure relates to an ionizable substituted piperazine lipid of formula VII having the following structure: [ka] or its salt or isomer [wherein L6 is -O(CO)-, -CH2-O(CO)-, or -O(CO)-NH-, n6 and n7 are independently 1, 2, 3, 4, 5, 6, 7, or 8, and G5 and G6 are independently bonded, -(CO)O- or -O(CO)-, R 11 and R 12 These are C5~C, which are substituted independently as needed. 20 Alkyl or C5-C 20 [Provides alkenil]

[0371] In some embodiments, n6 and n7 are independently 4, 5, 6, 7, or 8.

[0372] In some embodiments, R 11 and R 12 These are, independently, C8-C9 alkyl and C8-C 10 Alkyl, C8~C 11 Alkyl, C8~C 12 Alkyl, C8~C 13 Alkyl, C8~C 14 Alkyl, C8~C 15 Alkyl, C8~C 16 Alkyl, C8~C 17 Alkyl, C8~C 18 Alkyl, C8~C 19 Alkyl, C8~C 20 Alkyl, C8-C9 alkenyl, C8-C 10 Alkenil, C8~C 11 Alkenil, C8~C 12 Alkenil, C8~C 13 Alkenil, C8~C 14 Alkenil, C8~C 15 Alkenil, C8~C 16 Alkenil, C8~C 17 Alkenil, C8~C 18 Alkenil, C8~C 19 Alkenyl, and C8~C 20 In some embodiments, R 11 and R 12 They are the same. In some embodiments, R 11and R 12 They are different. In some embodiments, R 11 and R 12 Both are C 8~20 It is alkyl.

[0373] In some embodiments, R 11 and / or R 12 This corresponds to, for example, a cis double bond, a trans double bond, or a combination thereof, and is unsaturated R 11 and R 12 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites that can be located at specific positions on one or both of the side chains. For unsaturated side chains where a double bond exists between a hydrogen atom and an alkyl or alkylene chain, the chemical notation "E" refers to a trans double bond configuration, and the chemical notation "Z" refers to a cis double bond configuration. As a non-restrictive example, R 11 and R 12 One or both of are C8 alkyl groups containing any combination of cis and / or trans double bonds at one or more positions, and / or any structure shown in the following examples. Similarly, as a non-limiting example, R 11 and R 12 One or both of the C molecules contain any combination of double bonds that can be characterized by either the "E" chemical notation and / or the "Z" chemical notation at one or more positions in the side chain. 12 It is an alkyl group. Similarly, as a non-limiting example, R 11 and R 12 One or both of the C atoms contain any combination of double bonds that can be characterized by either the chemical notation "E" and / or "Z" at one or more positions in the side chain. 15 It is an alkyl group. Similarly, as a non-limiting example, R 11 and R 12 One or both of the C atoms contain any combination of double bonds that can be characterized by either the chemical notation "E" and / or "Z" at one or more positions in the side chain. 17 It is an alkyl group. In some embodiments, R 11 and R12 The saturation point is the same.

[0374] In some embodiments, R 11 and R 12 It is an alkenil independently selected from the group consisting of hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca-2-ene, deca-3-ene, deca-4-ene, deca-5-ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, and dodeca-8-ene.

[0375] In some embodiments, the linker connecting the phosphate group and the amine group may contain one, two, three, four, five, six, or more unsaturated sites corresponding to, for example, cis double bonds, trans double bonds, or combinations thereof, and / or one or more triple bonds, and may be located at specific positions within the linker. In some embodiments, the present disclosure relates to an ionizable substituted piperazine lipid of formula VIII having the following structure: [ka] or its salt or isomer [in the formula, m5 is 4, 5, 6, 7, or 8. G7 is a bond, -(CO)O- or -O(CO)-, R 13 and R 14 These are C8~C, which are substituted independently as needed. 20 Alkyl or C8-C 20 [Provides alkenil]

[0376] In some embodiments, R13 and R 14 They are the same.

[0377] In some embodiments, R 13 and R 14 They are different.

[0378] In some embodiments, R 13 or R 14 C8~C are substituted as needed. 19 Alkyl and C8-C 19 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 19 Alkyl and C8-C 19 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 18 Alkyl and C8-C 18 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 18 Alkyl and C8-C 18 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 16 Alkyl and C8-C 16 Independently selected from the group consisting of alkenyls, and R as needed. 13and R 14 C8~C are substituted as needed. 16 Alkyl and C8-C 16 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C8~C are substituted as needed. 15 Alkyl and C8-C 15 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C8~C are substituted as needed. 15 Alkyl and C8-C 15 Independently selected from the group consisting of alkenyls. In some embodiments, R 13 or R 14 C9~C are substituted as needed. 12 Alkyl and C9-C 12 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C9~C are substituted as needed. 12 Alkyl and C9-C 12 It is independently selected from the group consisting of alkenils.

[0379] In some embodiments, m5 is 4, 5, 6, 7, or 8.

[0380] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0381] In some embodiments, m5 is 5, 6, or 7.

[0382] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0383] In some embodiments, R 13 or R14 C is replaced as needed. 10 ~C 17 Alkyl and C 10 ~C 17 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C is replaced as needed. 10 ~C 17 Alkyl and C 10 ~C 17 It is independently selected from the group consisting of alkenils.

[0384] In some embodiments, m5 is 5, 6, or 7.

[0385] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0386] In some embodiments, R 13 or R 14 C is replaced as needed. 12 ~C 17 Alkyl and C 12 ~C 17 Independently selected from the group consisting of alkenyls, and R as needed. 13 and R 14 C is replaced as needed. 12 ~C 17 Alkyl and C 12 ~C 17 It is independently selected from the group consisting of alkenils.

[0387] In some embodiments, m5 is 5, 6, or 7.

[0388] In some embodiments, R 13 or R 14 It contains 1, 2, 3, 4, 5, 6, or more unsaturated sites.

[0389] In some embodiments, R 13C8~C are substituted as needed. 17 Alkyl and C8-C 17 Selected from the group consisting of alkenyls, R 14 C8~C are substituted as needed. 17 Alkyl and C8-C 17 Selected from the group consisting of alkenyls.

[0390] In some embodiments, m5 is 5.

[0391] In some embodiments, G7 is a bond or -(CO)O-.

[0392] In some embodiments, R 13 or R 14 C8, C is substituted as needed. 12 , C 15 or C 17 Alkyl, and C8, C 12 , C 15 or C 17 C8, C 12 , C 15 or C 17 Alkyl, and C8, C 12 , C 15 or C 17 It is independently selected from the group consisting of alkenils.

[0393] In some embodiments, R 13 and R 14 These are alkyl groups independently selected from the group consisting of octane, nonane, decane, undecane, and dodecane, each of which is substituted as needed.

[0394] In some embodiments, R 13 and R 14 This is an alkyl group independently selected from the group consisting of octane, nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane, and eicosane.

[0395] In some embodiments, R 13 and R 14These are independently Octa-1-en, Octa-2-en, Octa-3-en, Octa-4-en, Nona-1-en, Nona-2-en, Nona-3-en, Nona-4-en, Nona-5-en, Deca-1-en, Deca-2-en, Deca-3-en, Deca-4-en, Deca-5-en, Deca-6-en, Undeka-1-en, Undeka-2-en, Undeka-3-en, Undeka-4-en, Undeka-5-en, Undeka-6-en, Undeka-7-en, Dodeka-1-en, Dodeka-2-en, Dodeka-3-en, Dodeka-4-en, Dodeka-5-en, Dodeka-6-en, Trideka-1-en, Trideka-2-en, Trideka-3-en, Trideka-4-en, Trideka-5-en, Trideka-6-en, Trideka-7-en, Tetradeca-1-en, Tetradeca-2-en, Tetradeca-3-en, Tetradeca-4-en, Tetradeca-5-en, Tetradeca-6-en, Tetradeca-7-en, Pentadeca-1-en, Pentadeca-2-en, Pentadeca-3-en, Pentadeca-4-en, Pentadeca-5-en, Pentadeca-6-en, Pentadeca- 7-yen, hexadeca-1-yen, hexadeca-2-yen, hexadeca-3-yen, hexadeca-4-yen, hexadeca-5-yen, hexadeca-6-yen, hexadeca-7-yen, hexadeca-8-yen, heptadeca-1-yen, heptadeca-2-yen, heptadeca-3-yen, heptadeca-4-yen, heptadeca-5-yen, heptadeca-6-yen, heptadeca-7-yen, heptadeca-8-yen, octadeca-1-yen, octadeca-2-yen, octadeca-3-yen, octadeca-4-yen, octadeca-5-yen, O The alkenil is selected from the group consisting of tatadeca-6-en, octadeca-7-en, octadeca-8-en, octadeca-9-en, nonadeca-1-en, nonadeca-2-en, nonadeca-3-en, nonadeca-4-en, nonadeca-5-en, nonadeca-6-en, nonadeca-7-en, nonadeca-8-en, nonadeca-9-en, icosa-1-en, icosa-2-en, icosa-3-en, icosa-4-en, icosa-5-en, icosa-6-en, icosa-7-en, icosa-8-en, and icosa-9-en.

[0396] In some embodiments, R13 and R 14 It independently includes one or more additional double bonds, and optionally R 13 and R 14 It contains one double bond independently.

[0397] In further embodiments, the present disclosure relates to aryl and heteroaryl lipid compounds of formula IX having the following structure: [ka] or its salt or isomer [in the formula, X2, X3, and X4 are independently CH or N. G8, G9, and G 10 These are independently O, -(CO)O-, -CH2O(CO)-, and -(CH2)2(CO)NR 15 -,-(CH2)O(CO)NR 15 -, CH2(CO)NR 15 -, -(CO)NR 15 -, -NR 15 (CO)-, or NR 15 (CO)O-, where X2, X3, and X4 are all CH, then G8, G9, and G 10 All of the following are -(CO)NR 15 -That is not the case, a1, b1, and c1 are independently 0, 1, 2, 3, or 4. m6, m7, and m8 are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. E5, E6, and E7 are independently -(CO)O- or -O(CO)-, T5, T6, and T7 are independently branched or unbranched C5~C 22 Alkyl, C5~C 22 Alkenyl, or C5~C 22 It is alkinyl, R 15 [These are H, or optionally functionalized C1-C6 alkyl, C2-C6 alkenyl, or C2-C6 alkynyl].

[0398] The compounds described herein can be prepared by known organic synthesis techniques, including the methods described in the following examples.

[0399] Lipid-based composition The techniques of this specification provide lipid-based compositions (e.g., LNPs and analogues) comprising one or more ionizable lipids or salts thereof as described herein. In some embodiments, the lipid-based compositions of this disclosure further comprise one or more substituted aryl and / or heteroaryl lipid-like compounds, one or more ionizable substituted piperazine lipids, and / or other ionizable aryl and / or heteroaryl lipid compounds. In some embodiments, the lipid-based compositions further comprise one or more complex lipids that can reduce or inhibit particle aggregation. In some embodiments, the lipid-based compositions further comprise one or more active agents or therapeutic agents, such as nucleic acids (e.g., siRNA, ASO, tRNA, miRNA, mRNA, DNA, and analogues), proteins, peptides, and other macromolecules.

[0400] As disclosed herein, lipid-based compositions include, but are not limited to, lipid nanoparticles, lipid vesicles (e.g., liposomes), and analogues. As used herein, lipid vesicles may include structures having a lipid-containing membrane surrounding an aqueous interior. In some embodiments, lipid-based compositions comprising one or more ionizable lipids described herein may be used to encapsulate therapeutic agents, such as nucleic acids, within lipid vesicles. In some embodiments, lipid vesicles comprising one or more ionizable lipids described herein may be complexed with nucleic acids.

[0401] The lipid-based compositions of this disclosure typically comprise a therapeutic agent, an ionizable lipid, a noncationic lipid, and a complex lipid (e.g., polyethylene glycol (PEG)-lipid) that inhibits particle aggregation. In some embodiments, the therapeutic agent is completely encapsulated within the lipid portion of the lipid-based composition to be resistant to enzymatic degradation by, for example, nucleases or proteases. In some embodiments, the lipid-based compositions described herein are substantially nontoxic to mammals such as humans.

[0402] The lipid-based compositions described herein are typically intended within the scope of this disclosure to have average diameters of about 30 nm to about 250 nm, about 40 nm to about 200 nm, about 50 nm to about 150 nm, about 60 nm to about 130 nm, about 70 nm to about 110 nm, or about 70 nm to about 90 nm. In some embodiments, the lipid-based compositions disclosed herein have lipid:therapeutic agent (e.g., lipid:nucleic acid) ratios (mass / mass ratios) of about 1:1 to about 1000:1, about 1:1 to about 500:1, about 2:1 to about 250:1, about 3:1 to about 200:1, about 5:1 to about 150:1, about 5:1 to about 100:1, about 5:1 to about 50:1, about 5:1 to about 25:1, about 5:1 to about 20:1, about 5:1 to about 10:1, or about 6:1 to about 9:1. Alternatively, the lipid-based compositions disclosed herein have lipid:therapeutic agent (e.g., lipid:nucleic acid) ratios (mol / mol ratios) of about 1:1 to about 30:1, about 2:1 to about 20:1, about 2:1 to about 15:1, about 3:1 to about 10:1, about 4:1 to about 9:1, about 5:1 to about 8:1, or about 6:1 to about 8:1.

[0403] In some embodiments, the lipid-based compositions of the present disclosure are nucleic acid-lipid particles comprising interfering RNA (e.g., dsRNA, e.g., siRNA, Dicer-substrate dsRNA, shRNA, aiRNA, and / or miRNA), ionizable lipids (e.g., one or more lipids of formulas I-XIX described herein or their salts), noncationic lipids (e.g., a mixture of one or more phospholipids and cholesterol), and complex lipids that inhibit particle aggregation (e.g., one or more PEG-lipid complexes). The nucleic acid-lipid particles may contain at least one, two, three, four, five, six, seven, eight, nine, ten, or more unmodified and / or modified interfering RNA molecules (e.g., siRNA). Nucleic acid-lipid particles and methods for preparing them are described, for example, in U.S. Patents 5,753,613, 5,785,992, 5,705,385, 5,976,567, 5,981,501, 6,110,745, and 6,320,017, and PCT Publication No. WO96 / 40964, the contents of which these disclosures are incorporated herein by reference in their entirety for all purposes.

[0404] In the nucleic acid-lipid particles disclosed herein, the nucleic acid is completely encapsulated in the lipid portion of the particle, thereby protecting the nucleic acid from degradation by nucleases. In preferred embodiments, nucleic acid-lipid particles containing nucleic acids such as interfering RNA are completely encapsulated in the lipid portion of the particle, thereby protecting the nucleic acid from degradation by nucleases. In some embodiments, the nucleic acid may be complexed with the lipid portion of the particle. Within the scope of this disclosure, the lipid-based compositions disclosed herein are intended to be substantially nontoxic to mammals such as humans.

[0405] As used herein, the term “fully encapsulated” means that the nucleic acids in nucleic acid-lipid particles are not significantly degraded after exposure to serum or nuclease assays that significantly degrade free DNA or RNA. In fully encapsulated systems, in a process that would normally degrade 100% of free nucleic acids, preferably less than about 25% of the nucleic acids in the particles are degraded, more preferably less than about 10%, and most preferably less than about 5%.

[0406] In some embodiments, the present disclosure provides a nucleic acid-lipid particle composition comprising a plurality of nucleic acid-lipid particles.

[0407] In some cases, the nucleic acid-lipid particle composition is approximately 30% to 100%, 40% to 100%, 50% to 100%, 60% to 100%, 70% to 100%, 80% to 100%, 90% to 100%, 30% to 95%, 40% to 95%, 50% to 95%, 60% to 95%, 70% to 95%, 80% to 95%, 85% to 95%, 90% to 95%, 30% to 90%, 40% to 90%, 50% to Approximately 90%, approximately 60% to approximately 90%, approximately 70% to approximately 90%, approximately 80% to approximately 90%, or at least approximately 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% (or any fraction or range thereof) contain nucleic acids that are completely encapsulated within the lipid portion of the particle.

[0408] The techniques described herein allow for variations in the proportion of components in lipid-based compositions, and provide that the delivery efficiency of a particular formulation can be measured, for example, using an endosomal release parameter (ERP) assay. Within the scope of this disclosure, the lipid-based compositions disclosed herein are intended to have increased delivery efficiency by enhancing endosomal release, at least partially induced by the novel ionizable substituted aryl and / or heteroaryl lipid-like compounds, novel ionizable substituted piperazine lipids, and / or other novel ionizable aryl and / or heteroaryl lipid compounds disclosed herein.

[0409] In accordance with some of the techniques described herein, one or more of the novel ionizable substituted aryl and / or heteroaryllipid-like compounds of formula I, one or more of the novel ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryllipid compounds of formula IX may be used alone or in combination with one or more other cationic or non-cationic lipid species in the lipid-based compositions disclosed herein.

[0410] Obligate cationic lipids or salts thereof, and / or other ionizable lipids or salts thereof may also be included in the lipid-based compositions of this disclosure.

[0411] In some embodiments, the ionizable substituted aryl and heteroaryl lipid-like compounds disclosed herein, the ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryl lipid compounds of formula IX constitute about 40 mol% to about 90 mol%, about 40 mol% to about 85 mol%, about 40 mol% to about 80 mol%, about 40 mol% to about 75 mol%, about 40 mol% to about 70 mol%, about 40 mol% to about 65 mol%, about 40 mol% to about 60 mol%, about 40 mol% to about 55 mol%, about 50 mol% to about 90 mol%, about 50 mol% to about 85 mol%, about 50 mol% to about 80 mol%, about 50 mol% to about 75 mol%, about 50 mol% to about 70 mol%, about 50 mol% to about 65 mol%, and about 50 mol% to about 60 mol% of the total lipids present in the particles.

[0412] In some embodiments, the ionizable substituted aryl and heteroaryllipid-like compounds disclosed herein, the ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryllipid compounds constitute about 50 mol% to about 58 mol%, about 51 mol% to about 59 mol%, about 51 mol% to about 58 mol%, about 51 mol% to about 57 mol%, about 52 mol% to about 58 mol%, about 52 mol% to about 57 mol%, about 52 mol% to about 56 mol%, or about 53 mol% to about 55 mol% of the total lipids present in the particles. In some embodiments, ionizable substituted aryl and / or heteroaryllipid-like compounds, ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryllipid compounds constitute approximately 50 mol%, 51 mol%, 52 mol%, 53 mol%, 54 mol%, 55 mol%, 56 mol%, 57 mol%, 58 mol%, 59 mol%, 60 mol%, 61 mol%, 62 mol%, 63 mol%, 64 mol%, or 65 mol% (or any fraction or range thereof) of the total lipids present in the particles. In some embodiments, ionizable substituted aryl and / or heteroaryl lipid-like compounds, ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryl lipid compounds constitute at least about 66 mol%, 67 mol%, 68 mol%, 69 mol%, 70 mol%, 71 mol%, 72 mol%, 73 mol%, 74 mol%, 75 mol%, 76 mol%, 77 mol%, 78 mol%, 79 mol%, 80 mol%, 81 mol%, 82 mol%, 83 mol%, 84 mol%, 85 mol%, 86 mol%, 87 mol%, 88 mol%, 89 mol%, or 90 mol% of the total lipids present in the particles.In some embodiments, the ionizable substituted aryl and / or heteroaryllipid-like compounds disclosed herein, the ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryllipid compounds constitute approximately 45 mol%, 46 mol%, 47 mol%, 48 mol%, 49 mol%, 50 mol%, 51 mol%, 52 mol%, 53 mol%, 54 mol%, or 55 mol% of the total lipids present in the nucleic acid-lipid particles.

[0413] In some embodiments, the ionizable substituted aryl and / or heteroaryllipid-like compounds disclosed herein, the ionizable substituted piperazine lipids of formula VII, and / or other ionizable aryl and / or heteroaryllipid compounds constitute about 2 mol% to about 60 mol%, about 5 mol% to about 50 mol%, about 10 mol% to about 50 mol%, about 20 mol% to about 50 mol%, about 20 mol% to about 40 mol%, about 30 mol% to about 40 mol%, or about 40 mol% of the total lipids present in the particles.

[0414] Those skilled in the art will understand that the target amounts are the percentages of ionizable substituted aryl and / or heteroaryl lipid-like compounds, the ionizable substituted piperazine lipid of formula VII, and / or other ionizable aryl and / or heteroaryl lipid compounds present in the lipid-based compositions of the present disclosure, and that the actual amount of cationic lipids present in the formulation may vary, for example, by about ±5 mol%.

[0415] Lipid-based compositions disclosed herein include phospholipids, such as lecithin, phosphatidylethanolamine, lysolecithin, lysophosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, sphingomyelin, egg sphingomyelin (ESM), cephalin, cardiolipin, phosphatidic acid, cerebroside, dicetyl phosphate, distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoylphosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyloleoylphosphatidylethanolamine The lipids may also include, but are not limited to, various noncationic lipids, including (POPE), palmitoyl oleoyl phosphatidylglycerol (POPG), dioleoyl phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoyl phosphatidylethanolamine (DPPE), dimyristoyl phosphatidylethanolamine (DMPE), distearoyl phosphatidylethanolamine (DSPE), monomethyl phosphatidylethanolamine, dimethyl phosphatidylethanolamine, dierydoyl phosphatidylethanolamine (DEPE), stearoyl oleoyl phosphatidylethanolamine (SOPE), lysophosphatidylcholine, dilinoleoyl phosphatidylcholine, and mixtures thereof. Other diacylphosphatidylcholine and diacylphosphatidylethanolamine phospholipids may also be used. The acyl group in these lipids is preferably C 10 ~C 24 The acyl group is derived from a fatty acid having a carbon chain, such as lauroyl, myristoyl, palmitoyl, stearoyl, or oleoyl.

[0416] Other examples of noncationic lipids include, but are not limited to, sterols such as cholesterol and their derivatives. Non-limiting examples of cholesterol derivatives include polar analogs such as 5α-cholestanol, 5β-coprostanol, cholesteryl-(2'-hydroxy)-ethyl ether, cholesteryl-(4'-hydroxy)-butyl ether, and 6-ketocholestanol; non-polar analogs such as 5α-cholestan, cholestenone, 5α-cholestanone, 5β-cholestanone, and cholesteryl decanoate; and mixtures thereof. In preferred embodiments, the cholesterol derivative is a polar analog such as cholesteryl-(4'-hydroxy)-butyl ether.

[0417] In some embodiments, noncationic lipids constitute approximately 10 mol% to approximately 60 mol%, approximately 20 mol% to approximately 55 mol%, approximately 20 mol% to approximately 45 mol%, approximately 20 mol% to approximately 40 mol%, approximately 25 mol% to approximately 50 mol%, approximately 25 mol% to approximately 45 mol%, approximately 30 mol% to approximately 50 mol%, approximately 30 mol% to approximately 45 mol%, approximately 30 mol% to approximately 40 mol%, approximately 35 mol% to approximately 45 mol%, approximately 37 mol% to approximately 42 mol%, or approximately 35 mol%, 36 mol%, 37 mol%, 38 mol%, 39 mol%, 40 mol%, 41 mol%, 42 mol%, 43 mol%, 44 mol%, or 45 mol% (or any fraction or range thereof) of the total lipids present in the particles.

[0418] As discussed above with respect to cationic lipids, those skilled in the art will understand that the percentage of non-cationic lipids present in the lipid particles of this disclosure is the target amount, and that the actual amount of non-cationic lipids present in the formulation may vary, for example, by ±5 mol%.

