Cationic lipid compound for delivery system and use thereof

Abstract

The present application relates to a class of cationic lipid compounds for a delivery system and use thereof, and in particular, to a lipid compound, and a lipid carrier, a nucleic acid lipid nanoparticle composition, a pharmaceutical formulation, a cell, and a kit based thereon. The compound having the structure of formula (I) of the present application can be used alone to prepare a lipid carrier, or can be used together with other lipid compounds to prepare a lipid carrier. The lipid carrier has high encapsulation efficiency for the nucleic acid drug, which helps to improve the in vivo delivery efficiency of the nucleic acid drug. Moreover, the lipid carrier can also effectively deliver the nucleic acid drug to an organ where enrichment is desired, and thus has good application prospects.

Description

[0001] Cationic lipid compounds for delivery systems and their uses

[0002] Technical Field

[0003] This application relates to the field of biomedicine, specifically to a class of cationic lipid compounds for delivery systems and their uses. Background Art

[0004] The discovery of mRNA has opened new avenues for treating diseases, cancers, and infectious diseases caused by loss-of-function mutations. mRNA drugs can replace dysfunctional genes, express CRISPR-associated RNA or DNA nucleases, and generate antigens that induce protective immunity. However, delivering these large, multi-anionic molecules into the cytoplasm for translation remains a significant challenge. Furthermore, mRNA is highly unstable and readily degraded by ubiquitous nucleases. To address these issues, scientists have developed various non-viral nanoparticles for mRNA delivery, among which lipid nanoparticles (LNPs) are the most clinically feasible.

[0005] LNPs mainly consist of four components: cationic lipids, cholesterol, accessory lipids, and polyethylene glycol lipids. Cationic lipids play a crucial role in LNPs because they are essential for mRNA encapsulation within LNPs and subsequent escape from the endosome into the cytoplasm. Under acidic conditions, cationic lipids carry a positive charge, facilitating RNA loading into LNPs, but remain neutral at physiological pH to prevent harmful effects associated with cationic lipids. During endocytosis, these cationic lipids are protonated in the acidic environment of the endosome and bind to phospholipids, disrupting the stability of the endosome membrane and allowing mRNA to escape.

[0006] In 2018, the FDA approved the first LNP-based siRNA drug (Onpattro); two years later, two SARS-CoV-2 vaccines based on mRNA and LNPs (Comirnaty and Spikevax) entered clinical trials and saved millions of lives during the COVID-19 pandemic. Notably, the three FDA-approved LNP formulations each use a unique cationic lipid, while the other three components—cholesterol, cofactor lipids, and PEGylated lipids—are almost identical. Therefore, cationic lipids are crucial for mRNA delivery efficiency.

[0007] Although many studies have reported on various LNPs containing such cationic lipids, their safety and efficacy still need improvement. Therefore, there is an urgent need for lipid compounds with better safety and efficacy for delivery systems.

[0008] Summary of the Invention

[0009] This application aims to provide a class of novel cationic lipid compounds based on aminomalonic acid as a core, which can be used in delivery systems, including its stereoisomers, tautomers, solvates, or pharmaceutically acceptable salts thereof; and uses whereby the compound can be used alone to prepare a lipid carrier or co-prepared with other lipid compounds to prepare a lipid carrier for the delivery of therapeutic agents. Structurally, due to the close proximity of the two hydroxyl groups to nitrogen (N), they can conjugate with the lone pair electrons on N to enhance the electron cloud of the core.

[0010] Density, and the steric hindrance effect of the two Wienyl groups can also change the spatial orientation of the two branches, thus having unpredictable effects on LNP formation and in vivo efficacy. Based on these considerations and investigations, we designed the novel cationic lipid in this application, which can improve the encapsulation efficiency, delivery efficiency, and safety of LNPs. Furthermore, compared with current cationic lipids based on aminopropylene glycol, it has better delivery efficiency and can better exert its efficacy in vivo. This application increases the variety of cationic lipid compounds, providing more options for the effective delivery of nucleic acid drugs, vaccines, gene-editing drugs, epigenetic editing, and protein replacement drugs. This application provides lipid carriers containing the above compounds, nucleic acid lipid nanocomposites containing the above compounds or the above lipid carriers, and pharmaceutical formulations containing the above compounds, the above lipid carriers, or the above nucleic acid lipid nanocomposites.

