Ionizable lipid compound, lipid nanoparticle comprising same, and use of ionizable lipid compound

Abstract

The present invention provides a compound represented by formula (I) or a pharmaceutically acceptable salt, isomer, isotope compound or solvate of the compound represented by formula (I). The present invention also provides a lipid nanoparticle comprising the compound represented by formula I or the pharmaceutically acceptable salt, isomer, isotope compound or solvate of the compound represented by formula I. The lipid nanoparticle can be used for nucleic acid delivery and has high organ targeting efficiency.

Description

Ionizable lipid compound, lipid nanoparticles containing the same and applications thereof Technical Field

[0001] The present invention relates to the field of biomedicine technology, and in particular to an ionizable lipid compound, lipid nanoparticles containing the same, and applications thereof. Background Art

[0002] Lipid nanoparticles (LNPs) are lipid vesicles with a uniform lipid core and are widely used for the delivery of small molecules and nucleic acid drugs. Naked mRNA is inherently unstable and susceptible to rapid degradation by nucleases and autolysis. Encapsulation in LNPs can protect mRNA from extracellular RNases and facilitate its intracellular delivery.

[0003] In LNP formulations, ionizable lipids are neutral at physiological pH, but positively charged in the acidic environment of the body. Due to the significant improvement in effectiveness and toxicity characteristics, the pH-dependent ionization ability makes ionizable lipids a suitable material for nucleic acid delivery. Some existing LNPs can be observed to have significant organ toxicity after delivery and administration, which is mainly due to the current problem of low organ targeting of LNPs in nucleic acid delivery. Therefore, there is a need to develop more new cationic lipid compounds with higher organ targeting for nucleic acid delivery, and lipid nanoparticles containing them. Summary of the Invention

[0004] When targeting specific diseases, accurate and efficient LNP targeted delivery is particularly critical. For example, protein replacement therapy for the treatment of rare liver diseases requires LNP to be concentratedly targeted to the liver. The inventors found that the organ toxicity (such as hepatotoxicity) of existing LNPs is mainly due to the ionizable lipids of the core component of LNPs. Therefore, it is urgent to provide a class of compounds that can achieve accurate and efficient organ-targeted delivery while having excellent biosafety as ionizable lipids and LNPs containing them.

[0005] In order to solve one of the above-mentioned technical problems existing in the prior art, the present invention provides a novel compound that can be used to prepare lipid nanoparticles, which has organ-targeting properties for the delivery of nucleic acids such as mRNA.

[0006] The first aspect of the present invention provides a compound, which is a compound represented by Formula I or a pharmaceutically acceptable salt, pharmaceutically acceptable ester, isomer, isotope-labeled compound (especially deuterated compound) or solvate of the compound represented by Formula I:

[0007] Wherein, X1 and X2 are the same or different and are each independently selected from -O- or -NR a -;

[0008] R1, R2, R3 and R4 are the same or different and are each independently selected from a C2-C30 saturated or unsaturated aliphatic hydrocarbon group, wherein one or more methylene units in the C2-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; hydrogen atoms on carbon atoms in the C2-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from hydroxyl groups and C1-C6 alkoxy groups;

[0009] L1 and L2 are the same or different and are each independently selected from a C1-C20 saturated or unsaturated aliphatic alkylene group, a C6-C15 arylene group, a C3-C15 heteroarylene group, a 5-6 membered heterocycloalkylene group or a combination thereof, wherein one or more methylene units in the C1-C20 saturated or unsaturated aliphatic alkylene group are optionally and independently replaced by L;

[0010] L is selected from -O-, -(C=O)O-, -NR a -, -(C=O)NR a - or a combination thereof;

[0011] R a Selected from hydrogen or C1-C6 alkyl.

[0012] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C6-C30 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C6-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C6-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0013] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C30 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C8-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C8-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0014] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C6-C24 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C6-C24 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C6-C24 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from hydroxyl groups and C1-C6 alkoxy groups.

[0015] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C24 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C8-C24 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C8-C24 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0016] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C22 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C8-C22 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C8-C22 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0017] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C20 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C8-C20 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C8-C20 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0018] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C18 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C8-C18 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C8-C18 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0019] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C11-C21 saturated or unsaturated aliphatic hydrocarbon group, one or more methylene units in the C8-C21 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; the hydrogen atoms on the carbon atoms in the C11-C21 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from a hydroxyl group and a C1-C6 alkoxy group.

[0020] According to some embodiments of the present invention, the saturated or unsaturated aliphatic hydrocarbon group is a linear or branched saturated or unsaturated aliphatic hydrocarbon group.

[0021] According to some embodiments of the present invention, the saturated or unsaturated aliphatic hydrocarbon group is an unsaturated aliphatic hydrocarbon group.

[0022] According to some embodiments of the present invention, the unsaturated aliphatic hydrocarbon group has 1-3 carbon-carbon double bonds and / or carbon-carbon triple bonds. In some embodiments, the unsaturated aliphatic hydrocarbon group has 1 carbon-carbon double bond or carbon-carbon triple bond. In some embodiments, the unsaturated aliphatic hydrocarbon group has 2 carbon-carbon double bonds and / or carbon-carbon triple bonds. In some embodiments, the unsaturated aliphatic hydrocarbon group has 3 carbon-carbon double bonds and / or carbon-carbon triple bonds.

[0023] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl, C3-C20 cycloalkyl, C4-C24 alkenyl, C4-C24 alkynyl, and one or more methylene units in the C4-C24 alkyl, C3-C20 cycloalkyl, C4-C24 alkenyl, C4-C24 alkynyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on carbon atoms in the C4-C24 alkyl, C3-C20 cycloalkyl, C4-C24 alkenyl, C4-C24 alkynyl are optionally substituted by one or more hydroxyl groups.

[0024] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C6-C24 alkyl, C6-C24 alkenyl, one or more methylene units in the C6-C24 alkyl, C6-C24 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C6-C24 alkyl, C6-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0025] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C6-C22 alkyl, C6-C22 alkenyl, one or more methylene units in the C6-C22 alkyl, C6-C22 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C6-C22 alkyl, C6-C22 alkenyl are optionally substituted by one or more hydroxyl groups.

[0026] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C8-C24 alkyl, C8-C24 alkenyl, one or more methylene units in the C8-C24 alkyl, C8-C24 alkenyl are optionally and independently replaced by a group selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C8-C24 alkyl, C8-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0027] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C8-C22 alkyl, C8-C22 alkenyl, one or more methylene units in the C8-C22 alkyl, C8-C22 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C8-C22 alkyl, C8-C22 alkenyl are optionally substituted by one or more hydroxyl groups.

[0028] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C8-C20 alkyl, C8-C20 alkenyl, one or more methylene units in the C8-C20 alkyl, C8-C20 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C8-C20 alkyl, C8-C20 alkenyl are optionally substituted by one or more hydroxyl groups.

[0029] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C8-C18 alkyl, C8-C18 alkenyl, one or more methylene units in the C8-C18 alkyl, C8-C18 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C8-C18 alkyl, C8-C18 alkenyl are optionally substituted by one or more hydroxyl groups.

[0030] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C10-C16 alkyl, C10-C18 alkenyl, one or more methylene units in the C10-C16 alkyl, C10-C18 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C10-C16 alkyl, C10-C18 alkenyl are optionally substituted by one or more hydroxyl groups.

[0031] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C11-C21 alkyl, C11-C21 alkenyl, one or more methylene units in the C11-C21 alkyl, C11-C21 alkenyl are optionally and independently replaced by groups selected from -O(C=O)-, -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C11-C21 alkyl, C11-C21 alkenyl are optionally substituted by one or more hydroxyl groups.

[0032] According to some embodiments of the present invention, R1, R2, R3 and R4 are each independently selected from a C4-C20 alkyl group, a C4-C20 alkenyl group, a combination of a C4-C20 alkyl group and -(C=O)O-, a combination of a C4-C20 alkyl group and -(C=O)NH-, a combination of a C4-C20 alkenyl group and -(C=O)O-, and a combination of a C4-C20 alkenyl group and -(C=O)NH-; optionally, the C4-C20 alkyl group and the C4-C20 alkenyl group are substituted by one or more hydroxyl groups.

[0033] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C6-C22 alkyl group or a C4-C18 alkenyl group, and one or more methylene units in the C6-C22 alkyl group or the C4-C18 alkenyl group are optionally and independently replaced by a group selected from -(C=O)O- and -(C=O)NH-; and the hydrogen atoms on the carbon atoms in the C6-C22 alkyl group or the C4-C18 alkenyl group are optionally substituted by one or more hydroxyl groups.

[0034] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C11-C21 alkyl, C11-C21 alkenyl, and one methylene unit in the C11-C21 alkyl, C11-C21 alkenyl is optionally and independently replaced by a group selected from -(C=O)O- and -(C=O)NH-. According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C11-C21 alkyl, C11-C21 alkenyl.

[0035] In some embodiments, the C11-C21 alkenyl group has 1, 2, or 3 carbon-carbon double bonds. In some specific embodiments, the C11-C21 alkenyl group has 1 carbon-carbon double bond. In some embodiments, the C11-C21 alkenyl group has 2 or 3 carbon-carbon double bonds, preferably, the 2 or 3 carbon-carbon double bonds are not conjugated.

[0036] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C20 alkyl group or a C8-C20 alkenyl group. In some embodiments, the C8-C20 alkenyl group has 1, 2 or 3 carbon-carbon double bonds. In some specific embodiments, the C8-C20 alkenyl group has 2 carbon-carbon double bonds. In some specific embodiments, when the C8-C20 alkenyl group has more than 2 carbon-carbon double bonds, there is a spacing of 2-4 carbon atoms between (each) two carbon-carbon double bonds.

[0037] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are independently selected from C11-C21 linear or branched alkyl, C8-C18 linear or branched alkenyl.

[0038] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from C11-C21 linear alkyl and C8-C21 linear alkenyl.

[0039] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different, and are each independently selected from a C8-C18 linear or branched alkyl group, a C8-C18 linear or branched alkenyl group.

[0040] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different, and are each independently selected from C8-C18 linear alkyl and C8-C18 linear alkenyl.

[0041] In some specific embodiments, R1, R2, R3 and R4 are each independently selected from (straight-chain) alkenyl (such as C4-C24 (straight-chain) alkenyl, C4-C20 (straight-chain) alkenyl, C4-C18 (straight-chain) alkenyl, C6-C24 (straight-chain) alkenyl, C6-C22 (straight-chain) alkenyl, C8-C24 (straight-chain) alkenyl, C8-C22 (straight-chain) alkenyl, C8-C20 (straight-chain) alkenyl, C8-18 (straight-chain) alkenyl or C10-C18 (straight-chain) alkenyl), and when there are two or more carbon-carbon double bonds in the (straight-chain) alkenyl, (each) two carbon-carbon double bonds may be separated by 1 carbon atom, 2 carbon atoms or 3 carbon atoms, preferably by 1 carbon atom.

[0042] In some embodiments, the carbon-carbon double bond is at least 1, 2, 3, 4, 5, 6, 7, 8, or 9 carbon atoms away from the end of the molecule. In some embodiments, the number of methylene groups on the left and right sides of the carbon-carbon double bond differs by no more than 3, such as no more than 2, no more than 1, or preferably, the difference is 0.

[0043] In some specific embodiments, R1, R2, R3 and R4 are each independently selected from (straight chain) alkenyl (such as C4-C24 (straight chain) alkenyl, C4-C20 (straight chain) alkenyl, C4-C18 (straight chain) alkenyl, C6-C24 (straight chain) alkenyl, C6-C22 (straight chain) alkenyl, C8-C24 (straight chain) alkenyl, C8-C22 (straight chain) alkenyl, C8-C20 (straight chain) alkenyl, C8-18 (straight chain) alkenyl or C10-C18 (straight chain) alkenyl), and there is one carbon-carbon double bond in the (straight chain) alkenyl.

[0044] According to some embodiments of the present invention, R1, R2, R3 and R4 are each independently selected from the following groups:

[0045] wherein n1, n2, n5 and n6 are each independently selected from 1, 2, 3, 4, 5, 6, 7 or 8; n3, n4, n7, n8, n9 and n10 are each independently selected from 1, 2, 3, 4, 5, 6, 7 or 8; 10 Each independently selected from 0, 1, 2, 3, 4 or 5; n 11 Selected from 1, 2, 3, 4, 5, 6, 7 or 8; n 12 and n 13 Each is independently selected from 2, 3, 4, 5, 6 or 7.

[0046] According to some embodiments of the present invention, R1, R2, R3 and R4 are the same or different and are each independently selected from the following groups:

[0047] According to some embodiments of the present invention, L1 and L2 are the same or different and are each independently selected from C1-C15 alkylene, C3-C10 cycloalkylene, C6-C15 arylene, C3-C15 heteroarylene, 5-6 membered heterocycloalkylene or a combination thereof, wherein one or more methylene units in the C1-C15 alkylene and C3-C10 cycloalkylene are optionally and independently selected from -O- and -NR a -group replacement; the 5-6 membered heterocycloalkylene group has at least one heteroatom, and the heteroatom is selected from N, O and S.

[0048] According to some embodiments of the present invention, L1 and L2 are the same or different and are each independently selected from C2-C15 alkylene, C3-C10 cycloalkylene, C6-C15 arylene, C3-C15 heteroarylene, a combination of C2-C15 alkylene and 5-6 membered heterocycloalkylene, a combination of C2-C15 alkylene and C6-C15 arylene, a combination of C2-C15 alkylene and C3-C15 heteroarylene; 1 to 4 methylene units in the C2-C15 alkylene and C3-C10 cycloalkylene are optionally and independently selected from -O- and -NR a -group replacement; the 5-6 membered heterocycloalkylene group has 1-4 heteroatoms, and the heteroatoms are selected from N, O and S.

[0049] According to some embodiments of the present invention, L1 and L2 are each independently selected from C2-C15 alkylene, C6-C10 arylene, C3-C10 heteroarylene, a combination of C2-C10 alkylene and 5-6 membered heterocycloalkylene, a combination of C2-C10 alkylene and C6-C10 arylene, a combination of C2-C10 alkylene and C3-C10 heteroarylene; 1 to 4 methylene units in the C2-C15 alkylene are optionally and independently selected from -O- and -NR a -; the 5-6 membered heterocycloalkylene group has 1-3 hetero atoms.