[0419] Lipid nanoparticles of any size can be used in accordance with this disclosure. In certain embodiments of this disclosure, the lipid nanoparticles have a diameter in the range of about 0.02 microns to about 0.4 microns, between about 0.05 microns and about 0.2 microns, or between 0.07 microns and 0.12 microns.

[0420] In some embodiments, the LNP is a protonable tertiary amine (e.g., pH titrable) head group, C 18Other cationic lipids may also be included, but are not limited to, those containing alkyl chains (each alkyl chain independently having 0 to 3 (e.g., 0, 1, 2, or 3) double bonds) and ether, ester, or ketal links between the head group and the alkyl chain.Examples of such cationic lipids include 1,2-distearyloxy-N,N-dimethyl-3-aminopropane (DSDMA), N,N-dioleyl-N,N-dimethylammonium chloride ("DODAC"), 3-(N-(N',N'-dimethylaminoethane)-carbamoyl)cholesterol ("DC-Chol"), N-(1,2-dimyristyloxypropane-3-yl)-N,N-dimethyl-N-hydroxyethylammonium bromide ("DMRIE"), and 1,2-dioleyloxy-N,N-dimethyl-3-aminopropane (D ODMA), 1,2-dilinoleyloxy-N,N-dimethyl-3-aminopropane (DLinDMA), 1,2-dilinolenyloxy-N,N-dimethyl-3-aminopropane (DLenDMA), 1,2-di-γ-linolenyloxy-N,N-dimethylaminopropane (γ-DLenDMA), 1,2-dilinoleyloxy-keto-N,N-dimethyl-3-aminopropane (DLinK-DMA), 1,2-dilinoleyl-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (DLinKC2-DMA) (DLin-C2 (Also known as K-DMA, XTC2, and C2K), 2,2-dilinoleyl-4-(3-dimethylaminopropyl)[1,3]-dioxolane (DLin-K-C3-DMA), 2,2-dilinoleyl-4-(4-dimethylaminobutyl)[1,3]-dioxolane (DLin-K-C4-DMA), 1,2-dilinolenyloxy-4-(2-dimethylaminoethyl)-[1,3]-dioxolane (γ-DLen-C2K-DMA), 1,2-di-γ-linolenyloxy-4-(2-dimethylaminoethyl)-[1,3]-dioxolane This includes, but is not limited to, (γ-DLen-C2K-DMA), dilinoleylmethyl-3-dimethylaminopropionate (DLin-M-C2-DMA) (also known as MC2), (6Z,9Z,28Z,31Z)-heptatriaconta-6,9,28,31-tetraen-19-yl-4-(dimethylamino)butanoate (DLin-M-C3-DMA) (also known as MC3), and 3-(dilinoleylmethoxy)-N,N-dimethylpropane-1-amine (DLin-MP-DMA) (also known as 1-B11).

[0421] In some embodiments, the particles of the Disclosure may include neutral lipids, such as diacylphosphatidylcholine, diacylphosphatidylethanolamine, ceramide, sphingomyelin, cephalin, cholesterol, cerebroside, and diacylglycerol. In other embodiments, the LNP may include anionic lipids, including but not limited to phosphatidylglycerol, cardiolipin, diacylphosphatidylserine, diacylphosphatidic acid, N-dodecanoylphosphatidylethanolamine, N-succinylphosphatidylethanolamine, N-glutarylphosphatidylethanolamine, lysylphosphatidylglycerol, palmitoyloleoylphosphatidylglycerol (POPG), and other anionic modifying groups attached to neutral lipids. In some embodiments, the noncationic lipids used in the Disclosure are 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), and / or 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In some embodiments, one or more noncationic lipids of the particles are cholesterol (CHE), β-sitosterol, and / or derivatives thereof.

[0422] Cationic lipids disclosed herein may include, but are not limited to, the following exemplary cationic lipids: 1,2-dilinoleyloxy-N,N-dimethylaminopropane ("DLinDMA"), 1,2-dilinolenyloxy-N,N-dimethylaminopropane ("DLenDMA"), dioctadecyldimethylammonium ("DODMA"), distearyldimethylammonium ("DSDMA"), N,N-dioleyl-N,N-dimethylammonium chloride ("DODAC"), and N-(2,3-dioleyloxy) Ropyl-N,N,N-trimethylammonium chloride ("DOTMA"), N,N-distearyl-N,N-dimethylammonium bromide ("DDAB"), N-(2,3-dioleoyloxy)propyl-N,N,N-trimethylammonium chloride ("DOTAP"), 3-(N-(N',N'-dimethylaminoethane)-carbamoyl)cholesterol ("DC-Chol"), and N-(1,2-dimyristiloxypropane-3-yl)-N,N-dimethyl-N-hydroxyethylammonium bromide ("DMRIE"). For example, cationic lipids that have a positive charge below physiological pH include, but are not limited to, DODAP, DODMA, DMDMA, and SM-005. In some cases, cationic lipids have a protonable tertiary amine head group, C 18 It includes an alkyl chain, an ether bond between the head group and the alkyl chain, and 0 to 3 double bonds. Such lipids include, for example, DSDMA, DLinDMA, DLenDMA, and DODMA. In exemplary embodiments, such lipids may include SM-005, as well as its salts and isomers. The chemical structure of SM-005 is shown below. [ka]

[0423] In some embodiments using PEG complex lipids, the PEG complex lipid is one or more polyethylene glycol (PEG)-lipid complexes, polyamide (ATTA)-lipid complexes, and mixtures thereof. In one embodiment, the PEG-lipid complex is one or more PEG-dialkyloxypropyl (DAA), PEG-diacylglycerol (DAG), PEG-phospholipids, PEG-ceramide, and mixtures thereof. In one embodiment, the PEG-DAG complex is PEG-dilauroylglycerol (C 12 ), PEG-dimyristoylglycerol (C 14 ), PEG-Dipalmitoylglycerol (C 16 ), and PEG-distearoylglycerol (C 18 ) is one or more of the following. In one embodiment, the PEG-DAA complex is PEG-dilauryloxypropyl (C 12 ), PEG-Dimyristyloxypropyl (C 14 ), PEG-Dipalmityloxypropyl(C 16 ), and PEG-distearyloxypropyl (C 18 ) is one or more of the above. In some embodiments, PEG is 2-dimyristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG-DMG) and / or 1,2-distearoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (PEG-DSG).

[0424] In some embodiments, amphiphilic lipids are included in the particles of this disclosure. Amphiphilic lipids can refer to any suitable material in which the hydrophobic portion of the lipid material is oriented towards the hydrophobic phase, while the hydrophilic portion is oriented towards the aqueous phase. Such compounds include, but are not limited to, phospholipids, aminolipids, and sphingolipids. Representative phospholipids include sphingomyelin, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidylinositol, phosphatidic acid, palmitoyloleoylphosphatidylcholine, lysophosphatidylcholine, lysophosphatidylethanolamine, dipalmitoylphosphatidylcholine, dioleoylphosphatidylcholine, distearoylphosphatidylcholine, or dilinoleoylphosphatidylcholine. Other phosphorus-deficient compounds such as sphingolipids, the sphingoglycolipid family, diacylglycerols, and β-acyloxy acids may also be used. In addition, such amphiphilic lipids can be readily mixed with other lipids such as triglycerides and sterols.

[0425] Programmable fusion lipid formulations are also suitable for inclusion in the lipid particles of this disclosure. Such formulations have little tendency to fuse with cell membranes and deliver the formulation cargo until a given signaling event occurs. This allows the lipid formulation to be distributed more uniformly after injection into a biological or disease site before initiating fusion with cells. The signaling event may be, for example, a change in pH, temperature, ionic environment, or time. In the latter case, fusion-delaying or "cloaking" components, such as ATTA-lipid complexes or PEG-lipid complexes, can be readily replaced from the lipid nanoparticle membrane over time. By the time the formulation is suitably distributed in the body, it will have lost sufficient cloaking agents to become fusionable. For other signaling events, it is desirable to select signals related to the disease site or target cells, such as an increase in temperature at the site of inflammation.

[0426] In some embodiments, the lipid nanoparticles disclosed herein can be formulated by mixing two or three fluid streams containing nucleic acid cargo and lipid components, respectively, using a microfluidic mixer, a cross, or a T-junction.

[0427] In certain embodiments, it may be desirable to further target the lipid nanoparticles of this disclosure using cell type or tissue-specific targeting moieties. Targeting of lipid nanoparticles using various targeting moieties such as ligands, cell surface receptors, glycoproteins, vitamins (e.g., riboflavin), and monoclonal antibodies has been described previously (see, for example, U.S. Patents 4,957,773 and 4,603,044). The targeting moiety may include an entire protein or a fragment thereof.

[0428] The targeting mechanism generally requires that the targeting agent be positioned on the surface of lipid nanoparticles in a manner that allows the target portion to interact with the target, such as a cell surface receptor. Various different targeting agents and methods are known and available in the art, including, for example, those described in Sapra, P. and Allen, TM, Prog. Lipid Res. 42(5):439-62 (2003) and Abra, RM et al., J. Lipid nanoparticle Res. 12:1-3, (2002).

[0429] Standard methods for conjugating target drugs can be used. For example, phosphatidylethanolamine or derivatized lipophilic compounds such as lipid-derivatized bleomycin, which can be activated for target drug conjugation, can be used. Antibody-targeted lipid nanoparticles can be constructed, for example, using lipid nanoparticles incorporating protein A (see Renneisen, et al., J. Bio. Chem., 265:16337-16342 (1990) and Leonetti, et al., Proc. Natl. Acad. Sci. (USA), 87:2448-2451 (1990)). Other examples of antibody conjugation are disclosed in U.S. Patent No. 6,027,726, the teachings of which are incorporated herein by reference. Other proteins specific to cellular components, including antigens associated with neoplasms or tumors, can also be cited as examples of targeting moieties. The proteins used as the targeting moiety can be bound to lipid nanoparticles via covalent bonds (see Heath, Covalent Attachment of Proteins to Lipid nanoparticles, 149 Methods in Enzymology 111-119 (Academic Press, Inc. 1987)). Other targeting methods include biotin-avidin systems.

[0430] For example, Szoka, et al., Ann. Rev. Biophys. Bioeng., 9:467 (1980), U.S. Patents 4,186,183, 4,217,344, 4,235,871, 4,261,975, 4,485,054, No. 4,501,728, No. 4,774,085, No. 4,837,028, No. 4,946,787, PCT Publication No. WO 91 / 17424, Deamer and Bangham, Biochim. Biophys. Acta, 443:629-634 (1976), Fraley, et al., Proc. Natl. Acad. Sci. USA, 76:3348-3352 (1979), Hope, et al. Various methods for preparing lipid nanoparticles are known in the art, including those described in *Biochim. Biophys. Acta, 812:55-65 (1985)*, *Mayer, et al., Biochim. Biophys. Acta, 858:161-168 (1986)*, *Williams, et al., Proc. Natl. Acad. Sci., 85:242-246 (1988)*, ​​*Lipid nanoparticles*, Marc J. Ostro, ed., Marcel Dekker, Inc., New York, 1983, Chapter 1; *Hope, et al., Chem. Phys. Lip., 40:89 (1986)*, and *Lipid nanoparticles: A Practical Approach*, Torchilin, VP et al., ed., Oxford University Press (2003), as well as the references cited therein. Suitable methods include, but are not limited to, sonication, extrusion, high-pressure / homogenization, microfluidization, surfactant dialysis, calcium-induced fusion of small lipid nanoparticle vesicles, and ether injection, all of which are well known in the art.

[0431] Some embodiments of this disclosure include SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, and SM-131. LNPs including SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, and / or SM-173 were prepared using a microfluidic mixing process or a T-junction mixing process, which involved two fluid streams, one of which contained an aqueous solution of nucleic acid entities and the other containing an organic solution of lipid components and / or IC molecules.

[0432] Lipids / components are ionizable with 30-50 mol% ionizable substituted aryl and heteroaryl lipid-like compounds (e.g., SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM- Lipids were prepared by combining 20-40 mol% of helpers (such as those described herein), 25-35 mol% of structural lipids such as cholesterol (Chol or CHE), and 0.3-5 mol% of PEG-lipids (e.g., PEG-5K) in ethanol to a total concentration of approximately 10-30 mM. The lipid components were combined to the desired molar ratio (see, for example, Tables 4 and 6), and diluted with an aqueous nucleic acid solution to a final lipid concentration between 3 and 15 mM.

[0433] In other embodiments of the present disclosure, LNPs including SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, and SM-125 were prepared using a microfluidic mixing process or a T-joint mixing process, which included two fluid streams, one of which contained an aqueous solution of nucleic acid entities and the other containing an organic solution of lipid components and / or ic molecules.

[0434] Lipids / components were prepared by combining lipids according to a formulation of 35-55 mol% ionizable substituted piperazine lipids (e.g., SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, and SM-125), 15-35 mol% helper lipids such as those described herein, 25-40 mol% structural lipids such as cholesterol (Chol or CHE), and 0.3-2.5 mol% PEG-lipids (e.g., PEG-DMG) in ethanol at a total concentration of approximately 10-30 mM. The lipid components were combined to the desired molar ratio (see, for example, Tables 9 and 11), and diluted with an aqueous nucleic acid solution to a final lipid concentration between 3 and 15 mM.

[0435] In further embodiments of the present disclosure, LNPs including SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112 were prepared using a microfluidic mixing process or a T-joint mixing process, which included two fluid streams, one of which contained an aqueous solution of nucleic acid entities and the other containing an organic solution of lipid components and / or ic molecules.

[0436] Lipids / components were prepared by combining lipids in ethanol at a total concentration of approximately 10-30 mM, according to a formulation of 30-50 mol% of ionizable substituted aryl and heteroaryl lipid-like compounds (e.g., SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112), 20-40 mol% of helpers such as those described herein, 25-35 mol% of structural lipids such as cholesterol (Chol or CHE), and 0.3-5 mol% of PEG-lipids (e.g., PEG-5K). The lipid components were combined to the desired molar ratio (see, for example, Tables 12 and 13 below) and diluted with an aqueous nucleic acid solution to a final lipid concentration between 3 and 15 mM.

[0437] Nanoparticle compositions containing nucleic acids and lipid components are prepared by combining an organic solution containing lipids / components with an aqueous nucleic acid solution in a w / w ratio of total lipids to nucleic acids between approximately 10:1 and 100:1. The lipid solution is rapidly injected into the aqueous nucleic acid solution at a flow rate between approximately 8 and 12 mL / min using a NanoAssemblr microfluidic-based system, in a water:organic volume ratio between approximately 1:1 and 4:1. The mixture is then immediately diluted in a 1:1 volume ratio with nuclease-free water. The diluted mixture is then processed using a buffer exchange column or tangential flow filtration (TFF) system to exchange the solution with a desired final buffer, such as Tris-HCl or Tris / acetic acid buffer at a neutral pH between 7.0 and 7.5, containing up to 15% sucrose. The solution is then concentrated using a TFF or filtered centrifugation column. The concentrated solution is then sterile filtered and diluted to the desired nucleic acid concentration between approximately 0.1 mg / mL and 1.0 mg / mL, and then frozen for storage.

[0438] Lipid particles disclosed herein and prepared according to methods known in the art can, in certain embodiments, be loaded with drugs and stored for a considerable period of time before being administered to a patient. For example, lipid nanoparticles can be dehydrated before administration, stored, then rehydrated and loaded with one or more active agents. Lipid nanoparticles can also be dehydrated after being loaded with one or more active agents. Dehydration can be achieved by various methods available in the art, including dehydration and lyophilization procedures described in, for example, U.S. Patents 4,880,635, 5,578,320, 5,837,279, 5,922,350, 4,857,319, 5,376,380, 5,817,334, 6,355,267, and 6,475,517. In one embodiment, lipid nanoparticles are dehydrated using a standard freeze-drying apparatus, i.e., dehydrated under low-pressure conditions. Alternatively, the lipid nanoparticles may be frozen in liquid nitrogen, for example, before dehydration. Before dehydration, sugars can be added to the LNP environment, for example, a buffer containing the lipid nanoparticles, thereby improving the integrity of the lipid nanoparticles during dehydration. See, for example, U.S. Patent No. 5,077,056 or No. 5,736,155.

[0439] Lipid nanoparticles can be sterilized by conventional methods at any point during preparation, including, for example, after size adjustment or pH gradient generation.

[0440] Cargo-loaded lipid particle composition In various embodiments, the lipid particles of this disclosure can be used for many different applications, including the delivery of active agents to cells, tissues, organs, or targets. For example, the lipid nanoparticles of this disclosure can be used to deliver therapeutic agents systemically via the bloodstream or to deliver cosmetic substances to the skin. Thus, the lipid nanoparticles of this disclosure and one or more active agents as cargo are included in this disclosure.

[0441] Lipid particle cargo This disclosure describes lipid nanoparticles combined with active agents as cargo. Active agents include any molecule or compound that, when used herein, can exert a desired effect on cells, tissues, organs, or targets. Such effects may be, for example, biological, physiological, or cosmetic. Active agents may be any type of molecule or compound, including nucleic acids, e.g., single-stranded or double-stranded polynucleotides, plasmids, antisense RNA, e.g., DNA-DNA hybrids, DNA-RNA hybrids, RNA-DNA hybrids, RNA-RNA hybrids, small interfering RNA (siRNA), microRNA (mRNA), and small hairpin RNA (shRNA) and RNA interferants; peptides and polypeptides, e.g., antibodies, e.g., polyclonal antibodies, monoclonal antibodies, antibody fragments; humanized antibodies, recombinant antibodies, recombinant human antibodies, and Primatized® antibodies; cytokines, growth factors, apoptotic factors, differentiation-inducing factors, cell surface receptors and their ligands; hormones; and small molecules, including small organic molecules or compounds.

[0442] Therapeutic drugs As disclosed herein, therapeutic agents may include any molecule or compound capable of exerting a desired effect on cells, tissues, tumors, organs, or targets. Therapeutic agents may be any type of molecule or compound, including but not limited to nucleic acids, peptides, polypeptides, small molecules, and mixtures thereof.

[0443] In some embodiments, the therapeutic agent may be a salt or derivative thereof. The therapeutic agent may be therapeutically active on its own, or it may be a prodrug that becomes active upon further modification / alteration.

[0444] In some embodiments, the lipid-based compositions described herein can associate with nucleic acids such as siRNA, Dicer-substrate dsRNA, shRNA, aiRNA, miRNA, antisense oligonucleotides, ribozymes, and immunostimulatory oligonucleotides.

[0445] Nucleic acid therapy possesses a well-known and astonishing potential to treat diseases at the genetic level. However, a safe and effective delivery system is essential for nucleic acid therapy. Non-specific delivery to organs and tissues often results in effects and toxicity outside the target site. The delivery of therapeutic agents to specific organs of interest is a well-recognized need in lipid-nanoparticle development and in drug development in general. The concept of targeting only the cause of disease without harming other parts of the body was described by Ehrlich 120 years ago. However, existing methods do not provide a defined or well-known methodology for developing nanoparticles that target specific tissues without introducing additional ligand-based targeting strategies. Therefore, organ-specific targeting of lipid nanoparticles based on the structural affinity of lipids to tissues, as disclosed herein, satisfies a well-established need in reducing effects and toxicity outside the target site.

[0446] Nucleic acids associated with or encapsulated by LNPs include 2'-O-methyl modified nucleotides, nucleotides containing a 5'-phosphorothioate group, terminal nucleotides linked to cholesteryl derivatives, 2'-deoxy-2'-fluoro modified nucleotides, 5'-methoxy modified nucleotides (e.g., 5'-methoxyuridine), 2'-deoxy modified nucleotides, locked nucleotides, debasalized nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides, phosphoramidates, nucleotides containing unnatural bases; phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkylphosphotes Modifications may include, but are not limited to, those selected from the group of internucleoside links or skeletons, including methyl and other alkyl phosphonates, phosphinates, 3'-aminophosphoramides, and aminoalkylphosphoramides, thionophosphoramides, thionoalkylphosphonates, thionoalkylphosphotryesters, and boranophosphates having the usual 3'-5' linkage, their 2'-5' linked analogs, and those having inverted polarity, where pairs of adjacent nucleoside units are linked from 3'-5' to 5'-3' or 2'-5' to 5'-2'.

[0447] In certain embodiments, the active agent is mRNA or a vector capable of expressing mRNA within a cell.

[0448] In some embodiments, the active agent is a CRISPR / Cas system. If necessary, the LNPs of this disclosure can be formulated to include, for example, both a guide strand (gRNA) and a Cas enzyme as cargo, thereby providing a self-contained delivery vehicle capable of achieving and controlling CRISPR-mediated targeting of genes in target cells.

[0449] In certain characteristic embodiments, the active agent is a nucleic acid regulatory factor (for example, mRNA encoding a protein regulatory factor component, as described above).

[0450] In some embodiments, the active agent is a therapeutic agent, a salt thereof, or a derivative thereof. The therapeutic agent derivative may be therapeutically active on its own, or it may be a prodrug that becomes active upon further modification. Thus, in one embodiment, the therapeutic agent derivative retains some or all of the therapeutic activity compared to the unmodified agent, while in another embodiment, the therapeutic agent derivative is not therapeutically active.

[0451] In various embodiments, therapeutic agents include drugs and medications such as anti-inflammatory compounds, narcotics, sedatives, antidepressants, stimulants, hallucinogens, analgesics, antibiotics, contraceptives, antipyretics, vasodilators, anti-angiogenic agents, cellular angiogenesis agents, signaling inhibitors, vasoconstrictors, hormones, and steroids.

[0452] In certain embodiments, the active agent is an oncology drug, which may also be referred to as an antitumor drug, anticancer drug, tumor drug, anti-cancer agent, or similar name. Examples of oncology drugs that may be used in accordance with this disclosure include: Adriamycin, Alkeran, Allopurinol, Altretamine, Amifostine, Anastrozole, araC, Arsenic Trioxide, Azathioprine, Bexarotene, biCNU, Bleomycin, Intravenous Busulfan, Oral Busulfan, Capecitabine (Xeloda), Carboplatin, Carmustine, CCNU, Celecoxib, Chlorambucil, Cisplatin, Cladribine, Cyclospoli. N-A, cytarabine, cytosine arabinoside, daunorubicin, cytoxane, daunorubicin, dexamethasone, dexrazoxane, dodetaxel, doxorubicin, doxorubicin, DTIC, epirubicin, estramustine, etoposide phosphate, etoposide and VP-16, exemestane, FK506, fludarabine, fluorouracil, 5-FU, gemcitabine (Gemzar), gemtuzumab ozogamicin, goserelin acetate, ha Idrea, hydroxyurea, idarubicin, ifosfamide, imatinib mesylate, interferon, irinotecan (Camptostar, CPT-111), letrozole, leucovorin, leustatin, leuprolide, rebamisole, litretinoin, megastrol, melphalan, L-PAM, mesna, methotrexate, methoxsalen, mithramycin, mitomycin, mitoxantrone, nitrogen mustard, pacristoyl peroxide. Examples of oncological agents that may be used in accordance with this disclosure include, but are not limited to, taxel, pamidronate, pegademase, pentostatin, porfimer sodium, prednisone, rituxan, streptozocin, STI-571, tamoxifen, taxotere, temozolamide, teniposide, VM-26, topotecan (Hycamtin), toremifene, tretinoin, ATRA, barrubicin, vervan, vinblastine, vincristine, VP16, and vinorelbine. Other examples of oncological agents that may be used in accordance with this disclosure include ellipticin and its analogs or derivatives, epotilon, intracellular kinase inhibitors, and camptothecin.

[0453] The LNP compositions of this disclosure may generally comprise a single active agent, and in certain embodiments, may comprise more than one active agent.