[0011] On the one hand, this application provides a compound, said compound or its stereoisomers, tautomers or pharmaceutically acceptable salts, comprising the following structure (I):

[0012]

[0013] In this process, A3 is optionally substituted, and G1 and G2 are each independently selected from the optional substituted main chain consisting of 5 to 25 atoms.

[0014] In some embodiments, the compound or its stereoisomers, tautomers, or pharmaceutically acceptable salts are characterized by comprising the following structure (II):

[0015]

[0016] (II),

[0017] In this configuration, Li and L2 are each independently selected from optional heteroatoms, A3 is an optional substituent, R4 and R5 are each independently selected from H or optional substituents, or R4 and R5 are linked to form a ring, and G3 and G4 are each independently selected from optional substituents.

[0018] In some embodiments, the compound or its stereoisomers, tautomers, or pharmaceutically acceptable salts are characterized by comprising the following structure (III):

[0019]

[0020] In this context, Li and L2 are each independently selected from optionally substituted heteroatoms, L5 is a chemical bond or an optionally substituted substituent, g is 0 or 1, Yi and Y2 are each independently selected from optionally substituted heteroatoms or carbon atoms, and E is -O-, -S(O)b-, or -SS-, where b

[0021]

[0022] In some embodiments, L1 and L2 are each independently -O-, -S-, -N-, -NRa-, where Ra is H or a C1-C12 alkyl group.

[0023] In some embodiments, L5 is a chemical bond, -CH2-, -(C=O)-, -(C=O)S-, -(C=O)O-, or -(C=O)NRa-, wherein Ra is H or a C1-C12 alkyl group.

[0024] In some embodiments, Y1 and Y2 are each independently C, O, S, N, CRa or NRa, wherein Ra is H or C1-C12 alkyl.

[0025] In some embodiments, the compound, or its stereoisomers, tautomers, or pharmaceutically acceptable salts, is characterized by comprising the following structure (Illa):

[0026]

[0027] Wherein, L5 is -CH2- or -(C=O)-, Y1 and Y2 are each independently -O-, -C- or -CRa-, Ra is H or C1-C12 alkyl, A1, A2 and A3 are each independently selected from optional substituents, R3 is an optional alkyl group, R1 and R2 are each independently selected from optional substituents, R4 and R5 are each independently selected from H or optional substituents, or R4 and R5 are linked to form a ring.

[0028] In some embodiments, the compound, or its stereoisomers, tautomers, or pharmaceutically acceptable salts, is characterized by comprising the following structure (Illb):

[0029]

[0030] Where L5 is -CH2- or -(C=0)-, and Y1 and Y2 are independently -0-, -C-, or -CR. a -, where R 3 It is an H or C1-C12 alkyl group, A1, A2 and A3 are each independently selected from optional substituents, Ri and R2 are each independently selected from optional substituents, R4 and R5 are each independently selected from H or optional substituents, or R4 and R5 are linked together to form a ring.

[0031] In some embodiments, Ai, A2, and A3 are each independently an optionally substituted alkyl group, alicyclic group, and / or aliheterocyclic group.

[0032] In some embodiments, Ai, A2, and A3 are each independently an optionally substituted alkyl, alkenyl, and / or cycloalkyl group. In some embodiments, Ai, A2, and A3 are each independently an optionally substituted cycloalkyl, cycloalkenyl, and / or cyclocycloalkyl group.

[0033] In some embodiments, A1, A2, and A3 are each independently an optionally substituted C1-C24 alkylene, C1-C24 alkenylene, C3-C24 cycloalkylene, and / or C3-C24 cycloalkenylene.