[0050] According to some embodiments of the present invention, L1 and L2 are each independently selected from a C2-C15 alkylene group, a C3-C6 heteroarylene group, a combination of a C2-C8 alkylene group and a 5-6-membered heterocycloalkylene group, a combination of a C2-C8 alkylene group and a C6-C10 arylene group, and a combination of a C2-C8 alkylene group and a C3-C6 heteroarylene group; 1 to 4 methylene units in the C2-C15 alkylene group are optionally and independently replaced by a group selected from -O- and -NH-; the 5-6-membered heterocycloalkylene group has 1-3 (e.g., 1, 2 or 3) N atoms.

[0051] According to some embodiments of the present invention, L1 and L2 are each independently selected from C2-C10 alkylene groups. According to some embodiments of the present invention, L1 and L2 are each independently selected from C2-C8 alkylene groups, such as ethylene, propylene, butylene, pentylene, hexylene, heptylene, octylene, etc. According to some embodiments of the present invention, L1 and L2 are each independently selected from C2-C8 straight-chain alkylene groups.

[0052] According to some embodiments of the present invention, L1 and L2 are each independently selected from the following groups:

[0053] According to some embodiments of the present invention, in Formula I: X1 and X2 are the same or different and are each independently selected from -NR a -, R a Selected from hydrogen or C1-C6 alkyl.

[0054] According to some embodiments of the present invention, in Formula I: X1 and X2 are the same or different and are each independently selected from -NR a -, R a Selected from hydrogen or C1-C4 alkyl.

[0055] According to some embodiments of the present invention, in Formula I: X1 and X2 are the same.

[0056] According to some embodiments of the present invention, in Formula I: X1 and X2 are both -NH-.

[0057] According to some embodiments of the present invention, in Formula I:

[0058] X1 and X2 are the same or different and are each independently selected from -NR a -preferably -NH-; and / or

[0059] L1 and L2 are the same or different and are each independently selected from C2-C8 alkylene; for example, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene or C8 alkylene; preferably C2-C8 straight-chain alkylene, for example, C4 straight-chain alkylene, C5 straight-chain alkylene, C6 straight-chain alkylene, C7 straight-chain alkylene or C8 straight-chain alkylene; and / or

[0060] R1, R2, R3 and R4 are the same or different and are each independently selected from C11-C21 alkyl, C11-C21 alkenyl, preferably C11-C21 straight-chain alkyl, C11-C21 straight-chain alkenyl, such as C11 alkyl, C13 alkyl, C15 alkyl, C17 alkyl, C21 alkyl, C15 alkenyl, C17 alkenyl, C21 alkenyl; in the C11-C21 alkenyl, the carbon-carbon double bond is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 carbon atoms away from the end of the molecule.

[0061] R1, R2, R3 and R4 are the same or different and are each independently selected from C8-C18 alkyl, C8-C18 alkenyl, preferably C8-C18 straight-chain alkyl, C8-C18 straight-chain alkenyl, such as C10 alkyl, C12 alkyl, C14 alkyl, C16 alkyl, C18 alkyl, C10 alkenyl, C12 alkenyl, C14 alkenyl, C16 alkenyl, C18 alkenyl, in which the carbon-carbon double bond is at least 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14 or 15 carbon atoms away from the end of the molecule.

[0062] According to some embodiments of the present invention, in Formula I: X1 and X2 are the same or different and are each independently selected from -NR a -, R a is selected from hydrogen or C1-C6 alkyl; L1 and L2 are the same or different and are each independently selected from C2-C8 alkylene; R1, R2, R3 and R4 are the same or different and are each independently selected from C0-C16 alkyl, C10-C16 alkenyl.

[0063] According to some embodiments of the present invention, in Formula I: X1 and X2 are the same and are both selected from -NR a -, R a Selected from hydrogen or C1-C6 alkyl; L1 and L2 are the same and are both selected from C2-C8 alkylene; R1, R2, R3 and R4 are the same and are all selected from C8-C18 alkyl, C8-C18 alkenyl.

[0064] According to some embodiments of the present invention, in Formula I: X1 and X2 are the same and are both selected from -NH-; L1 and L2 are the same and are both selected from C2-C8 straight-chain alkylene; R1, R2, R3 and R4 are the same and are all selected from C8-C18 straight-chain alkyl and C8-C18 straight-chain alkenyl.

[0065] According to some embodiments of the present invention, in Formula I:

[0066] X1 and X2 are the same or different and are each independently selected from -NR a -, R a is selected from hydrogen or C1-C6 alkyl;

[0067] L1 and L2 are the same or different and are each independently selected from a C2-C15 alkylene group, a C6-C10 arylene group, a C3-C10 heteroarylene group, a combination of a C2-C10 alkylene group and a 5-6 membered heterocycloalkylene group, a combination of a C2-C10 alkylene group and a C6-C10 arylene group, or a combination of a C2-C10 alkylene group and a C3-C10 heteroarylene group; 1 to 4 methylene units in the C2-C15 alkylene group are independently selected from -O- and -NRa - group replacement; the 5-6 membered heterocycloalkylene group has 1-3 heteroatoms;

[0068] R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl and C4-C24 alkenyl, one or more methylene units in the C4-C24 alkyl and C4-C24 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C4-C24 alkyl and C4-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0069] According to some embodiments of the present invention, in Formula I:

[0070] X1 and X2 are the same or different and are each independently selected from -NR a -, R a is selected from hydrogen or C1-C6 alkyl;

[0071] L1 and L2 are the same or different and are each independently selected from a C2-C15 alkylene group, a C6-C10 arylene group, a C3-C10 heteroarylene group, a combination of a C2-C10 alkylene group and a 5-6-membered heterocycloalkylene group, a combination of a C2-C10 alkylene group and a C6-C10 arylene group, or a combination of a C2-C10 alkylene group and a C3-C10 heteroarylene group; 1 to 4 methylene units in the C2-C15 alkylene group are independently replaced by a group selected from -O- and -NH-; the 5-6-membered heterocycloalkylene group has 1 to 3 heteroatoms;

[0072] R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl and C4-C24 alkenyl, 1 to 4 methylene units in the C4-C24 alkyl and C4-C24 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C4-C24 alkyl and C4-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0073] According to some embodiments of the present invention, in Formula I:

[0074] X1 and X2 are both -NH-;

[0075] L1 and L2 are the same and are each independently selected from a C2-C15 alkylene group, a C3-C6 heteroarylene group, a combination of a C2-C8 alkylene group and a 5-6-membered heterocycloalkylene group, a combination of a C2-C8 alkylene group and a C6-C10 arylene group, and a combination of a C2-C8 alkylene group and a C3-C6 heteroarylene group; 1 to 4 methylene units in the C2-C15 alkylene group are optionally and independently replaced by a group selected from -O- and -NH-; the 5-6-membered heterocycloalkylene group has 1 to 3 N atoms;

[0076] R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl and C4-C24 alkenyl, 1 to 4 methylene units in the C4-C24 alkyl and C4-C24 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C4-C24 alkyl and C4-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0077] According to some embodiments of the present invention, in Formula I:

[0078] X1 and X2 are both -NH-;

[0079] L1 and L2 are the same and are each independently selected from C2-C15 alkylene, C6 heteroarylene, (C2-C8) alkylene-6-membered heterocycloalkylene-(C2-C8) alkylene, (C2-C8) alkylene-(C6-C10) arylene-(C2-C8) alkylene; 1 to 4 methylene units in the C2-C15 alkylene are optionally and independently replaced by a group selected from -O-, -NH- and -N(C1-C6 alkyl)-, preferably Preferably, when 2 to 4 methylene units in the C2-C15 alkylene group are independently replaced by -O- or -NH-, there are 1 to 6 (preferably 2, 3, 4, 5 or 6) methylene groups between -O- and / or -NH- in the replaced group; the C2-C15 alkylene group may be linear or branched; the (C2-C8) alkylene group may be linear or branched, preferably linear; the 6-membered heterocycloalkylene group has 1 to 3 (e.g., 2) N atoms;

[0080] R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl and C4-C24 alkenyl, 1 to 4 methylene units in the C4-C24 alkyl and C4-C24 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C4-C24 alkyl and C4-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0081] According to some embodiments of the present invention, in Formula I:

[0082] X1 and X2 are both -NH-;

[0083] L1 and L2 are the same and are independently selected from C2-C15 alkylene, pyridylene (such as 3,4-pyridylene ), (C2-C8) alkylene-piperazinylene-(C2-C8) alkylene (such as (C2-C8) alkylene-1,4-piperazinylene-(C2-C8) alkylene), (C2-C8) alkylene-C6 arylene-(C2-C8) alkylene (such as (C2-C8) alkylene-phenylene-(C2-C8) alkylene); 1 to 4 methylene units in the C2-C15 alkylene are optionally and independently selected from -O-, -NH- and -N(C1-C6 alkyl)- replacement groups. Preferably, when 2 to 4 methylene units in the C2-C15 alkylene group are independently replaced by -O- or -NH-, there are 1 to 6 (preferably 2, 3, 4, 5 or 6) methylene groups between -O- and / or -NH- in the replaced group. The C2-C15 alkylene group may be linear or branched. The (C2-C8) alkylene group may be linear or branched, preferably linear.

[0084] R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl and C4-C24 alkenyl, 1 to 4 methylene units in the C4-C24 alkyl and C4-C24 alkenyl are optionally and independently replaced by groups selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on the carbon atoms in the C4-C24 alkyl and C4-C24 alkenyl are optionally substituted by one or more hydroxyl groups.

[0085] According to some embodiments of the present invention, in Formula I: R1, R2, R3 and R4 are each independently selected from the following groups:

[0086] and

[0087] L1 and L2 are each independently selected from the following groups: For example, both L1 and L2 are and

[0088] X1 and X2 are both -NH-.

[0089] According to some embodiments of the present invention, in Formula I: R1, R2, R3 and R4 are each independently selected from the following groups:

[0090] and

[0091] L1 and L2 are each independently selected from the following groups:

[0092] and

[0093] X1 and X2 are both -NH-.

[0094] According to some embodiments of the present invention, the compound has a structure represented by Formula I-1 or Formula I-2:

[0095] wherein, X1, X2, R1, R2, R3, R4, L1 and L2 are as defined in Formula I.

[0096] According to some embodiments of the present invention, the compound has a structure represented by Formula I-3 or Formula I-4:

[0097] wherein, the definitions of R1, R2, R3, R4, L1 and L2 are the same as those in Formula I.

[0098] According to some embodiments of the present invention, in the above formula I, formula I-1 to formula I-4, R1 and R2 are the same. According to some embodiments of the present invention, in the above formula I, formula I-1 to formula I-4, R3 and R4 are the same.

[0099] According to some embodiments of the present invention, in the above Formula I, Formula I-1 to Formula I-4, R1, R2, R3 and R4 are all the same.

[0100] According to some embodiments of the present invention, in the above Formula I, Formula I-1 to Formula I-4, L1 and L2 are the same.

[0101] According to some embodiments of the present invention, in the above Formula I, Formula I-1 to Formula I-4, L1 and L2 are different.

[0102] According to some embodiments of the present invention, in the above formula I, formula I-1 to formula I-4, L1 and L2 are not propylene groups at the same time.

[0103] According to some embodiments of the present invention, the compound has a structure represented by Formula I-5:

[0104] wherein R1 to R4 are as defined in Formula I, m1 and m2 are the same or different and are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8.

[0105] In some embodiments, in Formula I-5, R1 to R4 are the same. In some embodiments, in Formula I-5, R1 to R4 are different. In some embodiments, in Formula I-5, R1 to R4 are the same and are independently selected from C8-C20 alkyl groups. In some embodiments, in Formula I-5, R1 to R4 are the same and are independently selected from C8-C20 straight-chain alkyl groups. In some embodiments, in Formula I-5, R1 to R4 are the same and are independently selected from C12-C16 straight-chain alkyl groups.

[0106] In some embodiments, in formula I-5, m1 and m2 are the same or different and are independently selected from 2, 4, 5, 6, 7 or 8. In some embodiments, in formula I-5, m1 and m2 are the same or different and are independently selected from 2, 4, 5, 6 or 8.

[0107] According to some embodiments of the present invention, the compound has a structure represented by Formula I-6:

[0108] wherein R1 to R4 are the same or different and are independently selected from C8-C20 alkyl groups; m1 and m2 are different and are independently selected from 2, 4, 5, 6 or 8.

[0109] In some embodiments, in Formula I-6, R1 to R4 are the same. In some embodiments, R1 to R4 are the same or different and are independently selected from C8-C20 straight chain alkyl groups. In some embodiments, R1 to R4 are the same or different and are independently selected from C12-C16 straight chain alkyl groups.

[0110] According to some embodiments of the present invention, the compound has a structure represented by Formula I-7:

[0111] Wherein, the definitions of L1 and L2 are the same as those described in Formula I; L3 to L6 are the same or different and are each independently selected from C2-C6 alkylene, preferably, C2-C6 straight-chain alkylene, such as ethylene, propylene, butylene, pentylene, hexylene; R5 to R8 are respectively defined the same as R1 to R4 in Formula I.

[0112] In some embodiments, R5 to R8 are the same or different and are independently selected from C5-C19 alkyl and C5-C19 alkenyl, preferably, C7-C17 straight chain alkyl and C7-C17 straight chain alkenyl, for example, C7 straight chain alkyl, C9 straight chain alkyl, C11 straight chain alkyl, C15 straight chain alkyl, C17 straight chain alkenyl.

[0113] In some embodiments, L1 and L2 are the same.

[0114] In some embodiments, L3 and L4 are the same as each other, and / or L5 and L6 are the same as each other. Preferably, L3 to L6 are the same as each other.

[0115] In some embodiments, R5 and R6 are the same as each other, and / or R7 and R8 are the same as each other. Preferably, R5 to R8 are the same as each other.

[0116] In some embodiments, L1 and L2 are the same; L3 to L6 are the same as each other; and R5 to R8 are the same as each other.

[0117] In some embodiments, L1 and L2 are different.

[0118] In some embodiments, L3 and L4 are different, and / or L5 and L6 are different. Preferably, at least two of L3 to L6 are different.

[0119] In some embodiments, R5 and R6 are different, and / or R7 and R8 are different. Preferably, at least two of R5 to R8 are different.

[0120] In some embodiments, L1 and L2 are independently selected from ethylene, butylene, and hexylene.

[0121] In some embodiments, L3 to L6 are ethylene.

[0122] In some embodiments, R5 to R8 are the same or different and are each independently selected from C7 linear alkyl, C9 linear alkyl, C11 linear alkyl, C15 linear alkyl, and C17 linear alkenyl.

[0123] According to some embodiments of the invention, the compound is not:

[0124] According to some embodiments of the present invention, L1 and L2 are not both propylene (-CH2CH2CH2-).