[0454] In other embodiments of the Disclosure, the lipid nanoparticles of the Disclosure have a plasma circulating half-life of at least 0.5, 0.8, 1.2, 1.5, 2.0, 4.0, 6.0, 8.0, or 12 hours. In some embodiments, the lipid nanoparticles have a plasma drug half-life of at least 0.5, 0.8, 1.2, 1.5, 2.0, 4.0, 6.0, 8.0, or 12 hours. The circulating and blood or plasma clearance half-lives can be determined, for example, as described in U.S. Patent Application Publication No. 2004-0071768-A1.

[0455] The techniques described herein further include lipid particles and / or pharmaceutical compositions in which therapeutic agents, such as nucleic acids (e.g., siRNA, ASO, tRNA, miRNA, mRNA, DNA, and analogues), proteins, peptides, and other macromolecules, are encapsulated within a lipid portion of a particle or composition so as to be protected from degradation. Such lipid particles and / or pharmaceutical compositions can be formed by any method known in the art, including but not limited to serial mixing, direct dilution, and in-line dilution.

[0456] In some embodiments, lipid particles and / or pharmaceutical compositions may contain any of the ionizable lipids disclosed herein or a salt thereof, alone or in combination with other cationic lipids and / or non-cationic lipids. In other embodiments, the non-cationic lipids include egg sphingomyelin (ESM), distearoyl phosphatidylcholine (DSPC), dioleoyl phosphatidylcholine (DOPC), 1-palmitoyl-2-oleoyl phosphatidylcholine (POPC), dipalmitoyl phosphatidylcholine (DPPC), monomethyl phosphatidylethanolamine, dimethyl phosphatidylethanolamine, 14:0PE (1,2-dimyristoyl phosphatidylethanolamine (DMPE)), 16:0PE (1,2-dipalmitoyl phosphatidylethanolamine (DPPE)), and 18:0PE (1,2-distearoyl phosphatidylethanolamine (DS These may be polyethylene glycol-based polymers (e.g., PEG2000, PEG5000, PEG-modified diacylglycerol, or PEG-modified dialkyloxypropyl), cholesterol, derivatives thereof, or combinations thereof.

[0457] Lipid particles and / or pharmaceutical compositions disclosed herein can be formed using techniques known in the art, such as continuous mixing, which has the effect of continuously diluting the lipid solution with the buffer solution by continuously introducing a lipid solution and a buffer solution into a mixing area, thereby generating lipid vesicles almost immediately after mixing. By mixing an aqueous solution containing a therapeutic agent with an organic lipid solution, the organic lipid solution can be continuously and stepwise diluted in the presence of a buffer solution to generate therapeutic agent-lipid particles. Such particles are approximately 30nm to 250nm, 40nm to 200nm, 50nm to 150nm, 60nm to 130nm, 70nm to 110nm, 70nm to 100nm, 80nm to 100nm, 90nm to 100nm, 70nm to 90nm, 80nm to 90nm, 70nm to 80nm, less than 120nm, less than 110nm, less than 100nm, less than 90nm or less than 80nm, or approximately 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, 65nm, 70nm, 75nm, 8 The particles can have sizes of 0 nm, 85 nm, 90 nm, 95 nm, 100 nm, 105 nm, 110 nm, 115 nm, 120 nm, 125 nm, 130 nm, 135 nm, 140 nm, 145 nm, 150 nm, 155 nm, 160 nm, 165 nm, 170 nm, 175 nm, 180 nm, 185 nm, 190 nm, 195 nm, 200 nm, 205 nm, 210 nm, 215 nm, 220 nm, 225 nm, 230 nm, 235 nm, 240 nm, 245 nm, or 250 nm, or any intermediate or partial range thereof. The particles do not aggregate once formed. According to the art of this specification, the particles can be sized to achieve a uniform particle size.

[0458] It is also intended within the scope of this disclosure that such particles may be prepared by direct dilution processes, such as those described in U.S. Patent Application Publication No. 20070042031 (e.g., forming a lipid vesicle solution and introducing it directly into a container with a controlled amount of dilution buffer), the disclosure of which is incorporated herein by reference in whole for all purposes. Particles formed using the direct dilution process typically fall within the following ranges: approximately 30nm to 250nm, 40nm to 200nm, 50nm to 150nm, 60nm to 130nm, 70nm to 110nm, 70nm to 100nm, 80nm to 100nm, 90nm to 100nm, 70nm to 90nm, 80nm to 90nm, 70nm to 80nm, less than 120nm, less than 110nm, less than 100nm, less than 90nm or less than 80nm, or approximately 30nm, 35nm, 40nm, 45nm, 50nm, 55nm, 60nm, and 65nm. The particles have sizes of 70nm, 75nm, 80nm, 85nm, 90nm, 95nm, 100nm, 105nm, 110nm, 115nm, 120nm, 125nm, 130nm, 135nm, 140nm, 145nm, 150nm, 155nm, 160nm, 165nm, 170nm, 175nm, 180nm, 185nm, 190nm, 195nm, 200nm, 205nm, 210nm, 215nm, 220nm, 225nm, 230nm, 235nm, 240nm, 245nm, or 250nm, or any intermediate or partial range thereof. The particles do not aggregate once formed. According to the art of this specification, the particles can be sized to achieve a uniform particle size.

[0459] In some embodiments, non-lipid polycations useful for cell lipofection can be added to the compositions of the present invention. Suitable examples of non-lipid polycations include hexadimethrin bromide (marketed under the brand name POLYBRENE® by Aldrich Chemical Co., Milwaukee, Wis., USA) or other salts of hexadimethrin. Other suitable polycations include, for example, salts of poly-L-ornithine, poly-L-arginine, poly-L-lysine, poly-D-lysine, polyallylamine, and polyethyleneimine. The addition of these salts is preferably carried out after the particles have been formed.

[0460] kit This disclosure also provides lipid nanoparticles and variant forms thereof in kit form. The kit may include a ready-made formulation or a formulation that requires mixing before administration. The kit typically includes a container that is compartmentalized to house the various elements of the kit. The kit contains the lipid nanoparticle composition or its components of this disclosure in hydrated or dehydrated form, along with instructions for use for their rehydration and administration. In certain embodiments, the kit includes at least one compartment containing the lipid nanoparticles of this disclosure loaded with an active agent. In other embodiments, the kit includes at least two compartments, one containing the lipid nanoparticles of this disclosure and the other containing an active agent. Naturally, it is understood that any of these kits may include additional compartments, such as those containing buffers, as described in U.S. Patent Application Publication No. 2004-0228909-A1. Ionizable lipids (e.g., SM-066, SM-078, SM-80, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, S M-101, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, SM-173, SM A kit of the present disclosure comprising lipid nanoparticles (including SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, SM-125, SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112) may also contain other features of the kit described in U.S. Patent Application Publication No. 2004-0228909A1. Furthermore, the kit may contain drug-loaded lipid nanoparticles in one compartment and empty lipid nanoparticles in a second compartment.Alternatively, the kit may contain the lipid nanoparticles of the Disclosure, an active agent to be loaded onto the lipid nanoparticles of the Disclosure in a second compartment, and empty lipid nanoparticles in a third compartment.

[0461] In certain embodiments, the kit of the present disclosure comprises a therapeutic compound encapsulated in lipid nanoparticles, the lipid nanoparticles being SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-14 1, including SM-152, SM-155, SM-158, SM-163, SM-170, SM-173, SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, SM-125, SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112, where SM-066, S M-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, SM -114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, SM-173, SM-048, SM-074, SM- SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, SM-125, SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112 constitute at least 20%, at least 50%, or at least 70% (on a molar basis) of the total phospholipids present in the lipid nanoparticles, and the kit also includes empty lipid nanoparticles.In some embodiments, SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-10 1, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, SM-173 SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, SM-125, SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112 constitute at least 45-55% (molar basis) of the total phospholipids present in the lipid nanoparticles, and the kit also includes empty lipid nanoparticles. In some embodiments, SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-10 1, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, SM-173 SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, SM-125, SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112 constitute at least 48-52% (molar basis) of the total phospholipids present in the lipid nanoparticles, and the kit also includes empty lipid nanoparticles.In some embodiments, SM-066, SM-078, SM-081, SM-082, SM-084, SM-089, SM-090, SM-091, SM-092, SM-093, SM-094, SM-095, SM-096, SM-098, SM-101, SM-103, SM-106, SM-114, SM-115, SM-117, SM-120, SM-129, SM-131, SM-138, SM-141, SM-152, SM-155, SM-158, SM-163, SM-170, SM-173, SM-048, SM-074, SM- SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, SM-125, SM-016, SM-062, SM-065, SM-067, SM-068, SM-070, SM-072, SM-073, SM-107, SM-111, and / or SM-112 constitute at least 48%, at least 49%, at least 50%, at least 51%, or at least 52% (on a molar basis) of the total phospholipids present in the lipid nanoparticles, and the kit also includes empty lipid nanoparticles. In one embodiment, lipid nanoparticles containing the therapeutic compound and empty lipid nanoparticles are located in different compartments of the kit.

[0462] Treatment method The LNP compositions of this disclosure can be used to treat any of the wide variety of diseases or disorders, including but not limited to inflammatory diseases, cardiovascular diseases, neurological diseases, tumors, demyelinating diseases, gastrointestinal diseases, endocrine diseases, reproductive diseases, blood and lymphatic diseases, immune diseases, mental diseases, musculoskeletal diseases, neurological diseases, neuromuscular diseases, metabolic diseases, sexually transmitted infections, skin and connective tissue diseases, urinary tract diseases, and infections.

[0463] In certain embodiments, the LNP composition can be used to treat or prevent lung diseases or disorders selected from lung cancer, pneumonia, pulmonary fibrosis, COPD, asthma, bronchiectasis, sarcoidosis, pulmonary hypertension, emphysema, alpha-1 antitrypsin deficiency, aspergillosis, bronchiolitis, bronchitis, pneumoconiosis, coronavirus, Middle East respiratory syndrome, severe acute respiratory syndrome, cystic fibrosis, Legionnaires' disease, influenza, pertussis, pulmonary embolism, and tuberculosis.

[0464] In other embodiments, the LNP compositions of this disclosure can be used to treat or prevent joint diseases or disorders, including but not limited to diseases or disorders selected from rheumatoid arthritis, psoriatic arthritis, gout, tendinitis, bursitis, carpal tunnel syndrome, and osteoarthritis.

[0465] In other embodiments, the LNP composition of this disclosure is used for inflammatory bowel disease, peritonitis, osteomyelitis, cachexia, pancreatitis, trauma-induced shock, bronchial asthma, allergic rhinitis, cystic fibrosis, acute bronchitis, acute severe bronchitis, osteoarthritis, rheumatoid arthritis, infectious arthritis, post-infectious arthritis, gonococcal arthritis, tuberculous arthritis, arthritis, osteoarthritis, gout, spondyloarthritis, ankylosing spondylitis, arthritis associated with vasculitis syndrome, neurogenic polyarteritis nodosa, hypersensitivity vasculitis, neoplastic granulomatosis, myalgia due to rheumatic polyposis, arthritis cell arteritis, and calcium dysplasia. It can be used to treat or prevent inflammatory diseases or disorders, including but not limited to diseases or disorders selected from among: polycystic arthropathy, corrosive gout, non-arthritis, bursitis, hay fever, suppurative inflammation (e.g., tennis elbow), neurogenic arthropathy, intra-articular hemorrhage, Henoch-Schlein purpura, hypertrophic osteoarthritis, multiple hemorrhoids, scoliosis, hemochromatosis, hyperlipoproteinemia, hypogammaglobulinemia, COPD, acute respiratory distress syndrome, acute lung injury, bronchopulmonary dysplasia, and systemic lupus erythematosus (SLE).

[0466] In other embodiments, the LNP compositions of the present disclosure can be used to treat or prevent epidermal diseases or disorders, including but not limited to psoriasis, atopic dermatitis, scleroderma, eczema, rosacea, seborrheic dermatitis, melanoma, actinic keratosis, ichthyosis, Glover's disease, verruca vulgaris, keratosacral cell tumor, and seborrheic keratosis.

[0467] In one embodiment, the LNP compositions of this disclosure can be used to treat or prevent certain types of cancer. In particular, these methods can be applied to cancers of the blood and lymphatic system, including lymphoma, leukemia, and myeloma. Examples of specific cancers that may be treated in accordance with this disclosure include, but are not limited to, Hodgkin lymphoma and non-Hodgkin lymphoma (NHL), including any type of NHL defined according to one of various classification systems such as Working Formulation, Rappaport classification, and preferably REAL classification. Such lymphomas include, but are not limited to, low-grade, intermediate-grade, and high-grade lymphomas, as well as both B-cell lymphoma and T-cell lymphoma. These categories include various types of lymphoma, including small cell, large cell, cleaved cell, lymphoid, follicular, diffuse, Burkitt, mantle cell, NK cell, CNS, AIDS-related, lymphoblastic, adult lymphoblastic, painless, invasive, transformative, and other types of lymphoma. The methods described herein can be used for adult-onset or pediatric lymphomas, and for lymphomas of any stage, such as stage I, II, III, or IV. Various types of lymphomas are well known to those skilled in the art and are described, for example, by the American Cancer Society (see, for example, www3.cancer.org).

[0468] The compositions and methods described herein can also be applied to all forms of leukemia, including adult and childhood forms of leukemia. For example, any acute, chronic, myeloid, or lymphoid form of the disease can be treated using the methods of this disclosure. In preferred embodiments, these methods are used to treat acute lymphoblastic leukemia (ALL). Further information on various types of leukemia can be found, among other things, at the American Leukemia Association (see, for example, www.leukemia.org). Further types of tumors, such as neuroblastoma, myeloma, prostate cancer, small cell lung cancer, colon cancer, ovarian cancer, non-small cell lung cancer, brain tumors, breast cancer, and other tumors, can also be treated using the methods described herein. The LNP compositions of this disclosure can be administered as first-line or second-line treatment. In addition, they can be administered as primary chemotherapy or as adjuvant or neoadjuvant chemotherapy. For example, treatment for relapsed, painless, transformative, and invasive forms of non-Hodgkin lymphoma can be administered after at least one course of primary anti-cancer treatment, such as chemotherapy and / or radiotherapy.

[0469] Administration of LNP composition The LNP compositions of this disclosure are administered by any of several methods, including parenteral, intravenous, systemic, topical, oral, intratumor, intramuscular, subcutaneous, intraperitoneal, inhalation, or any such delivery method. In one embodiment, the composition is administered parenterally, i.e., intra-articular, intravenous, intraperitoneal, subcutaneous, or intramuscular. In specific embodiments, the LNP composition is administered by intravenous infusion or intraperitoneal by bolus injection. For example, in one embodiment, the patient is given an intravenous infusion of the active agent encapsulated in lipid nanoparticles through an intravenous line for, for example, 5-10 minutes, 15-20 minutes, 30 minutes, 60 minutes, 90 minutes, or longer. In one embodiment, a 60-minute infusion is used. In other embodiments, infusions ranging from 6-10 minutes or 15-20 minutes are used. Such injections can be administered regularly, for example, once every 1, 3, 5, 7, 10, 14, 21, or 28 days, or for longer periods, preferably once every 7 to 21 days, preferably once every 7 or 14 days.

[0470] The LNP compositions of this disclosure can be formulated as pharmaceutical compositions suitable for delivery to a target. The pharmaceutical compositions of this disclosure often further include one or more buffers (e.g., neutral buffered saline or phosphate-buffered saline), carbohydrates (e.g., glucose, mannose, sucrose, dextrose or dextran), mannitol, proteins, polypeptides or amino acids such as glycine, antioxidants, bacteriostatic agents, chelating agents such as EDTA or glutathione, adjuvants (e.g., aluminum hydroxide), solutes to make the formulation isotonic, hypotonic or weakly hypertonic with the recipient's blood, suspending agents, thickeners, and / or preservatives. Alternatively, the compositions of this disclosure can be formulated as lyophilized products.

[0471] The concentrations of drug and lipid nanoparticles in pharmaceutical formulations can vary widely, i.e., less than about 0.05% by weight, typically around 2-5%, or at least 2-5%, up to 10-30%, and are selected according to the specific drug used, the condition of the disease being treated, and the physician's judgment. Furthermore, the concentrations of drug and lipid nanoparticles also take into account the volume of the fluid being administered, the osmotic pressure and gravimetric concentration of the solution being administered, and the tolerability of the drug and lipid nanoparticles. In some cases, it may be preferable to use lower drug or lipid nanoparticle concentrations to reduce the incidence or severity of side effects associated with the infusion.

[0472] Formulations suitable for use in this disclosure include, for example, Remington's Pharmaceutical Sciences, Mack Publishing Company, Philadelphia, Pa., 17 thThis can be found in Ed. (1985). In many cases, intravenous compositions contain a solution of lipid nanoparticles suspended in an acceptable carrier, such as an aqueous carrier. Any of the various aqueous carriers, e.g., water, buffered water, 0.4% saline, 0.9% isotonic saline, 0.3% glycine, 5% glucose, and analogues, can be used and may contain glycoproteins such as albumin, lipoprotein, and globulin to enhance stability. Often, ordinary buffered saline (135-150 mM NaCl) or 5% glucose is used. These compositions can be sterilized by conventional sterilization techniques such as filtration. The resulting aqueous solution can be packaged for use or filtered under sterile conditions and lyophilized, and the lyophilized preparation is combined with a sterile aqueous solution before administration. The composition may also contain pharmaceutically acceptable auxiliary substances such as pH adjusters and buffers, tonicity adjusters, and analogues, e.g., sodium acetate, sodium lactate, sodium chloride, potassium chloride, calcium chloride, etc., as needed to approximate physiological conditions. In addition, the composition may contain lipid protectants that protect lipids from damage caused by free radicals and lipid peroxidation during storage. Lipophilic free radical quenchers such as α-tocopherol and water-soluble iron-specific chelating agents such as ferrioxamine are preferred.

[0473] The amount of active agent administered per dose is selected to be above the minimum therapeutic dose but below the toxic dose. The selection of the dose per dose depends on several factors, including the patient's medical history, use of other treatments, and the nature of the disease. In addition, the amount of active agent administered may be adjusted throughout the treatment depending on the patient's response to the treatment and the presence or severity of any side effects associated with the treatment. In certain embodiments, the dose or frequency of administration of the LNP composition is approximately the same as the dose and schedule of the treatment using the corresponding free active agent. However, it is understood that the dose may be increased or the frequency increased compared to the free drug treatment, especially when the LNP composition exhibits low toxicity. It is also understood that the dose may be reduced or the frequency decreased compared to the free drug treatment, especially when the LNP composition exhibits increased efficacy compared to the free drug. Exemplary dosages and administrations of various chemotherapy compounds (free drugs) are known to those skilled in the art and are readily available, for example, as described in Physician's Cancer Chemotherapy Drug Manual, E. Chu and V. Devita (Jones and Bartlett, 2002).

[0474] Patients typically receive at least two courses of such treatment, but potentially more courses depending on their response to the treatment. With monotherapy regimens, the overall course of treatment is determined by the patient and physician based on the observed response and toxicity.

[0475] Combination therapy In certain embodiments, the LNP compositions of the Disclosure may be administered in combination with one or more additional compounds or treatments, such as surgery, radiotherapy, chemotherapy, or other active agents including any of the above. The LNP compositions may be administered in combination with a second active agent for a variety of reasons, including to increase efficacy or reduce undesirable side effects. The LNP compositions may be administered before, after, or concurrently with the additional treatment. Furthermore, when the LNP compositions of the Disclosure (containing a first active agent) are administered in combination with a second active agent, the second active agent may be administered as a free agent, as a separate LNP formulation, or as a component of the LNP composition containing the first agent. In certain embodiments, multiple active agents are loaded onto the same lipid nanoparticles. In other embodiments, lipid nanoparticles containing active agents are used in combination with one or more free agents. In certain embodiments, LNP compositions containing active agents are formed individually and then combined with other compounds for single-dose co-administration. Alternatively, certain treatments are administered sequentially in a predetermined order. Thus, the LNP compositions of the Disclosure may contain one or more active agents.

[0476] Other combination therapies known to those skilled in the art can be used in combination with the method of this disclosure.

[0477] Unless otherwise defined, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art to which this disclosure belongs. Similar or equivalent methods and materials to those described herein may be used in the implementation or testing of this disclosure, but preferred methods and materials are described below. All publications, patent applications, patents, and other references referenced herein are incorporated herein by reference in their entirety. In the event of any conflict, this specification, including definitions, shall prevail. In addition, materials, methods, and examples are illustrative and not intended to be limiting.

[0478] Hereinafter, exemplary embodiments of the present disclosure will be referred to in detail. While the present disclosure will be described in conjunction with these exemplary embodiments, it will be understood that the present disclosure is not intended to be limited to these embodiments. Rather, it is intended to encompass alternative forms, modifications, and equivalent forms that may fall within the spirit and scope of the present disclosure as defined by the appended claims. These utilize standard techniques well known in the art, or techniques specifically described below. [Examples]

[0479] (Example 1) Synthesis of SM-066 [ka] [ka]

[0480] Step 1: tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-chlorocarbonyl-benzoyl]amino]propyl]carbamate (2): (EC5059-482) [ka]

[0481] In 30 mL of DCM and 2.29 g of TEA (22.60 mmol, 3.15 mL, 2 equivalents), tert-butyl N-(3-aminopropyl)carbamate (3.54 g, 20.34 mmol, 3.55 mL, 1.8 equivalents) was added dropwise to a 50 mL solution of benzene-1,3,5-tricarbonyl chloride (3 g, 11.30 mmol, 1 equivalent) in DCM (50 mL) under N2 at 0°C. After addition, the mixture was stirred at 20°C for 16 hours. The compound tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-chlorocarbonylbenzoyl]aminopropyl]carbamate (6.1 g, crude) was used directly as a yellow liquid in the next step.

[0482] Step 2: tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[3-(dimethylamino)propylcarbamoyl]benzoyl]amino]propyl]carbamate(3):(EC5059-483 / 488) [ka]

[0483] TEA (2.85 g, 28.19 mmol, 3.92 mL, 2.5 equivalents) and N',N'-dimethylpropane-1,3-diamine (1.38 g, 13.53 mmol, 1.7 mL, 1.2 equivalents) were added to a solution of tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-chlorocarbonylbenzoyl]aminopropyl]carbamate (6.1 g, 11.27 mmol, 1 equivalent) in DCM (30 mL) at 0°C under N2. The mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure and the solvent was removed to obtain the crude product. The residue was purified by flash silica gel chromatography (80g SepaFlash® silica flash column, DCM:MeOH: 0-10%) to obtain the compound tert-butyl N-[[2-(2,6-dioxo-3-piperidyl)-1-oxo-isoindorin-5-yl]methyl]carbamate (2.1g, 5.62 mmol, yield 52.5%) as a white solid. LCMS:[M+H] + :607.6.

[0484] Step 3: N1,N5-bis(3-aminopropyl)-N3-[3-(dimethylamino)propyl]benzene-1,3,5-tricarboxamide(4):(EC5059-491) [ka]

[0485] HCl / dioxane (4M, 5mL, 9.33 equivalents) was added to a solution of tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[3-(dimethylamino)propylcarbamoyl]benzoyl]amino]propyl]carbamate (1.3g, 2.14 mmol, 1 equivalent) in DCM (5mL). The mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated directly under reduced pressure to obtain the crude product. The crude product was ground in (PE / HCl=3 / 1, 20mL) at 20°C for 1 hour, filtered appropriately, and dried to obtain compound N1,N5-bis(3-aminopropyl)-N3-[3-(dimethylamino)propyl]benzene-1,3,5-tricarboxamide (930mg, crude, HCl) as a yellow solid.

