Imidazole-based lipids and uses thereof

Abstract

This disclosure relates to an imidazole lipid and its applications. Specifically, it provides a compound as shown in Formula I or a salt thereof, as well as lipid particles or pharmaceutical compositions containing the same. The compound can be used as a delivery system for the preparation of medicaments for the prevention and / or treatment of cancer, infectious diseases, autoimmune diseases, neurodegenerative diseases, and inflammation, etc. The definitions of the various groups in Formula (I) are as described in the specification.

Description

Imidazole lipid and its application Technical Field

[0001] The present invention relates to the field of medicine and relates to an imidazole lipid and its application. Background Art

[0002] Gene therapy involves introducing exogenous, normal genes into target cells to correct or compensate for diseases caused by defective or abnormal genes. Since the COVID-19 pandemic, gene therapy has become widely known and has become another hot topic after immunotherapy for cancer, gradually becoming a reality.

[0003] However, naked therapeutic genes are easily degraded by nucleases and exhibit poor cellular uptake. Therefore, the preparation of safe and efficient gene vectors is essential for successful gene therapy. Mainstream nucleic acid drug delivery systems are divided into viral vector systems and non-viral systems, with lipid nanoparticle-mediated nucleic acid drug delivery being the predominant method in non-viral delivery systems.

[0004] Nanoparticles composed of cationic lipids and other auxiliary lipids, such as those formed from cholesterol, phospholipids, and PEGylated lipids, can encapsulate nucleic acids, protecting them from degradation while promoting cellular uptake and reducing immune responses. Furthermore, lipid nanoparticles offer other advantages for cellular delivery of bioactive ingredients, including good targeting, minimal side effects, excellent stability, and high transfection efficiency.

[0005] However, lipid nanoparticles are generally not very selective for specific organs. For example, most lipid nanoparticles accumulate through biological processes in the liver, thereby reducing the effectiveness of active substance delivery to the target organ.

[0006] Therefore, there is still a need to develop new lipid nanoparticles for preferential delivery to specific organs and specific cells, such as better delivery systems to enhance T cell engineering in vivo. Summary of the Invention

[0007] The present disclosure provides a compound represented by formula I or a salt thereof,

[0008] in,

[0009] H 1 Selected from C 1-6 Alkylene, C 2-6 Alkenylene or C 1-6 heteroalkylene;

[0010] H 2 Selected from C 1-12 heteroalkylene;

[0011] R 1 Selected from C1-24 Alkyl, C 1-24 Heteroalkyl, C 2-24 Alkenyl or C 2-24 heteroalkenyl;

[0012] R 2 Selected from C 1-24 Alkyl, C 2-24 Alkenyl or -R b -L 1 -R c ;

[0013] L 1 and L 2 Each independently selected from -C(O)O-, -OC(O)-, -C(O)-, -OC(O)O-, -O-, -S(O) q -, -SS-, -C(O)S-, -SC(O)-, -NR a C(O)-、-C(O)NR a -、-NR a C(O)NR a -、-NR a C(O)O-、-OC(O)NR a -、-P(O)(OR a )-or key, R a Selected from hydrogen or C 1-6 Alkyl, q is selected from 0, 1 or 2;

[0014] R b Selected from C 1-12 Alkylene, C 1-12 Heteroalkylene, C 1-12 Alkenylene or C 1-12 heteroalkenylene;

[0015] R c Selected from C 1-24 Alkyl, C 1-24 Heteroalkyl, C 2-24 Alkenyl or C 2-24 heteroalkenyl;

[0016] R 3 Each independently selected from halogen, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkoxy, C 3-7 Cycloalkyl or C 3-7 heterocycloalkyl;

[0017] r is selected from an integer between 0 and 3, such as 1 or 2.

[0018] In some embodiments, in the compound of Formula I or a salt thereof, H1 Selected from C 1-6 Alkylene, C 1-6 Alkenylene or C 1-6 heteroalkylene;

[0019] H 2 Selected from C 1-12 heteroalkylene;

[0020] R 1 Selected from C 1-24 Alkyl, C 1-24 Heteroalkyl, C 2-24 Alkenyl or C 2-24 heteroalkenyl;

[0021] R 2 Selected from C 1-24 Alkyl, C 2-24 Alkenyl or -R b -L 1 -R c ;

[0022] L 1 and L 2 Each independently selected from -C(O)O-, -OC(O)-, -C(O)-, -OC(O)O-, -O-, -S(O) q -, -SS-, -C(O)S-, -SC(O)-, -NR a C(O)-、-C(O)NR a -、-NR a C(O)NR a -、-NR a C(O)O-、-OC(O)NR a -、-P(O)(OR a )-or key, R a Selected from hydrogen or C 1-6 Alkyl, q is selected from 0, 1 or 2;

[0023] R b Selected from C 1-12 alkylene;

[0024] R c Selected from C 1-24 Alkyl, C 1-24 Heteroalkyl, C 2-24 Alkenyl or C 2-24 heteroalkenyl;

[0025] R 3 Each independently selected from halogen, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkoxy, C 3-7 Cycloalkyl or C3-7 heterocycloalkyl;

[0026] r is selected from an integer between 0 and 3, such as 1 or 2.

[0027] In some embodiments, in the compound of Formula I or a salt thereof, H 2 Selected from C 1-12 A heteroalkylene group, wherein the heteroalkylene group comprises at least one heteroatom selected from O, N and S.

[0028] In some embodiments, in the compound of Formula I or a salt thereof, H 2 Selected from C 1-12 Heteroalkylene, such as C 2-9 Heteroalkylene, C 2-7 Heteroalkylene, C 2-4 Heteroalkylene, C 3-5 Heteroalkylene or C 3-6 Heteroalkylene.

[0029] In some embodiments, in the compound of formula I or its salt, the heteroalkylene group is a heteroalkylene group containing at least one oxygen atom. In some embodiments, in the compound of formula I or its salt, the heteroalkylene group is a heteroalkylene group containing two oxygen atoms. In some embodiments, in the compound of formula I or its salt, the heteroalkylene group is a heteroalkylene group containing at least one nitrogen atom. In some embodiments, in the compound of formula I or its salt, the heteroalkylene group is a heteroalkylene group containing at least one oxygen atom.

[0030] In some embodiments, in the compound of Formula I or a salt thereof, R 1 Selected from C 1-24 Alkyl, including C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Alkyl, C 11 Alkyl, C 12 Alkyl, C 13 Alkyl, C 14 Alkyl, C 15 Alkyl, C 16 Alkyl, C 17 Alkyl, C 18 Alkyl, C 19 Alkyl, C 20 Alkyl, C 21 Alkyl, C 22 Alkyl or C 23 alkyl.

[0031] In other embodiments, in the compound of Formula I or a salt thereof, R 1 Selected from C 4-18 alkyl.

[0032] In some embodiments, in the compound of Formula I or a salt thereof, R 1 For straight chain C 4-18 alkyl.

[0033] In some embodiments, in the compound of Formula I or a salt thereof, R 1 For branched chain C 4-18 alkyl.

[0034] In some embodiments, in the compound of Formula I or a salt thereof, R 1 For branched chain C 17 alkyl.

[0035] In other embodiments, in the compound of Formula I or a salt thereof, R 1 Selected from C 2-24 Alkenyl, including C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C 10 Alkenyl, C 11 Alkenyl, C 12 Alkenyl, C 13 Alkenyl, C 14 Alkenyl, C 15 Alkenyl, C 16 Alkenyl, C 17 Alkenyl, C 18 Alkenyl, C 19 Alkenyl, C 20 Alkenyl, C 21 Alkenyl, C 22 Alkenyl, C 23 Alkenyl or C 24 Alkenyl.

[0036] In some other embodiments, the compound of formula I or its salt is provided as follows: 1 C 4-18 Alkenyl.

[0037] In some embodiments, in the compound of Formula I or a salt thereof, R 1 For straight chain C 4-18 Alkenyl.

[0038] In some embodiments, in the compound of Formula I or a salt thereof, R 1 For branched chain C 4-18 Alkenyl.

[0039] In other embodiments, the compound of formula I or its salt is provided, wherein R 2 -R b -L 2 -R c , where R b , L 2 or R c As defined above.

[0040] Furthermore, some embodiments provide compounds of formula I as compounds of formula II,

[0041] Wherein, X is -N(R d )- or -O-, o is selected from an integer between 1 and 11, p is selected from an integer between 1 and 5, and o+p is not greater than 12, R d Selected from hydrogen or C 1-6 Alkyl, R 4 、R 5 、R 6 、R 7 Each independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy, L 1 , L 2 、H 1 、R 1 、R 3 、R b and R c As defined in the compound represented by formula I.

[0042] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b Selected from C 1-12 Alkylene, including C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene, C 10 Alkylene, C 11 Alkylene or C 12 Alkylene.

[0043] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b For straight chain C 1-12 Alkylene.

[0044] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b For branched chain C 1-12 Alkylene.

[0045] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C7 alkylene group.

[0046] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C8 alkylene group.

[0047] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C9 alkylene group.