[0125] According to some embodiments of the present invention, the compound is selected from the following structures:

[0126] According to some embodiments of the present invention, the compound is selected from the following structures:

[0127] In the above structure, the value of each n is independently selected from 1, 2, 3, 4, 5, 6, 7 or 8. When n is 2-8, The group in the bracketed portion may contain an alkenyl group, preferably 1, 2, 3 or 4 alkenyl groups. In some preferred embodiments, in the above structure, the value of each n is independently selected from 1, 3, 4 or 5.

[0128] According to some embodiments of the present invention, the compound is selected from the following structures:

[0129] The second aspect of the present invention provides a method for preparing the compound of the first aspect, comprising the steps of:

[0130] (1) reacting a compound of formula A with an amine compound B to obtain an intermediate of formula C; wherein, in formula A, R X represents a C1-C10 straight chain or branched alkyl group, preferably a C1-C4 straight chain or branched alkyl group; the amine compound B includes H2 N -L1- X 1H and H2 N -L2- X 2 H , L1, L2, X1, X2 are as defined in Formula I;

[0131] (2) reacting the intermediate represented by formula C with an aldehyde compound R'CHO or an alcohol compound ROH to obtain a compound represented by formula I; wherein R' represents an R group that has lost a methylene unit, and R is defined as R1, R2, R3 and R4 in formula I;

[0132] According to some embodiments of the present invention, in step (1), the molar ratio of the compound of formula A to the amine compound B is 1:(2-6), preferably 1:(3-5).

[0133] According to some embodiments of the present invention, in step (1), the reaction temperature is 50-80°C.

[0134] According to some embodiments of the present invention, in step (2), the molar ratio of the intermediate represented by formula C to the aldehyde compound R'CHO or the alcohol compound ROH is 1:(4-10), preferably 1:(6-8).

[0135] According to some embodiments of the present invention, in step (2), the reaction temperature is 20-35°C.

[0136] According to some embodiments of the present invention, in step (2), the reaction is carried out in the presence of a solvent and a catalyst. In some embodiments, the solvent includes a halogenated hydrocarbon solvent and an alcohol solvent. In some specific embodiments, the solvent includes dichloromethane and methanol. In some embodiments, the volume ratio of the halogenated hydrocarbon solvent to the alcohol solvent is (5 to 15):1. In some embodiments, the catalyst includes NaBH(OAC)3. In some embodiments, the molar ratio of the catalyst to the aldehyde compound R'CHO or the alcohol compound ROH is 1:(0.5 to 1).

[0137] The third aspect of the present invention provides a lipid nanoparticle comprising an ionizable lipid compound, wherein the ionizable lipid compound comprises the compound described in the first aspect of the present invention.

[0138] According to some embodiments of the invention, the lipid nanoparticle further comprises one or more of a helper lipid, cholesterol, and a conjugate-linker lipid.

[0139] According to some embodiments of the invention, the lipid nanoparticle further comprises a helper lipid, cholesterol, and a conjugate-linker lipid.

[0140] In some embodiments, the helper lipid preferably comprises DOPE or DSPC. In some embodiments, the conjugate-linker lipid comprises a PEGylated lipid. In some embodiments, the conjugate-linker lipid comprises DMG-PEG 2000.

[0141] According to some embodiments of the invention, the molar content of the ionizable lipid compound is 20-70% based on the total moles of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid.

[0142] According to some embodiments of the present invention, the molar content of the helper lipid is 1-20% based on the total moles of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid.

[0143] According to some embodiments of the present invention, the molar content of cholesterol is 20-50% based on the total moles of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid.

[0144] According to some embodiments of the present invention, the molar content of the conjugate-linker lipid is 1-10% based on the total moles of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid.

[0145] According to some embodiments of the present invention, based on the total molar number of the ionizable lipid compound, auxiliary lipid, cholesterol and conjugate-linker lipid, the molar content of the ionizable lipid compound is 20-70%, the molar content of the auxiliary lipid is 1-20%, the molar content of the cholesterol is 20-50%, and the molar content of the conjugate-linker lipid is 1-10%.

[0146] According to some embodiments of the present invention, based on the total molar number of the ionizable lipid compound, auxiliary lipid, cholesterol and conjugate-linker lipid, the molar content of the ionizable lipid compound is 30-60%, the molar content of the auxiliary lipid is 5-20%, the molar content of the cholesterol is 25-45%, and the molar content of the conjugate-linker lipid is 1-5%.

[0147] According to some embodiments of the present invention, based on the total molar number of the ionizable lipid compound, auxiliary lipid, cholesterol and conjugate-linker lipid, the molar content of the ionizable lipid compound is 30-55%, the molar content of the auxiliary lipid is 5-20%, the molar content of the cholesterol is 35-45%, and the molar content of the conjugate-linker lipid is 1-5%.

[0148] The fourth aspect of the present invention provides a drug delivery system or drug carrier, which includes the compound described in the first aspect of the present invention or the lipid nanoparticles described in the third aspect of the present invention.

[0149] According to some embodiments of the present invention, the drug delivery system further comprises a therapeutic and / or prophylactic agent. In some embodiments, the therapeutic and / or prophylactic agent comprises one or more of a DNA molecule, an RNA molecule, a protein, a polypeptide, and a small molecule drug.

[0150] According to some embodiments of the present invention, the therapeutic and / or preventive agent includes a pharmaceutical active ingredient for treating or preventing one or more of liver disease, spleen disease, lung disease, heart disease, and kidney disease.

[0151] According to some embodiments of the present invention, the administration method of the drug delivery system includes intramuscular injection or intravenous injection.

[0152] The fifth aspect of the present invention provides a pharmaceutical composition comprising the lipid nanoparticles described in the third aspect of the present invention and a therapeutic and / or preventive agent. The therapeutic and / or preventive agent includes, but is not limited to, one or more of a DNA molecule, an RNA molecule, a protein, a polypeptide, and a small molecule drug.

[0153] According to some embodiments of the present invention, the therapeutic and / or preventive agent comprises an RNA molecule, including but not limited to one or more of siRNA, microRNA, mRNA, tRNA, and circRNA.

[0154] According to some embodiments of the present invention, the therapeutic and / or preventive agent includes a pharmaceutical active ingredient for treating or preventing one or more of liver disease, spleen disease, lung disease, heart disease, and kidney disease.

[0155] According to some embodiments of the present invention, the administration of the pharmaceutical composition comprises one or more of intramuscular injection, intradermal injection, intravenous injection, arterial injection, transdermal absorption, intraperitoneal injection, oral administration, and nasal spray. According to some embodiments of the present invention, the administration of the pharmaceutical composition comprises one or more of intramuscular injection or intravenous injection.

[0156] According to some embodiments of the present invention, the pharmaceutical composition further comprises any one or a combination of at least two of a pharmaceutically acceptable carrier, excipient, or auxiliary material.

[0157] The sixth aspect of the present invention provides use of the compound described in the first aspect or the lipid nanoparticles described in the third aspect in preparing a drug delivery system.

[0158] According to some embodiments of the present invention, the active ingredient of the drug includes one or more of a DNA molecule, an RNA molecule, a protein, a polypeptide and a small molecule drug.

[0159] According to some embodiments of the present invention, the drug delivery system comprises a nucleic acid delivery system. In some embodiments, the nucleic acid comprises RNA. The RNA of the present invention includes, but is not limited to, one or more of siRNA, microRNA, mRNA, tRNA, and circRNA.

[0160] According to some embodiments of the present invention, the ionizable lipid compound or the lipid nanoparticle is formulated to deliver the active ingredient of the drug to target cells; the target cells preferably include spleen cells, liver cells such as hepatocytes, bone marrow cells (such as bone marrow mononuclear cells), lung cells, immune cells, muscle cells such as myocytes, heart cells such as cardiomyocytes, kidney cells or central nervous system cells, hematopoietic stem cells or one or more.

[0161] The seventh aspect of the present invention provides use of the compound described in the first aspect or the lipid nanoparticles described in the third aspect in the preparation of organ-targeted drugs.

[0162] According to some embodiments of the present invention, the compound or the lipid nanoparticle is formulated to deliver the active ingredient of the drug to target cells.

[0163] According to some embodiments of the present invention, the target cells include one or more of spleen cells, liver cells such as hepatocytes, bone marrow cells (such as bone marrow mononuclear cells), lung cells, immune cells, muscle cells such as myocytes, heart cells such as cardiomyocytes, kidney cells or central nervous system cells, and hematopoietic stem cells.

[0164] In an eighth aspect, the present invention provides a transfection complex comprising the compound described in the first aspect of the present invention or the lipid nanoparticles described in the third aspect of the present invention.

[0165] According to some embodiments of the present invention, the transfection complex comprises at least one bioactive agent, including but not limited to one or more of a DNA molecule, an RNA molecule, a protein, a polypeptide, and a small molecule drug.

[0166] The ninth aspect of the present invention provides a method for administering a therapeutic agent and / or a preventive agent to an individual in need thereof, the method comprising administering the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention to the individual.

[0167] In the tenth aspect of the present invention, a method for treating a disease or condition in an individual in need thereof is provided, the method comprising administering to the individual the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention, wherein the therapeutic agent and / or preventive agent can effectively treat or prevent the disease or condition.

[0168] In the eleventh aspect of the present invention, a method for delaying and / or curbing the progression of a disease or condition in an individual in need thereof is provided, the method comprising administering to the individual the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention, wherein the therapeutic agent and / or preventive agent can effectively treat or prevent the disease or condition.

[0169] In a twelfth aspect of the present invention, a method for delivering a therapeutic agent and / or a preventive agent to mammalian cells derived from an individual is provided, the method comprising contacting the cells of the individual to which the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention has been administered.

[0170] The thirteenth aspect of the present invention provides a method for producing a polypeptide of interest in a mammalian cell, the method comprising contacting the cell with the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention, wherein the therapeutic agent and / or preventive agent is or includes mRNA, and wherein the mRNA encodes the polypeptide of interest, whereby the mRNA can be translated in the cell to produce the polypeptide of interest.

[0171] In the fourteenth aspect of the present invention, a method for inhibiting the production of a polypeptide of interest in a mammalian cell is provided, the method comprising contacting the cell with the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention, wherein the therapeutic agent and / or preventive agent is or includes RNA, whereby the RNA is capable of inhibiting the production of the polypeptide of interest.

[0172] In a fifteenth aspect, the present invention provides a method for specifically delivering a therapeutic agent and / or a prophylactic agent to a mammalian organ, the method comprising contacting the mammalian organ with the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention, thereby delivering the therapeutic agent and / or prophylactic agent to the organ.

[0173] The sixteenth aspect of the present invention provides a method of vaccination, which is performed by administering the drug delivery system described in the fourth aspect of the present invention or the pharmaceutical composition described in the fifth aspect of the present invention.

[0174] In the seventeenth aspect of the present invention, a method for inducing an adaptive immune response in an individual is provided, comprising administering to the individual an effective amount of a composition comprising at least one RNA; wherein the composition comprises the lipid nanoparticles described in the third aspect of the present invention.

[0175] Compared with the prior art, the present invention has the following beneficial effects:

[0176] The present invention provides a new class of compounds suitable for use as ionizable lipids in the preparation of lipid nanoparticles, which can be used for organ-targeted therapy or disease prevention. Specifically, lipid nanoparticles prepared using the compounds of the present invention exhibit high liver targeting after intravenous or intramuscular injection, particularly high liver targeting via intravenous injection. Furthermore, because the lipids of the present invention contain endogenous groups, LNPs containing them exhibit excellent biosafety. Experimental data demonstrate that the toxicity of the lipids of the present invention is significantly lower than that of conventional commercial lipids (with a safe dose two to three times that of conventional lipids). BRIEF DESCRIPTION OF THE DRAWINGS

[0177] FIG1 shows the size of LNPs synthesized in Example 2 of the present invention.

[0178] FIG2 shows the PDI of LNPs synthesized in Example 2 of the present invention.

[0179] FIG3 shows the mRNA encapsulation efficiency of the LNPs synthesized in Example 2 of the present invention.

[0180] FIG4 shows an in vivo imaging diagram after intramuscular injection of LNP in an in vivo experiment in Example 4 of the present invention.

[0181] FIG5 shows organ imaging after intramuscular injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0182] FIG6 shows the statistics of relative fluorescence intensity of the liver after intramuscular injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0183] FIG7 shows the percentage of total fluorescence values ​​of various organs after intramuscular injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0184] FIG8 shows the in vivo imaging diagram after intravenous injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0185] FIG9 shows organ imaging after intravenous injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0186] FIG10 shows a statistical graph of relative fluorescence intensity of the liver after intravenous injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0187] FIG11 shows the percentage of total fluorescence value of each organ after intravenous injection of LNP in the in vivo experiment in Example 4 of the present invention.

[0188] FIG12 shows the in vivo imaging after intravenous injection of LNPs of different configurations in the in vivo experiment in Example 4 of the present invention.

[0189] FIG13 shows a statistical graph of relative fluorescence intensity of in vivo imaging after intravenous injection of LNPs of different configurations in the in vivo experiment in Example 4 of the present invention.

[0190] FIG14 shows organ imaging after intravenous injection of LNPs of different configurations in the in vivo experiment in Example 4 of the present invention.

[0191] FIG15 shows a statistical graph of relative fluorescence intensity of organ imaging after intravenous injection of LNPs of different configurations in the in vivo experiment in Example 4 of the present invention.

[0192] FIG16 shows the survival rate of 293T cells in the in vitro test in Example 5 of the present invention.

[0193] FIG17 shows the survival rate of DC2.4 cells in the in vitro test in Example 5 of the present invention.

[0194] FIG18 shows the survival rate of BEAS-2B cells in the in vitro test in Example 5 of the present invention.

[0195] FIG. 19 shows the survival rate of LX-2 cells in the in vitro test in Example 5 of the present invention.

[0196] FIG20 shows the percentage of luciferase activity of 293T cells in the in vitro test in Example 5 of the present invention.

[0197] FIG21 shows the percentage of luciferase activity of DC2.4 cells tested in vitro in Example 5 of the present invention.

[0198] FIG22 shows the percentage of luciferase activity of BEAS-2B cells in the in vitro test in Example 5 of the present invention.

[0199] FIG. 23 shows the percentage of luciferase activity in LX-2 cells tested in vitro in Example 5 of the present invention.

[0200] 24A-24B respectively show the NMR spectrum and mass spectrum of compound LA-8-8.

[0201] 25A-25B respectively show the NMR spectrum and mass spectrum of compound LA-8-10.