[0486] Step 4: 1-[3-[4-[3-[bis(2-hydroxydodecyl)amino]propyl]piperazin-1-yl]propyl-(2-hydroxydodecyl)amino]dodecane-2-ol (SM-066): (EC5059-495 / 498 / 499) [ka]

[0487] NaBH3CN (680.91 mg, 10.84 mmol, 8 equivalents) was added to a solution of N1,N5-bis(3-aminopropyl)-N3-[3-(dimethylamino)propyl]benzene-1,3,5-tricarboxamide (600 mg, 1.35 mmol, 1 equivalent, HCl) and NaOAc (888.85 mg, 10.84 mmol, 8 equivalents) in MeOH (30 mL). After addition, the mixture was stirred at 25°C for 0.5 hours, and then 1-ethylhexyl 9-oxononanoate (2.31 g, 8.13 mmol, 6 equivalents) was added. The resulting mixture was stirred at 25°C for 15.5 hours. The reaction mixture was concentrated under reduced pressure to remove MeOH. The residue was diluted with H2O (60 mL) and extracted with RINKAN (50 mL x 3). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® silica flash column, DCM:MeOH: 0-10%, twice) to obtain SM-066 (645 mg, 428.50 μmol, yield 48.8%, purity 98.34%) as a colorless gum-like substance. LCMS:[M+H]+:1480.8

[0488] 1 H NMR (400 MHz, CDCl3) δ = 8.20 (s, 3H), 8.06 - 7.98 (m, 3H), 4.83 - 4.76 (m, 4H), 3.63 -3.52 (m, 6H), 3.38 - 3.26 (m, 6H), 3.17 - 3.07 (m, 8H), 2.99 (s, 6H), 2.28 (t, J = 7.2 Hz, 8H), 2.21 - 2.12 (m, 6H), 1.78 - 1.68 (m, 8H), 1.65 - 1.55 (m, 14H), 1.54 - 1.48 (m, 10H), 1.39 - 1.25 (m, 56H), 0.90 - 0.85 (m, 24H). Alternative synthesis of SM-066 Synthesis of SM-066 (Method 2) [ka] [ka]

[0489] Step 1: 3,5-Bis(methoxycarbonyl)benzoic acid

[0490] A solution of trimethyl benzene-1,3,5-tricarboxylic acid (27 g, 107.05 mmol, 1.0 equivalent) in MeOH (350 mL) was added dropwise to a solution of NaOH (4.28 g, 107.05 mmol, 1.0 equivalent) in H2O (70 mL). The mixture was stirred at 60°C for 2 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with DCM (300 mL) and extracted with saturated NaHCO3 solution (300 mL). The aqueous phase was acidified to pH=1 with 5% hydrochloric acid and extracted with ELISA (3 × 300 mL). The combined organic phase was dried over MgSO4 and concentrated under vacuum to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 330g SepaFlash® silica flash column, eluent 0-50% Â / PE, gradient at 100 mL / min) to obtain compound 3,5-bis(methoxycarbonyl)benzoic acid (8g, 33.59 mmol, yield 15.7%) as a white solid.

[0491] 1 H NMR (400 MHz, CD3OD-d4) δ = 8.73 - 8.68 (m, 2H), 8.66 - 8.62 (m, 1H), 3.96 (s, 6H).

[0492] Step 2: Dimethyl 5-[3-(dimethylamino)propylcarbamoyl]benzene-1,3-dicarboxylate

[0493] To a 100 mL solution of 3,5-bis(methoxycarbonyl)benzoic acid (6.9 g, 28.97 mmol, 1.0 equivalent) and N',N'-dimethylpropane-1,3-diamine (5.92 g, 57.94 mmol, 7.25 mL, 2.0 equivalents) in DCM, EDCI (8.33 g, 43.45 mmol, 1.5 equivalents) and HOBt (5.87 g, 43.45 mmol, 1.5 equivalents) were added. The mixture was stirred at 20°C for 12 hours. The reaction mixture was diluted with water (100 mL) and extracted with DCM (100 mL x 2). The combined organic layer was dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 80g SepaFlash® silica flash column, eluent 0-10% MeOH / DCM, gradient at 100 mL / min) to obtain the compound dimethyl 5-[3-(dimethylamino)propylcarbamoyl]benzene-1,3-dicarboxylate (6.1 g, 18.54 mmol, yield 64.0%, purity 98%) as a white solid.

[0494] 1 H NMR (400 MHz, CD3OD-d4) δ = 8.68 (d, J = 1.6 Hz, 1H), 8.64 (d, J = 1.6 Hz, 2H), 3.97 (s, 6H), 3.45 (t, J = 7.2 Hz, 2H), 2.53 - 2.40 (m, 2H), 2.29 (s, 6H), 1.90 - 175 (m, 2H).

[0495] Step 3: 5-[3-(dimethylamino)propylcarbamoyl]benzene-1,3-dicarboxylic acid

[0496] To a solution of dimethyl 5-[3-(dimethylamino)propylcarbamoyl]benzene-1,3-dicarboxylate (6.1 g, 18.92 mmol, 1.0 equivalent) in MeOH (100 mL), NaOH (1.67 g, 41.63 mmol, 2.2 equivalents) and H2O (20 mL) were added. The mixture was stirred at 60°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with DCM (300 mL) and extracted with saturated NaHCO3 solution (300 mL). The aqueous phase was acidified to pH=1 with 5% hydrochloric acid and extracted with ELISA (3 × 300 mL). The combined organic layers were dried over Mg2SO4 and concentrated under vacuum to obtain compound 5-[3-(dimethylamino)propylcarbamoyl]benzene-1,3-dicarboxylic acid (5 g, 15.12 mmol, yield 79.9%, HCl) as a white solid.

[0497] 1 H NMR (400 MHz, CD3OD-d4) δ = 8.73 (s, 1H), 8.50 (s, 2H), 3.52 (t, J = 6.4 Hz, 2H), 3.15 (t, J = 7.6 Hz, 2H), 2.84 (s, 6H), 2.16 - 2.00 (m, 2H).

[0498] Step 4: Tetra(octan-3-yl)9,9',9'',9'''-((((5-((3-(dimethylamino)propyl)carbamoyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate

[0499] 5-[3-(dimethylamino)propylcarbamoyl]benzene-1,3-dicarboxylic acid (600 mg, 2.04 mmol, 1.0 equivalent) and 1-ethylhexyl 9-[3-aminopropyl-[9-(1-ethylhexoxy)-9-oxononyl]amino]nonanoate (3.96 g, 6.12 mmol, 3.0 equivalent, HCl) were dissolved in pyridine (20 mL), to which EDCI (1.37 g, 7.14 mmol, 3.5 equivalents) was added. The mixture was stirred under N2 at 20°C for 16 hours. The reaction mixture was concentrated directly under reduced pressure, and the residual pyridine was co-evaporated with DCM to obtain the residue. The residue was diluted with H2O (100 mL) and extracted with DCM (60 mL x 3). The combined organic layers were washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (40g SepaFlash® silica flash column, DCM:MeOH: 0-10%) and then by preparative HPLC (column: Phenomenex luna C18 150×25mm×10um, mobile phase: [A / B water (FA)-ACN], gradient: 65%-95% B over 8 minutes) to obtain the compound tetra(octan-3-yl)9,9',9'',9'''-((((5-((3-(dimethylamino)propyl)carbamoyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate (1.1g, 739.62μmol, yield 49.8%, purity 99.53%) as a yellow gum-like substance. LCMS:[M+H] + :1480.6

[0500] 1H NMR (400 MHz, CDCl3) δ = 9.21 (s, 1H), 8.63 - 8.51 (m, 2H), 8.49 - 8.33 (m, 3H), 4.87 - 4.75 (m, 4H), 3.69 - 3.47 (m, 6H), 2.63 (t, J = 5.6 Hz, 4H), 2.58 - 2.45 (m, 10H), 2.38 (s, 6H), 2.31 - 2.22 (m, 8H), 1.78 (d, J = 2.8 Hz, 6H), 1.62 - 1.44 (m, 32H), 1.33 - 1.19 (m, 56H), 0.91 - 0.82 (m, 24H). Synthesis of SM-066 (Method 3)

change

change

[0501] (2,3,4,5,6-pentafluorophenyl)3,5-bis[3-[bis[9-(1-ethylhexoxy)-9-oxononyl]amino]propylcarbamoyl]benzoate (16.5 g, 10.56 mmol, 1.0 equivalent) was dissolved in THF (300 mL) and DIEA (6.83 g, 52.81 mmol, 9.20 mL, 5.0 equivalent) and N',N'-dimethylpropane-1,3-diamine (2.16 g, 21.12 mmol, 2.64 mL, 2.0 equivalent) were added. The mixture was stirred at 20°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was analyzed by flash silica gel chromatography (ISCO®, 330g SepaFlash® silica flash column, eluent 0-20% MeOH / DCM ether, gradient at 100 mL / min), preparative NP-HPLC (column: Welch Ultimate XB-CN 250×50×10um, mobile phase: [A / B hexane-EtOH (0.1% NH3.H2O)], gradient: 10%-40% B over 16 minutes), and preparative NP-HPLC (column: Welch Ultimate XB-Diol The mixture was purified over 16 minutes in a 250×50×10um mobile phase [A / B hexane-EtOH (0.1% NH3, H2O)] gradient with 8% to 35% of B) to obtain SM-066, also known as tetra(octane-3-yl)=9,9',9'',9'''-((((5-((3-(dimethylamino)propyl)carbamoyl)isophthaloyl)bis(azandiyl))=bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate (23g, 15.46 mmol, yield 76.3%, purity 99.51%) as a yellow oily substance. LCMS:[M+H] + :1480.3

[0502] 1H NMR (400 MHz, CDCl3) δ = 9.22 (s, 1H), 8.57 (t, J = 4.4 Hz, 2H), 8.44 - 8.28 (m, 3H), 4.87 - 4.80 (m, 4H), 3.65 - 3.50 (m, 6H), 2.62 (t, J = 6.0 Hz, 4H), 2.57 - 2.48 (m, 10H), 2.38 (s, 6H), 2.26 (t, J = 7.6 Hz, 8H), 1.77 - 1.74 (m, 6H), 1.61 - 1.44 (m, 32H), 1.33 - 1.21 (m, 56H), 0.94 - 0.82 (m, 24H).

[0503] (Example 2) Synthesis of SM-078 (also known as 1-ethylhexyl 9-[3-[[3-[3-[bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]-5-[4-(dimethylamino)butanoylamino]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate) [ka]

[0504] Tetra(octan-3-yl)9,9',9'',9'''-((((5-(4-(dimethylamino)butanamide)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate. [ka] [ka] [ka]

[0505] Step 1: tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-nitro-benzoyl]amino]propyl]carbamate (2) [ka]

[0506] To a solution of 5-nitrobenzene-1,3-dicarboxylic acid (7 g, 33.16 mmol, 1.0 equivalent) and tert-butyl N-(3-aminopropyl)carbamate (14.44 g, 82.89 mmol, 14.47 mL, 2.5 equivalents) in Py (100 mL), EDCI (15.89 g, 82.89 mmol, 2.5 equivalents) was added under N2 at 0°C. After addition, the mixture was stirred at 20°C for 16 hours. The reaction mixture was diluted with H2O (150 mL) and extracted with SiO (80 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (120 g SepaFlash® silica flash column, PE: Â: 0-40%) to obtain the compound tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-nitro-benzoyl]amino]propyl]carbamate (8.5 g, 16.20 mmol, yield 49.3%, purity 99.8%) as a white solid.

[0507] 1 H NMR (400 MHz, CDCl3) δ = 8.87 (s, 2H), 8.69 (s, 1H), 7.93 (s, 2H), 5.00 (s, 2H), 3.57 - 3.52 (m, 4H), 3.30 - 3.23 (m, 4H), 1.80 - 1.72 (m, 4H), 1.45 (s, 18H).

[0508] Step 2: tert-butyl N-[3-[[3-amino-5-[3-(tert-butoxycarbonylamino)propylcarbamoyl]benzoyl]amino]propyl]carbamate (3) [ka]

[0509] To a solution of tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-nitro-benzoyl]amino]propyl]carbamate (8.5 g, 16.23 mmol, 1 equivalent) in MeOH (150 mL), Pd / C (17.28 g, 16.23 mmol, purity 10%) was added under N2 conditions. The suspension was degassed under vacuum and purged with H2 several times. The mixture was stirred under H2 (15 psi) at 40°C for 16 hours. The reaction mixture was filtered, the filter cake was washed with MeOH (30 mL x 2), and the filtrate was concentrated under reduced pressure to obtain the crude product. The crude product was ground in (PE / Â=10 / 1, 50 mL) at 20°C for 0.5 hours and filtered. The filtered cake was dried to obtain the pure compound tert-butyl N-[3-[[3-amino-5-[3-(tert-butoxycarbonylamino)propylcarbamoyl]benzoyl]amino]propyl]carbamate (7.5 g, 15.19 mmol, yield 93.6%) as a white solid.

[0510] 1 H NMR (400 MHz, DMSO-d6) δ = 8.27 (t, J = 5.6 Hz, 2H), 7.33 (s, 1H), 7.09 (s, 2H), 6.79 (t, J = 5.2 Hz, 2H), 5.40 (s, 2H), 3.21 (q, J = 6.4 Hz, 4H), 2.96 (q, J = 6.4 Hz, 4H), 1.65 - 1.56 (m, 4H), 1.37 (s, 18H).

[0511] Step 3: tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[4-(dimethylamino)butanoylamino]benzoyl]amino]propyl]carbamate(4) [ka]

[0512] In 50 mL of DMF, a mixture of tert-butyl N-[3-[[3-amino-5-[3-(tert-butoxycarbonylamino)propylcarbamoyl]benzoyl]amino]propyl]carbamate (4 g, 8.10 mmol, 1.0 equivalent) and 4-(dimethylamino)butanoic acid (1.28 g, 9.72 mmol, 1.2 equivalents) was mixed with HATU (3.70 g, 9.72 mmol, 1.2 equivalents) and DIPEA (2.09 g, 16.21 mmol, 2.82 mL, 2.0 equivalents). The mixture was stirred under N2 at 20°C for 16 hours. The reaction mixture was diluted with H2O (120 mL) and extracted with siRNA (60 mL x 3). The combined organic layer was washed with brine (80 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (80g SepaFlash® silica flash column, DCM:MeOH: 0-15%) to obtain the compound tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[4-(dimethylamino)butanoylamino]benzoyl]amino]propyl]carbamate (3.7g, 6.10 mmol, yield 75.5%) as a yellow solid.

[0513] Step 4: N1,N3-Bis(3-aminopropyl)-5-[4-(dimethylamino)butanoylamino]benzene-1,3-dicarboxamide(5) [ka]

[0514] To a solution of tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[4-(dimethylamino)butanoylamino]benzoyl]amino]propyl]carbamate (1.5 g, 2.47 mmol, 1.0 equivalent) in DCM (10 mL), HCl / dioxane (4 M, 5 mL) was added. The mixture was stirred at 20 °C for 16 hours. The reaction mixture was directly concentrated under reduced pressure to obtain compound N1,N3-bis(3-aminopropyl)-5-[4-(dimethylamino)butanoylamino]benzene-1,3-dicarboxamide (1.05 g, crude, HCl) as a yellow solid. The crude product was used in the next step without further purification.

[0515] 1 H NMR (400 MHz, CD3OD-d4) δ = 8.29 - 8.26 (m, 2H), 8.18 - 8.15 (m, 1H), 3.55 (t, J = 6.4 Hz, 4H), 3.31 - 3.25 (m, 2H), 3.06 (t, J = 7.2 Hz, 4H), 2.96 (s, 6H), 2.65 (t, J = 6.4 Hz, 2H), 2.18 - 2.11 (m, 2H), 2.08 - 2.00 (m, 4H).

[0516] Step 5: 1-Ethylhexyl 9-[3-[[3-[3-[Bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]-5-[4-(dimethylamino)butanoylamino]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate (SM-078) [ka]

[0517] NaBH3CN (567.43 mg, 9.03 mmol, 8.0 equivalents) was added to a solution of N1,N3-bis(3-aminopropyl)-5-[4-(dimethylamino)butanoylamino]benzene-1,3-dicarboxamide (500 mg, 1.13 mmol, 1.0 equivalent, HCl) and NaOAc (740.71 mg, 9.03 mmol, 8.0 equivalents) in MeOH (15 mL). After addition, the mixture was stirred at 20°C for 0.5 hours, and then 1-ethylhexyl 9-oxononanoate (2.57 g, 9.03 mmol, 8.0 equivalents) was added. The resulting mixture was stirred at 20°C for 15.5 hours. The reaction mixture was concentrated under reduced pressure to remove MeOH. The residue was diluted with H2O (60 mL) and extracted with RINKAN (50 mL x 3). The combined organic layers were washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® silica flash column, DCM:MeOH: 0-10%, 2% NH3·H2O in MeOH) and preparative HPLC (column: Welch Xtimate C1100 × 30 mm × 5 μm, mobile phase: [water (FA)-MeOH], gradient: 63%-93% B over 8 minutes) to obtain SM-078 (655 mg, 441.47 μmol, yield 38.44%, purity 99.77%) as a pale yellow oily substance. LCMS:[M+H] + :1480.6

[0518] 1H NMR (400 MHz, CDCl3) δ = 10.28 (s, 1H), 8.19 (s, 2H), 8.16 - 8.08 (m, 2H), 8.05 (s, 1H), 4.88 - 4.76 (m, 4H), 3.58 - 3.45 (m, 4H), 2.76 - 2.67 (m, 4H), 2.65 - 2.58 (m, 6H), 2.51 (br t, J = 6.8 Hz, 4H), 2.37 (s, 6H), 2.28 (t, J = 7.2 Hz, 8H), 1.92 - 1.84 (m, 4H), 1.63 - 1.48 (m, 32H), 1.35 - 1.23 (m, 60H), 0.90 - 0.85 (m, 24H).

[0519] (Example 3) Synthesis of SM-081 [ka]

[0520] Tetra(octan-3-yl)9,9',9'',9'''-((((4-((3-(dimethylamino)propyl)carbamoyl)pyridine-2,6-dicarbonyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate. [ka] [ka] [ka]

[0521] Step 1: 2,6-Dichloro-N-(3-(dimethylamino)propyl)isonicotinamide (2)

[0522] To a solution of 2,6-dichloropyridine-4-carboxylic acid (30 g, 156.25 mmol, 1.0 equivalent) in DCM (300 mL), EDCI (44.93 g, 234.38 mmol, 1.5 equivalent), HOBt (31.67 g, 234.38 mmol, 1.5 equivalent), and TEA (47.43 g, 468.75 mmol, 65.24 mL, 3 equivalents) were added. Then, N',N'-dimethylpropane-1,3-diamine (23.95 g, 234.38 mmol, 29.31 mL, 1.5 equivalent) was added to the mixture, and the resulting mixture was stirred at 20°C for 12 hours. The reaction mixture was diluted with water (300 mL) and extracted with DCM / IPA (300 mL x 3). The combined organic layers were combined and concentrated under reduced pressure to obtain the residue. The residue was purified by reverse-phase HPLC (0.1% FA conditions) to obtain compound 2,6-dichloro-N-(3-(dimethylamino)propyl)isonicotinamide (9.5 g, 34.29 mmol, yield 21.9%, purity 99.7%) as a yellow, gum-like substance.

[0523] 1 H NMR (400 MHz, DMSO-d6) δ = 8.90 (s, 1H), 7.87 (s, 2H), 3.31 - 3.26 (m, 2H), 2.24 (t, J = 7.2 Hz, 2H), 2.12 (s, 6H), 1.70 - 1.58 (m, 2H).

[0524] Step 2: Dimethyl 4-[3-(dimethylamino)propylcarbamoyl]pyridine-2,6-dicarboxylate (3)

[0525] To a solution of 2,6-dichloro-N-[3-(dimethylamino)propyl]pyridine-4-carboxamide (9 g, 32.59 mmol, 1 equivalent) in DMF (60 mL) and MeOH (40 mL), TEA (9.89 g, 97.77 mmol, 13.61 mL, 3.0 equivalents) and Pd(dppf)Cl2 (2.38 g, 3.26 mmol, 0.1 equivalents) were added under N2. The suspension was degassed under vacuum and purged with CO three times. The mixture was stirred under CO (50 psi) at 80°C for 12 hours. The reaction mixture was concentrated under reduced pressure to remove MeOH. The residue was diluted with water (150 mL) and extracted with siRNA (100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the crude product. The residue was purified by flash silica gel chromatography (ISCO®, 120g SepaFlash® silica flash column, eluent 0-20% MeOH / DCM, 100 mL / min) to obtain the crude product. The crude product was then ground with RINKAN at 20°C for 30 minutes to obtain the desired compound, dimethyl 4-[3-(dimethylamino)propylcarbamoyl]pyridine-2,6-dicarboxylate (3.3 g, 10.01 mmol, yield 31.3%, purity 98.04%), as a yellow solid.

[0526] 1 H NMR (400 MHz, DMSO-d6) δ= 9.28 (t, J = 5.2 Hz, 1H), 8.64 (s, 2H), 3.95 (s, 6H), 3.41 - 3.33 (m, 3H), 2.63 (s, 2H), 2.40 (s, 6H), 1.85 - 1.75 (m, 2H).

[0527] Step 3: tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[3-(dimethylamino)propylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate(4)

[0528] A mixture of dimethyl 4-[3-(dimethylamino)propylcarbamoyl]pyridine-2,6-dicarboxylate (2 g, 6.19 mmol, 1.0 equivalent) and tert-butyl N-(3-aminopropyl)carbamate (3.23 g, 18.56 mmol, 3.24 mL, 3.0 equivalent) was stirred in MeOH (20 mL) at 50°C for 6 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The crude product was ground with EtOA at 20°C for 30 minutes to obtain the compound tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[3-(dimethylamino)propylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate (3.03 g, 4.54 mmol, yield 73.3%, purity 90.9%) as an off-white solid.

[0529] 1 H NMR (400 MHz, DMSO-d6) δ = 9.33 (t, J = 4.8 Hz, 2H), 9.26 - 9.16 (m, 1H), 8.56 (s, 2H), 7.00 - 6.75 (m, 2H), 3.41 - 3.35 (m, 4H), 3.30 (s, 4H), 3.04 - 2.95 (m, 4H), 2.14 (s, 6H), 1.74 - 1.62 (m, 6H), 1.37 (s, 18H).

[0530] Step 4: N2,N6-bis(3-aminopropyl)-N4-[3-(dimethylamino)propyl]pyridine-2,4,6-tricarboxamide(5)

[0531] To a solution of tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[3-(dimethylamino)propylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate (500 mg, 822.72 μmol, 1.0 equivalent) in DCM (2.5 mL), HCl / dioxane (4 M, 2.5 mL) was added, and the mixture was stirred at 20°C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain the crude product N2,N6-bis(3-aminopropyl)-N4-[3-(dimethylamino)propyl]pyridine-2,4,6-tricarboxamide (590 mg, crude, 8HCl) as a white solid. The crude product was used in the next step without further purification.