[0048] In other embodiments, in the compound of Formula I or II or a salt thereof, R b Selected from C 2-12 Alkenylene, including C3 alkenylene, C4 alkenylene, C5 alkenylene, C6 alkenylene, C7 alkenylene, C8 alkenylene, C9 alkenylene, C 10 Alkenylene, C 11 Alkenylene or C 12 Alkenylene.

[0049] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b For branched chain C 2-14 Alkenylene.

[0050] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C7 alkenylene.

[0051] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight-chain C8 alkenylene group.

[0052] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight-chain C9 alkenylene group.

[0053] In other embodiments, in the compound of Formula I or II or a salt thereof, R c Selected from C 1-24 Alkyl, including C2 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C 10 Alkyl, C 11 Alkyl, C 12 Alkyl, C 13 Alkyl, C 14 Alkyl, C 15 Alkyl, C 16 Alkyl, C 17 Alkyl, C 18 Alkyl, C 19 Alkyl, C 20 Alkyl, C 21 Alkyl, C 22 Alkyl, C 23 Alkyl or C 24 alkyl.

[0054] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For straight chain C 1-12 alkyl.

[0055] In some embodiments, in the compound represented by Formula I or II or a salt thereof, Rc For branched chain C 1-12 alkyl.

[0056] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For branched chain C 11 alkyl.

[0057] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For branched chain C 17 alkyl.

[0058] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c It is a straight chain C9 alkyl group.

[0059] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For branched chain C 11-18 alkyl.

[0060] In other embodiments, in the compound of Formula I or II or a salt thereof, R c Selected from C 2-24 Alkenyl, including C2 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C 10 Alkenyl, C 11 Alkenyl, C 12 Alkenyl, C 13 Alkenyl, C 14 Alkenyl, C 15 Alkenyl, C 16 Alkenyl, C 17 Alkenyl, C 18 Alkenyl, C 19 Alkenyl, C 20 Alkenyl, C 21 Alkenyl, C 22 Alkenyl, C 23 Alkenyl or C 24 Alkenyl.

[0061] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For straight chain C 2-12 Alkenyl.

[0062] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For branched chain C 2-12 Alkenyl.

[0063] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For branched chain C 11 Alkenyl.

[0064] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c For branched chain C 17 Alkenyl.

[0065] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c It is a straight chain C9 alkenyl.

[0066] In some embodiments, in the compound of Formula I or II or a salt thereof, p is selected from 1 or 2.

[0067] In some embodiments, in the compound of Formula I or II or a salt thereof, p is selected from 3 or 4.

[0068] In some embodiments, in the compound represented by Formula I or II or a salt thereof, o is selected from an integer between 4 and 7, such as 5 or 6.

[0069] In some embodiments, in the compound of Formula I or II or a salt thereof, X is -O-.

[0070] In some embodiments, in the compound of Formula I or II or a salt thereof, p is selected from 1 or 2; X is -O-. In some embodiments, in the compound of Formula I or II or a salt thereof, p is selected from 3 or 4, o is selected from 5 or 6, and X is -O-.

[0071] In some embodiments, R 4 、R 5 Each independently selected from C 1-6 Alkyl groups such as methyl, ethyl or propyl.

[0072] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R 6 、R 7 Each independently selected from C 1-6 Alkyl groups such as methyl, ethyl or propyl.

[0073] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R 4 、R 5 、R 6 、R 7 are each independently selected from hydrogen.

[0074] In some embodiments, in the compound of formula I or II or a salt thereof, p is selected from 3 or 4, o is selected from 5 or 6, X is -O-, R 4 、R 5 、R 6 、R 7 are each independently selected from hydrogen.

[0075] In some embodiments, in the compound of formula I or II or a salt thereof, p is selected from 1 or 2, o is selected from 5 or 6, X is -O-, R 4 、R 5 、R 6 、R 7 are each independently selected from hydrogen.

[0076] In some embodiments, in the compound of formula I or II or a salt thereof, p is selected from 1 or 2, o is selected from 5 or 6, X is -O-, R 4 、R 5 、R 6 、R 7 are each independently selected from hydrogen, L 1 and L 2 Each is independently selected from -C(O)O- or -OC(O)-.

[0077] In other embodiments, in the compound represented by Formula I or Formula II or a salt thereof, R 3 Each independently selected from C 1-6 Alkyl or C 1-6 Alkylamino.

[0078] In other embodiments, in the compound represented by Formula I or Formula II or a salt thereof, R 3 Each independently selected from C 1-6 Alkyl groups, such as methyl or ethyl.

[0079] In certain embodiments, in the compound represented by Formula I or Formula II or a salt thereof, R 3 Selected from C 1-6 Alkylamino, for example methylamino or dimethylamino.

[0080] In certain embodiments, in the compound represented by Formula I or Formula II or a salt thereof, R 3 are each independently selected from halogen, such as fluorine or bromine.

[0081] In certain embodiments, in the compound represented by Formula I or Formula II or a salt thereof, R 3 Each independently selected from C 1-6 Alkoxy groups, such as methoxy or ethoxy.

[0082] In certain embodiments, in the compound represented by Formula I or Formula II or a salt thereof, R 3 Each independently selected from C 3-7 Cycloalkyl or C 3-7 Heterocycloalkyl, for example cyclopropyl or oxiranyl.

[0083] In some embodiments, in the compound represented by Formula I or II or a salt thereof, L 1 and L 2are each independently selected from -C(O)O-, -OC(O)- or a bond.

[0084] Furthermore, the compound represented by formula I or formula II in the present disclosure is a compound represented by formula III-a or a compound represented by formula III-b

[0085] Among them, X, H 1 、R 1 、R 4 ~R 7 、R b 、R c , o, p are as defined above.

[0086] In some embodiments, in the compound represented by Formula I or II or a salt thereof, L 1 and L 2 Each independently selected from -S(O) q -, -SS-, -C(O)S-, -SC(O)- or a bond.

[0087] In certain embodiments, in the compound of formula I or II or a salt thereof, L 1 and L 2 Each independently selected from -NR a C(O)-、-C(O)NR a -、-NR a C(O)NR a -、-NR a C(O)O- or bond, R a Selected from hydrogen or C 1-6 alkyl.

[0088] In certain embodiments, in the compound of formula I or II or a salt thereof, L 1 and L 2 Each independently selected from -NR a C(O)NR a -、-NR a C(O)O-、-OC(O)NR a - or key, R a Selected from hydrogen or C 1-6 alkyl.

[0089] On the other hand, in some embodiments, in the compound represented by Formula I or II, H 1 Selected from C 1-6 The alkylene group includes a C2 alkylene group, a C3 alkylene group, a C4 alkylene group, a C5 alkylene group or a C6 alkylene group.

[0090] In some embodiments, in the compound of Formula I or II, H 1 Selected from straight chain C 1-6Alkylene.

[0091] In some embodiments, in the compound of Formula I or II, H 1 is selected from C3 alkylene, such as linear C3 alkylene.

[0092] In some embodiments, in the compound of Formula I or II, H 1 Selected from C4 alkylene, such as linear C3 alkylene.

[0093] In some embodiments, in the compound of Formula I or II, H 1 Selected from C5 alkylene, such as linear C3 alkylene.

[0094] In some embodiments, in the compound of formula I or II or a salt thereof, p is selected from 3 or 4, o is selected from 5 or 6, X is -O-, R 4 、R 5 、R 6 、R 7 are each independently selected from hydrogen, H 1 Selected from straight chain C3 alkylene.

[0095] In some embodiments, in the compound of formula I or II or a salt thereof, p is selected from 3 or 4, o is selected from 5 or 6, X is -O-, R 4 、R 5 、R 6 、R 7 are each independently selected from hydrogen, H 1 Selected from linear C3 alkylene, R b For straight chain C 6-8 Alkylene.

[0096] In some embodiments, in the compounds of formula I or II, R c Selected from:

[0097] On the other hand, some embodiments provide compounds of formula I or II, wherein R 1 Selected from:

[0098] On the other hand, in some embodiments, in the compound represented by Formula I or II or a salt thereof, R b Selected from C 1-12 Heteroalkylene, including C3 heteroalkylene, C4 heteroalkylene, C5 heteroalkylene, C6 heteroalkylene, C7 heteroalkylene, C8 heteroalkylene, C9 heteroalkylene, C 10 Heteroalkylene, C 11 Heteroalkylene or C 12 Heteroalkylene.

[0099] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b For straight chain C 1-12 Heteroalkylene.

[0100] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b For branched chain C 1-12 Heteroalkylene.

[0101] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C7 heteroalkylene.

[0102] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C8 heteroalkylene.

[0103] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b It is a straight chain C9 heteroalkylene group.

[0104] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R b Selected from:

[0105] On the other hand, in some embodiments, in the compound represented by Formula I or II or a salt thereof, R c Selected from C 1-24 Heteroalkyl, including C2 heteroalkyl, C4 heteroalkyl, C5 heteroalkyl, C6 heteroalkyl, C7 heteroalkyl, C8 heteroalkyl, C9 heteroalkyl, C 10 Heteroalkyl, C 11 Heteroalkyl, C 12 Heteroalkyl, C 13 Heteroalkyl, C 14 Heteroalkyl, C 15 Heteroalkyl, C 16 Heteroalkyl, C 17 Heteroalkyl, C 18 Heteroalkyl, C 19 Heteroalkyl, C 20 Heteroalkyl, C 21 Heteroalkyl, C 22 Heteroalkyl, C 23 Heteroalkyl or C 24 Heteroalkyl.