[0202] 26A-26B respectively show the NMR spectrum and mass spectrum of compound LA-8-12.

[0203] 27A-27B respectively show the NMR spectrum and mass spectrum of compound LA-8-14.

[0204] 28A-28B respectively show the NMR spectrum and mass spectrum of compound LA-8-18D.

[0205] 29A-29B respectively show the NMR spectrum and mass spectrum of compound LA-13-8.

[0206] 30A-30B respectively show the NMR spectrum and mass spectrum of compound LA-13-10.

[0207] 31A-31B respectively show the NMR spectrum and mass spectrum of compound LA-13-12.

[0208] 32A-32B respectively show the NMR spectrum and mass spectrum of compound LA-13-14.

[0209] 33A-33B respectively show the NMR spectrum and mass spectrum of compound LA-13-18D.

[0210] 34A-34B respectively show the NMR spectrum and mass spectrum of compound LA-18-8.

[0211] 35A-35B respectively show the NMR spectrum and mass spectrum of compound LA-18-10.

[0212] 36A-36B respectively show the NMR spectrum and mass spectrum of compound LA-18-12.

[0213] 37A-37B respectively show the NMR spectrum and mass spectrum of compound LA-18-14.

[0214] 38A-38B respectively show the NMR spectra and mass spectra of compound LA-18-18D. DETAILED DESCRIPTION

[0215] In order to make the purpose, technical solutions and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments and drawings. The specific embodiments described herein are only used to illustrate the present invention and are not intended to constitute any limitation to the present invention.

[0216] Unless otherwise specified, the reagents used in the examples are all commercially available products or reagents prepared according to conventional methods. Unless otherwise specified, the methods used in the examples are all conventional experimental methods. Unless otherwise specified, the instruments used in the examples are all commercially available.

[0217] The compounds described herein may exist as isomers, such as stereoisomeric forms (enantiomers, diastereomers, cis-trans isomers), or tautomers, depending on their structure. Thus, the present invention relates to enantiomers or diastereomers and their respective mixtures. Stereoisomerically pure components can be isolated from such enantiomer and / or diastereomeric mixtures in a known manner. When the compounds described herein may exist as optical isomers, the compositions provided herein generally contain substantially pure optical isomers.

[0218] Where compounds described herein exist in tautomeric forms, the present invention encompasses all tautomeric forms.

[0219] As used herein, the phrase "tautomer" is used to describe different isomeric forms of organic compounds that can be easily converted into each other. Tautomers can be characterized by the migration of hydrogen atoms or protons, accompanied by the conversion of single bonds and adjacent double bonds. In some embodiments, tautomers can be generated by proton tautomerism (i.e., the relocation of protons). In some embodiments, tautomers can be generated by valence tautomerism (i.e., the rapid reorganization of bonding electrons). All such tautomers are intended to be included within the scope of this application. In some embodiments, the tautomers of a compound exist in dynamic equilibrium with each other, so a mixture is formed when attempting to prepare a separate substance. In some embodiments, the tautomers of a compound are separable and separable compounds. In some embodiments of the present application, a chemical composition can be provided as follows: it is or contains a pure preparation of a single tautomer of a compound. In some embodiments, a chemical composition can be provided as a mixture of two or more tautomers of a compound. In certain embodiments, such a mixture contains equal amounts of different tautomers; in certain embodiments, such a mixture contains different amounts of at least two different tautomers of a compound. In some embodiments of the application, chemical composition can contain all tautomers of compound.In some embodiments of the application, chemical composition can contain less than whole tautomers of compound.In some embodiments of the application, chemical composition can contain one or more tautomers of compound, and its amount changes over time because of mutual conversion.In some embodiments of the application, tautomerism is keto-enol tautomerism. It will be appreciated by those skilled in the art of chemistry that keto-enol tautomerism can use any suitable reagent " capture " (that is, chemical modification is so that it keeps " enol " form) known in the art to provide the enol derivatives that can use one or more suitable technology separations known in the art subsequently. Unless otherwise indicated, all tautomers of related compounds are contained in the application, no matter it is pure form or mixed with each other.

[0220] As used herein, the term "solvate" refers to an association formed between one or more solvent molecules and a compound of the present invention. Solvents that form solvates include, but are not limited to, water, isopropanol, ethanol, methanol, dimethyl sulfoxide, ethyl acetate, acetic acid, and aminoethanol. Hydrates are preferred as solvates within the scope of the present invention. The term "hydrate" refers to an association formed when the solvent molecule is water.

[0221] The term "isotopic labeling" refers to the process of replacing an element in a molecule with an isotope. An "isotope" is an element that has different masses but the same chemical properties. The process of replacing an atom in a molecule with an isotope of that atom is called isotopic labeling. Examples of isotopes that can be incorporated into the compounds described herein include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, chlorine, bromine, and iodine, for example 2 H (deuterium), 3 H (tritium), 13 C. 14 C. 15 N. 17 O. 18 O. 32 P. 33 P. 33 S. 34 S. 35 S. 36 S. 18 F. 36 Cl, 82 Br, 123 I. 124 I. 129 I and 131 I.

[0222] As described herein, the compounds of the present application may contain optionally substituted and / or substituted moieties. In general, the term "substituted by" or "substituted", whether preceded by the term "optionally", means that one or more hydrogens of the specified moiety are replaced by suitable substituents. "Substitution" applies to one or more hydrogens that are explicitly or implicitly indicated in the structure. Unless otherwise indicated, an "optionally substituted by" group may have a suitable substituent at each substitutable position of the group, and when more than one position in any given structure may be substituted by one or more substituents selected from a specified group, the substituents at each position may be the same or different. The substituent combinations contemplated by the present disclosure are preferably substituent combinations that result in the formation of stable or chemically feasible compounds. Groups described as "substituted by" preferably have between 1 and 4 substituents, more preferably 1 or 2 substituents. Groups described as "optionally substituted by" may be unsubstituted or "substituted by", as described above.

[0223] As used herein, the term "saturated or unsaturated aliphatic hydrocarbon group" means a straight chain (i.e., unbranched) or branched substituted or unsubstituted hydrocarbon chain that is completely saturated or contains one or more unsaturated units, or a monocyclic or bicyclic hydrocarbon (also known as "carbocycle," "carbocyclyl," or "cycloaliphatic") that is completely saturated or contains one or more unsaturated units but is not aromatic. Unless otherwise specified, a saturated or unsaturated aliphatic hydrocarbon group contains 1 to 30 aliphatic carbon atoms. In some embodiments, a saturated or unsaturated aliphatic hydrocarbon group contains 1 to 20 carbon atoms. In some embodiments, a saturated or unsaturated aliphatic hydrocarbon group contains 2 to 20 carbon atoms. In some embodiments, a saturated or unsaturated aliphatic hydrocarbon group contains 4 to 20 carbon atoms. In some embodiments, a saturated or unsaturated aliphatic hydrocarbon group contains 8 to 20 carbon atoms. Suitable saturated or unsaturated aliphatic hydrocarbon groups include, but are not limited to, linear or branched, substituted or unsubstituted alkyl, alkenyl, alkynyl groups and mixtures thereof, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or (cycloalkyl)alkenyl.

[0224] As used herein, the term "alkyl" refers to a straight or branched chain hydrocarbon group having a specified number of carbon atoms (i.e., a C1-C20 alkyl group refers to a straight or branched chain hydrocarbon group having 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, or 20 carbon atoms). In some embodiments, the alkyl group has 1 to 30 carbon atoms. In certain embodiments, the straight or branched chain alkyl group has about 1 to 20 carbon atoms in its backbone (e.g., C1-C20 for a straight chain alkyl group and C2-C20 for a branched chain alkyl group), and alternatively, has about 1 to 10 carbon atoms in its backbone. Examples of alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, n-pentyl, n-hexyl, and the like. When an alkyl group is a linking group, the alkyl group represents the alkylene group that is connected.

[0225] The term "cycloalkyl" as used herein includes saturated monocyclic (e.g., C3-C20), bicyclic (e.g., C5-C20 fused bicyclic, C5-C20 spirobicyclic), or polycyclic cyclic alkyl groups. "C3-C20 cycloalkyl" refers to a group containing 3 to 20 carbon atoms. Examples of cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. When a cycloalkyl group is a linking group, the cycloalkyl group represents a linked cycloalkylene group.

[0226] As used herein, the term "heterocycloalkyl" refers to a stable monocyclic, bicyclic, or polycyclic heterocyclic ring, including fused, spirocyclic, and / or bridged ring structures, which is saturated, partially unsaturated, and contains carbon atoms and 1, 2, 3, or 4 heteroatoms independently selected from N, O, and S. When heterocycloalkyl is a linking group, heterocycloalkyl refers to the linked heterocycloalkylene group. Heterocycloalkyl includes, but is not limited to, tetrahydropyrrolyl, tetrahydrofuranyl, tetrahydrothiophenyl, piperidinyl, piperazinyl, and the like.

[0227] As used herein, the term "alkenyl" refers to a straight or branched hydrocarbon group containing one or more double bonds and having a specified number of carbon atoms. For example, "C2-C20 alkenyl" refers to an alkenyl group containing 2 to 20 carbon atoms. Alkenyl includes, but is not limited to, ethenyl, propenyl, butenyl, 1-methyl-2-butene-1-yl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, undecenyl, dodecenyl, tridecenyl, tetradecenyl, pentadecenyl, hexadecenyl, heptadecenyl, octadecenyl, etc.

[0228] As used herein, the term "alkynyl" refers to a straight or branched hydrocarbon group containing one or more triple bonds and having a specified number of carbon atoms. For example, "C2-C20 alkynyl" refers to an alkynyl group containing 2 to 20 carbon atoms. Alkenyl groups include, but are not limited to, ethynyl, propynyl, butynyl, pentynyl, hexynyl, heptynyl, octynyl, nonynyl, decynyl, undecynyl, dodecynyl, tridecynyl, tetradecynyl, pentadecynyl, hexadecynyl, heptadecenyl, octadecynyl, and the like.

[0229] As used herein, the term "aryl" refers to an aromatic monocyclic or polycyclic ring system composed of carbon atoms. In some embodiments, the polycyclic ring system is bicyclic or tricyclic. In some embodiments, the aryl group is a 6-15 membered aryl group; in some embodiments, the aryl group is a 6-12 membered aryl group; in other embodiments, the aryl group is a 6-10 membered aryl group; in other embodiments, the aryl group is a phenyl or naphthyl group. When the aryl group is a linking group, the aryl group represents a linking arylene group.

[0230] As used herein, the term "heteroaryl" refers to an aromatic monocyclic or polycyclic ring system composed of at least one carbon atom and one or more heteroatoms as ring atoms. Such heteroatoms include, but are not limited to, O, S, and N. A heteroaryl group may contain 1, 2, 3, or 4 ring heteroatoms. Representative examples of heteroaryl groups include, but are not limited to, pyrrolyl, furanyl, thiophenyl, imidazolyl, oxazolyl, thiazolyl, triazolyl, pyrazolyl, pyridyl, pyrazinyl, pyridazinyl, and pyrimidinyl. When a heteroaryl group is a linking group, the heteroaryl group represents the linked heteroarylene group.

[0231] It should also be understood that some of the composite groups described herein are not limited to a particular orientation in the structures shown. For example, when referring to -(C=O)O- or -(C=O)NR a -, in the structure shown, the C=O part can be closer to the distal end, or O or NR a For another example, when it is mentioned that one or more methylene units in an alkyl group are optionally and independently selected from -(C=O)O-, it can mean It can also be expressed

[0232] As used herein, the term "in vitro" refers to events that occur in an artificial environment, such as in a test tube or reactor, in cell culture, etc., rather than in a multicellular organism.

[0233] As used herein, the term "in vivo" refers to events that occur within multicellular organisms, such as humans and non-human animals. In the context of cell-based systems, the term can be used to refer to events that occur within living cells (as opposed to, for example, in vitro systems).

[0234] The term "nanoparticle" as used herein refers to a particle having a diameter of less than 1000 nanometers (nm). In some embodiments, as defined by the National Science Foundation of the United States, the diameter of the nanoparticle is less than 300 nm. In some embodiments, as defined by the National Institutes of Health of the United States, the diameter of the nanoparticle is less than 100 nm. In some embodiments, the nanoparticle is a micelle, which is that it includes a closed compartment separated from the bulk solution by a micelle membrane, the membrane typically including an amphiphilic entity that surrounds and encloses a space or compartment (e.g., to define an inner cavity). In some embodiments, the micelle membrane includes at least one polymer, such as a biocompatible and / or biodegradable polymer. In some embodiments, the average diameter of the lipid nanoparticles described herein may be from about 30 to about 160 nm. In some embodiments, the lipid nanoparticles described herein have an average hydrodynamic diameter of about 30 nm, 35 nm, 40 nm, 45 nm, 50 nm, 55 nm, 60 nm, 65 nm, 70 nm, 75 nm, 80 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, or any range having endpoints defined by any two of the aforementioned values. For example, in some embodiments, the lipid nanoparticles described herein have an average hydrodynamic diameter of between 50 nm and 100 nm.