[0532] Step 5: SM-081 (also known as: 1-ethylhexyl 9-[3-[[6-[3-[bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]-4-[3-(dimethylamino)propylcarbamoyl]pyridine-2-carbonyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate)

[0533] NaBH3CN (530.28 mg, 8.44 mmol, 10.0 equivalent) was added to a solution of N2,N6-bis(3-aminopropyl)-N4-[3-(dimethylamino)propyl]pyridine-2,4,6-tricarboxamide (590 mg, 843.83 μmol, 1.0 equivalent, 8HCl) and NaOAc (830.67 mg, 10.13 mmol, 12.0 equivalents) in MeOH (15 mL). After addition, the mixture was stirred at 25°C for 0.5 hours, and then 1-ethylhexyl 9-oxononanoate (1.92 g, 6.75 mmol, 8.0 equivalents) was added. The resulting mixture was stirred at 25°C for 15.5 hours. The reaction mixture was diluted with water (30 mL) and extracted with RINKAN (30 mL x 3). The combined organic layers were washed with 30 mL of brine, dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 40 g SepaFlash® silica flash column, eluent 0-10% DCM / MeOH, gradient at 80 mL / min) to obtain the crude product. The crude product was then purified by preparative HPLC (column: Welch Xtimate C1 100 × 30 mm × 5 μm, mobile phase: [water (FA)-MeOH], gradient: 63%-93% B over 8 minutes) to obtain SM-081 (305.92 mg, 204.71 μmol, yield 24.3%, purity 99.12%) as a pale yellow oily substance. LCMS:[M+H] + :1481.7

[0534] 1H NMR (400 MHz, CDCl3) δ = 9.75 (d, J = 3.6 Hz, 1H), 8.68 (s, 2H), 8.34 - 8.24 (m, 1H), 4.86 - 4.77 (m, 4H), 3.66 - 3.52 (m, 6H), 2.63 - 2.52 (m, 6H), 2.49 - 2.42 (m, 4H), 2.41 (s, 6H), 2.28 (t, J = 7.5 Hz, 8H), 1.87 - 1.76 (m, 12H), 1.64 - 1.48 (m, 28H), 1.30 - 1.21 (m, 58H), 0.92 - 0.84 (m, 24H).

[0535] (Example 4) Synthesis of SM-082 [ka]

[0536] Tetra(octan-3-yl)9,9',9'',9'''-((((5-((3-(4-methylpiperazine-1-yl)propyl)carbamoyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate. [ka] [ka]

[0537] Step 1: 3,5-Bis(methoxycarbonyl)benzoic acid (2)

[0538] To a solution of benzene-1,3,5-tricarboxylic acid trimethyl (20 g, 79.30 mmol, 1.0 equivalent) in MeOH (300 mL) and H2O (50 mL), NaOH (3.49 g, 87.23 mmol, 1.1 equivalent) was added. The mixture was stirred at 60°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove EtOH. The residue was diluted with H2O (150 mL) and extracted with siRNA (50 mL x 3). The pH of the aqueous phase was adjusted to 2 with 4 M HCl and extracted with siRNA (100 mL x 3). The combined organic layers were washed with brine (100 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain compound 3,5-bis(methoxycarbonyl)benzoic acid (14.6 g, 61.29 mmol, yield 77.3%) as a white solid. The crude product was used in the next step without further purification.

[0539] 1 H NMR (400 MHz, DMSO-d6) δ = 13.64 - 13.50 (m, 1H), 8.59 - 8.5 (d, J = 1.6 Hz, 2H), 8.54 (d, J = 1.6 Hz, 1H), 3.91 (s, 6H).

[0540] Step 2: Dimethyl 5-[3-(4-methylpiperazine-1-yl)propylcarbamoyl]benzene-1,3-dicarboxylate(3)

[0541] To a solution of 3,5-bis(methoxycarbonyl)benzoic acid (14.5 g, 60.87 mmol, 1.0 equivalent) in DCM (200 mL), HATU (27.78 g, 73.05 mmol, 1.2 equivalents) and DIPEA (9.44 g, 73.05 mmol, 12.72 mL, 1.2 equivalents) were added at 0°C. After addition, the reaction mixture was stirred at 20°C for 0.5 hours. Next, 3-(4-methylpiperazin-1-yl)propan-1-amine (11.49 g, 73.05 mmol, 1.2 equivalents) was added. The resulting mixture was stirred under N2 at 20°C for 16 hours. The reaction mixture was diluted with H2O (150 mL) and extracted with DCM (100 mL x 3). The combined organic layers were washed with brine (150 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (330 g SepaFlash® silica flash column, DCM:MeOH: 0-10%, 2% NH3·H2O in MeOH) to obtain the compound dimethyl 5-[3-(4-methylpiperazin-1-yl)propylcarbamoyl]benzene-1,3-dicarboxylate (15.6 g, 39.77 mmol, yield 65.3%, purity 96.2%) as a white solid.

[0542] 1 H NMR (400 MHz, DMSO-d6) δ = 8.95 (t, J = 5.2 Hz, 1H), 8.67 - 8.64 (m, 2H), 8.57- 8.53 (m, 1H), 3.93 (s, 6H), 3.36 - 3.30 (m, 2H), 2.74 - 2.50 (m, 8H), 2.46 (t, J = 7.2 Hz, 2H), 2.39 (s, 3H), 1.80 - 1.67 (m, 2H).

[0543] Step 3: 5-[3-(4-methylpiperazine-1-yl)propylcarbamoyl]benzene-1,3-dicarboxylic acid (4)

[0544] Dimethyl 5-[3-(4-methylpiperazin-1-yl)propylcarbamoyl]benzene-1,3-dicarboxylate (10 g, 26.49 mmol, 1.0 equivalent) was dissolved in MeOH (60 mL) and H2O (15 mL), to which NaOH (3.18 g, 79.48 mmol, 3.0 equivalent) was added. The mixture was stirred at 70°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the MeOH. The residue was diluted with H2O (30 mL) and extracted with ELISA (30 mL x 3). The pH of the aqueous phase was adjusted to 2 with 4 M HCl, and the residue was concentrated under reduced pressure to obtain the residue. The residue was ground in (PE / EA = 1 / 1, 50 mL) at 20°C for 0.5 hours, properly filtered, and the filtered cake was dried to obtain compound 5-[3-(4-methylpiperazin-1-yl)propylcarbamoyl]benzene-1,3-dicarboxylic acid (9.2 g, crude) as a white solid.

[0545] Step 4: tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoyl]benzoyl]amino]propyl]carbamate(5)

[0546] In 50 mL of Py, a mixture of 5-[3-(4-methylpiperazine-1-yl)propylcarbamoyl]benzene-1,3-dicarboxylic acid (5 g, 14.31 mmol, 1.0 equivalent) and tert-butyl N-(3-aminopropyl)carbamate (7.48 g, 42.93 mmol, 7.50 mL, 3.0 equivalent) was added, to which EDCI (8.23 g, 42.93 mmol, 3.0 equivalent) was added. The mixture was stirred under N2 at 20°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove most of the Py. The residue was diluted with saturated NH4Cl (50 mL) and extracted with DCM / MeOH (20 / 1, 50 mL x 5). The combined organic layers were washed with brine (30 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (120g SepaFlash® silica flash column, DCM:MeOH:0~15%, 2% NH3·H2O in MeOH) to obtain the compound tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoyl]benzoyl]amino]propyl]carbamate (6.9g, 10.43 mmol, yield 72.6%) as a yellow oily substance.

[0547] Step 5: N1,N5-Bis(3-aminopropyl)-N3-[3-(4-methylpiperazine-1-yl)propyl]benzene-1,3,5-tricarboxamide(6)

[0548] To a solution of tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[3-(4-methylpiperazine-1-yl)propylcarbamoyl]benzoyl]amino]propyl]carbamate (4 g, 6.04 mmol, 1.0 equivalent) in DCM (15 mL), HCl / dioxane (4 M, 15 mL) was added. The mixture was stirred at 20 °C for 2 hours. The reaction mixture was directly concentrated under reduced pressure to obtain compound N1,N5-bis(3-aminopropyl)-N3-[3-(4-methylpiperazine-1-yl)propyl]benzene-1,3,5-tricarboxamide (3.1 g, crude, HCl) as a dark brown solid. The crude product was used in the next step without further purification.

[0549] Step 6: (Alternate name) 1-Ethylhexyl 9-[3-[[3-[3-[Bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]-5-[3-(4-methylpiperazin-1-yl)propylcarbamoyl]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate (SM-082)

[0550] To a 15 mL solution of N1,N5-bis(3-aminopropyl)-N3-[3-(4-methylpiperazin-1-yl)propyl]benzene-1,3,5-tricarboxamide (600 mg, 1.20 mmol, 1.0 equivalent, HCl) in MeOH (1), NaBH3CN (757.04 mg, 12.05 mmol, 10.0 equivalent) and NaOAc (988.24 mg, 12.05 mmol, 10.0 equivalent) were added. After addition, the mixture was stirred at 25°C for 0.5 hours, and then 1-ethylhexyl 9-oxononanoate (2.74 g, 9.64 mmol, 8.0 equivalent) was added to the mixture. The resulting mixture was stirred at 25°C for 15.5 hours. The combined reaction mixture was concentrated under reduced pressure to remove the MeOH. The residue was diluted with H2O (80 mL) and extracted with ₹ (50 mL x 3). The combined organic layer was washed with brine (60 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® silica flash column, DCM:MeOH: 0-10%, 2% NH3·H2O in MeOH) and preparative HPLC (column: Welch Xtimate C1100 x 30 mm x 5 μm, mobile phase: [water (FA)-MeOH], gradient: 60%-90% B over 8 minutes) to obtain SM-082 (651 mg, 418.92 μmol, yield 35.7%, purity 98.80%) as a yellow oily substance. LCMS:[M+H] + :1535.8

[0551] 1H NMR (400 MHz, CDCl3) δ = 8.62 - 8.55 (m, 1H), 8.54 - 8.52 (m, 1H), 8.46 - 8.43 (m, 1H), 8.33 (t, J = 4.8 Hz, 1H), 4.84 - 4.78 (m, 4H), 3.58 - 3.52 (m, 4H), 2.90 - 2.75 (m, 4H), 2.70 - 2.61 (m, 8H), 2.56 (t, J = 6.0 Hz, 2H), 2.36 (s, 3H), 2.30 - 2.22 (m, 10H), 1.92 - 1.88 (m, 2H), 1.84 - 1.78 (m, 2H), 1.65 - 1.47 (m, 36H), 1.34 - 1.21 (m, 62H), 0.89 - 0.85 (m, 24H).

[0552] (Example 5) Synthesis of SM-084 [ka]

[0553] Tetra(octan-3-yl)9,9',9'',9'''-((((5-(2-((3-(dimethylamino)propyl)amino)-2-oxoethyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate. [ka] [ka]

[0554] Step 1: 2-(3,5-dibromophenyl)-N-[3-(dimethylamino)propyl]acetamide(2)

[0555] To a solution of 2-(3,5-dibromophenyl)acetic acid (500 mg, 1.70 mmol, 1.0 equivalent) in DMF (5 mL), HATU (711.46 mg, 1.87 mmol, 1.1 equivalent), N',N'-dimethylpropane-1,3-diamine (260.71 mg, 2.55 mmol, 319.11 μL, 1.5 equivalent), and DIEA (659.54 mg, 5.10 mmol, 888.87 μL, 3.0 equivalent) were added. The mixture was stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was quenched by adding H2O (20 mL) and filtered to obtain compound 2-(3,5-dibromophenyl)-N-[3-(dimethylamino)propyl]acetamide (435 mg, 1.13 mmol, yield 66.4%, purity 98.2%) as a white solid.

[0556] Step 2: Dimethyl 5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxylate(3)

[0557] To a solution of 2-(3,5-dibromophenyl)-N-[3-(dimethylamino)propyl]acetamide (13 g, 34.38 mmol, 1.0 equivalent) in DMF (78 mL), MeOH (52 mL), TEA (10.44 g, 103.15 mmol, 14.36 mL, 3.0 equivalent), and Pd(dppf)Cl2 (2.52 g, 3.44 mmol, 0.1 equivalent) were added under an N2 atmosphere. The suspension was degassed and purged with CO three times. The mixture was stirred at 80°C for 16 hours under CO (2 MPa). The reaction mixture was poured into water (500 mL) and extracted with SiO (300 mL x 2). The combined organic layers were washed with brine (300 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (80g SepaFlash® silica flash column, siRNA:MeOH: 0-20%) to obtain the compound dimethyl 5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxylate (3g, 8.66 mmol, yield 25.2%, purity 97.0%) as a dark brown solid.

[0558] 1 H NMR (400 MHz, DMSO-d6) δ = 8.36 (s, 1H), 8.26 (s, 1H), 8.13 (s, 2H), 3.90 (s, 6H), 3.60 (s, 2H), 3.08 (q, J = 6.0 Hz, 2H), 2.41 (t, J = 6.8 Hz, 2H), 2.26 (s, 6H), 1.65 - 1.55 (m, 2H).

[0559] Step 3: 5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxylic acid (4)

[0560] To a solution of dimethyl 5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxylate (3 g, 8.92 mmol, 1.0 equivalent) in EtOH (4.75 mL), NaOH (1.78 g, 44.59 mmol, 5.0 equivalents) was added in H2O (0.25 mL). The mixture was stirred at 60°C for 3 hours. The reaction mixture was poured into water (5 mL) and extracted with SiO2 (5 mL x 2). The water was concentrated under reduced pressure to obtain compound 5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxylic acid (1.7 g, crude) as a dark brown solid. The crude product was used in the next step without further purification.

[0561] Step 4: tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzoyl]amino]propyl]carbamate (5)

[0562] In 15 mL of DMF, a mixture of 5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxylic acid (1.5 g, 4.86 mmol, 1.0 equivalent), tert-butyl N-(3-aminopropyl)carbamate (2.54 g, 14.59 mmol, 2.55 mL, 3.0 equivalents), HATU (4.07 g, 10.70 mmol, 2.2 equivalents), and DIEA (3.77 g, 29.19 mmol, 5.08 mL, 6 equivalents) was degassed and purged with N2 three times. The mixture was then stirred under an N2 atmosphere at 20°C for 12 hours. The reaction mixture was diluted with H2O (150 mL) and extracted with siRNA (50 mL x 3). The combined organic layers were washed with brine (50 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (80g SepaFlash® silica flash column, Depositphotos:MeOH:0-20% Depositphotos and 0.15% NH3H2O ​​in MeOH) to obtain the compound tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzoyl]amino]propyl]carbamate (1.8g, 2.88 mmol, yield 59.3%, purity 99.4%) as a yellow solid.

[0563] 1 H NMR (400 MHz, DMSO-d6) δ = 8.52 (t, J = 5.2 Hz, 2H), 8.17 - 8.10 (m, 2H), 7.84 (s, 2H), 6.81 (t, J = 5.2 Hz, 2H), 3.50 (s, 2H), 3.26 (q, J = 6.4 Hz, 4H), 3.08 (q, J = 6.4 Hz, 2H), 2.97 (q, J = 6.4 Hz, 4H), 2.43 (t, J = 7.2 Hz, 2H), 2.28 (s, 6H), 1.68 - 1.57 (m, 6H), 1.37 (s, 18H).

[0564] Step 5: N1,N3-Bis(3-aminopropyl)-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxamide(6)

[0565] To a solution of tert-butyl N-[3-[[3-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzoyl]amino]propyl]carbamate (1 g, 1.61 mmol, 1.0 equivalent) in DCM (10 mL), HCl / dioxane (4 M, 5.00 mL) was added. The mixture was stirred at 20 °C for 1 hour. The reaction mixture was concentrated under reduced pressure to obtain compound N1,N3-bis(3-aminopropyl)-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxamide (1.25 g, crude, 10 HCl) as a pink solid. The crude product was used in the next step without further purification.

[0566] Step 6: (Alternate name) 1-Ethylhexyl 9-[3-[[3-[3-[bis[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]propylcarbamoyl]-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxo-nonyl]amino]nonanoate (SM-084)

[0567] To a solution of N1,N3-bis(3-aminopropyl)-5-[2-[3-(dimethylamino)propylamino]-2-oxo-ethyl]benzene-1,3-dicarboxamide (1.25 g, 1.59 mmol, 1.0 equivalent, 10HCl) in MeOH (31 mL), NaOAc (1.83 g, 22.29 mmol, 14.0 equivalents) and NaBH3CN (1.00 g, 15.92 mmol, 10.0 equivalents) were added. The mixture was stirred at 20°C for 0.5 hours, then 1-ethylhexyl 9-oxononanoate (3.62 g, 12.74 mmol, 8.0 equivalents) was added, and the mixture was stirred at 20°C for 14 hours. The reaction mixture was diluted with water (50 mL) and extracted with ELISA (30 mL x 2). The combined organic layers were washed with brine (30 mL), dried over anhydrous Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (40 g SepaFlash® silica flash column, DCM:MeOH: 0-15%) and preparative HPLC (column: Welch Xtimate C1 100 × 30 mm × 5 μm, mobile phase: [water (FA)-MeOH], gradient: 63%-93% B over 8 minutes) to obtain SM-084 (448.31 mg, 293.86 μmol, yield 18.5%, purity 97.95%) as a yellow oily substance. LCMS:[M+H] + :1494.8

[0568] 1H NMR (400 MHz, CDCl3) δ = 8.31 (t, J = 4.8 Hz, 2H), 8.19 (s, 1H), 7.84 (d, J = 0.8 Hz, 2H), 7.37 (t, J = 4.8 Hz, 1H), 4.87 - 4.76 (m, 4H), 3.59 (s, 2H), 3.54 (q, J = 6.0 Hz, 4H), 3.31 (q, J = 6.0 Hz, 2H), 2.63 (t, J = 6.0 Hz, 4H), 2.55 - 2.45 (m, 8H), 2.28 (t, J = 7.6 Hz, 10H), 1.99 (s, 6H), 1.81 - 1.74 (m, 4H), 1.60 - 1.54 (m, 12H), 1.53 - 1.43 (m, 18H), 1.33 - 1.20 (m, 60H), 0.90 - 0.85 (m, 24H).

[0569] (Example 6) Synthesis of SM-089 [ka] [ka]

[0570] Step 1: Dimethyl 5-[3-(dimethylamino)propoxy]benzene-1,3-dicarboxylate (2)

[0571] To a solution of dimethyl 5-hydroxybenzene-1,3-dicarboxylate (8 g, 38.06 mmol, 1.0 equivalent) and 3-chloro-N,N-dimethylpropan-1-amine (5.55 g, 45.67 mmol, 1.2 equivalents) in MeCN (200 mL), K2CO3 (7.89 g, 57.09 mmol, 1.5 equivalents), 18-C-6 (503.02 mg, 1.90 mmol, 0.05 equivalents), and KI (631.84 mg, 3.81 mmol, 0.1 equivalents) were added. The mixture was stirred at 80°C for 16 hours. The reaction mixture was filtered, and the filter cake was washed with Â(50 mL x 2). The filtrate was concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (120 g SepaFlash® silica flash column, DCM:MeOH: 0-10%) to obtain the compound dimethyl 5-[3-(dimethylamino)propoxy]benzene-1,3-dicarboxylate (14.8 g, 50.11 mmol, yield 66.1%) as a yellow oily substance.

[0572] 1 H NMR (400 MHz, CDCl3) δ = 8.24 (s, 1H), 7.71 (s, 2H), 4.09 (t, J = 6.0 Hz, 2H), 3.92 (s, 6H), 2.56 (t, J = 7.6 Hz, 2H), 2.32 (s, 6H), 2.07 - 1.98 (m, 2H).

[0573] Step 2: Compound 5-[3-(dimethylamino)propoxy]benzene-1,3-dicarboxylic acid (3)

[0574] Dimethyl 5-[3-(dimethylamino)propoxy]benzene-1,3-dicarboxylate (7.5 g, 25.40 mmol, 1.0 equivalent) was dissolved in MeOH (60 mL) and H2O (15 mL) to which NaOH (3.05 g, 76.19 mmol, 3.0 equivalents) was added. The mixture was stirred at 60°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the MeOH. The residue was diluted with H2O (30 mL) and extracted with HCl (30 mL x 3). The pH of the aqueous phase was adjusted to 2 with 4 M HCl, and the residue was concentrated under reduced pressure to obtain the residue. The residue was ground in (PE / HCl = 5 / 1, 50 mL x 2) at 20°C for 0.5 hours and properly filtered. The filtered cake was dried to obtain compound 5-[3-(dimethylamino)propoxy]benzene-1,3-dicarboxylic acid (5.1 g, 19.08 mmol, yield 75.1%) as a yellow solid.

[0575] Step 3: SM-089, also known as tetra(octane-3-yl)9,9',9'',9'''-((((5-(3-(dimethylamino)propoxy)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate

[0576] 5-[3-(dimethylamino)propoxy]benzene-1,3-dicarboxylic acid (500 mg, 1.22 mmol, 1.0 equivalent) and 1-ethylhexyl 9-[3-aminopropyl-[9-(1-ethylhexoxy)-9-oxononyl]amino]nonanoate (1.97 g, 3.04 mmol, 2.5 equivalents, HCl) were dissolved in pyridine (15 mL), to which EDCI (582.75 mg, 3.04 mmol, 2.5 equivalents) was added. The mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated directly under reduced pressure, and the residual pyridine was co-evaporated with DCM to obtain the residue. The residue was diluted with H2O (50 mL) and extracted with DCM (35 mL x 3). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (SepaFlash® silica flash column 20g, DCM:MeOH:0~10%) and preparative HPLC (column: Welch Xtimate C1100×30mm×5um, mobile phase: [A / B water (FA)-MeOH], gradient: 65%~95%B over 8 minutes) to obtain SM-089 (430mg, 293.26μmol, yield 25.07%, purity 99.11%) as a pale yellow oily substance. LCMS:[M+H] + :1454.4

[0577] 1 H NMR (400 MHz, CDCl3) δ = 8.25 (s, 1H), 7.97 - 7.82 (m, 1H), 7.64 (d, J = 8.0 Hz, 1H), 7.49 (s, 1H), 4.81 (t, J = 6.0 Hz, 4H), 4.16 - 4.03 (m, 2H), 3.70 - 3.47 (m, 4H), 2.75 - 2.62 (m, 4H), 2.58 - 2.42 (m, 8H), 2.31 - 2.21 (m, 14H), 2.04 - 1.93 (m, 2H), 1.88 - 1.73 (m, 4H), 1.64 - 1.45 (m, 32H), 1.35 - 1.21 (m, 58H), 0.91 - 0.85 (m, 24H).

[0578] (Example 7) Synthesis of SM-090 [ka] [ka] [ka]

[0579] Step 1: 2,6-Dichloro-N-[2-(dimethylamino)ethyl]pyridine-4-carboxamide

[0580] To a solution of 2,6-dichloropyridine-4-carboxylic acid (5 g, 26.04 mmol, 1.0 equivalent) in DCM (50 mL), SOCl2 (6.20 g, 52.08 mmol, 3.78 mL, 2.0 equivalents) was added at 0°C. The mixture was stirred at 25°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain compound 2,6-dichloropyridine-4-carbonyl chloride (5.4 g, 25.66 mmol, yield 98.5%) as a brown oily substance. To a 50 mL solution of N',N'-dimethylethane-1,2-diamine (2.71 g, 30.79 mmol, 3.36 mL, 1.2 equivalents) and TEA (7.79 g, 76.98 mmol, 10.71 mL, 3.0 equivalents) in DCM, 2,6-dichloropyridine-4-carbonyl chloride (5.4 g, 25.66 mmol, 1.0 equivalent) was added at 0°C. The mixture was stirred at 25°C for 12 hours. The residue was diluted with 50 mL of water and extracted with DCM (100 mL x 3). The combined organic layers were washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 1 / 0 to 20 / 1) to obtain compound 2,6-dichloro-N-[2-(dimethylamino)ethyl]pyridine-4-carboxamide (6.6 g, 23.92 mmol, yield 93.2%, purity 94.9%) as a yellow solid.