[0106] In some embodiments, R c For straight chain C 1-12 Heteroalkyl.

[0107] In some embodiments, R cFor branched chain C 1-12 Heteroalkyl.

[0108] In some embodiments, R c For branched chain C 11 Heteroalkyl.

[0109] In some embodiments, R c For branched chain C 17 Heteroalkyl.

[0110] In some embodiments, R c It is a straight chain C9 heteroalkyl group.

[0111] In some embodiments, in the compound represented by Formula I or II or a salt thereof, R c Selected from:

[0112] Typical compounds of the present disclosure are selected from:

[0113] The present disclosure also provides an isotope substitution of the aforementioned compound or a salt thereof. In some embodiments, the isotope substitution is a deuterium atom substitution.

[0114] The present disclosure also provides a lipid particle comprising the aforementioned compound or a salt thereof or an isotope thereof. Furthermore, in some embodiments, the lipid particle further comprises an active agent.

[0115] In some embodiments, the active agent is selected from a polynucleotide or a nucleic acid (such as ribonucleic acid or deoxyribonucleic acid). In some embodiments, the active agent is selected from DNA, siRNA or mRNA.

[0116] The present disclosure also provides a pharmaceutical composition comprising the aforementioned lipid particles and a pharmaceutically acceptable excipient. In certain embodiments, based on the total weight of the composition, the pharmaceutical composition contains 0.01%-99.99% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 0.1%-99.9% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 0.5%-99.5% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 1%-99% of a pharmaceutically acceptable excipient. In certain embodiments, the pharmaceutical composition contains 2%-98% of a pharmaceutically acceptable excipient.

[0117] The present disclosure also provides a use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of a drug for inducing an immune response in a subject.

[0118] In vitro activity tests of the aforementioned compounds or pharmaceutical compositions disclosed herein show that they can specifically deliver active agents to immune cells such as T cells, and at the same time, the delivered active agents exhibit better protein expression ability in immune cells such as T cells.

[0119] In addition, specific delivery was also observed in other cell types, such as NK cells, macrophages, dendritic cells, and monocytes.

[0120] The present disclosure also provides use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating diseases or conditions associated with polypeptide overexpression.

[0121] The present disclosure also provides use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating a disease or condition associated with insufficient polypeptide expression.

[0122] In some embodiments, the disease or condition includes, but is not limited to, cancer, infectious disease, autoimmune disease, neurodegenerative disease, and inflammation.

[0123] The present disclosure also provides a method for preventing and / or treating a disease or condition associated with inducing an immune response in a subject, comprising administering the aforementioned compound or its salt or isotope substitute, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition to a subject in need thereof.

[0124] The present disclosure also provides a method for preventing and / or treating a disease or condition associated with polypeptide overexpression, comprising administering the aforementioned compound or its salt or isotope substitute, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition to a subject in need thereof.

[0125] The present disclosure also provides a method for preventing and / or treating a disease or condition associated with insufficient polypeptide expression, comprising administering to a subject in need thereof a composition containing the aforementioned compound or its salt or isotope substitute, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition.

[0126] On the other hand, the present disclosure also provides the aforementioned compound or its salt or isotope substitute, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition, for use in preventing and / or treating a disease or condition associated with inducing an immune response in a subject.

[0127] The present disclosure also provides the aforementioned compound or its salt or isotope substitute thereof, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition, for use in preventing and / or treating diseases or disorders associated with polypeptide overexpression.

[0128] The present disclosure also provides the aforementioned compound or its salt or isotope substitute thereof, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition, for use in preventing and / or treating diseases or conditions associated with insufficient polypeptide expression.

[0129] The present disclosure also provides use of the aforementioned compound or its salt, or isotope substitution, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition in the preparation of a medicament for preventing and / or treating cancer, infectious diseases, autoimmune diseases, neurodegenerative diseases or inflammation.

[0130] The present disclosure also provides a method for preventing and / or treating cancer, infectious diseases, autoimmune diseases, neurodegenerative diseases or inflammation, comprising administering the aforementioned compound or its salt or isotope substitute, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition to a subject in need.

[0131] The present disclosure also provides the aforementioned compound or its salt or isotope substitute, or the aforementioned lipid particle, or the aforementioned pharmaceutical composition, for use in preventing and / or treating cancer, infection, autoimmune disease, neurodegenerative disease or inflammation.

[0132] The salts of the compounds described in this disclosure include "acid" addition salts and "base" addition salts. For example, salts formed by acid-base reaction with a basic group (amino group), wherein the acid includes an organic acid or an inorganic acid. In addition, salts of the compounds also include salts formed by quaternization with a basic group (amino group), wherein the quaternizing agent includes a linear or branched chlorinated hydrocarbon.

[0133] The compounds of the present disclosure may exist in specific geometric or stereoisomeric forms. The present disclosure contemplates all such compounds, including cis and trans isomers, (-)- and (+)-enantiomers, (R)- and (S)-enantiomers, diastereomers, (D)-isomers, (L)-isomers, and racemic mixtures and other mixtures thereof, such as enantiomerically or diastereomerically enriched mixtures, all of which are within the scope of the present disclosure. Additional asymmetric carbon atoms may be present in substituents such as alkyl groups. All of these isomers and their mixtures are included within the scope of the present disclosure. The compounds of the present disclosure containing asymmetric carbon atoms can be isolated in optically pure form or in racemic form. Optically pure forms can be resolved from racemic mixtures or synthesized by using chiral starting materials or chiral reagents.

[0134] Optically active (R)- and (S)-isomers, as well as D and L isomers, can be prepared by chiral synthesis or chiral reagents or other conventional techniques. If one enantiomer of a compound of the present disclosure is desired, it can be prepared by asymmetric synthesis or derivatization with a chiral auxiliary, wherein the resulting diastereomeric mixture is separated and the auxiliary groups are cleaved to provide the pure desired enantiomer. Alternatively, when the molecule contains a basic functional group (such as an amino group) or an acidic functional group (such as a carboxyl group), diastereomeric salts are formed with an appropriate optically active acid or base, followed by diastereomeric resolution by conventional methods known in the art, and then the pure enantiomers are recovered. In addition, separation of enantiomers and diastereoisomers is typically accomplished using chromatography using a chiral stationary phase, optionally combined with chemical derivatization (e.g., to form carbamates from amines).

[0135] In the chemical structures of the compounds disclosed herein, the bond Indicates that the configuration is not specified, that is, if chiral isomers exist in the chemical structure, the bond Can be or or include both and Two configurations. In the chemical structure of the compound disclosed in the present invention, the bond The configuration is not specified, that is, it can be Z configuration or E configuration, or contain both configurations.

[0136] The compounds and intermediates of the present disclosure may also exist in different tautomeric forms, and all such forms are included within the scope of the present disclosure. The term "tautomer" or "tautomeric form" refers to structural isomers of different energies that can interconvert via a low energy barrier. For example, proton tautomers (also known as prototropic tautomers) include interconversions via proton migration, such as keto-enol and imine-enamine, lactam-lactim isomerizations. For example, between A and B as shown below:

[0137] All tautomeric forms are within the scope of the present disclosure. The naming of compounds does not exclude any tautomers.

[0138] The present disclosure also includes isotopically labeled compounds of the present disclosure that are identical to those described herein, but where one or more atoms are replaced by an atom having an atomic mass or mass number different from the atomic mass or mass number usually found in nature. Examples of isotopes that can be incorporated into the compounds of the present disclosure include isotopes of hydrogen, carbon, nitrogen, oxygen, phosphorus, sulfur, fluorine, iodine, and chlorine, such as 2 H. 3 H. 11 C. 13 C.14 C. 13 N. 15 N. 15 O. 17 O. 18 O. 31 P. 32 P. 35 S. 18 F. 123 I. 125 I and 36 Cl et al.

[0139] Unless otherwise stated, when a position is specifically designated as deuterium (D), the position is understood to have deuterium (i.e., at least 10% deuterium incorporation) at least 1000 times greater than the natural abundance of deuterium (which is 0.015%). In the example, the compound has a natural abundance greater than deuterium of at least 1000 times the abundance of deuterium, at least 2000 times the abundance of deuterium, at least 3000 times the abundance of deuterium, at least 4000 times the abundance of deuterium, at least 5000 times the abundance of deuterium, at least 6000 times the abundance of deuterium, or more. The present disclosure also includes compounds in various deuterated forms. Each available hydrogen atom connected to a carbon atom can be independently replaced by a deuterium atom. Those skilled in the art can synthesize compounds in deuterated forms with reference to relevant literature. In preparing deuterated forms of the compounds, commercially available deuterated starting materials may be used, or they may be synthesized using conventional techniques with deuterated reagents including, but not limited to, deuterated borane, trideuterated borane in tetrahydrofuran, deuterated lithium aluminum hydride, deuterated iodoethane, deuterated iodomethane, and the like.