[0235] The term "nucleic acid" as used herein, in its broadest sense, refers to any compound and / or substance that is or can be incorporated into an oligonucleotide chain. In some embodiments, a nucleic acid is a compound and / or substance that is or can be incorporated into an oligonucleotide chain through a phosphodiester bond. As will be apparent from the context, in some embodiments, "nucleic acid" refers to individual nucleic acid residues (e.g., nucleotides and / or nucleosides); in some embodiments, "nucleic acid" refers to an oligonucleotide chain comprising individual nucleic acid residues. In some embodiments, "nucleic acid" is or includes RNA (e.g., mRNA, tRNA, rRNA); in some embodiments, "nucleic acid" is or includes DNA. In some embodiments, a nucleic acid is one or more natural nucleic acid residues, includes one or more natural nucleic acid residues, or consists of one or more natural nucleic acid residues. In some embodiments, a nucleic acid is one or more nucleic acid analogs, includes one or more nucleic acid analogs, or consists of one or more nucleic acid analogs. In some embodiments, nucleic acid analogs differ from nucleic acids in that they do not utilize a phosphodiester backbone. For example, in some embodiments, the nucleic acid is, includes, or consists of one or more "peptide nucleic acids," which are known in the art and have peptide bonds in the backbone instead of phosphodiester bonds and are considered to be within the scope of the present invention. Alternatively or additionally, in some embodiments, the nucleic acid has one or more phosphorothioate and / or 5'-N-phosphoamidate bonds instead of phosphodiester bonds. In some embodiments, the nucleic acid is, includes, or consists of one or more natural nucleosides (e.g., adenosine, thymidine, guanosine, cytidine, uridine, deoxyadenosine, deoxythymidine, deoxyguanosine, and deoxycytidine). In some embodiments, a "nucleic acid" is, comprises, or consists of one or more nucleoside analogs (e.g., 2-aminoadenosine, 2-thiothymidine, inosine, pyrrolopyrimidine, 3-methyladenosine, 5-methylcytidine, C-5 propynyl-cytidine, C-5 propynyl-uridine, 2-aminoadenosine, C5-bromouridine, C5-fluorouridine, C5-iodouridine, C5-propynyl-uridine, C5-propynyl-cytidine, C5-methylcytidine, 2-aminoadenosine, 7-deazaadenosine, 7-deazaguanosine, 8-oxoadenosine, 8-oxoguanosine, O(6)-methylguanine, 2-thiocytidine, methylated bases, embedded bases, and combinations thereof). In some embodiments, the nucleic acid comprises one or more modified sugars (e.g., 2'-fluororibose, ribose, 2'-deoxynucleosides, arabinose, and hexoses) compared to sugars in natural nucleic acids. In some embodiments, the nucleic acid has a nucleotide sequence that encodes a functional gene product such as an RNA or a protein. In some embodiments, the nucleic acid comprises one or more introns.In some embodiments, the nucleic acid is prepared by one or more of: isolation from a natural source, enzymatic synthesis (in vivo or in vitro) based on polymerization of a complementary template, propagation in a recombinant cell or system, and chemical synthesis. In some embodiments, the nucleic acid is at least 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 20, 225, 250, 275, 300, 325, 350, 375, 400, 425, 450, 475, 500, 600, 700, 800, 900, 1000, 1500, 2000, 2500, 3000, 3500, 4000, 4500, 5000, or more residues in length. In some embodiments, the nucleic acid is partially or completely single-stranded; in some embodiments, the nucleic acid is partially or completely double-stranded. In some embodiments, the nucleic acid has a nucleotide sequence that includes at least one element that encodes a polypeptide, or is the complement of a sequence that encodes a polypeptide. In some embodiments, the nucleic acid has enzymatic activity.

[0236] The term "polypeptide" as used herein generally has the art-recognized meaning of a polymer of at least three amino acids. It will be understood by those skilled in the art that the term "polypeptide" is intended to be general enough to encompass polypeptides having the complete sequences listed herein, while also encompassing polypeptides representing functional fragments of such complete polypeptides (e.g., fragments retaining at least one activity). In addition, it will be understood by those skilled in the art that protein sequences can generally undergo some substitutions without destroying activity. Thus, the related term "polypeptide" as used herein encompasses polypeptides that retain activity and share at least about 30 to 40% (generally greater than about 50%, 60%, 70% or 80%) overall sequence identity with another polypeptide of the same class, and further generally comprises at least one region with higher identity (in one or more highly conserved regions, the identity is generally greater than 90% or even 95%, 96%, 97%, 98% or 99%), generally encompassing any polypeptide of at least 3 to 4 and generally up to 20 or more amino acids. The polypeptide may contain L-amino acids, D-amino acids, or both thereof, and may contain any of a variety of amino acid modifications or analogs known in the art. Suitable modifications include, for example, terminal acetylation, amidation, methylation, and the like. In some embodiments, the protein may include natural amino acids, non-natural amino acids, synthetic amino acids, and combinations thereof. The term "peptide" is generally used to refer to polypeptides having a length of less than about 100 amino acids, less than about 50 amino acids, less than 20 amino acids, or less than 10 amino acids. In some embodiments, the protein is an antibody, an antibody fragment, a biologically active portion thereof, and / or a characteristic portion thereof.

[0237] As used herein, the term "pharmaceutical composition" refers to an active agent formulated with one or more pharmaceutically acceptable carriers. In some embodiments, the active agent is present in an amount suitable for administration in a unit dose in a treatment regimen that, when administered to a relevant population, shows a statistically significant probability of achieving a predetermined therapeutic effect. In some embodiments, the pharmaceutical composition can be specifically formulated for administration in solid or liquid form, including pharmaceutical compositions suitable for the following modes of administration: oral, such as drenches (aqueous or non-aqueous solutions or suspensions), tablets, such as tablets targeted for buccal, sublingual, and systemic absorption, boluses applied to the tongue, powders, granules, pastes; parenteral, such as by subcutaneous, intramuscular, intravenous, or epidural injection in the form of, for example, a sterile solution or suspension or sustained-release formulation; topical, such as in the form of a cream, ointment, or controlled-release patch or spray applied to the skin, lungs, or oral cavity; intravaginally or rectally, such as as a pessary, cream, or foam; sublingually; ophthalmically; transdermally; or nasally, pulmonary, and other mucosal surfaces.

[0238] As used herein, the term "protein" refers to one or more polypeptides that function as discrete units. If a single polypeptide is a discrete functional unit and does not need to be physically associated with other polypeptides permanently or temporarily to form a discrete functional unit, the terms "polypeptide" and "protein" can be used interchangeably. If a discrete functional unit includes more than one polypeptide physically associated with each other, the term "protein" can be used to refer to multiple polypeptides that are physically associated and function together as discrete units. In some embodiments, a protein may include parts other than amino acids (e.g., glycoproteins, proteoglycans, etc.) and / or may be processed or modified in other ways. One of ordinary skill in the art will appreciate that in some embodiments, the term "protein" may refer to a complete polypeptide chain produced by a cell (e.g., with or without a signal sequence), and / or to a form that is active in the cell (e.g., a truncated or complex form). In some embodiments where a protein includes multiple polypeptide chains, such chains may be covalently bound to each other, for example, by one or more disulfide bonds, or may be associated in other ways.

[0239] The term "pharmaceutically acceptable" as used herein refers to those compounds, materials, compositions and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problems or complications, commensurate with a reasonable benefit / risk ratio.

[0240] As used herein, "pharmaceutically acceptable carrier" means a pharmaceutically acceptable material, composition or vehicle, such as a liquid or solid filler, diluent, excipient or solvent encapsulating material, which participates in carrying or transporting the compound of the invention from one organ or part of the body to another organ or part of the body. In the sense of being compatible with the other ingredients of the formulation and not harmful to the patient

[0241] In general, each carrier must be "acceptable." Some examples of materials that can be used as pharmaceutically acceptable carriers include: sugars such as lactose, glucose, and sucrose; starches such as corn starch and potato starch; cellulose and its derivatives such as sodium carboxymethylcellulose, ethylcellulose, and cellulose acetate; powdered tragacanth; malt; gelatin; talc; excipients such as cocoa butter and suppository waxes; oils such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil, and soybean oil; glycols such as propylene glycol; polyols such as glycerol, sorbitol, mannitol, and polyethylene glycol; esters such as ethyl oleate and ethyl laurate; agar; buffers such as magnesium hydroxide and aluminum hydroxide; alginic acid; pyrogen-free water; isotonic saline; Ringer's solution; ethanol; pH buffered solutions; polyesters, polycarbonates, and / or polyanhydrides; and other nontoxic, compatible substances used in pharmaceutical formulations.

[0242] The term "pharmaceutically acceptable salt" as used herein refers to salts of such compounds suitable for use in a pharmaceutical context, i.e., salts suitable for contact with the tissues of humans and lower animals without excessive toxicity, irritation, allergic reactions, etc., and that are compatible with a reasonable benefit / risk ratio, within the scope of reasonable medical judgment. Pharmaceutically acceptable salts are well known in the art. For example, SM Berge et al. describe pharmaceutically acceptable salts in detail in J. Pharmaceutical Sciences, 66: 1-19 (1977). In some embodiments, pharmaceutically acceptable salts include, but are not limited to, non-toxic acid addition salts, which are salts of an amino group with an inorganic acid or with an organic acid, such as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and perchloric acid, an organic acid such as acetic acid, maleic acid, tartaric acid, citric acid, succinic acid, or malonic acid, or by using other methods used in the art, such as salts formed by ion exchange. In some embodiments, pharmaceutically acceptable salts include, but are not limited to, adipate, alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate, borate, butyrate, camphorate, camphorsulfonate, citrate, cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate, formate, fumarate, glucoheptonate, glycerophosphate, gluconate, hemisulfate, heptanoate, hexanoate, hydroiodide, 2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, lauryl sulfate, malate, maleate, malonate, methanesulfonate, 2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate, pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, picrate, pivalate, propionate, stearate, succinate, sulfate, tartrate, thiocyanate, p-toluenesulfonate, undecanoate, valerate, and the like. Representative alkali metal salts or alkaline earth metal salts include sodium salts, lithium salts, potassium salts, calcium salts, magnesium salts, etc. In some embodiments, pharmaceutically acceptable salts include non-toxic ammonium, quaternary ammonium, and amine cations formed using counterions such as halides, hydroxides, carboxylates, sulfates, phosphates, nitrates, alkyl groups having 1 to 6 carbon atoms, sulfonates, and arylsulfonates, as appropriate.

[0243] As used herein, an "excipient" refers to a pharmaceutically acceptable material, mixture, or vehicle that contributes to the consistency of a dosage form or pharmaceutical composition. Each excipient must be compatible with the other ingredients of the pharmaceutical composition when mixed to avoid interactions that could significantly reduce the efficacy of the disclosed compounds when administered to a patient and interactions that could result in a pharmaceutical composition that is not pharmaceutically acceptable. Furthermore, each excipient must be pharmaceutically acceptable, for example, possessing a sufficiently high purity. Suitable pharmaceutically acceptable excipients will vary depending on the specific dosage form selected. Furthermore, pharmaceutically acceptable excipients may be selected based on their specific function in the composition. For example, certain pharmaceutically acceptable excipients may be selected to facilitate the production of a uniform dosage form. Certain pharmaceutically acceptable excipients may be selected to facilitate the production of a stable dosage form. Certain pharmaceutically acceptable excipients may be selected to facilitate the carrying or transport of the disclosed compounds from one organ or part of the body to another organ or part of the body when administered to a patient. Certain pharmaceutically acceptable excipients may be selected to enhance patient compliance. Some examples of suitable excipients include lactose, glucose, sucrose, sorbitol, mannitol, starch, gum arabic, calcium phosphate, alginates, tragacanth gum, gelatin, calcium silicate, microcrystalline cellulose, polyvinyl pyrrolidone, cellulose, water, syrup and methylcellulose. Suitable pharmaceutically acceptable excipients also include the following types of excipients: solvents, propellants, solubilizers, cosolvents, emulsifiers, colorants, binders, disintegrants, fillers, lubricants, wetting agents, osmotic pressure regulators, stabilizers, glidants, flavoring agents, preservatives, suspending agents, coating materials, fragrances, anti-adherents, antioxidants, chelating agents, penetration enhancers, pH adjusters, plasticizers, surfactants, foaming agents, defoaming agents, thickeners, inclusion agents, humectants, absorbents, diluents, flocculants and deflocculating agents and filter aids. Those skilled in the art will recognize that certain pharmaceutically acceptable excipients can serve more than one function, and can provide alternative functions, depending on how much of the excipient is present in the formulation and which other excipients are present in the formulation. The compounds of the invention can be formulated using methods known in the art so as to provide quick, sustained, or delayed release of the active ingredient after administration to a patient.

[0244] As used herein, the term "lipid nanoparticle (LNP)" includes compositions, formulations, and / or nanomaterials of lipid nanoparticles. In some embodiments, the lipid nanoparticles include one or more components. In some embodiments, the lipid nanoparticles include one or more components, such as compounds, ionizable lipids, sterols, conjugate-linker lipids, and phospholipids. In addition, the present disclosure describes that the selection and combination of one or more of the components described herein will affect the characteristics of the lipid nanoparticles, such as diameter, pKa, stability, and ionizability. In addition, the present disclosure describes that the selection and combination of one or more of the components described herein will affect the functional activity of the lipid nanoparticles, such as tropism, stability, and drug delivery efficacy. For example, the present disclosure describes that the combination of components can be better suited for the delivery of siRNA. As another example, the present disclosure describes that the combination of components can be better suited for the delivery of mRNA. As another example, the present disclosure describes that the combination of components can be better suited for the delivery of DNA. In some embodiments, the lipid nanoparticles include one or more compounds as described herein. In some embodiments, the lipid nanoparticles include one or more ionizable lipid compounds as described herein. In some embodiments, the lipid nanoparticles include one or more sterols as described herein. In some embodiments, the lipid nanoparticles include one or more conjugate-linker lipids as described herein. In some embodiments, the lipid nanoparticles include one or more phospholipids as described herein.

[0245] As used herein, the term "sterol" refers to cholesterol or a variant or derivative thereof. In some embodiments, the cholesterol is modified. In some embodiments, the cholesterol is oxidized cholesterol. In some embodiments, the cholesterol is esterified cholesterol. Unmodified cholesterol can be enzymatically formed into side chain or epoxidized variants. In some embodiments, the cholesterol can be oxidized at the β-ring structure or the hydrocarbon tail structure. In some embodiments, the sterol is a phytosterol. Exemplary sterols considered suitable for use in the disclosed lipid nanoparticles include, but are not limited to, 25-hydroxycholesterol (25-OH), 20α-hydroxycholesterol (20α-OH), 27-hydroxycholesterol, 6-keto-5α-hydroxycholesterol, 7-ketocholesterol, 7β-hydroxycholesterol, 7α-hydroxycholesteryl, 7β-25-dihydroxycholesterol, β-sitosterol, stigmasterol, brassicasterol, campesterol, or combinations thereof. In some embodiments, side chain oxidized cholesterol can enhance the delivery of preventive / therapeutic agents relative to other cholesterol variants. In some embodiments, the cholesterol is unmodified cholesterol. In some embodiments, the LNP composition includes about 20 mol% to about 50 mol% cholesterol. In some embodiments, the LNP composition includes about 25 mol% to about 45 mol% cholesterol. In some embodiments, the LNP composition includes about 35 mol% to about 45 mol% cholesterol.