[0581] 1H NMR (400 MHz, CDCl3) δ = 9.08 - 9.02 (m, 1H), 7.63 (s, 2H), 3.27 - 3.20 (m, 2H), 2.70 - 2.61 (m, 2H), 2.42 - 2.30 (m, 6H).

[0582] Step 2: Dimethyl 4-[2-(dimethylamino)ethylcarbamoyl]pyridine-2,6-dicarboxylate

[0583] To a solution of 2,6-dichloro-N-[2-(dimethylamino)ethyl]pyridine-4-carboxamide (6.6 g, 25.18 mmol, 1.0 equivalent) in MeOH (70 mL), TEA (7.64 g, 75.53 mmol, 10.51 mL, 3.0 equivalents) and Pd(dppf)Cl2 (1.84 g, 2.52 mmol, 0.1 equivalent) were added under N2. The suspension was degassed under vacuum and purged with CO three times. The mixture was stirred at 80°C for 12 hours under CO (50 psi). The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 80g SepaFlash® silica flash column, eluent 0-5.3% MeOH / DCM, gradient at 60 mL / min) to obtain the compound dimethyl 4-[2-(dimethylamino)ethylcarbamoyl]pyridine-2,6-dicarboxylate (7g, 22.63 mmol, yield 89.9%) as a brown solid.

[0584] 1 H NMR (400 MHz, DMSO-d6) δ = 9.56 (s, 1H), 8.69 (s, 2H), 3.95 (s, 6H), 3.70 - 3.67 (m, 2H), 3.28 - 3.26 (m, 2H), 2.81 - 2.78 (m, 6H).

[0585] Step 3: tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[2-(dimethylamino)ethylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate

[0586] To a solution of dimethyl 4-[2-(dimethylamino)ethylcarbamoyl]pyridine-2,6-dicarboxylate (1 g, 3.23 mmol, 1.0 equivalent) in MeOH (10 mL), tert-butyl N-(3-aminopropyl)carbamate (1.69 g, 9.70 mmol, 1.69 mL, 3.0 equivalent) was added. The mixture was stirred at 50°C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 20g SepaFlash® silica flash column, eluent 0-5.3% MeOH / DCM, gradient at 60 mL / min) to obtain the compound tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[2-(dimethylamino)ethylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate (1.9 g, 2.72 mmol, yield 84.1%, purity 85%) as a brown solid.

[0587] Step 4: N2,N6-Bis(3-aminopropyl)-N4-[2-(dimethylamino)ethyl]pyridine-2,4,6-tricarboxamide

[0588] To a solution of tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[2-(dimethylamino)ethylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate (1.9 g, 3.20 mmol, 1.0 equivalent) in DCM (15 mL), HCl / dioxane (4 M, 15 mL) was added. The mixture was stirred at 25 °C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was ground with PE / HCl = 10 / 1 for 30 minutes to obtain compound N2,N6-bis(3-aminopropyl)-N4-[2-(dimethylamino)ethyl]pyridine-2,4,6-tricarboxamide (1.3 g, 2.79 mmol, yield 87.1%, 2HCl) as a gray solid.

[0589] Step 5: Tetra(octane-3-yl)9,9',9'',9'''-((((4-((2-(dimethylamino)ethyl)carbamoyl)pyridine-2,6-dicarbonyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate: SM-090

[0590] NaBH3CN (331.41 mg, 5.27 mmol, 6.0 equivalents) was added to a solution of N2,N6-bis(3-aminopropyl)-N4-[2-(dimethylamino)ethyl]pyridine-2,4,6-tricarboxamide (409.94 mg, 878.94 μmol, 1.0 equivalent, 2HCl) and NaOAc (432.62 mg, 5.27 mmol, 6.0 equivalents) in MeOH (20 mL). After addition, the mixture was stirred at 25°C for 0.5 hours, and then 1-ethylhexyl 9-oxononanoate (1.5 g, 5.27 mmol, 6.0 equivalents) was added to the mixture. The mixture was stirred at 25°C for 11.5 hours. The reaction mixture was diluted with water (30 mL) and extracted with DCM (60 mL x 2). The combined organic layers were washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 40 / 1 to 20 / 1) and preparative TLC (SiO2, DCM / MeOH = 10 / 1), and further purified by preparative HPLC (column: Welch Xtimate C1 100 × 30 mm × 5 μm, mobile phase: [A / B water (FA)-MeOH], gradient: 63% to 93% B over 8 minutes) to obtain the compound tetra(octan-3-yl)9,9',9'',9'''-((((4-((2-(dimethylamino)ethyl)carbamoyl)pyridine-2,6-dicarbonyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate (265.37 mg, 179.55 μmol, yield 20.43%, purity 99.27%) as a colorless gum-like substance. LCMS:[M+H] + :1468.2

[0591] 1H NMR (400 MHz, CDCl3) δ = 9.75 (s, 2H), 8.67 (s, 1H), 8.65 (s, 2H), 4.85 - 4.75 (m, 4H), 3.76 - 3.50 (m, 8H), 2.94 - 2.90 (m, 4H), 2.78 - 2.72 (m, 6H), 2.55 - 2.50 (m, 2H), 2.37 - 2.19 (m, 14H), 2.13 - 1.96 (m, 4H), 1.66 - 1.43 (m, 32H), 1.32 - 1.25 (m, 56H), 0.93 - 0.81 (m, 24H).

[0592] (Example 8) Synthesis of SM-091 [ka] [ka]

[0593] Step 1: 2,6-Dichloro-N-[4-(dimethylamino)butyl]pyridine-4-carboxamide

[0594] To a solution of 2,6-dichloropyridine-4-carboxylic acid (10 g, 52.08 mmol, 1.0 equivalent) in DCM (100 mL), SOCl2 (12.39 g, 104.17 mmol, 7.57 mL, 2.0 equivalents) was added at 0°C. The mixture was stirred at 40°C for 2 hours. The reaction mixture was concentrated under reduced pressure to obtain compound 2,6-dichloropyridine-4-carbonyl chloride (10.9 g, 51.79 mmol, 99.5% yield) as a brown oily substance. To a 100 mL solution of N',N'-dimethylbutane-1,4-diamine (7.22 g, 62.15 mmol, 1.2 equivalents) and TEA (15.72 g, 155.38 mmol, 21.63 mL, 3.0 equivalents) in DCM, 2,6-dichloropyridine-4-carbonyl chloride (10.9 g, 51.79 mmol, 1.0 equivalent) was added at 0°C. The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 1 / 0~20 / 1) to obtain compound 2,6-dichloro-N-[4-(dimethylamino)butyl]pyridine-4-carboxamide (12.2 g, 42.04 mmol, yield 81.2%) as a yellow solid.

[0595] 1 H NMR (400 MHz, DMSO-d6) δ = 9.26 - 9.21 (m, 1H), 7.97 (s, 2H), 3.31 - 3.25 (m, 2H), 3.03 - 2.96 (m, 2H), 2.67 (s, 6H), 1.75 - 1.50 (m, 4H).

[0596] Step 2: Dimethyl 4-[4-(dimethylamino)butylcarbamoyl]pyridine-2,6-dicarboxylate

[0597] To a solution of 2,6-dichloro-N-[4-(dimethylamino)butyl]pyridine-4-carboxamide (12.2 g, 42.04 mmol, 1.0 equivalent) in MeOH (150 mL), TEA (12.76 g, 126.13 mmol, 17.55 mL, 3.0 equivalents) and Pd(dppf)Cl2 (3.08 g, 4.20 mmol, 0.1 equivalents) were added under N2. The suspension was degassed under vacuum and purged with CO three times. The mixture was stirred at 80°C for 12 hours under CO (50 psi). The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 40 / 1 to 10 / 1) and ground with ethyl acetate for 1 hour to obtain the compound dimethyl 4-[4-(dimethylamino)butylcarbamoyl]pyridine-2,6-dicarboxylate (9.4 g, 27.86 mmol, yield 66.8%) as a gray solid.

[0598] 1 H NMR (400 MHz, DMSO-d6) δ = 9.34 (s, 1H), 8.66 (s, 2H), 3.95 (s, 6H), 3.35 - 3.30 (m, 2H), 3.07 -3.00 (m, 2H), 2.70 (s, 6H), 1.75 - 1.54 (m, 4H).

[0599] Step 3: tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[4-(dimethylamino)butylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate

[0600] To a solution of dimethyl 4-[4-(dimethylamino)butylcarbamoyl]pyridine-2,6-dicarboxylate (2 g, 5.93 mmol, 1.0 equivalent) in MeOH (30 mL), tert-butyl N-(3-aminopropyl)carbamate (5.16 g, 29.64 mmol, 5.18 mL, 5.0 equivalent) was added. The mixture was stirred at 50°C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 40g SepaFlash® silica flash column, eluent 0-5.3% MeOH / DCM, gradient at 60 mL / min) to obtain the compound tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[4-(dimethylamino)butylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate (3.6 g, 5.56 mmol, yield 93.8%, purity 96%) as a brown, gum-like substance.

[0601] Step 4: N2,N6-Bis(3-aminopropyl)-N4-[4-(dimethylamino)butyl]pyridine-2,4,6-tricarboxamide

[0602] To a solution of tert-butyl N-[3-[[6-[3-(tert-butoxycarbonylamino)propylcarbamoyl]-4-[4-(dimethylamino)butylcarbamoyl]pyridine-2-carbonyl]amino]propyl]carbamate (2 g, 3.22 mmol, 1.0 equivalent) in DCM (20 mL), HCl / dioxane (4 M, 20 mL) was added. The mixture was stirred at 25°C for 12 hours. The reaction mixture was concentrated under reduced pressure to obtain the crude product. The crude product was ground with PE / siRNA=10 / 1 for 30 minutes to obtain compound N2,N6-bis(3-aminopropyl)-N4-[4-(dimethylamino)butyl]pyridine-2,4,6-tricarboxamide (1.5 g, 3.03 mmol, yield 94.3%, HCl salt) as a gray solid.

[0603] Step 5: Tetra(octane-3-yl)9,9',9'',9'''-((((4-((4-(dimethylamino)butyl)carbamoyl)pyridine-2,6-dicarbonyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate: SM-091

[0604] NaBH3CN (441.88 mg, 7.03 mmol, 8.0 equivalents) was added to a solution of N2,N6-bis(3-aminopropyl)-N4-[4-(dimethylamino)butyl]pyridine-2,4,6-tricarboxamide (434.60 mg, 878.94 μmol, 1.0 equivalent, 2HCl) and NaOAc (576.82 mg, 7.03 mmol, 8 equivalents) in MeOH (15 mL). After addition, the mixture was stirred at 25°C for 0.5 hours, and then 1-ethylhexyl 9-oxononanoate (2 g, 7.03 mmol, 8.0 equivalents) was added to the mixture. The mixture was stirred at 25°C for 11.5 hours. The reaction mixture was diluted with water (30 mL) and extracted with DCM (60 mL x 2). The combined organic layers were washed with water, dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by column chromatography (SiO2, DCM / MeOH = 40 / 1 to 20 / 1), and then purified by preparative HPLC (column: Welch Xtimate C1 100 × 30 mm × 5 μm, mobile phase: [A / B water (FA)-MeOH], gradient: 63% to 93% B over 8 minutes) to obtain SM-091, also known as tetra(octane-3-yl)9,9',9'',9'''-((((4-((4-(dimethylamino)butyl)carbamoyl)pyridine-2,6-dicarbonyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate (312.02 mg, 205.42 μmol, yield 23.4%, purity 98.44%) as a pale yellow oily substance. LCMS:[M+H] + :1495.3

[0605] 1H NMR (400 MHz, CDCl3) δ = 9.29 (s, 1H), 8.66 (s, 2H), 4.85 - 4.76 (m, 4H), 3.62 - 3.55 (m, 4H), 3.53 - 3.45 (m, 2H), 2.67 (s, 4H), 2.61 - 2.42 (m, 8H), 2.35 - 2.33 (m, 2H), 2.31 - 2.24 (m, 14H), 1.89 (s, 4H), 1.80 - 1.71 (m, 2H), 1.69 (s, 2H), 1.64 - 1.43 (m, 32H), 1.31 - 1.25 (m, 56H), 0.97 - 0.81 (m, 24H).

[0606] (Example 9) Synthesis of SM-092 [ka] [ka]

[0607] Step 1: Dimethyl 5-chlorocarbonylbenzene-1,3-dicarboxylate (2)

[0608] To a solution of 3,5-bis(methoxycarbonyl)benzoic acid (3 g, 12.59 mmol, 1.0 equivalent) in DCM (30 mL), SOCl2 (4.50 g, 37.78 mmol, 2.74 mL, 3.0 equivalents) was added under N2 at 0°C. After the addition, the reaction mixture was stirred at 40°C for 3 hours. The reaction mixture was directly concentrated under reduced pressure to obtain the compound dimethyl 5-chlorocarbonylbenzene-1,3-dicarboxylate (3.2 g, crude) as a yellow solid. The crude product was used in the next step without further purification.

[0609] Step 2: Dimethyl 5-[(1-methyl-4-piperidyl)methylcarbamoyl]benzene-1,3-dicarboxylate (3)

[0610] To a 30 mL solution of (1-methyl-4-piperidyl)methanamine (1.92 g, 14.96 mmol, 1.2 equivalents) and TEA (3.15 g, 31.17 mmol, 4.34 mL, 2.5 equivalents) in DCM (20 mL), dimethyl 5-chlorocarbonylbenzene-1,3-dicarboxylate (3.2 g, 12.47 mmol, 1.0 equivalent) was added dropwise under N2 at 0°C. After addition, the resulting mixture was stirred at 20°C for 3 hours. The reaction mixture was directly concentrated under reduced pressure to obtain the residue. The residue was ground in (PE / siRNA = 10 / 1, 50 mL x 3) at 20°C for 0.5 hours and filtered. The filtered cake was dried to obtain the compound dimethyl 5-[(1-methyl-4-piperidyl)methylcarbamoyl]benzene-1,3-dicarboxylate (4.1 g, 11.77 mmol, yield 94.4%) as a yellow solid.

[0611] Step 3: 5-[(1-methyl-4-piperidyl)methylcarbamoyl]benzene-1,3-dicarboxylic acid (4)

[0612] Dimethyl 5-[(1-methyl-4-piperidyl)methylcarbamoyl]benzene-1,3-dicarboxylate (4.1 g, 11.77 mmol, 1.0 equivalent) was dissolved in MeOH (60 mL) and H2O (15 mL) and NaOH (1.41 g, 35.30 mmol, 3.0 equivalents) was added. The mixture was stirred at 60°C for 16 hours. The reaction mixture was concentrated under reduced pressure to remove the MeOH. The residue was diluted with H2O (30 mL) and extracted with HCl (30 mL x 3). The pH of the aqueous phase was adjusted to 2 with 4 M HCl and concentrated under reduced pressure to obtain the residue. The residue was ground in (PE / HCl = 1 / 1, 50 mL x 3) at 20°C for 0.5 hours and properly filtered. The filtered cake was dried to obtain compound 5-[(1-methyl-4-piperidyl)methylcarbamoyl]benzene-1,3-dicarboxylic acid (2.9 g, 9.05 mmol, yield 76.9%) as a white solid.

[0613] Step 4: SM-092, also known as tetra(octane-3-yl)9,9',9'',9'''-((((5-(((1-methylpiperidine-4-yl)methyl)carbamoyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate

[0614] 5-[(1-methyl-4-piperidyl)methylcarbamoyl]benzene-1,3-dicarboxylic acid (400 mg, 1.25 mmol, 1.0 equivalent) and 1-ethylhexyl 9-[3-aminopropyl-[9-(1-ethylhexoxy)-9-oxononyl]amino]nonanoate (2.02 g, 3.12 mmol, 2.5 equivalents, HCl) were dissolved in pyridine (15 mL), to which EDCI (598.43 mg, 3.12 mmol, 2.5 equivalents) was added. The mixture was stirred at 20°C for 16 hours. The reaction mixture was concentrated directly under reduced pressure, and the residual pyridine was co-evaporated with DCM to obtain the residue. The residue was diluted with water (50 mL) and extracted with DCM (35 mL x 3). The combined organic layers were washed with brine (40 mL), dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (20g SepaFlash® silica flash column, DCM:MeOH: 0-10%) and preparative HPLC (column: Welch Xtimate C1100 × 30 mm × 5 μm, mobile phase: [A / B water (FA)-MeOH], gradient: 60-90% MeOH over 8 minutes) to obtain SM-092 (430 mg, 0.29 mmol, yield 25.1%, purity 99.11%) as a pale yellow oily substance. LCMS:[M+H] + :1506.6

[0615] 1H NMR (400 MHz, CDCl3) δ = 8.65 - 8.57 (m, 2H), 8.49 - 8.41 (m, 3H), 7.10 - 6.85 (m, 1H), 4.84 - 4.75 (m, 4H), 3.63 - 3.52 (m, 4H), 3.37 (t, J = 6.0 Hz, 2H), 2.87 (d, J = 11.2 Hz, 2H), 2.75 - 2.66 (m, 4H), 2.62 - 2.48 (m, 8H), 2.30 - 2.24 (m, 10H), 1.95 (t, J = 10.8 Hz, 2H), 1.89 - 1.72 (m, 6H), 1.64 - 1.46 (m, 32H), 1.35 - 1.20 (m, 60H), 0.90 - 0.83 (m, 24H).

[0616] (Example 10) Synthesis of SM-093 [ka] [ka]

[0617] Step 1: 9-[3-(tert-butoxycarbonylamino)propyl-[9-(2-hexyloctanoyloxy)nonyl]amino]nonyl2-hexyloctanoate

[0618] To a solution of tert-butyl N-(3-aminopropyl)carbamate (1.5 g, 8.61 mmol, 1.50 mL, 1.0 equivalent) in MeOH (90 mL), HOAc (1.5 g, 25.8 mmol, 1.5 mL, 3.0 equivalents) and NaBH3CN (1.62 g, 25.83 mmol, 3.0 equivalents) were added. The mixture was stirred at 20°C for 0.5 hours. Then, 9-oxononyl 2-hexyl octanoate (9.52 g, 25.83 mmol, 3.0 equivalents) [see Int_5 for preparation] was added to the mixture and stirred at 20°C for 11.5 hours. The reaction mixture was concentrated under reduced pressure to remove the solvent. The residue was diluted with water (100 mL) and extracted with DCM (100 mL x 2). The combined organic layers were dried over Na2SO4, filtered, and concentrated under reduced pressure to obtain the residue. The residue was purified by flash silica gel chromatography (ISCO®, 120g SepaFlash® silica flash column, eluent 0-50% siRNA / PE, gradient: 100 mL / min) to obtain compound 9-[3-(tert-butoxycarbonylamino)propyl-[9-(2-hexyloctanoyloxy)nonyl]amino]nonyl 2-hexyloctanoate (6.9 g, 7.34 mmol, yield 85.2%, purity 93.5%) as a yellow, gum-like substance.

[0619] 1 H NMR (400 MHz, CDCl3) δ = 5...

Claims

1. Compound of formula I: 【Chemical 305】 or its salt or isomer [in the formula, X is CH or N, a and b are independently 2 to 5. I understand １ , m ２ , m ３ , and m ４ These are independently 4 to 10, E １ , E ２ , E ３ , and E ４ These are independently -O(CO)O-, -(CO)O-, or -O(CO)-, T １ 、 T ２ 、 T ３ 、 and T ４ are, independently, branched or unbranched C ５ -C ２２ alkyl, C ５ -C ２２ alkenyl, or C ５ -C ２２ alkynyl, and R １ and R ２ H and C are independent of each other. １ ~C ６ Alkyl, C ２ ~C ６ Alkenyl, or C ２ ~C ６ It is alkinyl, R ３ H and C are independent of each other. １ ~C ６ Alkyl, C ２ ~C ６ Alkenil, C ２ ~C ６ Alkinyl, or 【Chemical 306】 And, G is O, -(CO)NR ３ -, -NR ３ (CO)-, -(CO)O-, or -CH ２ (CO)NR ３ - and L is bonded to or, if necessary, substituted with C. １ ~C ４ It is alkyl, Z is CH or N, R ４ and R ５ H and C are independent of each other. １ ~C ３ Alkyl, C ２ ~C ３ Alkenil, C ２ ~C ３ Alkinil, 【Chemical 307】 Is it, or R ４ Or R ５ It forms a 3-7 membered ring which contains 0-2 heteroatoms and is optionally substituted, or R ４ and R ５ [It forms a substituted 3- to 7-membered ring together with L.]

2. R １ and R ２ a and b, m １ ~m ４ , E １ ~E ４ , and T １ ~T ４ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein one or more sets of the same elements are identical.

3. R １ and R ２ a and b, m １ ~m ４ , E １ ~E ４ , and T １ ~T ４ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein one or more of the compounds are different.

4. R １ and R ２ a and b, m １ ~m ４ , E １ ~E ４ , and T １ ~T ４ The compound of formula I according to claim 1, or a salt or isomer thereof, wherein all of the elements are the same.

5. R １ and R ２ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein is H.

6. A compound of formula I according to claim 1, or a salt or isomer thereof, wherein a and b are independently 3, 4, or 5.

7. A compound of formula I according to claim 1, or a salt or isomer thereof, wherein a and b are 3.

8. I understand １ , m ２ , m ３ , and m ４ The compound of formula I according to claim 1, or a salt or isomer thereof, wherein independently, the compound is 6, 7, 8, or 9.

9. I understand １ , m ２ , m ３ , and m ４ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein is 8.

10. E １ , E ２ , E ３ , and E ４ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein is -(CO)O-.

11. E １ 、E ２ 、E ３ 、and E ４ is -O(CO)- or -O(CO)O-, the compound of formula I according to claim 1, or a salt or isomer thereof.

12. T １ 、 T ２ 、 T ３ 、 or T ４ is independently selected from the group consisting of C ５ ~C １８ alkyl, C ５ ~C １８ alkenyl, and C ５ ~C １８ alkynyl, and optionally T １ 、 T ２ 、 T ３ 、 and T ４ is independently selected from the group consisting of C ５ ~C １８ alkyl, C ５ ~C １８ alkenyl, and C ５ ~C １８ alkynyl, the compound of formula I according to claim 1, or a salt or isomer thereof.

13. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ A compound of formula I according to claim 1, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

14. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ A compound of formula I according to claim 1, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

15. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ A compound of formula I according to claim 1, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

16. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ A compound of formula I according to claim 1, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

17. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ７ or C ８ Alkyl, C ７ or C ８ Alkenyl and C ７ or C ８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ７ or C ８ Alkyl, C ７ or C ８ Alkenyl and C ７ or C ８ A compound of formula I according to claim 1, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

18. G is O, -(CO)NR ３ -, and -NR ３ A compound of formula I according to claim 1, or a salt or isomer thereof, which is (CO)-.

19. R ３ A compound of the formula according to claim 1, or a salt or isomer thereof, wherein the compound is H or methyl.

20. R ３ but, 【Chemical 308】 A compound of the formula described in claim 1, or a salt or isomer thereof.

21. A compound of formula I according to claim 1, or a salt or isomer thereof, wherein L is a bond.

22. L is C １ A compound of formula I as described in claim 1, or a salt or isomer thereof, which is alkyl.

23. L is C ２ A compound of formula I as described in claim 1, or a salt or isomer thereof, which is alkyl.

24. L is C ３ A compound of formula I as described in claim 1, or a salt or isomer thereof, which is alkyl.

25. L is C ４ A compound of formula I as described in claim 1, or a salt or isomer thereof, which is alkyl.

26. R ４ , R ５ , or R ４ and R ５ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein the element is absent or is H.

27. R ４ , R ５ , or R ４ and R ５ but, 【Chemical 309】 The compound of formula I as described in claim 1, or a salt or isomer thereof.