[0140] "Optionally" or "optionally" means that the subsequently described event or circumstance may but need not occur, and the description includes instances where the event or circumstance occurs or does not occur. For example, "optionally substituted C 1-6 The term "alkyl" means that halogen or cyano may but need not be present, and the description includes both the case where the alkyl is substituted by halogen or cyano and the case where the alkyl is not substituted by halogen and cyano.

[0141] Explanation of terms

[0142] A "pharmaceutical composition" refers to a mixture containing one or more compounds described herein, or their physiologically acceptable salts or prodrugs, together with other chemical components, as well as other components such as physiologically acceptable carriers and excipients. The purpose of a pharmaceutical composition is to facilitate administration to an organism, facilitating absorption of the active ingredients and thereby exerting their biological activity.

[0143] "Pharmaceutically acceptable excipients" or "pharmaceutically acceptable excipients" include, but are not limited to, any adjuvant, carrier, glidant, sweetener, diluent, preservative, dye / colorant, flavoring agent, surfactant, wetting agent, dispersant, suspending agent, stabilizer, isotonic agent, solvent or emulsifier that has been approved by the U.S. Food and Drug Administration for use in humans or domestic animals.

[0144] As used herein, an "effective amount" or "therapeutically effective amount" encompasses an amount sufficient to ameliorate or prevent the symptoms or conditions of a medical condition. An effective amount also refers to an amount sufficient to permit or facilitate diagnosis. The effective amount for a particular patient or veterinary subject may vary depending on factors such as the condition being treated, the patient's overall health, the route and dosage of administration, and the severity of side effects. An effective amount can be the maximum dose or dosage regimen that avoids significant side effects or toxic effects.

[0145] The term "nucleic acid" is a polymer composed of nucleotides, such as deoxyribonucleotides (DNA) or ribonucleotides (RNA).

[0146] The term "oligonucleotide" refers to a single-stranded or double-stranded nucleotide polymer of 2 to 100 nucleotides in length. "Polynucleotide" refers to a single-stranded or double-stranded polymer composed of nucleotide monomers. In some embodiments, a polynucleotide is composed of more than 100 nucleotides in length.

[0147] In other embodiments, exemplary polynucleotides include, but are not limited to, deoxyribonucleotides (DNA), ribonucleic acids (RNA), including messenger RNA, RNAi-inducing agents, shRNA, siRNA, miRNA, antisense RNA, and the like.

[0148] The terms "polypeptide," "peptide," and "protein" are used interchangeably herein to refer to polymers of amino acid residues. The terms apply to amino acid polymers in which one or more amino acid residues is an artificial chemical analogue of a corresponding naturally occurring amino acid, as well as to naturally occurring amino acid polymers.

[0149] "Alkyl" refers to a saturated aliphatic hydrocarbon group, including straight-chain and branched-chain alkyl groups. In some embodiments, the alkyl group has 1-4 carbon atoms, also known as C 1-4 In some embodiments, an alkyl group has 10-22 carbon atoms, also known as a C 10-22 In some embodiments, the alkyl group has 4-22 carbon atoms, also known as C 4-22 alkyl.

[0150] The alkyl group may be unsubstituted or substituted by one or more groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the alkyl group is substituted with one or more alkyl groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 Alkoxy, 3 to 7 membered cycloalkyl group.

[0151] "Alkoxy" refers to -O-(alkyl), where alkyl is as defined above. Non-limiting examples of alkoxy include methoxy and ethoxy. Alkoxy may be unsubstituted or substituted with one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the alkoxy group is substituted by one or more halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0152] "Alkylamino" refers to -NH(alkyl) or -N(alkyl)2, where alkyl is as defined above. Exemplary alkylamino groups include C 1-6 Alkyl NH- or di(C 1-6 Alkyl) N-, for example methylamino or dimethylamino. Alkylamino may be unsubstituted or substituted by one or more groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the alkylamino group is substituted by one or more alkyl groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0153] "Heteroalkyl" refers to a straight or branched chain alkyl group preferably having 1 to 14 carbons, more preferably 2 to 10 carbons in the chain, wherein one or more carbons are substituted with a heteroatom selected from S, O and N, including -OO- or -SS- moieties. Exemplary heteroalkyl groups include wait.

[0154] Heteroalkyl may be unsubstituted or substituted by one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the heteroalkyl group is substituted with one or more halogen, hydroxy, oxo, cyano, amino, C 1-6Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0155] "Alkenyl" refers to an unsaturated aliphatic hydrocarbon group, including straight-chain and branched alkenyl groups. In some embodiments, the alkenyl group has 2-4 carbon atoms, also known as C 2-4 In some embodiments, the alkenyl group has 10-22 carbon atoms and is also referred to as a C 10-22 In some embodiments, the alkenyl group has 4-22 carbon atoms and is also referred to as a C 4-22 Alkenyl. Exemplary alkenyl groups include ethenyl, propenyl, n-butenyl, isobutenyl, 3-methylbut-2-enyl, n-pentenyl, heptenyl, octenyl, cyclohexyl-butenyl, and decenyl.

[0156] Alkenyl may be unsubstituted or substituted by one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the alkenyl group is substituted with one or more halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0157] "Heteroalkenyl" refers to the definition of "heteroalkyl", which refers to a straight or branched alkenyl group in which one or more carbon atoms are replaced by a heteroatom selected from S, O and N, including -OO- or -SS- moieties.

[0158] A "monovalent group" is a compound formed by formally eliminating a monovalent atom or group. A "subunit" is a compound formed by formally eliminating two monovalent or one divalent atom or group.

[0159] The term "alkylene" refers to the portion of an alkane molecule that remains after removing two hydrogen atoms. In some embodiments, an alkylene group has 1-4 carbon atoms, also known as a C 1-4 In some embodiments, an alkyl group has 10-22 carbon atoms, also known as a C 10-22 In some embodiments, the alkyl group has 4-22 carbon atoms, also known as C 4-22 Alkylene.

[0160] The alkylene group may be unsubstituted or substituted by one or more groups selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the alkylene group is substituted by one or more halogen, hydroxy, oxo, cyano, amino, C1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0161] The term "heteroalkylene" refers to the portion of a heteroalkane molecule that remains after removing two hydrogen atoms. In some embodiments, a heteroalkylene has 1-14 carbon atoms and is also referred to as a C 1-14 In some embodiments, the heteroalkylene group has 2-10 carbon atoms and is also referred to as a C 2-10 In other embodiments, heteroalkylene groups have one or more carbons in the chain substituted with heteroatoms selected from S, O, and N, including -OO- or -SS- moieties. Exemplary heteroalkylene groups include wait.

[0162] Heteroalkylene may be unsubstituted or substituted by one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0163] Similarly, the definitions of "alkyleneoxy" and "alkenylene" are the same as those of "alkylene".

[0164] The term "cycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent, wherein the cycloalkyl ring contains 3 to 8 carbon atoms. Non-limiting examples of monocyclic cycloalkyls include cyclopropyl, cyclobutyl, cyclopentyl, cyclopentenyl, cyclohexyl, cyclohexenyl, cyclohexadienyl, and the like; polycyclic cycloalkyls include spirocyclic, fused, and bridged cycloalkyls. Cycloalkyls may be unsubstituted or substituted with one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the cycloalkyl group is substituted by one or more halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0165] The term "heterocycloalkyl" refers to a saturated or partially unsaturated monocyclic or polycyclic hydrocarbon substituent containing 3 to 7 ring atoms, one or more of which is selected from nitrogen, oxygen or S(O) m (wherein m is an integer from 0 to 2) heteroatoms, excluding the ring portion of -OO-, -OS- or -SS-, the remaining ring atoms being carbon. Preferably, it contains 3 to 7 ring atoms, of which 1 to 4 are heteroatoms. Non-limiting examples of "heterocycloalkyl" include: Heterocycloalkyl may be unsubstituted or substituted by one or more radicals selected from halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 In some embodiments, the heterocycloalkyl group is substituted by one or more halogen, hydroxy, oxo, cyano, amino, C 1-6 Alkyl, C 1-6 The alkyl group may be substituted with an alkoxy group or a 3- to 7-membered cycloalkyl group.

[0166] The terms "branched alkyl" and "branched alkenyl" refer to an alkyl or alkenyl group in which one carbon atom is bonded to at least two other carbon atoms, and not to form a cyclic group, such as a spirocycle. Examples: C6 alkyl groups include: (straight chain) or (branched chain).

[0167] The term "hydroxy" refers to an -OH group.

[0168] The term "halogen" refers to fluorine, chlorine, bromine or iodine.

[0169] The term "amino" refers to -NH2.

[0170] The term "cyano" refers to -CN.

[0171] The term "oxo" refers to a =0 substituent.

[0172] "Substituted" means that one or more hydrogen atoms, preferably up to 5, more preferably 1 to 3 hydrogen atoms in a group are replaced independently of one another by a corresponding number of substituents. It goes without saying that the substituents are only in their possible chemical positions, and a person skilled in the art can determine (by experiment or theory) whether substitution is possible or not without undue effort. BRIEF DESCRIPTION OF THE DRAWINGS

[0173] FIG1 : Fluorescence intensity of the transfection efficiency of non-immune cells in vitro using the lipid nanoparticle composition of the present disclosure (ns indicates P>0.05, ** indicates P<0.01, *** indicates P<0.001).