[0246] The term "conjugate-linker lipid" as used in the present invention is or includes polyethylene glycol (PEG)-lipid or PEG-modified lipid. In some embodiments, PEG or PEG-modified lipids may alternatively be referred to as PEGylated lipids or PEG-lipids. The inclusion of PEGylated lipids can be used to enhance the colloidal stability and in vivo circulation time of lipid nanoparticles in vitro. In some embodiments, because the PEG moiety is gradually released in the blood circulation, PEGization is reversible. Exemplary PEG lipids include, but are not limited to, PEG conjugated to saturated or unsaturated alkyl chains of length C6-C20. PEG-modified phosphatidylethanolamine, PEG-modified phosphatidic acid, PEG-modified ceramide (PEG-CER), PEG-modified dialkylamine, PEG-modified diacylglycerol (PEG-DAG), PEG-modified dialkylglycerols, and mixtures thereof. For example, in some embodiments, the PEG lipid can be PEG-c-DOMG, PEG-DMG, PEG-DLPE, PEG-DMPE, PEG-DPPE, PEG-DSG, or PEG-DSPE lipid. In some embodiments, the conjugate-linker lipid includes a polyethylene glycol lipid. In some embodiments, the conjugate-linker lipid includes dimyristyl glycerol (DMG), 1,2-dipalmitoyl-rac-glycerol, methoxypolyethylene glycol (DPG-PEG) or 1,2-distearoyl-rac-glycerol-3-methylpolyoxyethylene (DSG-PEG). In some embodiments, the average molecular weight of the conjugate-linker lipid is about 500Da to about 5000Da. In some embodiments, the average molecular weight of the conjugate-linker lipid is about 2000Da. In some embodiments, the LNP composition includes about 0mol% to about 5mol% of the conjugate-linker lipid. In some embodiments, the LNP composition includes about 1-5mol% conjugate-linker lipid.

[0247] As used herein, the term "phospholipid" may be assembled into one or more lipid bilayers. In some embodiments, the one or more phospholipids may comprise a phospholipid moiety. In some embodiments, the one or more phospholipids may comprise one or more fatty acid moieties. In some embodiments, the one or more phospholipids may comprise a phospholipid moiety and one or more fatty acid moieties. In some embodiments, the phospholipid moiety includes, but is not limited to, phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, phosphatidylserine, phosphatidic acid, 2-lysophosphatidylcholine, and sphingomyelin. In some embodiments, the fatty acid moiety includes, but is not limited to, lauric acid, myristic acid, myristoleic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linolenic acid, α-linolenic acid, erucic acid, phytanic acid, arachidic acid, arachidonic acid, eicosapentaenoic acid, behenic acid, docosapentaenoic acid, and docosahexaenoic acid. Non-natural substances are also contemplated, including natural substances with modifications and substitutions including branching, oxidation, cyclization, and alkyne. For example, phospholipids can be functionalized or cross-linked with one or more alkynes (e.g., alkenyl groups in which one or more double bonds are replaced by triple bonds). Under appropriate reaction conditions, the alkynes can undergo copper-catalyzed cycloadditions when exposed to azides. Such reactions can be useful for functionalizing the lipid bilayer of nanoparticle compositions to facilitate membrane penetration or cell recognition, or for conjugating nanoparticle compositions to useful components, such as targeting or imaging moieties (e.g., dyes).Exemplary phospholipids include, but are not limited to, 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dilinoleoyl-sn-glycero-3-phosphocholine (DLPC), 1,2-dimyristoyl-sn-glycerophosphocholine (DMPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), 1,2-dipalmitoyl-sn-glycero-3- Phosphocholine (DPPC), 1,2-diondecanoyl-sn-glycerophosphocholine (DUPC), l-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-di-O-octadecenyl-sn-glycero-3-phosphocholine (18:0 diether PC), 1-oleoyl-2-cholestyrene hemisuccinyl l-sn-glycero-3-phosphocholine (OChemsPC), 1-hexadecyl-sn-glycero-3-phosphocholine (C16 Lyso PC), 1,2-dialinolenoyl-sn-glycero-3-phosphocholine, 1,2-arachidonoyl-sn-glycero-3-phosphocholine, 1,2-didocosahexaenoyl-sn-glycero-3-phosphocholine, 1,2-diphytanoyl-sn-glycero-3-phosphoethanolamine (ME 16.0PE), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinoleoyl-sn-glycero-3-phosphoethanolamine, 1,2-dilinolenoyl-sn-glycero-3-phosphoethanolamine, 1,2-diarachidonoyl-sn-glycero-3-phosphoethanolamine, 1,2-bisdocosahexaenoyl-sn-glycero-3-phosphoethanolamine, 1,2-dioleoyl-sn-glycero-3-phospho-rac-(1-glycerol) sodium salt (DOPG), dipalmitoylphosphatidylglycerol (DPPG), palmitoyl In some embodiments, the phospholipid is DSPC. In some embodiments, the phospholipid is DMPC.In some embodiments, the phospholipids include 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl) (succinyl PE), 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE), 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine-N-(succinyl) (succinyl-DPPE), 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), or a combination thereof. In some embodiments, the LNP composition includes about 1 mol% to about 20 mol% phospholipids. In some embodiments, the LNP composition includes about 5 mol% to about 15 mol% phospholipids.

[0248] The prophylactic / therapeutic agent delivered by LNP in the present invention can be a bioactive agent. In some embodiments, the prophylactic / therapeutic agent is or includes one or more bioactive agents, such as mRNA, guide RNA (gRNA), nucleic acid, RNA-guided DNA binder, expression vector, template nucleic acid, antibody (e.g., monoclonal antibody, chimeric antibody, humanized antibody, nanobody and fragments thereof, etc.), cholesterol, hormone, peptide, protein, chemotherapeutic agent and other types of anti-tumor agents, low molecular weight drugs, vitamins, cofactors, nucleosides, nucleotides, oligonucleotides, enzymatic nucleic acids, antisense nucleic acids, triplex forming oligonucleotides, antisense DNA or RNA compositions, chimeric DNA: RNA compositions, allozymes, aptamers, ribozymes, traps (decoy) and its analogs, plasmids and other types of bodies, and small nucleic acid molecules, RNAi agents, short interfering nucleic acids (siNA), short interfering RNA (siRNA), double-stranded RNA (dsRNA), microRNA (miRNA), short hairpin RNA (shRNA) and "self-replicating RNA" (encoding replicase activity and capable of guiding its own replication or amplification in vivo) molecules, peptide nucleic acids (PNA), locked nucleic acid ribonucleotides (LNA), morpholino nucleotides, threose nucleic acids (TNA), glycol nucleic acids (GNA), sisiRNA (small internal segmented interfering RNA), and RNA (asymmetric interfering RNA). The above list of bioactive agents is exemplary only and is not intended to be limiting. Such compounds can be purified or partially purified and can be naturally occurring or synthetic and can be chemically modified. The prophylactic / therapeutic agent delivered by the LNP composition can be RNA, such as an mRNA molecule encoding a protein of interest. For example, in some embodiments, mRNA for expressing proteins such as green fluorescent protein (GFP), RNA-guided DNA binders, or Cas nucleases is described herein. LNP compositions are provided, which include Cas nuclease mRNA, for example, 2 types of Cas nuclease mRNAs that are allowed to be expressed in cells of 2 types of Cas nucleases (such as Cas9 or Cpf1 proteins). In addition, the prophylactic / therapeutic agent may contain one or more guide RNAs or nucleic acids encoding guide RNAs. Template nucleic acids (for example, for repair or recombinant) may also be included in the composition, or template nucleic acids may be used in the methods described herein. In some embodiments, the prophylactic / therapeutic agent includes mRNA encoding optional Streptococcus pyogenes Cas9 and Streptococcus pyogenes gRNA. In some embodiments, the prophylactic / therapeutic agent includes mRNA encoding optional Neisseria meningitidis Cas9 and nme gRNA.

[0249] As used herein, the term "mRNA" refers to a polynucleotide and includes an open reading frame that can be translated into a polypeptide (i.e., can be used as a substrate for translation by ribosomes and aminoacylated tRNA). mRNA can include a phosphate-sugar backbone comprising a ribose residue or an analog thereof, such as a 2'-methoxyribose residue. In some embodiments, the sugar of the mRNA phosphate-sugar backbone consists essentially of a ribose residue, a 2'-methoxyribose residue, or a combination thereof. In general, mRNA does not contain a large amount of thymidine residues (e.g., 0 residues or less than 30, 20, 10, 5, 4, 3, or 2 thymidine residues; or less than 10%, 9%, 8%, 7%, 6%, 5%, 4%, 4%, 3%, 2%, 1%, 0.5%, 0.2%, or 0.1% thymidine content). mRNA can contain modified uridines at some or all of its uridine positions.

[0250] The methods of delivering preventive / therapeutic agents to cells, tissues or organs described in the present invention include in vivo and / or in vitro delivery. In some embodiments, such methods include in vivo delivery. In some embodiments, such methods include in vitro delivery. In some implementations, the present disclosure provides methods for delivering one or more therapeutic and / or preventive nucleic acids to individuals in need thereof as described herein. In some embodiments, the drug delivery system, lipid nanoparticle, and pharmaceutical composition include therapeutic agents and / or preventive agents that can be specifically delivered to the liver cells of an individual. Exemplary liver cells include, but are not limited to, hepatocytes. In some embodiments, the drug delivery system, lipid nanoparticle, and pharmaceutical composition include therapeutic agents and / or preventive agents that can be specifically delivered to the spleen cells of an individual. Exemplary spleen cells include, but are not limited to, spleen mononuclear cells, spleen T cells, spleen memory B cells, or spleen B cells. In some embodiments, the drug delivery system, lipid nanoparticle, and pharmaceutical composition include therapeutic agents and / or preventive agents that can be specifically delivered to the bone marrow cells of an individual. Exemplary bone marrow cells include, but are not limited to, bone marrow mononuclear cells, bone marrow B cells, bone marrow memory B cells, or bone marrow T cells. In some embodiments, the drug delivery system, lipid nanoparticles, pharmaceutical compositions include therapeutic agents and / or preventatives that can be specifically delivered to individual immune cells. Exemplary immune cells include, but are not limited to, CD8+, CD4+, or CD8+CD4+ cells. In some embodiments, the drug delivery system, lipid nanoparticles, pharmaceutical compositions include therapeutic agents and / or preventatives that can be specifically delivered to individual hematopoietic stem cells. In some embodiments, the drug delivery system, lipid nanoparticles, pharmaceutical compositions include therapeutic agents and / or preventatives that can be specifically delivered to cardiac cells (e.g., cardiomyocytes). In some embodiments, the drug delivery system, lipid nanoparticles, pharmaceutical compositions include therapeutic agents and / or preventatives that can be specifically delivered to muscle cells (e.g., myocytes).

[0251] In some embodiments, lipid nanoparticle can be deployed to and be delivered to mammal liver hepatocyte, liver immune cell, spleen T cell or lung endothelial cell in the absence of targeting ligand.Specificity is delivered to the cell of particular classification or type and shows that the lipid nanoparticle of higher ratio is delivered to the cell of target type or classification.In some embodiments, specificity is delivered and can cause than using conventional nanoparticle system (for example, the LNP containing MC3) delivery 2 times, 5 times, 10 times, 15 times or 20 times.

[0252] The methods of using the drug delivery systems, ionizable lipid compounds, lipid nanoparticles, and pharmaceutical compositions disclosed herein are used in methods for producing polypeptides. In addition, in some embodiments, the lipid nanoparticles described herein can be used to produce polypeptides in target cells of an individual in need. For example, in some embodiments, the lipid nanoparticles described herein can be used to produce polypeptides in target cells of an individual in need. In some embodiments, the drug delivery systems, ionizable lipid compounds, lipid nanoparticles, and pharmaceutical compositions disclosed herein include one or more nucleic acid sequences to be delivered to a cell. In some embodiments, the one or more nucleic acids are expressed in a cell. In some embodiments, expression of the nucleic acid sequence involves one or more of the following: (1) generating an RNA template from a DNA sequence (e.g., by transcription); (2) processing the RNA transcript (e.g., by splicing, editing, 5' cap formation and / or 3' end formation); (3) translating the RNA into a polypeptide or protein; and / or (4) post-translational modification of the polypeptide or protein.

[0253] The methods of using the drug delivery systems, ionizable lipid compounds, lipid nanoparticles, and pharmaceutical compositions disclosed herein are used for gene regulation. In addition, in some embodiments, the lipid nanoparticles described herein can be used to reduce and / or increase gene expression in target cells of an individual in need thereof. For example, in some embodiments, the lipid nanoparticles described herein can deliver one or more nucleic acids to an individual's target cells in the absence of a targeting ligand. In some embodiments, the nucleic acid is an inhibitor nucleic acid. In some embodiments, the inhibitory nucleic acid is an siRNA. In some embodiments, the nucleic acid is a nucleic acid described herein. In some embodiments, the lipid nanoparticles described herein can deliver a prophylactic / therapeutic agent to an individual's target cells in the absence of a targeting ligand.

[0254] In addition, methods for editing genes in cells of individuals in need thereof using the drug delivery systems, ionizable lipid compounds, lipid nanoparticles, and pharmaceutical compositions disclosed herein. In some embodiments, the cells targeted for gene regulation are immune cells. The immune cells can be T cells, such as CD8+ T cells, CD4+ T cells, or T regulatory cells. Other exemplary immune cells for gene editing include, but are not limited to, macrophages, dendritic cells, B cells, or natural killer cells. In some embodiments, cells targeted for gene regulation in hepatocytes. Exemplary genes that can be targeted include, but are not limited to, T cell receptor, B cell receptor, CTLA4, PD1, FOXO1, FOXO3, AKTs, CCR5, CXCR4, LAG3, TIM3, killer immunoglobulin-like receptor, GITR, BTLA, LFA-4, T4, LFA-1, Bp35, CD27L receptor, TNFRSF8, TNFRSF5, CD47, CD52, ICAM-1, LFA-3, L-selectin, Ki-24, MB1, B7, B70, M-CSFR, TNFR-II, IL-7R, OX-4 β, CD137, CD137L, CD30L, CD40L, FasL, TRAIL, CD257, LIGHT, TRAIL-R1, TRAILR2, TRAIL-R4, TWEAK-R, TNFR, BCMA, B7DC, BTLA, B7-H1, B7-H2, B7-H3, ICOS, VEGFR2, NKG2D, JAG1, GITR, CD4, CCR2, GATA-3, MTORC1, MTORC2, RAPTOR, GATOR, FOXP3, NFAT, IL2R, and IL7. Other exemplary genes that can be targeted include, but are not limited to, OCT, G6Pase, Mut, PCCA, PCCB, and PAH. Exemplary tumor-associated antigens that can be recognized by T cells and are contemplated for targeting include, but are not limited to, MAGE1, MAGE3, MAGE6, BAGE, GAGE, NYESO-1, MART1 / Melan A, MC1R, GP100, tyrosinase, TRP-1, TRP-2, PSA, CEA, Cyp-B, Her2 / Neu, hTERT, MUC1, PRAME, WT1, RAS, CDK-4, MUM-1, KRAS, MSLN, and β-catenin.