28. R ４ and R ５ However, independently, C １ , C ２ , or C ３ A compound of formula I as described in claim 1, or a salt or isomer thereof, which is alkyl.

29. R ４ and R ５ The compound of formula I according to claim 1, or a salt or isomer thereof, which forms a six-membered ring that is optionally substituted and contains one or two heteroatoms.

30. The compound of formula I according to claim 29, or a salt or isomer thereof, wherein at least one of the one or two heteroatoms is nitrogen.

31. The compound of formula I according to claim 29, or a salt or isomer thereof, wherein at least one hydrogen atom of the six-membered ring is substituted with a methyl group.

32. The compound of formula I according to claim 29, or a salt or isomer thereof, wherein at least one of the atoms is a nitrogen atom.

33. The compound of formula I according to claim 29, or a salt or isomer thereof, wherein the six-membered ring contains one, two, or three double bonds, and optionally the six-membered ring contains three double bonds.

34. R ４ and R ５ A compound of formula I according to claim 1, or a salt or isomer thereof, wherein is methyl.

35. R ４ and R ５ A compound of the formula according to claim 1, or a salt or isomer thereof, which optionally forms a five-membered ring comprising one or two heteroatoms and one or two double bonds.

36. The compound of formula I according to claim 35, or a salt or isomer thereof, wherein at least one of the one or two heteroatoms is N.

37. T １ , T ２ , T ３ , and T ４ The compound of formula I according to claim 1, or a salt or isomer thereof, wherein each is an alkyl selected from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as necessary.

38. T １ , T ２ , T ３ , and T ４ The compound of formula I according to claim 1, or a salt or isomer thereof, wherein the compound is octane or tridecane.

39. T １ , T ２ , T ３ , and T ４ However, independently, each is substituted as needed: buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca-2-ene, deca-3-ene, deca-4-ene, deca- A compound of formula I according to claim 1, or a salt or isomer thereof, which is an alkenyl selected from the group consisting of 5-ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and an alkenyl group having two or more double bonds.

40. T １ , T ２ , T ３ , and T ４ However, independently, each is substituted as needed: buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, deca-2-ine, deca-3-ine, deca-4-ine, deca- A compound of formula I according to claim 1, or a salt or isomer thereof, which is an alkynyl selected from the group consisting of 5-yne, deca-6-yne, undeca-1-yne, undeca-2-yne, undeca-3-yne, undeca-4-yne, undeca-5-yne, undeca-6-yne, undeca-7-yne, dodeca-1-yne, dodeca-2-yne, dodeca-3-yne, dodeca-4-yne, dodeca-5-yne, dodeca-6-yne, dodeca-8-yne, and an alkynyl group containing two or more triple bonds.

41. A compound of formula I according to claim 1, or a salt or isomer thereof, wherein X is N.

42. Compounds selected from the group consisting of the following: 【Chemical 310】 【Chemical 311】 【Chemical 312】 【Chemistry 313】 【Chemical 314】 【Chemical Industry 315】 【Chemical 316】 【Chemical 317】 【Chemical 318】

43. Lipids in formula I: 【Chemical 319】 or its salt or isomer [in the formula, X is CH or N, a and b are independently 2 to 5. I understand １ , m ２ , m ３ , and m ４ These are independently 4 to 10, E １ , E ２ , E ３ , and E ４ These are independently -O(CO)O-, -(CO)O-, or -O(CO)-, T １ , T ２ , T ３ , and T ４ Independently, branched or unbranched C ５ ~C ２２ Alkyl, C ５ ~C ２２ Alkenyl, or C ５ ~C ２２ It is alkinyl, R １ and R ２ These are, independently, H, or C １ ~C ６ Alkyl, C ２ ~C ６ Alkenil or C ２ ~C ６ It is alkinyl, R ３ These are, independently, H, or C １ ~C ６ Alkyl, C ２ ~C ６ Alkenil or C ２ ~C ６ Alkinyl, or 【Chem.320】 And, G is O, -(CO)NR ３ -, -NR ３ (CO)-, -(CO)O-, or -CH ２ (CO)NR ３ - and L is bonded to or, if necessary, substituted with C. １ ~C ４ It is alkyl, Z is CH or N, R ４ and R ５ H and C are independent of each other. １ ~C ３ Alkyl, C ２ ~C ３ Alkenil or C ２ ~C ３ Alkinyl, or 【Chemistry 321】 Is it, Or R ４ or R ５ It forms a 3-7 membered ring containing 0-2 heteroatoms, which may be substituted as needed, or R ４ and R ５ A pharmaceutical composition comprising [which, together with L, forms a substituted 3- to 7-membered ring].

44. R １ and R ２ a and b, m １ ~m ４ , E １ ~E ４ , and T １ ~T ４ The pharmaceutical composition according to claim 43, comprising one or more sets of the same formula I or a salt or isomer thereof.

45. R １ and R ２ a and b, m １ ~m ４ , E １ ~E ４ , and T １ ~T ４ The pharmaceutical composition according to claim 43, comprising one or more different salts or isomers of formula I or therein.

46. R １ and R ２ a and b, m １ ~m ４ , E １ ~E ４ , and T １ ~T ４ The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof, wherein all of the elements are the same.

47. R １ and R ２ The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof, wherein is H.

48. The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or the formula, wherein a and b are independently 3, 4, or 5.

49. The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or the formula I, wherein a and b are 3.

50. I understand １ , m ２ , m ３ , and m ４ The pharmaceutical composition according to claim 43, comprising, independently, 6, 7, 8, or 9, a salt or isomer of formula I or therein.

51. I understand １ , m ２ , m ３ , and m ４ The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof, wherein is 8.

52. E １ , E ２ , E ３ and E ４ The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof, wherein is -(CO)O-.

53. E １ , E ２ , E ３ , and E ４ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or the formula I, wherein the formula is -O(CO)- or -O(CO)O-.

54. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C １８ Alkyl, C ５ ~C １８ Alkenyl and C ５ ~C １８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C １８ Alkyl, C ５ ~C １８ Alkenyl and C ５ ~C １８ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or any of the alkynyl group, independently selected from the group consisting of alkynyls.

55. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or any of the alkynyl group, independently selected from the group consisting of alkynyls.

56. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or any of the alkynyl group, independently selected from the group consisting of alkynyls.

57. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or any of the alkynyl group, independently selected from the group consisting of alkynyls.

58. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or any of the alkynyl group, independently selected from the group consisting of alkynyls.

59. T １ , T ２ , T ３ , or T ４ However, each is replaced as needed in C ７ or C ８ Alkyl, C ７ or C ８ Alkenyl and C ７ or C ８ Independently selected from the group consisting of alkynnyls, and T as needed. １ , T ２ , T ３ , and T ４ However, each is replaced as needed in C ７ or C ８ Alkyl, C ７ or C ８ Alkenyl and C ７ or C ８ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or any of the alkynyl group, independently selected from the group consisting of alkynyls.

60. G is O, -(CO)NR ３ -, and -NR ３ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or the (CO)-.

61. R ３ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or the same, wherein the compound is H or methyl.

62. R ３ but, 【Chemistry 322】 The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof.

63. The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof, wherein L is a bond.

64. L is C １ The pharmaceutical composition according to claim 43, comprising an alkyl group, a salt or isomer of formula I or thereof.

65. L is C ２ The pharmaceutical composition according to claim 43, comprising an alkyl group, a salt or isomer of formula I or thereof.

22. L is C ３ A compound of formula I as described in claim 1, or a salt or isomer thereof, which is alkyl.

66. L is C ４ The pharmaceutical composition according to claim 43, comprising an alkyl group, a salt or isomer of formula I or thereof.

67. R ４ , R ５ , or R ４ and R ５ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or thereof, which is absent or H.

68. R ４ , R ５ , or R ４ and R ５ but, 【Chemical 323】 The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof.

69. R ４ and R ５ However, independently, C １ , C ２ , or C ３ The pharmaceutical composition according to claim 43, comprising an alkyl group, a salt or isomer of formula I or thereof.

70. R ４ and R ５ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or thereof, which optionally forms a six-membered ring containing one or two heteroatoms.

71. The pharmaceutical composition according to claim 70, comprising a salt or isomer of formula I or the same, wherein at least one of the one or two heteroatoms is nitrogen.

72. The pharmaceutical composition according to claim 70, comprising a salt or isomer of formula I or the formula I, wherein at least one hydrogen atom of the six-membered ring is substituted with a methyl group.

73. The pharmaceutical composition according to claim 72, comprising a salt or isomer of formula I or the formula I, wherein at least one of the atoms is a nitrogen atom.

74. The pharmaceutical composition according to claim 70, wherein the six-membered ring comprises one, two, or three double bonds, and optionally comprises a salt or isomer of formula I or the same, wherein the six-membered ring comprises three double bonds.

75. R ４ and R ５ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or thereof, wherein is methyl.

76. R ４ and R ５ The pharmaceutical composition according to claim 43, comprising a salt or isomer of formula I or thereof, which optionally forms a five-membered ring containing one or two heteroatoms and one or two double bonds.

77. The pharmaceutical composition according to claim 76, comprising a salt or isomer of formula I or the same, wherein at least one of the one or two heteroatoms is nitrogen.

78. T １ , T ２ , T ３ , and T ４ The pharmaceutical composition according to claim 43, comprising, independently, an alkyl selected from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as necessary, a salt or isomer of formula I or thereof.

79. T １ , T ２ , T ３ , and T ４ The pharmaceutical composition according to claim 43, comprising octane or tridecane, which is a salt or isomer of formula I or thereof.

80. T １ , T ２ , T ３ , and T ４ However, independently, each is substituted as needed: buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca-2-ene, deca-3-ene, deca-4-ene, deca-5-ene The pharmaceutical composition according to claim 43, comprising an alkenyl selected from the group consisting of n, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and a salt or isomer of formula I or an alkenyl group having two or more double bonds.

81. T １ , T ２ , T ３ , and T ４ However, independently, each is substituted as needed: buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, deca-2-ine, deca-3-ine, deca-4-ine, deca-5-ine The pharmaceutical composition according to claim 43, comprising an alkynyl selected from the group consisting of n, deca-6-in, undeca-1-in, undeca-2-in, undeca-3-in, undeca-4-in, undeca-5-in, undeca-6-in, undeca-7-in, dodeca-1-in, dodeca-2-in, dodeca-3-in, dodeca-4-in, dodeca-5-in, dodeca-6-in, dodeca-8-in, and a salt or isomer of formula I or an alkynyl group having two or more triple bonds.

82. The pharmaceutical composition according to claim 43, comprising formula I or a salt or isomer thereof, wherein X is N.

83. Lipid particles comprising the compounds described in claims 1 to 82.

84. Lipid particles according to claim 83, further comprising a therapeutic agent.

85. The lipid particles according to claim 84, wherein the therapeutic agent is nucleic acid.

86. A pharmaceutical composition comprising the lipid particles described in claim 83, and a pharmaceutically acceptable additive, carrier, or diluent.

87. Compounds of formula VII: 【Chemical 324】 or its salt or isomer [in the formula, L ６ is -O(CO)-, -CH ２ -O(CO)-, -CH ２ (CO)O- or -O(CO)-NH-, n ６ and n ７ These are independently 1, 2, 3, 4, 5, 6, 7, or 8, G ５ and G ６ These are independently a bond, -(CO)O-, or -O(CO)-, R １１ and R １２ C is independently substituted as needed. ５ ~C ２０ Alkyl or C ５ ~C ２０ It is Alkenil.

88. R １１ and R １２ The compound according to claim 87, or a salt or isomer thereof, which is the same.

89. R １１ and R １２ The compound according to claim 87, or a salt or isomer thereof, which differs in that respect.

90. R １１ or R １２ However, C is replaced as needed. ８ ~C ２０ Alkyl and C ８ ~C ２０ Independently selected from the group consisting of alkenyls, and R as needed. １１ and R １２ However, C is replaced as needed. ８ ~C ２０ Alkyl and C ８ ~C ２０ A compound according to claim 87, or a salt or isomer thereof, independently selected from the group consisting of alkenyls.

91. n ６ and n ７ The compound according to claim 90, or a salt or isomer thereof, wherein independently, it is 4, 5, 6, 7, or 8.

92. R １１ or R １２ The compound according to claim 91, or a salt or isomer thereof, comprising one, two, three, four, five, six, or more unsaturated sites.

93. R １１ or R １２ However, C is replaced as needed. ８ ~C １７ Alkyl and C ８ ~C １７ Independently selected from the group consisting of alkenyls, and R as needed. １１ and R １２ However, C is replaced as needed. ８ ~C １７ Alkyl and C ８ ~C １７ A compound according to claim 87, or a salt or isomer thereof, independently selected from the group consisting of alkenyls.

94. n ６ and n ７ The compound according to claim 93, or a salt or isomer thereof, wherein independently, it is 5, 6, or 7.

95. R １１ or R １２ The compound according to claim 94, or a salt or isomer thereof, comprising one, two, three, four, five, six, or more unsaturated sites.

96. R １１ or R １２ However, C is replaced as needed. １０ ~C １７ Alkyl and C １０ ~C １７ Independently selected from the group consisting of alkenyls, and R as needed. １１ and R １２ However, C is replaced as needed. １０ ~C １７ Alkyl and C １０ ~C １７ A compound according to claim 87, or a salt or isomer thereof, independently selected from the group consisting of alkenyls.

97. n ６ and n ７ The compound according to claim 96, or a salt or isomer thereof, wherein independently, it is 5, 6, or 7.

98. R １１ or R １２ The compound according to claim 96, or a salt or isomer thereof, comprising one, two, three, four, five, six, or more unsaturated sites.

99. R １１ or R １２ However, C is replaced as needed. １２ ~C １７ Alkyl and C １２ ~C １７ Independently selected from the group consisting of alkenyls, and R as needed. １１ and R １２ However, C is replaced as needed. １２ ~C １７ Alkyl and C １２ ~C １７ A compound according to claim 87, or a salt or isomer thereof, independently selected from the group consisting of alkenyls.

100. n ６ and n ７ The compound according to claim 99, or a salt or isomer thereof, wherein independently, it is 5, 6, or 7.

101. R １１ or R １２ The compound according to claim 99, or a salt or isomer thereof, comprising one, two, three, four, five, six, or more unsaturated sites.

102. R １１ However, C is replaced as needed. ８ ~C １７ Alkyl and C ８ ~C １７ Selected from the group consisting of alkenyls, R １２ However, C is replaced as needed. ８ ~C １７ Alkyl and C ８ ~C １７ A compound according to claim 87, or a salt or isomer thereof, selected from the group consisting of alkenyls.

103. n ６ and n ７ The compound according to claim 102, or a salt or isomer thereof, wherein the coefficient is 5.

104. G ５ and G ６ The compound according to claim 87, wherein it is the same.

105. G ５ and G ６ The compound according to claim 87, which is different in that respect.

106. G ５ G is a bond, ６ The compound according to claim 102, or a salt or isomer thereof, wherein is -(CO)O-.

107. R １１ or R １２ However, C is replaced as needed. ８ , C １２ , C １５ or C １７ Alkyl and C ８ , C １２ , C １５ or C １７ Independently selected from the group consisting of alkenyls, and R as needed. １１ and R １２ However, C is replaced as needed. ８ , C １２ , C １５ or C １７ Alkyl and C ８ , C １２ , C １５ or C １７ A compound according to claim 87, or a salt or isomer thereof, independently selected from the group consisting of alkenyls.

108. R １１ and R １２ The compound according to claim 87, or a salt or isomer thereof, wherein each is an alkyl selected from the group consisting of octane, nonane, decane, undecane, and dodecane, each of which is substituted as necessary.

109. R １１ and R １２ The compound according to claim 87, or a salt or isomer thereof, wherein the alkyl is independently selected from the group consisting of octane, nonane, decane, undecane, dodecane, tridecane, tetradecane, pentadecane, hexadecane, heptadecane, octadecane, nonadecane, and eicosane.

110. R １１ and R １２ However, independently, Octa-1-en, Octa-2-en, Octa-3-en, Octa-4-en, Nona-1-en, Nona-2-en, Nona-3-en, Nona-4-en, Nona-5-en, Deca-1-en, Deca-2-en, Deca-3-en, Deca-4-en, Deca-5-en, Deca-6-en, Undeka-1-en, Undeka-2-en, Undeka-3-en, Undeka-4-en, Undeka-5-en, Undeka-6-en, Undeka-7-en, Dodeka-1-en, Dodeka-2-en, Dodeka-3-en, Dodeka-4-en, Dodeka-5- En, Dodeca-6-en, Trideca-1-en, Trideca-2-en, Trideca-3-en, Trideca-4-en, Trideca-5-en, Trideca-6-en, Trideca-7-en, Tetradeca-1-en, Tetradeca-2-en, Tetradeca-3-en, Tetradeca-4-en, Tetradeca-5-en, Tetradeca-6-en, Tetradeca-7-en, Pentadeca-1-en, Pentadeca-2-en, Pentadeca-3-en, Pentadeca-4-en, Pentadeca-5-en, Pentadeca-6-en, Pentadeca-7-en, Hexadeca-1- En, Hexadeca-2-en, Hexadeca-3-en, Hexadeca-4-en, Hexadeca-5-en, Hexadeca-6-en, Hexadeca-7-en, Hexadeca-8-en, Heptadeca-1-en, Heptadeca-2-en, Heptadeca-3-en, Heptadeca-4-en, Heptadeca-5-en, Heptadeca-6-en, Heptadeca-7-en, Heptadeca-8-en, Octadeca-1-en, Octadeca-2-en, Octadeca-3-en, Octadeca-4-en, Octadeca-5-en, Octadeca-6-en, Octadeca-7-en The compound according to claim 87, or a salt or isomer thereof, which is an alkenyl selected from the group consisting of octadeca-8-ene, octadeca-9-ene, nonadeca-1-ene, nonadeca-2-ene, nonadeca-3-ene, nonadeca-4-ene, nonadeca-5-ene, nonadeca-6-ene, nonadeca-7-ene, nonadeca-8-ene, nonadeca-9-ene, icosa-1-ene, icosa-2-ene, icosa-3-ene, icosa-4-ene, icosa-5-ene, icosa-6-ene, icosa-7-ene, icosa-8-ene, and icosa-9-ene.

111. R １１ and R １２ The compound according to claim 110, or a salt or isomer thereof, wherein at least one of the compounds comprises one or more additional double bonds.

112. Compounds selected from the following group: 【Chemical 325】 (9,9'-di(heptadecan-9-yl)=O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate),SM-048); 【Chemistry 326】 ((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=dioleate, SM-074); 【Chemistry 327】 (9,9'-bis(2-butyloctyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate),SM-076); 【Chemical 328】 (1-(heptadecan-9-yl)=9-(4-(2-(2-(1-(2-(4-(2-(2-(2-(4-(oleoyloxy)phenyl)acetoxy)ethyl)piperidine-1-yl)acetyl)piperazine-1-yl)-2-oxoethyl)piperidine-4-yl)ethoxy)-2-oxoethyl)phenyl)=nonanedioate, SM-077); 【Chemistry 329】 ((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=bis(9-((2-heptylnonanoyl)oxy)nonanoate),SM-079); 【Chemistry 330】 (9,9'-bis(2-ethylhexyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate),SM-083); 【Chemistry 331】 (Di(heptadecan-9-yl)9,9'-(((((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(oxy))bis(carbonyl))bis(azandiyl))dinonanoate;SM-085); and 【Chemistry 332】 (9,9'-di(heptadecan-9-yl)O'1,O1-(((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(methylene))=di(nonanedioate);SM-088), 【Chemical 333】 (7,7'-Dinonyl O'1,O1-((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(heptanedioate); SM-100) 【Chemistry 334】 ((di(heptadecan-9-yl)9,9'-((2,2'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(acetyl))bis(oxy))dinonanoete); SM-109) 【Chemistry 335】 (Di(heptadecan-9-yl)O,O'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=diglutarate); SM-113) 【Chemistry 336】 (Bis(9-(heptadecane-9-yloxy)-9-oxononyl)O,O'-(((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))=diterephthalate); SM-125), and their salts and isomers.

113. Lipid particles containing the compounds described in claims 87 to 112.

114. Lipid particles according to claim 113, further comprising a therapeutic agent.

115. The lipid particles according to claim 114, wherein the therapeutic agent is nucleic acid.

116. A pharmaceutical composition comprising the lipid particles described in claim 113, and a pharmaceutically acceptable additive, carrier, or diluent.

117. A nucleic acid-lipid particle for delivering nucleic acid cargo to a target, wherein the nucleic acid-lipid particle is a compound selected from the group consisting of the following: 【Chemistry 337】 (9,9'-di(heptadecan-9-yl)=O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate),SM-048); 【Chemical 338】 ((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=dioleate, SM-074); 【Chemistry 339】 (9,9'-bis(2-butyloctyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate),SM-076); 【Chemistry 340】 (1-(heptadecan-9-yl)=9-(4-(2-(2-(1-(2-(4-(2-(2-(2-(4-(oleoyloxy)phenyl)acetoxy)ethyl)piperidine-1-yl)acetyl)piperazine-1-yl)-2-oxoethyl)piperidine-4-yl)ethoxy)-2-oxoethyl)phenyl)=nonanedioate, SM-077); 【Chemistry 341】 ((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene)=bis(9-((2-heptylnonanoyl)oxy)nonanoate),SM-079); 【Chemistry 342】 (9,9'-bis(2-ethylhexyl)O'1,O1-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(nonanedioate),SM-083); 【Transformation 343】 (Di(heptadecan-9-yl)9,9'-(((((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(oxy))bis(carbonyl))bis(azandiyl))dinonanoate; SM-085); 【Transformation 344】 (9,9'-di(heptadecan-9-yl)O'1,O1-(((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(methylene))=di(nonanedioate);SM-088); 【Chemistry 345】 (7,7'-Dinonyl O'1,O1-((((((Piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=di(heptanedioate); SM-100), 【Transformation 346】 ((di(heptadecan-9-yl)9,9'-((2,2'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))bis(acetyl))bis(oxy))dinonanoete); SM-109), 【Transformation 347】 (Di(heptadecan-9-yl)O,O'-((((((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))bis(oxy))bis(2-oxoethane-2,1-diyl))bis(4,1-phenylene))=diglutarate); SM-113), and 【Transformation 348】 (Bis(9-(heptadecane-9-yloxy)-9-oxononyl)O,O'-(((piperazine-1,4-diylbis(2-oxoethane-2,1-diyl))bis(piperidine-1,4-diyl))bis(ethane-2,1-diyl))=diterephthalate); SM-125) Includes, Nucleic acid-lipid particles wherein the compound further constitutes about 30 to 70 mol%, or about 40 to 60 mol%, or about 50 mol%, of the total lipids present in the nucleic acid-lipid particles, and optionally constitutes 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, or 55 mol% of the total lipids present in the nucleic acid-lipid particles.