[0174] FIG2 : Relative fluorescence intensity of the in vitro transfection efficiency of non-immune cells by the lipid nanoparticle composition of the present disclosure (ns indicates P>0.05, ** indicates P<0.01, *** indicates P<0.001).

[0175] FIG3 : Fluorescence intensity of the immune cell transfection efficiency detected by the lipid nanoparticle composition of the present disclosure in vitro (** indicates P<0.01).

[0176] FIG4 : Relative fluorescence intensity of immune cell transfection efficiency in vitro of the lipid nanoparticle composition of the present disclosure (** indicates P<0.01).

[0177] Figure 5: Flow cytometry analysis of the positive cell ratio of T cells (** indicates p < 0.01, *** indicates p < 0.001).

[0178] Figure 6: Flow cytometry analysis of the positive cell ratio of NK cells (** indicates p < 0.01, *** indicates p < 0.001).

[0179] Figure 7: Flow cytometry analysis of the positive cell ratio of macrophages (** indicates p < 0.01, *** indicates p < 0.001).

[0180] Figure 8: Flow cytometry analysis of the positive cell ratio of dendritic cells (** indicates p < 0.01, *** indicates p < 0.001).

[0181] Figure 9: Flow cytometry analysis of the positive cell ratio of monocytes (** indicates p < 0.01, *** indicates p < 0.001).

[0182] Figure 10: Fluorescence intensity of mouse spleen tissue detected by IVIS (ns indicates P>0.05, * indicates P<0.05, ** indicates P<0.01).

[0183] Figure 11: Fluorescence intensity of mouse liver tissue detected by IVIS (ns indicates P>0.05, * indicates P<0.05, ** indicates P<0.01). DETAILED DESCRIPTION

[0184] The present disclosure is further described below with reference to examples, but these examples are not intended to limit the scope of the present disclosure.

[0185] Experimental methods in the examples of this disclosure that do not specify specific conditions are generally based on conventional conditions or the conditions recommended by the raw material or product manufacturers. Reagents without specific sources are conventional reagents purchased from the market.

[0186] The structures of the compounds were determined by nuclear magnetic resonance (NMR) and / or mass spectrometry (MS). -6 The unit of ppm is given. NMR measurements were performed using a Bruker AVANCE-400 NMR spectrometer. The solvents used were deuterated dimethyl sulfoxide (DMSO-d6), deuterated chloroform (CDCl3), or deuterated methanol (Methanol-d4), and tetramethylsilane (TMS) was used as the internal standard.

[0187] HPLC determination used Agilent1100 high pressure liquid chromatograph, GAS15B DAD UV detector, Water Vbridge C18 150*4.6mm 5um chromatographic column.

[0188] MS was determined using an Agilent 6120 triple quadrupole mass spectrometer, a G1315D DAD detector, and a Waters Xbridge C18 4.6*50mm, 5um column. The samples were scanned in positive / negative ion mode with a mass scan range of 80-1200.

[0189] The thin layer chromatography silica gel plate used was Yantai Huanghai HSGF254 silica gel plate. The specification of the silica gel plate used in thin layer chromatography (TLC) was 0.2 mm ± 0.03 mm. The specification used for thin layer chromatography separation and purification products was 0.4 mm - 0.5 mm.

[0190] The flash column purification system used was Combiflash Rf150 (TELEDYNE ISCO) or Isolara one (Biotage).

[0191] Forward column chromatography generally uses Yantai Huanghai silica gel 200-300 mesh or 300-400 mesh silica gel as the carrier, or uses Changzhou Santai pre-packed ultra-pure normal phase silica gel column (40-63μm, 60g, 24g, 40g, 120g or other specifications).

[0192] The known starting materials in the present disclosure can be synthesized by methods known in the art, or can be purchased from Shanghai Titan Technology, ABCR GmbH & Co. KG, Acros Organics, Aldrich Chemical Company, Accela ChemBio Inc, Bid Pharmaceuticals, etc.

[0193] Unless otherwise specified in the examples, all reactions were carried out under a nitrogen atmosphere.

[0194] Nitrogen atmosphere means that the reaction bottle is connected to a nitrogen balloon with a capacity of about 1L.

[0195] Hydrogen atmosphere means that the reaction bottle is connected to a hydrogen balloon with a capacity of about 1L.

[0196] Hydrogen was produced by a QPH-1L hydrogen generator from Shanghai Quanpu Scientific Instrument Co., Ltd.

[0197] The nitrogen atmosphere or hydrogen atmosphere is usually evacuated and filled with nitrogen or hydrogen, and the operation is repeated three times.

[0198] Unless otherwise specified in the examples, the solution refers to an aqueous solution.

[0199] Unless otherwise specified in the examples, the reaction temperature is room temperature, 20°C to 30°C.

[0200] The reaction progress in the examples was monitored by thin layer chromatography (TLC). The developing solvent used in the reaction, the column chromatography eluent system used to purify the compound, and the developing solvent system for thin layer chromatography, the volume ratio of the solvent were adjusted according to the polarity of the compound, and a small amount of alkaline or acidic reagents such as triethylamine and acetic acid could be added for adjustment.

[0201] Example 1. Preparation of Compound 1

[0202] Step 1: Preparation of compound 1c

[0203] Compound 1a (5 g, 32.85 mmol, purchased from Adamas) and compound 1b (14.66 g, 65.71 mmol, purchased from Leyan) were dispersed in a 50% aqueous NaOH solution (8 M, 69.81 mL) and stirred. The mixture was cooled to 0°C, tetrabutylammonium hydrogen sulfate (1.12 g, 3.29 mmol) was added, and the mixture was heated to 60°C for 16 hours. After cooling to 0°C, water (100 mL) was added, followed by ethyl acetate (100 mL). The aqueous phase was extracted with ethyl acetate (50 mL x 2). The organic phases were combined, washed with saturated brine (50 mL), dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was purified by column chromatography (PE:EA = 0-20%) to obtain 7.69 g of compound 1c (yield 79.51%).

[0204] Step 2: Preparation of compound 1d

[0205] Compound 1c (7.69 g, 26.12 mmol) was dissolved in MeOH (80 mL) and stirred. Palladium on carbon (10 wt.% 50% water, 1.83 g, 13.06 mmol) was added, and the air was replaced three times with hydrogen. The mixture was pressurized to 60 psi and heated to 45°C for 16 hours. Filtered through celite, the filter cake was washed with methanol, and the filtrate was concentrated to dryness to yield 5.54 g of crude compound 1d, which was used directly in the next step.

[0206] Step 3: Preparation of Compound 1f

[0207] Compound 1d (5.34 g, 26.14 mmol), compound 1e (8.04 g, 31.37 mmol, purchased from Bismuth), and DMAP (3.19 g, 26.14 mmol) were dissolved in DCM (60 mL) and stirred at 0°C. EDCI (7.52 g, 39.21 mmol) was added, and the mixture was allowed to warm to room temperature and react for 16 hours. DCM (50 mL) and water (50 mL) were added, and the organic phase was washed with saturated sodium bicarbonate (25 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was purified by column chromatography (PE:EA = 0-10%) to obtain 10.01 g of compound 1f (yield 86.50%).

[0208] 1 H NMR (400MHz, CDCl3): δ4.59-4.54(m,1H),4.24-4.17(m,2H),3.89-3.76(m,2H),3.63-3.59(m,2H),3.58-3.52(m,2H),3.51-3.43(m,2H) ,2.39-2.29(m,1H),1.91-1.75(m,3H),1.74-1.66(m,1H),1.65-1.47(m,6H),1.46-1.36(m,2H),1.33-1.15(m,20H),0.92-0.80(m,6H).

[0209] Step 4: Preparation of compound 1g

[0210] Compound 1f (1 g, 2.26 mmol) was dissolved in hydrochloric acid / dioxane (4 M, 10.05 mL) and stirred at room temperature for 2 hours. The reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE:EA = 0-20%) to obtain 203 mg of compound 1g (yield 25.06%).

[0211] 1 H NMR (400MHz, CDCl3): δ4.27-4.19(m,2H),3.76(t,J=5.6Hz,2H),3.68-3.61(m,4H),2.40-2.30(m,1H),2.29-2 .20(brs,1H),1.87-1.78(m,2H),1.66-1.51(m,3H),1.50-1.35(m,2H),1.35-1.15(m,19H),0.92-0.80(m,6H).

[0212] Step 5: Preparation of compound 1h

[0213] Compound 1g (0.16g, 446.24μmol) and triphenylphosphine (175.56mg, 669.36μmol) were dissolved in dichloromethane (2mL) and stirred at 0°C. Carbon tetrabromide (221.98mg, 669.36μmol) was added portionwise, and the mixture was allowed to warm to room temperature for 2 hours. The reaction solution was concentrated to dryness, and the residue was purified by column chromatography (PE:EA = 0-5%) to obtain 149mg of compound 1h (yield 79.23%).

[0214] 1 H NMR (400MHz, CDCl3): δ4.25-4.19(m,2H),3.66-3.61(m,2H),3.58(t,J=6Hz,2H),3.49(t,J=6.4Hz,2H),2.40- 2.29(m,1H),2.13-2.04(m,2H),1.65-1.52(m,2H),1.50-1.36(m,2H),1.34-1.15(m,20H),0.92-0.79(m,6H).