[0255] As used herein, the terms "tissue" and / or "organ" refer to living cell material in aggregated form, such as a small portion of an organ, as well as dispersed cells, such as cells dispersed, separated and / or grown from muscle, myocardium, liver or kidney, including bone marrow cells and progeny cells, hematogenous stem cells and progeny, and various other blood elements. In some embodiments, the tissue and / or organ refers to kidney, heart, liver, stomach, spleen, pancreas, lung, brain, eye, intestine, bladder, skin or skin tissue, blood vessels, veins, arteries, heart valves, sperm and oocytes. As used herein, the term "organ" encompasses solid organs, such as kidney, heart, liver, lung, and functional parts of organs, such as skin segments, arterial segments, venous segments, transplantable liver lobes, kidney lobes, lung lobes, etc.

[0256] As used herein, the term "prevent" refers to delaying the onset of one or more symptoms of a particular disease, disorder, or condition, and / or reducing their frequency and / or severity. In some embodiments, prevention is assessed on a population basis, such that an agent is considered to "prevent" a particular disease, disorder, or condition if a statistically significant reduction in the development, frequency, and / or intensity of one or more symptoms of the disease, disorder, or condition is observed in a population susceptible to the disease, disorder, or condition. Prevention can be considered accomplished when the onset of the disease, disorder, or condition has been delayed for a predetermined period of time.

[0257] As used herein, the term "treatment" refers to the administration of a therapy that partially or completely alleviates, ameliorates, alleviates, inhibits one or more symptoms, features, and / or causes of a particular disease, disorder, and / or condition, delays its onset, reduces its severity, and / or reduces its incidence. In some embodiments, treatment may be directed to individuals who do not develop signs of the relevant disease, disorder, and / or condition and / or individuals who only develop early signs of the disease, disorder, and / or condition. Alternatively or in addition, such treatment may be directed to individuals who develop one or more confirmed symptoms of the relevant disease, disorder, and / or condition. In some embodiments, treatment may be directed to individuals who have been diagnosed with the relevant disease, disorder, and / or condition. In some embodiments, treatment may be directed to individuals who are known to have one or more susceptibility factors that are statistically associated with an increased risk of developing the relevant disease, disorder, and / or condition. Therefore, in some embodiments, treatment may be preventive; in some embodiments, treatment may be therapeutic.

[0258] The "treated individual" of the present invention is a mammal suffering from cancer, autoimmune disease, infectious disease, organ transplantation, organ failure, protein deficiency or a combination thereof. In some embodiments, the individual is a human. In some embodiments, the methods described herein can cause liver cells to translate certain proteins. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA or siRNA to liver cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA or siRNA to spleen T cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA or siRNA to spleen B cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA or siRNA to spleen mononuclear cells. In some embodiments, the methods described herein can be used to deliver one or more DNA, mRNA, sgRNA or siRNA to bone marrow cells.

[0259] As used herein, the phrase "therapeutic agent" refers to an agent that has a therapeutic effect and / or induces a desired biological and / or pharmacological effect when administered to a subject. In some embodiments, a therapeutic agent is any substance that can be used to alleviate, ameliorate, alleviate, inhibit, prevent, delay the onset of, reduce the severity of, and / or reduce the incidence of one or more symptoms or features of a disease, disorder, and / or condition.

[0260] As used herein, the term "therapeutically effective amount" means an amount of a substance (e.g., a therapeutic agent, composition, and / or formulation) that elicits a desired biological response when administered as part of a treatment regimen. In some embodiments, a therapeutically effective amount of a substance is an amount sufficient to treat, diagnose, inhibit, alleviate, prevent, and / or delay the onset of a disease, disorder, and / or condition when administered to an individual suffering from or susceptible to the disease, disorder, and / or condition. As will be understood by one of ordinary skill in the art, the effective amount of a substance can vary depending on factors such as the desired biological endpoint, the substance to be delivered, the target cell or tissue, and the like. For example, an effective amount of a compound in a formulation for treating a disease, disorder, and / or condition is an amount that alleviates, ameliorates, mitigates, inhibits, prevents, delays the onset of, reduces the severity of, and / or reduces the incidence of, one or more symptoms or characteristics of the disease, disorder, and / or condition. In some embodiments, the therapeutically effective amount is administered in a single dose; in some embodiments, multiple unit doses are required to deliver the therapeutically effective amount. The exact dosage will vary depending on various factors, such as individual variables (e.g., age, immune system health, etc.), the disease, and the treatment being performed.

[0261] As used herein, "administering a pharmaceutical composition" refers to administering a pharmaceutical composition disclosed herein to an individual in a therapeutically effective amount as described herein. In some embodiments, a person of ordinary skill will be able to design appropriate dosage levels and dosing regimens using the pharmaceutical compositions described herein to treat a variety of conditions in a variety of patients, taking into account the recipient's therapeutic environment, age, and general health. For example, in some embodiments, the selected dosage depends on the desired therapeutic effect, route of administration, and duration of treatment desired. In some embodiments, a dosage level of about 0.001 mg to about 5 mg of nucleic acid per kilogram of body weight is typically administered to a mammal. More specifically, in some embodiments, the preferred dosage of nucleic acid in the disclosed compositions is about 0.1 mg / kg to about 1.0 mg / kg. For the disclosed pharmaceutical compositions, a dosage level of about 0.2 mg to about 100 mg of the four components (ionizable lipid, cholesterol, conjugate-linked conjugate, and phospholipid) is typically applied to a mammal. More specifically, in some embodiments, the preferred dosage of the disclosed nanoparticles is about 0.5 mg / kg to about 5 mg / kg of the four components per kilogram of body weight.

[0262] The synthetic route of the ionizable lipid molecules in the examples of this application is as follows:

[0263] In the above intermediate or product structures, each group L1, R', R, R1, R2, R3 and R4 is as defined above in this application.

[0264] Example 1: Synthesis of ionizable lipid molecule LA-3

[0265] (1) Synthesis of tetradecanal

[0266] The synthetic route is as follows:

[0267] The amount of each material used in the reaction is shown in Table 1 below.

[0268] Table 1

[0269] ① Process: Dissolve tetradecanol in a mixed solution of DCM and DMSO, place at 0°C, add Et3N, then add SO3-pyridine, stir for 10 minutes, naturally warm to room temperature, stir for 0.5 hours, and detect the completion of the reaction by TLC.

[0270] ② Post-treatment: The reaction solution was poured into saturated NH4Cl (30 mL), separated using a separatory funnel, the aqueous phase was extracted with DCM (3×50 mL), the organic phases were combined, dried over anhydrous MgSO4, concentrated, and subjected to silica gel column chromatography.

[0271] ③ Purification: PE:EA=100:1 to obtain the product.

[0272] (2) Synthesis of Intermediate 003

[0273] The synthetic route is as follows:

[0274] The amount of each material used in the reaction is shown in Table 2 below.

[0275] Table 2

[0276] Procedure: Stir the two reactants together and react at 60°C for 1 h (without adding solvent).

[0277] Post-treatment: Use an oil pump (70℃) to rotate for 2 hours to remove 1,3-propylenediamine. Do not pass through the column and directly proceed to the next step.

[0278] (3) Synthesis of LA-3

[0279] The synthetic route is as follows:

[0280] The amount of each material used in the reaction is shown in Table 3 below.

[0281] Table 3

[0282] Procedure: Dissolve intermediate 003 in DCM:MeOH = 10:1, then add tetradecanal, stir for 10 min, then add NaBH(OAC)3, and react at room temperature overnight.

[0283] Post-treatment: add saturated NaHCO3 to make it alkaline, extract the aqueous phase with DCM (3×50 mL), combine the organic phases, dry over anhydrous MgSO4, concentrate, and chromatograph on a silica gel column.

[0284] Purification: DCM:MeOH=15:1 to obtain the product.

[0285] The same method as LA-3 in Example 1 was used to prepare the corresponding ionizable lipid molecules in Tables 7 and 8 below, except that the head portion raw material in Table 4 was replaced with L-dimethyl malate, the linker portion raw material in Table 5 was replaced with 1,3-propylenediamine, and the tail portion raw material in Table 6 was replaced with tetradecane.

[0286] Table 4 Head part raw materials

[0287] Table 5 Linker raw materials

[0288] Table 6 Raw materials for tail

[0289] Table 7: Ionizable lipid molecules

[0290] Table 8: Ionizable lipid molecules

[0291] The structural formula, NMR and mass spectrometry data of the ionizable lipid molecules in Table 7 above are shown in Table 9, and the structural formula, NMR and mass spectrometry data of the ionizable lipid molecules in Table 8 above are shown in Figures 24A-38B, where the NMR and mass spectrometry characterization methods are as follows:

[0292] H NMR spectrum:

[0293] 10-20 mg of the compound was dissolved in deuterated chloroform, and 1H NMR data were collected using a Bruker AVANCE 400 (400 MHz).

[0294] Mass spectrometry characterization: 10-20 mg of the compound was dissolved in anhydrous methanol as solvent and characterized in positive ion mode.

[0295] Table 9

[0296] The same method as LA-3 in Example 1 was used to prepare the corresponding ionizable lipid molecules shown in Table 13 below, except that the head portion raw material in Table 10 was replaced with dimethyl L-malate, the linker portion raw material in Table 11 was replaced with 1,3-propylenediamine, and the tail portion raw material in Table 12 was replaced with tetradecane.

[0297] Table 10: Raw materials for head

[0298] Table 11: Some raw materials of linker

[0299] Table 12: Raw materials for tail

[0300] Table 13: Ionizable lipid molecules

[0301] Example 2 mRNA-LNP Preparation

[0302] 1. The ionizable lipid molecules prepared in Example 1 or the ionizable lipid molecules C12-200 (purchased from MedChemExpress, product number HY-145405) as a control were dissolved in ethanol to obtain a lipid mixture according to the molar ratio of ionizable lipid molecules: DSPC (purchased from Jiangsu Southeast Nanomaterials Co., Ltd.): cholesterol (purchased from Japan Seika Co., Ltd.): DMG-PEG2000 (purchased from Ailiying Pharmaceutical Group (Tianjin) Co., Ltd.) = 48.5:10:40:1.5, with a total concentration of 10 mg / mL.

[0303] 2. mRNA was dissolved in 100 mM sodium acetate buffer (pH = 4.0).

[0304] 3. The lipid mixture prepared in step 1 was mixed with sodium acetate buffer (pH 4.0) containing mRNA in a ratio of 1:3 using a microfluidic device (Ignite, PNI) at a flow rate of 16 mL / min to obtain a mixed solution.

[0305] 4. Add the mixed solution from step 3 to a 10K MVCO ultrafiltration centrifuge tube (Merck Millipore), add 0.8× TBS to balance, and centrifuge at 4000 g for 25 minutes using a high-speed centrifuge.

[0306] 5. Add 0.8× TBS again, centrifuge at 4000 g for 25 minutes, and collect the liquid.

[0307] Example 3 In vitro test - LNP physicochemical property test

[0308] The size test results of representative LNPs prepared in Example 2 are shown in Figure 1 , the PDI test results are shown in Figure 2 , and the mRNA encapsulation efficiency is shown in Figure 3 . Liposome particle size and PDI were measured using a Zetasizer Nano ZS (Malvern Panalytical, USA), and data were analyzed using Zetasizer Software. Encapsulation efficiency was measured using the Quant-iT RiboGreen RNA Quantification Kit (Invitrogen, USA).

[0309] Results: The size of the prepared LNP is usually around 100 nm; the PDI of the LNP is less than 0.2; and the mRNA encapsulation efficiency of the LNP is at least greater than 60% (preferably greater than 70%).

[0310] Example 4 In vivo test - targeting test (muscular injection and intravenous injection)

[0311] Experimental operation:

[0312] Babl / c mice aged 8 weeks were adaptively fed for 2 weeks and administered the representative mRNA-LNPs prepared in Example 2 via tail vein / intramuscular injection. Six hours after this, 200 μl of luciferase substrate (15 mg / ml) was intraperitoneally injected. Under gas anesthesia, in vivo imaging (IVIS Spectrum, PerkinElmer, USA) was performed. Following in vivo imaging, organ samples (heart, liver, spleen, lung, kidney, lymph node, and muscle) were harvested and imaged (IVIS Spectrum, PerkinElmer, USA). Dosages are shown in Table 14, and dosing frequency was once. Images and data were analyzed using Living Image 4.4 software.

[0313] Table 14

[0314] As shown in Figures 4 to 7, for intramuscular injection, the ionizable lipid compounds LA-8, LA-9, LA-17, LA-18, LA-19 and LA-20 prepared by the present invention have significantly better liver targeting compared with control compound 1 (referred to as "control 1" in this application) and control compound 2 (referred to as "control 2" in this application).

[0315] Control compound 1

[0316] Control compound 2

[0317] As shown in Figures 8 to 11, for intravenous injection, the ionizable lipid compounds LA-2, LA-3, LA-5, LA-8, LA-9, LA10, LA-12, LA-13, LA-14, LA-17, LA-18, LA-19 and LA-20 prepared by the present invention have significantly better liver targeting compared with Control 1 and Control 2, especially LA-18, which almost completely (up to 96.74%) enters the liver after intravenous injection.

[0318] The above results indicate that the lipid nanoparticles prepared from the ionizable lipid molecules provided by the present invention have good liver targeting properties.

[0319] In addition, the present invention found that the difference in the chiral configuration of ionizable lipid molecules has a significant impact on the efficiency of LNPs containing them in delivering mRNA and translating it into protein. In particular, the efficiency of S-configuration LA-18(S) in delivering and translating mRNA is significantly better than that of R-configuration LA-18(R).

[0320] Following intravenous injection, the in vivo fluorescence intensity of the S-configuration compound group was significantly higher than that of the R-configuration compound group, particularly LA-18, whose fluorescence intensity was several times that of the R-configuration. The fluorescence intensity of the S-configuration compound group in vitro organ imaging was also significantly higher than that of the R-configuration compound group, particularly LA-18, whose fluorescence intensity was several times that of the R-configuration compound. The in vivo imaging results for compounds LA-8(S) and LA-8(R) (i.e., compound DA-38), LA-9(S) and LA-9(R) (i.e., compound DA-39), and LA-18(S) and LA-18(R) (i.e., compound DA-40) are shown in Table 15, the cell assay results are shown in Figures 12-13, and the in vitro organ imaging results are shown in Figures 14-15 and Table 16.

[0321] Table 15

[0322] Table 16

[0323] Example 5 In vitro test - cell test (cell survival rate and mRNA transfection effect)

[0324] Cell types: 293T (purchased from ATCC), BEAS-2B (purchased from Shanghai Meiyan Biotechnology Co., Ltd.), LX-2 (purchased from Shanghai Meiyan Biotechnology Co., Ltd.), DC2.4 (purchased from Millpore).