118. It contains a complex lipid that inhibits particle aggregation, constituting 0.01 to 2% of the total lipids present, and optionally the complex lipid contains a polyethylene glycol (PEG)-lipid complex, and optionally the PEG of the PEG-lipid complex has an average molecular weight of 550 daltons to 5000 daltons, and optionally the PEG-lipid complex is a PEG5000-lipid complex, and optionally the PEG-lipid complex is a PEG2000-lipid complex, and optionally the PEG2000-lipid complex contains one or more of 1,2-dimiristoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DMG-PEG2k) and 1,2-distearoyl-rac-glycero-3-methoxypolyethylene glycol-2000 (DSG-PEG2k), and optionally the PEG2000-lipid complex contains 1,2-dimiristoyl-rac-glycero- The nucleic acid-lipid particle according to claim 116, wherein the nucleic acid-lipid particle comprises cello-3-methoxypolyethylene glycol-2000 (DMG-PEG2k), and optionally the PEG2000-lipid complex is 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)-polyethylene glycol methoxy (DOPE-mPEG2k), and optionally the PEG2000-lipid complex is 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE-mPEG2k) complexed with methoxylpoly(ethylene glycol), and optionally the nucleic acid-lipid particle comprises a PEG-lipid complex at a concentration selected from the group consisting of about 0.5 mol% of the total lipids present in the nucleic acid-lipid particle, about 1.0 mol% of the total lipids present in the nucleic acid-lipid particle, and about 0.5 to 3.0 mol% of the total lipids present in the nucleic acid-lipid particle.

119. The nucleic acid-lipid particle according to claim 118, wherein the PEG-lipid complex is DMG-PEG2k constituting about 1.5 mol% of the total lipids present in the nucleic acid-lipid particle.

120. The nucleic acid-lipid particles according to claims 117 to 119, comprising one or more noncationic lipids constituting 20 mol% to 80 mol% of the total lipids present in the lipid-nucleic acid particles, wherein the one or more noncationic lipids optionally comprise cholesterol or a derivative thereof.

121. The nucleic acid-lipid particle according to claim 120, comprising cholesterol or a derivative in a concentration range selected from the group consisting of 35 mol% to 45 mol% of the total lipids present in the nucleic acid-lipid particle, 45 mol% to 55 mol% of the total lipids present in the nucleic acid-lipid particle, and 55 mol% to 65 mol% of the total lipids present in the nucleic acid-lipid particle, wherein the cholesterol or derivative may optionally constitute about 35%, about 36%, about 37%, about 38%, about 39%, about 40%, about 41%, about 42%, about 43%, about 44%, or about 45% of the total lipids present in the nucleic acid-lipid particle, and wherein the cholesterol or derivative may optionally constitute about 40% of the total lipids present in the nucleic acid-lipid particle.

122. The nucleic acid-lipid particle according to claims 117 to 121, comprising one or more noncationic lipids other than cholesterol or its derivatives, wherein the one or more noncationic lipids other than cholesterol or its derivatives optionally constitute 5 mol% to 20 mol% of the total lipids present in the lipid-nucleic acid particle, and wherein the one or more noncationic lipids other than cholesterol or its derivatives optionally constitute about 10 mol% of the total lipids present in the nucleic acid-lipid particle.

123. The nucleic acid-lipid particle according to claim 122, wherein the one or more noncationic lipids other than cholesterol or its derivatives include a noncationic lipid selected from the group consisting of 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), and β-sitosterol, and optionally the one or more noncationic lipids other than cholesterol or its derivatives are DOPE.

124. The nucleic acid-lipid particles according to claims 117 to 123, wherein the nucleic acid cargo comprises synthetic or naturally occurring RNA or DNA, or derivatives thereof, and optionally the nucleic acid cargo is modified RNA, and optionally the modified RNA is selected from the group consisting of modified mRNA, modified antisense oligonucleotides, and modified siRNA, and optionally the modified mRNA encodes a nucleic acid regulatory factor.

125. The nucleic acid cargo includes 2'-O-methyl modified nucleotides, nucleotides containing a 5'-phosphorothioate group, terminal nucleotides linked to cholesteryl derivatives, 2'-deoxy-2'-fluoro modified nucleotides, 5'-methoxy modified nucleotides (e.g., 5'-methoxyuridine), 2'-deoxy modified nucleotides, locked nucleotides, debased nucleotides, 2'-amino modified nucleotides, 2'-alkyl modified nucleotides, morpholino nucleotides, phosphoramidates, nucleotides containing unnatural bases; phosphorothioates, chiral phosphorothioates, phosphorodithioates, phosphotriesters, aminoalkyl phosphotriesters, and 3'-alkylenephosphonates. Nucleic acid-lipid particles according to claims 117 to 124, comprising one or more modifications selected from the group consisting of internucleoside links or skeletons, including methyl and other alkylphosphonates, phosphinates, 3'-aminophosphoramides and aminoalkylphosphoramides, thionophosphoramides, thionoalkylphosphonates, thionoalkylphosphotryesters, and boranophosphates having the usual 3'-5' linkage, their 2'-5' linked analogs, and internucleoside links or skeletons having inverted polarity, such as those in which pairs of adjacent nucleoside units are linked from 3'-5' to 5'-3' or 2'-5' to 5'-2'.

126. The nucleic acid-lipid particles according to claims 117 to 125, wherein the nucleic acid-lipid particles contain approximately 50 mol% of SM-048, SM-074, SM-076, SM-077, SM-079, SM-083, SM-085, SM-088, SM-100, SM-109, SM-113, or SM-125 of the total lipids present in the nucleic acid-lipid particles, approximately 38.5 mol% of cholesterol of the total lipids present in the nucleic acid-lipid particles, approximately 10 mol% of DOPE of the total lipids present in the nucleic acid-lipid particles, and approximately 1.5 mol% of DMG-PEG2k of the total lipids present in the nucleic acid-lipid particles.

127. A compound selected from the following group. 【Chemistry 349】

128. Compounds of formula IX: [Chemical 350] or its salt or isomer [in the formula, X ２ , X ３ , and X ４ These are independently CH or N, G ８ G ９ , and G １０ These are independently O, -(CO)O-, -CH ２ O(CO)-,-(CH ２ ) ２ (CO)NR １５ -, - (CH ２ ) O(CO)NR １５ - CH ２ (CO)NR １５ -, -(CO)NR １５ -, -NR １５ (CO)-, or NR １５ (CO)O-, where X ２ , X ３ , and X ４ If all are CH, then G ８ G ９ , and G １０ All of the following - (CO)NR １５ - That is not the case, a １ , b １ , and c １ These are independently 0, 1, 2, 3, or 4. I understand ６ , m ７ , and m ８ These are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. E ５ , E ６ , and E ７ These are independently -(CO)O- or -O(CO)-, T ５ , T ６ , and T ７ Independently, C ５ ~C ２２ Alkyl, C ５ ~C ２２ Alkenyl, or C ５ ~C ２２ It is alkinyl, R １ H, C １ ~C ６ Alkyl, C ２ ~C ６ Alkenyl, or C ２ ~C ６ It is Alkinnil.

129. I understand ６ ~m ８ , E ５ ~E ７ , and T ５ ~T ７ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein two or more sets of these sets are the same.

130. I understand ６ ~m ８ , E ５ ~E ７ , and T ５ ~T ７ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein one or more of the following are different.

131. I understand ６ ~m ８ , E ５ ~E ７ , and T ５ ~T ７ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein two of the compounds are the same.

132. a １ , b １ , or c １ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the compound is 2, 3, or 4.

133. a １ , b １ , and c １ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the compound is 2, 3, or 4.

134. a １ , b １ , or c １ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the compound is 3 or 4.

135. a １ and b １ 3, c １ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein is 4.

136. a １ , b １ , and c １ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein is 3.

137. I understand ６ , m ７ , or m ８ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the ratio is 1 to 8.

138. I understand ６ , m ７ , or m ８ The range is 5 to 8, and m as needed. ６ , m ７ , and m ８ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the ratio is 5 to 8.

139. I understand １ , m ２ , or m ３ The number is 5-6, and m as needed. ６ , m ７ , and m ８ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the ratio is 5 to 6.

140. I understand １ , m ２ , or m ３ It is 6, and m as needed ６ , m ７ , and m ８ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein is 6.

141. X ２ , X ３ , or X ４ One of them is N, and X as needed. ２ , X ３ , or X ４ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein two of the elements are N.

142. X ２ , X ３ , and X ４ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein CH is present.

143. G ８ G ９ , or G １０ Two of them are the same, and G as needed ８ G ９ , and G １０ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein all of the above are the same.

144. G ８ G ９ , or G １０ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein one or more of the following are different.

145. X ２ , X ３ , or X ４ One of them is N, and G ８ G ９ , and G １０ ga-(CO)NR １５ - The compound of formula IX according to claim 128, or a salt or isomer thereof.

146. G ８ G ９ , and G １０ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein is -(CO)O-.

147. G ８ G ９ , and G １０ Two of them are -(CO)NR １５ - The compound of formula IX according to claim 128, or a salt or isomer thereof.

148. G ８ G ９ , and G １０ One of them is -NR １５ A compound of formula IX according to claim 128, or a salt or isomer thereof, which is (CO)-.

149. G ８ G ９ , and G １０ All of them -NR １５ A compound of formula IX according to claim 128, or a salt or isomer thereof, which is (CO)-.

150. G ８ G ９ , and G １０ One of them is -(CO)NR １５ - and G ８ G ９ , and G １０ Two of them are -NR １５ A compound of formula IX according to claim 128, or a salt or isomer thereof, which is (CO)-.

151. G ８ G ９ , and G １０ One of them is -(CO)NR １５ - and G ８ G ９ , and G １０ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein two of the elements are O.

152. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C １８ Alkyl, C ５ ~C １８ Alkenyl and C ５ ~C １８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C １８ Alkyl, C ５ ~C １８ Alkenyl and C ５ ~C １８ A compound of formula IX according to claim 128, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

153. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ A compound of formula IX according to claim 128, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

154. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ A compound of formula IX according to claim 128, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

155. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ A compound of formula IX according to claim 128, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

156. T ５ , T ６ , or T ７ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ A compound of formula IX according to claim 128, or a salt or isomer thereof, independently selected from the group consisting of alkynyl compounds.

157. T ５ 、 T ６ 、 or T ７ is independently selected from the group consisting of C ７ or C ８ alkyl, C ７ or C ８ alkenyl, and C ７ or C ８ alkynyl, and optionally T ５ 、 T ６ 、 and T ７ is independently selected from the group consisting of C ７ or C ８ alkyl, C ７ or C ８ alkenyl, and C ７ or C ８ alkynyl, the compound of formula IX according to claim 128, or a salt or isomer thereof.

158. T ５ 、 T ６ 、 or T ７ is independently C ８ alkyl, C ８ alkenyl, or C ８ alkynyl, optionally T ５ 、 T ６ 、 and T ７ are each independently C ８ alkyl, C ８ alkenyl, or C ８ alkynyl, a compound of formula IX according to claim 128, or a salt or isomer thereof.

159. E ５ , E ６ , or E ７ is -(CO)O-, and E as needed ５ , E ６ , and E ７ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein is -(CO)O-.

160. T ５ , T ６ , and T ７ The compound of formula IX according to claim 128, or a salt or isomer thereof, wherein each is an alkyl selected from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as necessary.

161. T ５ , T ６ , and T ７ The compound of formula IX according to claim 128, or a salt or isomer thereof, wherein the compound is octane or tridecane.

162. T ５ , T ６ , and T ７ However, independently, each is substituted as needed: buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca-2-ene, deca-3-ene, deca-4-ene, deca-5-ene A compound of formula IX according to claim 128, or a salt or isomer thereof, which is an alkenyl selected from the group consisting of ene, deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and an alkenyl group having two or more double bonds.

163. T ５ , T ６ , and T ７ However, independently, each is substituted as needed: buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, deca-2-ine, deca-3-ine, deca-4-ine, deca-5-ine A compound of formula IX according to claim 128, or a salt or isomer thereof, which is an alkynyl selected from the group consisting of yin, deca-6-yin, undeca-1-yin, undeca-2-yin, undeca-3-yin, undeca-4-yin, undeca-5-yin, undeca-6-yin, undeca-7-yin, dodeca-1-yin, dodeca-2-yin, dodeca-3-yin, dodeca-4-yin, dodeca-5-yin, dodeca-6-yin, dodeca-8-yin, and an alkynyl group containing two or more triple bonds.

164. R １ A compound of formula IX according to claim 128, or a salt or isomer thereof, wherein is H.

165. Compounds selected from the following group: 【Chemistry 351】 (SM-016; 1-ethylhexyl 9-[3-[[3,5-bis[3-[bis[9-(1-ethylhexoxy)-9-oxononyl]amino]propylcarbamoyl]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxononyl]amino]nonanoate) 【Chemistry 352】 (SM-062; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-((((pyridine-2,4,6-tricarbonyl)tris(azandiyl))tris(propane-3,1-diyl))tris(azantriyl))hexanonaate), 【Chemistry 353】 (SM-065; Tris(3-(bis(9-(octan-3-yloxy)-9-oxononyl)amino)propyl)=benzene-1,3,5-tricarboxylate), 【Chemistry 354】 (SM-067; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(4-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)butanamide)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemical 355】 (SM-68; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-(((benzene-1,3,5-triyltris(azandiyl))tris(4-oxobutane-4,1-diyl))tris(azantriyl))hexanonaate), 【Transformation 356】 (SM-070; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)aminopropyl)carbamoyl)-1,3-phenylene)bis(azandiyl))bis(4-oxobutan-4,1-diyl))bis(azantriyl))tetranonanoate), 【Chemistry 357】 (SM-072; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)carbamoyl)-1,3-phenylene)bis(oxy))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemical 358】 (SM-073; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(2-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-2-oxoethyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemistry 359】 (SM-107; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(3-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-3-oxopropyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemical 360】 (SM-111; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(((5-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)pentanoyl)oxy)methyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), and 【Chemical 361】 (SM-112; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(19-ethyl-8-(9-(octane-3-yloxy)-9-oxononyl)-3,17-dioxo-2,18-dioxa-4,8-diazatetracosyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate).

166. Lipids in formula IX: 【Chemical 362】 or its salt or isomer [in the formula, X ２ , X ３ , and X ４ These are independently CH or N, G ８ G ９ , and G １０ These are independently O, -(CO)O-, -CH ２ O(CO)-,-(CH ２ ) ２ (CO)NR １５ -, - (CH ２ ) O(CO)NR １５ - CH ２ (CO)NR １５ -, -(CO)NR １５ -, -NR １５ (CO)-, or NR １５ (CO)O-, where X ２ , X ３ , and X ４ If all are CH, then G ８ G ９ , and G １０ All of the following - (CO)NR １５ - That is not the case, a １ , b １ , and c １ These are independently 0, 1, 2, 3, or 4. I understand ６ , m ７ , and m ８ These are independently 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. E ５ , E ６ , and E ７ These are independently -(CO)O- or -O(CO)-, T ５ 、T ６ 、and T ７ are, independently, C ５ to C ２２ alkyl, C ５ to C ２２ alkenyl, or C ５ to C ２２ alkynyl, and R １ H, C １ ~C ６ Alkyl, C ２ ~C ６ Alkenyl, or C ２ ~C ６ A pharmaceutical composition containing alkynyl.

167. I understand ６ ~m ８ , E ５ ~E ７ , and T ５ ~T ７ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein two or more sets of these sets are the same.

168. I understand ６ ~m ８ , E ５ ~E ７ , and T ５ ~T ７ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein one or more of the above are different.

169. I understand ６ ~m ８ , E ５ ~E ７ , and T ５ ~T ７ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein two of the components are the same.

170. a １ , b １ , or c １ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the lipid is 2, 3, or 4.

171. a １ , b １ , and c １ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the lipid is 2, 3, or 4.

172. a １ , b １ , or c １ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the lipid is 3 or 4.

173. a １ and b １ 3, c １ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the ratio is 4.

174. a １ , b １ , and c １ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the ratio is 3.

175. I understand １ , m ２ , or m ３ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the ratio is 1 to 8.

176. I understand １ , m ２ , or m ３ The range is 5 to 8, and m as needed. ６ , m ７ , and m ８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the ratio is 5 to 8.

177. I understand １ , m ２ , or m ３ The number is 5-6, and m as needed. ６ , m ７ , and m ８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the ratio is 5 to 6.

178. I understand １ , m ２ , or m ３ It is 6, and m as needed ６ , m ７ , and m ８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein is 6.

179. X ２ , X ３ , or X ４ One of them is N, and X as needed. ２ , X ３ , or X ４ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein two of the molecules are N.

180. X ２ , X ３ , and X ４ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein CH is present.

181. G ８ G ９ , or G １０ Two of them are the same, and G as needed ８ G ９ , and G １０ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein all of the above are the same.

182. G ８ G ９ , or G １０ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein one or more of the above are different.

183. X ２ , X ３ , or X ４ One of them is N, and G ８ G ９ , and G １０ ga-(CO)NR １５ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof.

184. G ８ G ９ , and G １０ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein is -(CO)O-.

185. G ８ G ９ , and G １０ Two of them are -(CO)NR １５ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof.

186. G ８ G ９ , and G １０ One of them is -NR １５ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, which is (CO)-, or a salt or isomer thereof.

187. G ８ G ９ , and G １０ All of them -NR １５ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, which is (CO)-, or a salt or isomer thereof.

188. G ８ G ９ , and G １０ One of them is -(CO)NR １５ - and G ８ G ９ , and G １０ Two of them are -NR １５ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, which is (CO)-, or a salt or isomer thereof.

189. G ８ G ９ , and G １０ One of them is -(CO)NR １５ - and G ８ G ９ , and G １０ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein two of the components are O.

190. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C １８ Alkyl, C ５ ~C １８ Alkenyl and C ５ ~C １８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C １８ Alkyl, C ５ ~C １８ Alkenyl and C ５ ~C １８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, independently selected from the group consisting of alkynyls.

191. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C １２ Alkyl, C ５ ~C １２ Alkenyl and C ５ ~C １２ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, independently selected from the group consisting of alkynyls.

192. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C １０ Alkyl, C ５ ~C １０ Alkenyl and C ５ ~C １０ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, independently selected from the group consisting of alkynyls.

193. T ５ , T ６ , or T ７ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ５ ~C ８ Alkyl, C ５ ~C ８ Alkenyl and C ５ ~C ８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, independently selected from the group consisting of alkynyls.

194. T ５ , T ６ , or T ７ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ６ ~C ８ Alkyl, C ６ ~C ８ Alkenyl and C ６ ~C ８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, independently selected from the group consisting of alkynyls.

195. T ５ , T ６ , or T ７ However, each is replaced as needed in C ７ or C ８ Alkyl, C ７ or C ８ Alkenyl and C ７ or C ８ Independently selected from the group consisting of alkynnyls, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ７ or C ８ Alkyl, C ７ or C ８ Alkenyl and C ７ or C ８ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, independently selected from the group consisting of alkynyls.

196. T ５ , T ６ , or T ７ However, C is independently substituted as needed. ８ Alkyl, C ８ Alkenyl, or C ８ It is an alkynyl, and T as needed. ５ , T ６ , and T ７ However, each is replaced as needed in C ８ Alkyl, C ８ Alkenyl, or C ８ A pharmaceutical composition comprising an alkynyl lipid of formula IX as described in claim 166, or a salt or isomer thereof.

197. E ５ , E ６ , or E ７ is -(CO)O-, and E as needed ５ , E ６ , and E ７ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein is -(CO)O-.

198. T ５ , T ６ , and T ７ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein each is independently an alkyl selected from the group consisting of butane, pentane, hexane, heptane, octane, nonane, decane, undecane, dodecane, and tridecane, each of which is substituted as necessary.

199. T ５ , T ６ , and T ７ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein the lipid is octane or tridecane.

200. T ５ , T ６ , and T ７ However, independently, each is substituted as needed: buta-1-ene, buta-2-ene, penta-1-ene, penta-2-ene, hexa-1-ene, hexa-2-ene, hexa-3-ene, hepta-1-ene, hepta-2-ene, hepta-3-ene, octa-1-ene, octa-2-ene, octa-3-ene, octa-4-ene, nona-1-ene, nona-2-ene, nona-3-ene, nona-4-ene, nona-5-ene, deca-1-ene, deca-2-ene, deca-3-ene, deca-4-ene, deca-5-ene, A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, which is an alkenyl selected from the group consisting of deca-6-ene, undeca-1-ene, undeca-2-ene, undeca-3-ene, undeca-4-ene, undeca-5-ene, undeca-6-ene, undeca-7-ene, dodeca-1-ene, dodeca-2-ene, dodeca-3-ene, dodeca-4-ene, dodeca-5-ene, dodeca-6-ene, dodeca-8-ene, and an alkenyl group having two or more double bonds.

201. T ５ , T ６ , and T ７ However, independently, each is substituted as needed: buta-1-ine, buta-2-ine, penta-1-ine, penta-2-ine, hexa-1-ine, hexa-2-ine, hexa-3-ine, hepta-1-ine, hepta-2-ine, hepta-3-ine, octa-1-ine, octa-2-ine, octa-3-ine, octa-4-ine, nona-1-ine, nona-2-ine, nona-3-ine, nona-4-ine, nona-5-ine, deca-1-ine, deca-2-ine, deca-3-ine, deca-4-ine, deca-5-ine, A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, which is an alkynyl selected from the group consisting of deca-6-in, undeca-1-in, undeca-2-in, undeca-3-in, undeca-4-in, undeca-5-in, undeca-6-in, undeca-7-in, dodeca-1-in, dodeca-2-in, dodeca-3-in, dodeca-4-in, dodeca-5-in, dodeca-6-in, dodeca-8-in, and an alkynyl having two or more triple bonds.

202. R １ A pharmaceutical composition comprising a lipid of formula IX according to claim 166, or a salt or isomer thereof, wherein H is present.

203. A pharmaceutical composition containing a lipid selected from the following group: 【Chemical 363】 (SM-016; 1-ethylhexyl 9-[3-[[3,5-bis[3-[bis[9-(1-ethylhexoxy)-9-oxononyl]amino]propylcarbamoyl]benzoyl]amino]propyl-[9-(1-ethylhexoxy)-9-oxononyl]amino]nonanoate) 【Chemical 364】 (SM-062; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-((((pyridine-2,4,6-tricarbonyl)tris(azandiyl))tris(propane-3,1-diyl))tris(azantriyl))hexanonaate), 【Chemical 365】 (SM-065; Tris(3-(bis(9-(octan-3-yloxy)-9-oxononyl)amino)propyl)=benzene-1,3,5-tricarboxylate), 【Chemical 366】 (SM-067; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(4-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)butanamide)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemical 367】 (SM-068; Hexa(octan-3-yl)9,9',9'',9''',9'''',9'''''-(((benzene-1,3,5-triyltris(azandiyl))tris(4-oxobutane-4,1-diyl))tris(azantriyl))hexanonaate), 【Chemical 368】 (SM-070; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)aminopropyl)carbamoyl)-1,3-phenylene)bis(azandiyl))bis(4-oxobutan-4,1-diyl))bis(azantriyl))tetranonanoate), 【Chemical 369】 (SM-072; Tetra(octane-3-yl)9,9',9'',9'''-((((5-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)carbamoyl)-1,3-phenylene)bis(oxy))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemistry 370】 (SM-073; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(2-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-2-oxoethyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemistry 371】 (SM-107; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(3-((3-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)propyl)amino)-3-oxopropyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), 【Chemistry 372】 (SM-111; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(((5-(bis(9-(octane-3-yloxy)-9-oxononyl)amino)pentanoyl)oxy)methyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate), and 【Chemistry 373】 (SM-112; Tetra(octane-3-yl)9,9',9'',9'''-((((5-(19-ethyl-8-(9-(octane-3-yloxy)-9-oxononyl)-3,17-dioxo-2,18-dioxa-4,8-diazatetracosyl)isophthaloyl)bis(azandiyl))bis(propane-3,1-diyl))bis(azantriyl))tetranonanoate).

204. Lipid particles comprising the compounds described in claims 128 to 165.

205. Lipid particles according to claim 204, further comprising a therapeutic agent

206. Lipid particles according to claim 204, wherein the pretreatment agent is nucleic acid.

207. A pharmaceutical composition comprising lipid particles according to claim 204, and a pharmaceutically acceptable additive, carrier, or diluent.

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