[0215] Step 6: Preparation of compound 1j

[0216] Compound 1i (5 g, 27.61 mmol, purchased from Shaoyuan), compound 1e (8.50 g, 33.14 mmol), and DMAP (3.37 g, 27.61 mmol) were dissolved in DCM (50 mL) and stirred at 0°C. EDCI (7.94 g, 41.42 mmol) was added and the mixture was allowed to warm to room temperature for 16 hours. 2M HCl (10 mL) was added, and the aqueous phase was extracted with dichloromethane (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was purified by column chromatography (PE:EA = 0-10%) to obtain 5.96 g of compound 1j (yield 51.45%).

[0217] 1 H NMR (400MHz, CDCl3): δ4.06(t,J=6.4Hz,2H),3.40(t,J=6.4Hz,2H),2.35-2.25(m,1H),1.91 -1.81(m,2H),1.68-1.52(m,5H),1.51-1.34(m,5H),1.33-1.18(m,20H),0.92-0.79(m,6H).

[0218] Step 7: Preparation of compound 1k

[0219] Compound 1j (2 g, 4.77 mmol) and compound G (2.98 g, 23.84 mmol, purchased from Adamas) were dissolved in ethanol (20 mL) and stirred at 78°C for 16 hours. The reaction solution was concentrated to dryness, and the residue was purified by column chromatography (DCM:MeOH = 0-15%) to obtain 1.13 g of compound 1k (yield 51.11%).

[0220] 1 H NMR (400MHz, CDCl3): δ7.46(s,1H),7.04(s,1H),6.91(s,1H),4.11-3.98(m,4H),2.64-2.52(m,4H),2.35-2 .24(m,1H),1.96-1.86(m,2H),1.68-1.51(m,4H),1.50-1.32(m,9H),1.31-1.17(m,20H),0.92-0.81(m,6H).

[0221] Step 8: Preparation of Compound 1

[0222] Compound 1k (0.5 g, 1.08 mmol) and compound 1h (413.10 mg, 980.18 μmol) were dissolved in acetonitrile (4.5 mL) and cyclopentyl methyl ether (4.5 mL). Potassium iodide (178.98 mg, 1.08 mmol) and potassium carbonate (541.88 mg, 3.92 mmol) were added, and the mixture was heated to 86°C for 16 hours. The mixture was filtered, the filter cake washed with acetonitrile, and the filtrate concentrated to dryness. The product was purified by reverse-phase C4 column chromatography (H2O / ACN = 0-100%) to obtain 156.9 mg of compound 1 (yield 19.90%).

[0223] 1 H NMR (400MHz, CDCl3): δ7.45(s,1H),7.04(s,1H),6.91(s,1H),4.24-4.18(m ,2H),4.05(t,J=6.8Hz,2H),3.97(t,J=7.2Hz,2H),3.62-3.57(m,2H),3.46( t,J=6.4Hz,2H),2.49-2.41(m,2H),2.40-2.25(m,6H),1.92-1.83(m,2H),1. 75-1.50(m,8H),1.49-1.33(m,8H),1.32-1.17(m,42H),0.91-0.81(m,12H).

[0224] Example 2 Preparation of Compound 2

[0225] Step 1: Preparation of compound 2a

[0226] Add water (46.8 mL) to a 500 mL three-necked flask. Cool to below 5°C in an ice-water bath, then slowly add sodium hydroxide (46.8 g, 1170 mmol), maintaining the internal temperature below 15°C. Once the sodium hydroxide is completely dissolved, add toluene (46.8 mL) and compound E (9.71 g, 50.0 mmol, purchased from Adamas), and stir at room temperature for 1 hour. Cool again to below 5°C in an ice-water bath, and slowly add compound F (34.1 g, 175 mmol, purchased from Adamas) dropwise. Heat to an external temperature of 25°C in an oil bath, and stir for 24 hours. The reaction mixture was cooled to below 5°C, water (200 mL) was added, and the mixture was extracted with methyl tert-butyl ether (100 mL × 3). The aqueous phase was cooled to below 5°C with an ice-water bath, and concentrated hydrochloric acid was slowly added until the pH reached about 3. The mixture was then extracted with DCM (100 mL × 5). The DCM phases were combined, washed with saturated brine, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. The residue was purified by column chromatography (EA / PE = 0-100%) to give 11.97 g of compound 2a (yield 94%).

[0227] 1 H NMR (400MHz, CDCl3): δ8.50(brs,1H),7.36-7.26(m,5H),4.51(s,2H),4.13~ 4.10(m,2H),3.56(t,2H),3.48(t,2H),1.67~1.61(m,4H),1.50~1.43(m,2H).

[0228] Step 2: Preparation of compound 2c

[0229] Compound 2a (9.3 g, 36.86 mmol), compound 2b (8.51 g, 33.17 mmol, purchased from Admas), and DCM (186 mL) were added to a 100 mL single-necked flask. The mixture was cooled to 0°C under a nitrogen atmosphere. DMAP (4.50 g, 36.86 mmol) was added and stirred for 10 minutes. EDCI (8.48 g, 44.23 mmol) was slowly added to the reaction mixture. After the addition was complete, the mixture was naturally warmed to room temperature and reacted for 16 hours. The mixture was cooled to 0°C, water (186 mL) was added, and the mixture was stirred for 5 minutes. The aqueous phase was extracted with DCM (100 mL x 2). The organic phases were combined, washed with saturated NaHCO₃ solution (200 mL), washed with NaCl solution (200 mL), dried, and concentrated under reduced pressure. The residue was separated and purified by column chromatography (PE:EA = 100:0 to 95:5) to obtain 13.5 g of compound 2c (yield 74.63%).

[0230] 1H NMR (400MHz, CDCl3): δ (ppm) 7.34-7.27 (m, 4H), 4.99-4.94 (m, 1H), 4.49 (s, 2H), 4.03 (s, 2H), 3 .54-3.46(m,4H),1.68-1.61(m,4H),153-1.42(m,8H),1.25-1.24(m,21H),0.89-0.86(m,8H).

[0231] Step 3: Preparation of compound 2d

[0232] Compound 2c (13.5 g, 27.51 mmol) was dissolved in THF (108 mL), and Pd(OH)2 / C (3.7 g, 50% wt) was added. The atmosphere was replaced with hydrogen three times, and the mixture was heated to 30°C for 48 h. The reaction was complete, and the mixture was filtered and concentrated to dryness. The residue was separated and purified by column chromatography (PE:EA = 100:0 to 60:10) to obtain 8.07 g of compound 2d (yield 74.63%).

[0233] MS m / z(ESI):423.3[M+23] + .

[0234] Step 4: Preparation of compound 2e

[0235] Compound 2d (3 g, 7.49 mmol) and DCM (24 mL) were added to a 100 mL single-necked flask. The mixture was cooled to 0°C under a nitrogen atmosphere, and carbon tetrabromide (4.37 g, 13.18 mmol) was added and stirred for 10 min. Triphenylphosphine (3.14 g, 11.98 mmol) was added portionwise to the reaction mixture. The mixture was allowed to warm to room temperature and react for 16 h. Upon completion of the reaction, silica gel (18 g, 100-200 mesh) was added and the mixture was dried to a powder. The mixture was then purified by column chromatography (PE:EA = 100:0 to 90:10) to obtain 3.2 g of compound 2e (92.19% yield).

[0236] 1 H NMR (400MHz, CDCl3): δ (ppm) 4.99-4.93 (m, 1H), 4.04 (s, 2H), 3.53 (t, J = 6Hz, 2H), 3.41 (t, J = 6.8Hz,2H),1.93-1.86(m,2H),1.64-1.53(m,8H),1.29-1.25(m,24H),0.87(t,J=6.4Hz,6H).

[0237] Step 5: Preparation of compound 2g

[0238] Compound 2e (3.1 g, 6.69 mmol), compound G (2.51 g, 20.06 mmol), and ethanol (31 mL) were added to a 100 ml single-necked flask. The atmosphere was purged with nitrogen three times and heated to 65°C for 16 h. The reaction solution was concentrated to dryness, and the residue was purified by column chromatography (DCM:MeOH = 100:0 to 90:10) to obtain 1.2 g of compound 2g (yield 35.34%).

[0239] MS m / z(ESI):508.4[M+1] + .

[0240] Step 6: Preparation of compound 2j

[0241] Compound 2h (1.0 g, 5.80 mmol, purchased from Bidex), compound 2i (1.94 g, 8.71 mmol, purchased from Shaoyuan), and DCM (20 mL) were added to a 100 mL single-necked flask and cooled to 0°C. DMAP (70.90 mg, 580.36 μmol), EDCI (1.67 g, 8.71 mmol), and DIEA (2.25 g, 17.41 mmol) were added sequentially. The mixture was allowed to warm to room temperature and react for 16 h. After cooling to 0°C, H2O (20 mL) was added and stirred. The organic phase was washed with water, dried, and concentrated under reduced pressure. The residue was isolated and purified by column chromatography (PE:EA = 100:0 to 90:10) to obtain 1.4 g of compound 2j (yield 63.92%).