[0325] Experimental operation:

[0326] 1. Cell Culture: 293T cells were cultured in DMEM supplemented with 10% fetal bovine serum; BEAS-2B cells were cultured in DMEM supplemented with 10% fetal bovine serum; LX-2 cells were cultured in DMEM supplemented with 10% fetal bovine serum; DC2.4 cells were cultured in RPMI 1640 supplemented with 10% fetal bovine serum.

[0327] 2. mRNA transfection: Cells in the logarithmic growth phase were inoculated into 96-well plates. After overnight culture, cells were transfected with lipo max (Invitrogen) and mRNA-LNP. Lipo max transfected 100 ng per well, and LNP samples transfected 200 ng and 400 ng, and incubated at 37 ° C under 95% relative humidity and 5% CO2 for 24 h. RNA expression was detected on a microplate reader (Varioskan LUX, Thermo Scientific, USA) using a luciferase reporter gene detection reagent (Vazyme, DD1203-02). The results are shown in Figures 20 to 23. As can be seen from the figure, the LNPs prepared using the lipids of the present invention can achieve transfection and mRNA expression for different cell types. In addition, the experimental results show that the in vitro mRNA expression of different lipid LNPs is different from the in vivo expression, which is consistent with the literature reports.

[0328] 3. Cytotoxicity: Cells in the logarithmic growth phase were seeded in 96-well plates. After overnight culture, cells were transfected with lipomax and mRNA-LNPs, as before, and incubated at 37°C, 95% relative humidity, and 5% CO2 for 24 hours. Detection was performed using a CCK8 assay kit (GLPBIO), and OD values ​​were measured using a microplate reader (Varioskan LUX, Thermo Scientific, USA). After removing the maximum and minimum values ​​of each group, cell viability was calculated according to the formula: I (%) = [(A experimental group - A blank group) / (A control group - A blank group)] × 100%. The results are shown in Figures 16 to 19. As can be seen from the figures, at doses of 200 ng and 400 ng, the LNPs prepared using the lipids of the present invention had no significant effect on cell viability. The dose of conventional lipid cytotoxicity experiments is typically set at around 100 ng. This shows that the developed lipids have superior biosafety compared to conventional lipids.

[0329] The technical solution of the present invention is not limited to the above-mentioned specific embodiments. Any technical variations made according to the technical solution of the present invention fall within the protection scope of the present invention.

Claims

1. A compound, which is a compound of Formula I or a pharmaceutically acceptable salt, pharmaceutically acceptable ester, isomer, isotope-labeled compound (especially a deuterated compound) or solvate of a compound of Formula I: in, X1 and X2 are the same or different and are each independently selected from -O- or -NR a -; R1, R2, R3 and R4 are the same or different and are each independently selected from a C2-C30 saturated or unsaturated aliphatic hydrocarbon group, wherein one or more methylene units in the C2-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally and independently replaced by L; hydrogen atoms on carbon atoms in the C2-C30 saturated or unsaturated aliphatic hydrocarbon group are optionally substituted by one or more substituents selected from hydroxyl groups and C1-C6 alkoxy groups; L1 and L2 are the same or different and are each independently selected from a C1-C20 saturated or unsaturated aliphatic alkylene group, a C6-C15 arylene group, a C3-C15 heteroarylene group, a 5-6 membered heterocycloalkylene group or a combination thereof, wherein one or more methylene units in the C1-C20 saturated or unsaturated aliphatic alkylene group are optionally and independently replaced by L; L is selected from -O-, -(C=O)O-, -NR a -, -(C=O)NR a - or a combination thereof; R a Selected from hydrogen or C1-C6 alkyl.

2. The compound according to claim 1, characterized in that R1, R2, R3 and R4 are the same or different and are each independently selected from C4-C24 alkyl, C3-C20 cycloalkyl, C4-C24 alkenyl, C4-C24 alkynyl, one or more methylene units in the C4-C24 alkyl, C3-C20 cycloalkyl, C4-C24 alkenyl, C4-C24 alkynyl are optionally and independently replaced by a group selected from -(C=O)O- and -(C=O)NH-; the hydrogen atoms on carbon atoms in the C4-C24 alkyl, C3-C20 cycloalkyl, C4-C24 alkenyl, C4-C24 alkynyl are optionally substituted by one or more hydroxyl groups.

3. The compound according to claim 1, characterized in that R1, R2, R3 and R4 are each independently selected from the following groups: wherein n1, n2, n5 and n6 are each independently selected from 1, 2, 3, 4, 5, 6, 7 or 8; n3, n4, n7, n8, n9 and n10 are each independently selected from 1, 2, 3, 4, 5, 6, 7 or 8; 10 Each is independently selected from 0, 1, 2, 3, 4 or 5; n 11 Selected from 1, 2, 3, 4, 5, 6, 7 or 8; n 12 and n 13 Each is independently selected from 2, 3, 4, 5, 6 or 7.

4. The compound according to claim 1, characterized in that R1, R2, R3 and R4 are each independently selected from the following groups:

5. The compound according to claim 1, characterized in that L1 and L2 are the same or different and are each independently selected from C1-C15 alkylene, C3-C10 cycloalkylene, C6-C15 arylene, C3-C15 heteroarylene, 5-6 membered heterocycloalkylene or a combination thereof, wherein one or more methylene units in the C1-C15 alkylene and C3-C10 cycloalkylene are optionally and independently selected from -O- and -NR a - group replacement; the 5-6 membered heterocycloalkylene group has at least one heteroatom, and the heteroatom is selected from N, O and S; Preferably, L1 and L2 are each independently selected from the following groups:

6. The compound according to claim 1, characterized in that In Formula 1: X1 and X2 are the same or different and are each independently selected from -NR a -; and / or, L1 and L2 are the same or different and are each independently selected from a C2-C8 straight chain alkylene group; and / or, R1, R2, R3 and R4 are the same or different and are each independently selected from a C8-C18 straight-chain alkyl group or a C8-C18 straight-chain alkenyl group; Preferably, in Formula I: R1, R2, R3 and R4 are each independently selected from the following groups: And L1 and L2 are each independently selected from the following groups: and X1 and X2 are both -NH-; Alternatively, in Formula I: R1, R2, R3 and R4 are each independently selected from the following groups: and L1 and L2 are each independently selected from the following groups: and X1 and X2 are both -NH-.

7. The compound according to claim 1, characterized in that The compound has a structure represented by Formula I-1 or Formula I-2: wherein, the definitions of X1, X2, R1, R2, R3, R4, L1 and L2 are the same as those in Formula I.

8. The compound according to claim 1, characterized in that The compound has a structure represented by Formula I-3 or Formula I-4: wherein, the definitions of R1, R2, R3, R4, L1 and L2 are the same as those in Formula I.

9. The compound according to claim 1, characterized in that The compound has a structure represented by formula I-5: wherein R1 to R4 are as defined in Formula I, m1 and m2 are the same or different and are independently selected from 1, 2, 3, 4, 5, 6, 7 or 8.

10. The compound according to claim 1, characterized in that The compound has a structure represented by formula I-6: wherein R1 to R4 are the same or different and are each independently selected from a C8-C20 straight chain alkyl group; m1 and m2 are the same or different and are each independently selected from 2, 4, 5, 6 or 8.

11. The compound according to claim 1, characterized in that The compound has a structure represented by formula I-7: Wherein, L1 and L2 are defined as described in Formula I; L3 to L6 are the same or different and are independently selected from C2-C6 alkylene; R5 to R8 are the same or different and are independently selected from C5-C19 alkyl and C5-C19 alkenyl.

12. The compound according to claim 1, characterized in that The compound is not:

13. The compound according to claim 1, characterized in that The compound is selected from the following structures:

14. A method for preparing the compound according to any one of claims 1 to 13, characterized in that The steps include: (1) reacting a compound of formula A with an amine compound B to obtain an intermediate of formula C; wherein, in formula A, R X represents a C1-C10 straight-chain or branched alkyl group, preferably a C1-C4 straight-chain or branched alkyl group; the amine compound B includes H2N-L1-X1H and H2N-L2-X2H, where L1, L2, X1, and X2 are as defined in Formula I for L1, L2, X1, and X2; (2) reacting the intermediate represented by formula C with an aldehyde compound R'CHO or an alcohol compound ROH to obtain a compound represented by formula I; wherein R' represents an R group that has lost a methylene unit, and R is defined as R1, R2, R3 and R4 in formula I; 15. The method according to claim 14, characterized in that In step (1), the molar ratio of the compound of formula A to the amine compound B is 1:(2-6), preferably 1:(3-5); and / or, In step (1), the reaction temperature is 50-80°C; and / or, In step (2), the molar ratio of the intermediate represented by formula C to the aldehyde compound R'CHO or the alcohol compound ROH is 1:(4-10), preferably 1:(6-8); and / or, In step (2), the reaction temperature is 20-35°C; and / or, In step (2), the reaction is carried out in the presence of a solvent and a catalyst; preferably, the solvent comprises a halogenated hydrocarbon solvent and an alcohol solvent, preferably comprises dichloromethane and methanol; preferably, the volume ratio of the halogenated hydrocarbon solvent to the alcohol solvent is (5-15):1; preferably, the catalyst comprises NaBH(OAC)3; preferably, the molar ratio of the catalyst to the aldehyde compound R'CHO or the alcohol compound ROH is 1:(0.5-1).

16. A lipid nanoparticle comprising an ionizable lipid compound, wherein the ionizable lipid compound comprises the compound according to any one of claims 1 to 13.

17. The lipid nanoparticle according to claim 16, characterized in that The lipid nanoparticles further comprise a helper lipid, cholesterol and a conjugate-linker lipid; the helper lipid preferably comprises DOPE or DSPC; the conjugate-linker lipid preferably comprises a PEGylated lipid and more preferably comprises DMG-PEG 2000.

18. The lipid nanoparticle according to claim 17, characterized in that Based on the total molar number of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid, the molar content of the ionizable lipid compound is 20-70%, the molar content of the helper lipid is 1-20%, the molar content of the cholesterol is 20-50%, and the molar content of the conjugate-linker lipid is 1-10%; Preferably, based on the total molar number of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid, the molar content of the ionizable lipid compound is 30-60%, the molar content of the helper lipid is 5-20%, the molar content of the cholesterol is 25-45%, and the molar content of the conjugate-linker lipid is 1-5%; More preferably, based on the total molar number of the ionizable lipid compound, the helper lipid, cholesterol and the conjugate-linker lipid, the molar content of the ionizable lipid compound is 30-55%, the molar content of the helper lipid is 5-20%, the molar content of the cholesterol is 35-45%, and the molar content of the conjugate-linker lipid is 1-5%.

19. A drug delivery system or drug carrier comprising the ionizable lipid compound according to any one of claims 1 to 13 or the lipid nanoparticle according to any one of claims 15 to 17.

20. The drug delivery system according to claim 19, wherein Also included are therapeutic and / or preventive agents, preferably comprising one or more of DNA molecules, RNA molecules, proteins, polypeptides, and small molecule drugs; and / or, The therapeutic and / or preventive agent includes a pharmaceutical active ingredient for treating or preventing one or more of liver disease, spleen disease, lung disease, heart disease, and kidney disease; and / or, The administration mode of the drug delivery system includes one or more of intramuscular injection, intradermal injection, intravenous injection, arterial injection, transdermal absorption, intraperitoneal injection, oral administration, and nasal spray, preferably intramuscular injection or intravenous injection.

21. A pharmaceutical composition comprising: The lipid nanoparticles according to any one of claims 16 to 18, and therapeutic and / or preventive agents; the therapeutic and / or preventive agents preferably include one or more of DNA molecules, RNA molecules, proteins, polypeptides and small molecule drugs.

22. The pharmaceutical composition according to claim 21, characterized in that The therapeutic and / or preventive agent comprises an RNA molecule; the RNA molecule preferably comprises one or more of siRNA, microRNA, mRNA, tRNA and circRNA.

23. The pharmaceutical composition according to claim 21, characterized in that The therapeutic and / or preventive agent includes a pharmaceutical active ingredient for treating or preventing one or more of liver disease, spleen disease, lung disease, heart disease, and kidney disease.

24. The pharmaceutical composition according to claim 21, characterized in that The administration of the pharmaceutical composition includes one or more of intramuscular injection, intradermal injection, intravenous injection, arterial injection, transdermal absorption, intraperitoneal injection, oral administration, and nasal spray, preferably intramuscular injection or intravenous injection.

25. The pharmaceutical composition according to claim 21, characterized in that The pharmaceutical composition further comprises any one or a combination of at least two of pharmaceutically acceptable carriers, excipients, and adjuvants.

26. Use of the compound according to any one of claims 1 to 13 or the lipid nanoparticle according to any one of claims 16 to 18 in preparing a drug delivery system.

27. The use according to claim 26, characterized in that The active ingredients of the drug include one or more of DNA molecules, RNA molecules, proteins, polypeptides and small molecule drugs; Preferably, the drug delivery system comprises a nucleic acid delivery system; the nucleic acid preferably comprises an RNA molecule, more preferably comprises one or more of siRNA, microRNA, mRNA, tRNA and circRNA; Preferably, the compound or the lipid nanoparticle is formulated to deliver the active ingredient of the drug to target cells; the target cells preferably include spleen cells, liver cells such as hepatocytes, bone marrow cells (such as bone marrow mononuclear cells), lung cells, immune cells, muscle cells such as myocytes, heart cells such as cardiomyocytes, kidney cells or central nervous system cells, hematopoietic stem cells or one or more.

28. Use of the compound according to any one of claims 1 to 13 or the lipid nanoparticle according to any one of claims 16 to 18 in the preparation of organ-targeted drugs.

29. The use according to claim 28, characterized in that The compound or the lipid nanoparticle is formulated to deliver the active ingredient of the drug to target cells; Preferably, the target cells include one or more of spleen cells, liver cells such as hepatocytes, bone marrow cells (such as bone marrow mononuclear cells), lung cells, immune cells, muscle cells such as myocytes, heart cells such as cardiomyocytes, kidney cells or central nervous system cells, and hematopoietic stem cells.

30. A transfection complex comprising the compound of any one of claims 1 to 13 or the lipid nanoparticle of any one of claims 16 to 18; Optionally, the transfection complex comprises at least one bioactive agent; Optionally, the bioactive agent comprises one or more of a DNA molecule, an RNA molecule, a protein, a polypeptide and a small molecule drug.

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