[0242] 1 H NMR (400MHz, CDCl3): δ (ppm) 4.84-4.78 (m, 1H), 3.4 (t, J = 7.2Hz, 2H), 2.34 (t, J = 7.6H) z,2H),1.87-1.83(m,2H),1.63-1.43(m,10H),1.35-1.26(m,14H),0.89-0.85(m,6H).

[0243] Step 7: Preparation of Compound 2

[0244] Compound 2g (592.02 mg, 1.17 mmol), compound 2j (400.00 mg, 1.06 mmol), potassium iodide (193.54 mg, 1.17 mmol, 62.03 μL), and potassium carbonate (585.95 mg, 4.24 mmol, 255.87 μL) were added to a 25 ml single-necked flask. Acetonitrile (6 mL) and cyclopentyl methyl ether (6 mL) were added under a nitrogen atmosphere. The atmosphere was replaced with nitrogen three times and the reaction was heated to 86°C. The reaction was detected to be complete, and the mixture was cooled to room temperature. The insoluble material was removed by filtration and concentrated to dryness. The residue was purified by reverse-phase C4 column chromatography (water:acetonitrile = 100:0 to 0:100) to obtain 115 mg of compound 2 (yield 13.49%).

[0245] 1 H NMR (400MHz, CDCl3): δ (ppm) 7.46 (s, 1H), 7.04 (s, 1H), 6.91 (s, 1H), 4.99-4.93 (m, 1H), 4.84-4.78 (m, 1H), 4.04 (s, 2H), 3.98 (t, J = 6.8Hz, 2H), 3 .53(t,J=6.2Hz,2H),2.37-2.34(m,6H),2.29(t,J=7.6Hz,2H),1.91-1. 84(m,2H),1.67-1.52(m,14H),1.38-1.25(m,46H),0.89-0.86(m,12H).

[0246] Example 3 Preparation of Compound 3

[0247] Step 1: Preparation of compound 3c

[0248] Compound 2b (1.92 g, 7.47 mmol), compound 3a (2 g, 8.96 mmol, purchased from Shaoyuan), and DMAP (912.64 mg, 7.47 mmol) were dissolved in DCM (20 mL) and stirred at 0°C. EDCI (2.15 g, 11.21 mmol) was added and the mixture was allowed to warm to room temperature for 16 hours. 2M HCl (10 mL) was added, and the aqueous phase was extracted with dichloromethane (20 mL x 2). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and concentrated to dryness. The residue was purified by column chromatography (PE:EA = 0-10%) to obtain 2.78 g of compound 3c (yield 80.63%).

[0249] Step 2: Preparation of compound 3

[0250] Compound 2g (0.1g, 196.93μmol) and compound 3c (82.63mg, 179.03μmol) were dissolved in acetonitrile (1mL) and cyclopentyl methyl ether (1mL). Potassium iodide (32.69mg, 196.93μmol) and potassium carbonate (98.97mg, 716.12μmol) were added, and the mixture was heated to 86°C for 16 hours. After cooling to room temperature, the mixture was filtered, and the filter cake was washed with acetonitrile. The filtrate was concentrated to dryness, and the residue was purified by reverse-phase C4 column chromatography (H2O:ACN = 0-100%) to obtain 32.10mg of compound 3 (yield 20.18%).

[0251] 1 H NMR (400MHz, CDCl3): δ7.47(s,1H),7.05(s,1H),6.91(s,1H),5.00-4.91(m,1H) ),4.90-4.82(m,1H),4.04(s,2H),3.99(t,J=6.8Hz,2H),3.52(t,J=6.4Hz,2H), 2.42-2.31(m,6H),2.28(t,J=7.2Hz,2H),1.93-1.81(m,2H),1.68-1.60(m,4H) ,1.56-1.46(m,8H),1.45-1.34(m,6H),1.33-1.18(m,54H),0.91-0.80(m,12H).

[0252] Comparative Example 1

[0253] Prepared according to the method in WO2021113777.

[0254] Comparative Example 2

[0255] Prepared according to the method in WO2021178396.

[0256] Comparative Example 3

[0257] Prepared according to the method in WO2023125738.

[0258] Example 5: Preparation of lipid nanoparticle composition

[0259] 1.1 Preparation method

[0260] Compound 1, compound 2, compound 3, and comparative compound 1, comparative compound 2, and comparative compound 3 were dissolved in ethanol solution and mixed with DSPC, cholesterol, and DMG-PEG solution dissolved in ethanol at a molar ratio of 50:10:38.5:1.5 to prepare an ethanol lipid solution. mRNA (Luciferase, GenBank: MN728548.1) was dissolved in 50mM pH 5 acetate buffer to prepare an mRNA aqueous solution (0.2 mg / mL). The ethanol lipid solution and the mRNA aqueous solution were mixed by microfluidics, with the ethanol phase and the aqueous phase volume ratio being 1:3, and liposomes were prepared with a total lipid to mRNA weight ratio of approximately 20:1. Ethanol was removed by dialysis in a 20mM Tris pH 7.5 solution, and the solution was finally replaced in a 20mM Tris, 8% sucrose solution and frozen to obtain a lipid nanoparticle composition encapsulating mRNA.

[0261] 1.2 Characterization of lipid nanoparticle composition

[0262] The nanoparticle size and polydispersity index (PDI) were determined by dynamic light scattering using a Malvern Zetasizer Pro in 173° backscattering detection mode.

[0263] The liposome encapsulation efficiency was determined using the Quant-iT RiboGreen RNA Assay Kit.

[0264] The results are shown in Table 1 below.

[0265] Table 1

[0266] Test Example 1: Comparison of the delivery efficiency of lipid nanoparticle compositions in immune cells and non-immune cells

[0267] Non-immune cells were taken as examples of A549 cells (human non-small cell lung cancer cells), and immune cells were taken as examples of Jurkat cells (human T cells) (cell source ATCC), and they were cultured in 96-well plates. The lipid nanoparticle compositions encapsulating luciferase mRNA (Luciferase, GenBank: MN728548.1) were added to the cell well plates. After incubation for 24 hours, luciferase substrate (D-luciferin potassium salt, catalog number: abs42075819) was added, and the fluorescence intensity was detected using a microplate reader. The level of fluorescence intensity represents the level of luciferase protein expression, which indicates the delivery efficiency of lipid nanoparticles corresponding to each compound on different cells. Each group was repeated at least 3 times, and the average value was calculated. ns indicates no statistically significant difference, i.e., statistical P>0.5. ** indicates a statistically significant difference, i.e., statistical P<0.01, and *** indicates a statistically significant difference, i.e., P<0.001.

[0268] Conclusion: As shown in Tables 2 and 3 and Figures 1 and 2, in non-immune cells, the delivery efficiency of the lipid nanoparticle compositions corresponding to compounds 1-3 was not significantly different from that of the lipid nanoparticle compositions corresponding to comparative compounds 1 and 2, or was significantly lower than that of the lipid nanoparticle compositions corresponding to comparative compounds 1 and 2. The relative fluorescence intensity of each group was calculated using the fluorescence intensity of the lipid nanoparticle composition corresponding to comparative compound 2 as 1. The relative fluorescence intensity of the lipid nanoparticle compositions corresponding to compounds 1-3 was around 1, indicating that the expression levels of the lipid nanoparticles corresponding to compounds 1-3 on non-immune cells were comparable to or lower than those of the lipid nanoparticle compositions corresponding to the comparative compounds.

[0269] In immune cells, the fluorescence intensity of the lipid nanoparticle compositions corresponding to compounds 1-3 delivered into the cells was significantly higher than that of the lipid nanoparticle compositions corresponding to comparative compound 1 and comparative compound 2. The fluorescence intensity of the lipid nanoparticle compositions corresponding to compounds 1, 2, and 3 was 9 times, 13 times, and 6 times that of the lipid nanoparticle composition corresponding to comparative compound 2, respectively.

[0270] It can be seen that the lipid nanoparticle compositions corresponding to compounds 1-3 have comparable or significantly lower protein expression in non-immune cells compared to the lipid nanoparticle compositions corresponding to comparative compounds 1-2, while significantly higher protein expression in immune cells, indicating better targeting and higher delivery efficiency for immune cells.

[0271] Table 2 Non-immune cell transfection efficiency detection

[0272] Table 3 Immune cell transfection efficiency detection

[0273] Test Example 2: Evaluation of the delivery efficiency of lipid nanoparticle compositions to mRNA in vivo immune cells via tail vein injection

[0274] 6-8 week old male CAG-LSL-tdTomato model mice (Shanghai Southern Model Organisms Science Co., Ltd.) were injected with a lipid nanoparticle composition (prepared according to the preparation method of Example 5) encapsulating Cre recombinant mRNA (GenBank: JQ033384.1) via the tail vein at a dose of 0.5 mg / kg. In cells in the mouse body, if Cre mRNA is delivered into the cells, the mouse will be gene-edited to express tdTomato fluorescent protein. Therefore, by detecting the fluorescent signal of tdTomato protein, the proportion of positive cells, that is, the delivery efficiency of the lipid nanoparticle composition in different cells in the body, was analyzed. 48 hours after administration, the spleen of the mouse was dissected and the single cell suspension was prepared after grinding and lysis of the red blood cells and resuspended in PBS pH 7.4 buffer. Different immune cells were labeled using fluorescently labeled antibodies (BioLegend), respectively. Cells were grouped using flow cytometry, and the proportion of positive signal tdTomato fluorescence in the corresponding cells was detected, which reflected the delivery efficiency of the nanoparticle lipid composition in the corresponding immune cells. The blank control group was injected with an equal volume of PBS.

[0275] The lipid nanoparticle composition corresponding to each compound was subjected to at least three biological replicates, and the average percentage was calculated. The data are shown in Table 4 and Figures 5 to 9.

[0276] The lipid nanoparticle composition corresponding to the comparative compound 1 and the blank control group were used as controls to detect the delivery efficiency of mRNA in the immune cells in vivo after tail vein injection of the liposome nanoparticle composition corresponding to the compound 1 in mice.

[0277] Table 4 In vivo immune cell delivery efficiency

[0278] Conclusion: As shown in Table 4 and Figures 5 to 9, the percentage of fluorescence-positive cells indicates that the lipid nanoparticle composition corresponding to Compound 1 can effectively deliver mRNA to T cells, NK cells, macrophages, dendritic cells, and mononuclear cells. Compared with the blank control group and the lipid nanoparticle composition corresponding to Compound 1, the percentage of positive cells corresponding to Compound 1 was significantly increased. This indicates that the lipid nanoparticle composition corresponding to Compound 1 can achieve efficient transfection of immune cells in vivo.

[0279] Test Example 3: Evaluation of mRNA tissue expression in vivo via tail vein injection of lipid nanoparticle compositions

[0280] Female Balb / c mice aged 6-8 weeks were injected via the tail vein with a lipid nanoparticle composition (prepared according to the preparation method of Example 5) encapsulating luciferase mRNA (Luciferase, GenBank: MN728548.1) at a dose of 0.5 mg / kg. Six hours after administration, each mouse was intraperitoneally injected with luciferase substrate (D-luciferin potassium salt, Catalog No. abs42075819). The liver and spleen were dissected and imaged using an IVIS Small Animal Optical In vivo Imaging System (PerkinElmer). Fluorescence intensity of each tissue was calculated using at least three biological replicates for each lipid nanoparticle composition. Fluorescence intensity represents luciferase protein expression. The relative fluorescence intensity (i.e., relative protein expression) of Compounds 1, 2, and 3, and Comparative Compounds 1 and 3 was calculated, with the fluorescence intensity corresponding to the lipid nanoparticle composition of Comparative Compound 1 as the unit of 1. For example, the ratio of the tissue fluorescence intensity of compound 1 to the tissue fluorescence intensity of reference compound 1 is the relative protein expression level of compound 1.

[0281] As shown in Table 5, Figures 10 and 11, the relative protein expression was calculated by taking statistical fluorescence intensities of the spleen and liver of mice using an IVIS small animal optical in vivo imager. This indicates the efficiency of the lipid nanoparticle composition in delivering mRNA in different tissue cells. In Figures 10 and 11, * indicates statistical P < 0.05, ** indicates statistical P < 0.01, and *** indicates statistical P < 0.001, indicating that there is a statistically significant difference between the groups; ns indicates statistical P ≥ 0.05, indicating that there is no statistically significant difference between the groups.

[0282] Table 5

[0283] Conclusion: The spleen is mainly composed of immune cells. The protein expression levels of the lipid nanoparticle compositions corresponding to compounds 1, 2, and 3 in the spleen were significantly higher than those of the control compound 1, and the protein expression levels of the lipid nanoparticle compositions corresponding to compounds 2 and 3 in the spleen were significantly higher than those of the control compound 3. In the liver, the protein expression levels of the lipid nanoparticle compositions corresponding to compounds 1 and 3 were significantly lower than those of the control compound 1, and the protein expression levels of the lipid nanoparticle compositions corresponding to compounds 1, 2, and 3 were significantly lower than those of the control compound 3. This indicates that compounds 1, 2, and 3 have better immune cell delivery capabilities, can be expressed more in the spleen, and can reduce the expression of non-immune cells such as in the liver.

Claims

1. A compound represented by formula I or a salt thereof, in, H 1 Selected from C 1-6 Alkylene, C 2-6 Alkenylene or C 1-6 heteroalkylene; H 2 Selected from C 1-12 heteroalkylene; R 1 Selected from C 1-24 Alkyl, C 1-24 Heteroalkyl, C 2-24 Alkenyl or C 2-24 heteroalkenyl; R 2 Selected from C 1-24 Alkyl, C 2-24 Alkenyl or -R b -L 1 -R c ; L 1 and L 2 Each is independently selected from -C(O)O-, -OC(O)-, -C(O)-, -OC(O)O-, -O-, -S(O) q -, -SS-, -C(O)S-, -SC(O)-, -NR a C(O)-、-C(O)NR a -、-NR a C(O)NR a -、-NR a C(O)O-、-OC(O)NR a -、-P(O)(OR a )-or key, R a Selected from hydrogen or C 1-6 Alkyl, q is selected from 0, 1 or 2; R b Selected from C 1-12 Alkylene; R c Selected from C 1-24 Alkyl, C 1-24 Heteroalkyl, C 2-24 Alkenyl or C 2-24 heteroalkenyl; R 3 Each independently selected from halogen, amino, hydroxyl, C 1-6 Alkyl, C 1-6 Alkylamino, C 1-6 Alkoxy, C 3-7 Cycloalkyl or C 3-7 Heterocycloalkyl; r is selected from an integer between 0 and 3, such as 1 or 2.

2. The compound of formula I according to claim 1 or a salt thereof, wherein H 2 Selected from C 1-12 Heteroalkylene, preferably C 2-9 Heteroalkylene.

3. The compound of formula I or a salt thereof according to claim 1 or 2, wherein R 1 Selected from C 1-24 Alkyl, preferably C 4-18 Alkyl; or R 1 Selected from C 2-24 Alkenyl, preferably C 4-18 Alkenyl.

4. The compound of formula I or a salt thereof according to any one of claims 1 to 3, wherein R 2 Selected from -R b -L 2 -R c , R b , L 2 or R c As defined in claim 1.

5. The compound of formula I or a salt thereof according to any one of claims 1 to 4, which is a compound of formula II or a salt thereof, in, X is -N(R d )- or -O-, o is selected from an integer between 1 and 11, p is selected from an integer between 1 and 5, and o+p is not greater than 12, R d Selected from hydrogen or C 1-6 Alkyl, R 4 , R 5 , R 6 , R 7 are independently selected from hydrogen, halogen, hydroxyl, C 1-6 Alkyl or C 1-6 Alkoxy, L 1 , L 2 , H 1 , R 1 , R 3 , R b and R c As defined in claim 1.

6. The compound according to claim 5, wherein p is selected from 1 or 2.

7. A compound according to claim 5 or a salt thereof, wherein o is selected from an integer between 4 and 7, such as 5 or 6.

8. according to any one of claims 1 to 7, the compound shown in formula I or its salt, wherein H 1 Selected from C 1-6 Alkylene, preferably C 3-6 Alkylene, more preferably straight chain C 3-6 Alkylene.

9. The compound or salt thereof of formula I according to any one of claims 1 to 8, wherein R 3 Each independently selected from C 1-6 Alkyl or C 1-6 Alkylamino. 10 . The compound of formula I or a salt thereof according to claim 1 , wherein r=0.

11. according to any one of claims 1 to 10, the compound shown in formula I or its salt, wherein L 1 and L 2 Each is independently selected from -C(O)O-, -OC(O)- or a bond, preferably -C(O)O-.

12. The compound of formula I or a salt thereof according to any one of claims 1 to 11, which is a compound of formula III-a or III-b or a salt thereof, in, X, H 1 , R 1 , R 4 ~R 7 , R b , R c , o, p as defined in claim 5.

13. according to any one of claims 1 to 12, the compound shown in formula I or its salt, wherein R c Selected from C 1-24 Alkyl, preferably branched C 11-18 alkyl.

14. according to any one of claims 1 to 13, the compound shown in formula I or its salt, wherein R b Selected from C 7-9 Alkylene.

15. according to any one of claims 1 to 14, the compound shown in formula I or its salt, wherein R 1 or R c Selected from:

16. A compound of formula I or a salt thereof according to any one of claims 1 to 15, wherein the compound is selected from:

17. An isotope-substituted compound of the compound of formula I according to any one of claims 1 to 16, preferably, the isotope-substituted compound is a deuterium atom-substituted compound.

18. A lipid particle comprising a compound shown in formula I according to any one of claims 1 to 16 or a salt thereof or an isotope substituted product according to claim 17, and an activating agent, wherein the activating agent is preferably a polynucleotide or a nucleic acid, such as DNA, siRNA or mRNA.

19. A pharmaceutical composition comprising the lipid particles according to claim 18 and a pharmaceutically acceptable excipient.

20. Use of the compound of formula I or its salt as described in any one of claims 1 to 16, or the isotope substitution as described in claim 17, or the lipid particle as described in claim 18, or the pharmaceutical composition as described in claim 19 in the preparation of a medicament for preventing and / or treating cancer, infectious diseases, autoimmune diseases, neurodegenerative diseases and inflammation.

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