[0034] In some embodiments, A1, A2, and A3 are each independently an optionally substituted C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, and / or C8 alkylene.

[0035] In some embodiments, R1, R2, R4 and R5 are each independently an optionally substituted alkyl, alicyclic, aliheterocyclic, heterocyclic, aryl, and / or heteroaryl group.

[0036] In some embodiments, R1, R2, R4 and R5 are each independently an optionally substituted alkyl, alkenyl, and / or alkyl group.

[0037] In some embodiments, R1, *, R4, and R5 are each independently an optionally substituted cycloalkyl, cycloalkenyl, and...

[0038] / or ring block base.

[0039] In some embodiments, R1 and R2 are each independently an optionally substituted C6-C24 alkyl, C6-C24 alkenyl, and / or C6-C24 hydroxyl group.

[0040] In some embodiments, R3 is an optionally substituted C2-C24 alkyl, C2-C24 alkenyl, and / or C2-C24 alkyl group. In some embodiments, R4 and R5 are each independently an optionally substituted C1-C12 alkyl, C2-C12 alkenyl, C2-C12 cycloalkyl, C3-C8 cycloalkyl, C3-C8 cycloalkenyl, C3-C8 cyclocycloalkyl, 4- to 8-membered heterocyclic group, C6-C10 aryl, and / or 5- to 10-membered heteroaryl group.

[0041] In some embodiments, Gi, G2, G3 and G4 are each independently an optionally substituted alkoxy, thioalkyl, alicyclic and / or heterocyclic group consisting of 5 to 25 atoms.

[0042] In some embodiments, Gi, G2, G3 and G4 are each independently an optionally substituted alkyl, alkenyl, and / or bulk group consisting of 5 to 25 atoms.

[0043] In some embodiments, one, two, or three atoms of Gi, Ch, G3, and G4 are each independently replaced by N, O, or S, wherein when replaced by an N atom, the N atom may be replaced by Ra, where R is H or a C1-C12 alkyl group.

[0044] In some embodiments, Li and L2 are each independently -O-, -S-, -NR?-, and when g is 1, Yi and Y2 are each independently C, O, S, NRa or CRa, wherein Ra is H or C1-C12 alkyl.

[0045] In some embodiments, L5 is -(C=O)O-, g is 0, f is 1, E is -0-, -S(O>-, or -SS-, where b is 0, 1, or 2, Ai is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, and A3 is an optionally substituted C1-C24 alkyl group.

[0046]

[0047] In this context, b can be 0, 1, or 2; Ai represents an optional substituted C1-C24 group; A? represents an optional substituted C1-C24 group; and A3 represents an optional substituted C1-C24 group.

[0048]

[0049] In some embodiments, L5 is -(C=O)NRa-, g is 0, f is 1, E is -0-, -S(o)b-, or -SS-, where b is 0, 1, or 2, Ai is an optionally substituted C1-C24 sodium group, A2 is an optionally substituted C1-C24 sodium group, A3 is an optionally substituted C1-C24 sodium group, R1 is an optionally substituted C6-C24 sodium group, R2 is an optionally substituted C6-C24 sodium group, and R3 is an optionally substituted R4 sodium group.

[0050] The substituted C2-C24 alkyl group or, R4 is an optional substituted C1-C12 ® group, and R5 is an optional substituted C1-C12 alkyl group.

[0051] In some embodiments, L5 is a chemical bond, -CH2- or -(C=0)-, g is 0, f is 1, E is -0-, -S(O)b- or -SS-, where b is 0, 1 or 2, Ai is an optionally substituted C1-C24 sodium group, and A2 is an optionally substituted C1-C24 sodium group.

[0052]

[0053] In some embodiments, L5 is -(C=O)O-, g is 1, Y1 and Y2 are each independently C, CR\O, S or NRa, f is O, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, and A3 is an optionally substituted C1-C24 alkyl group.

[0054]

[0055] In some embodiments, L5 is -(C=O)S-, g is 1, Y1 and Y2 are each independently C, C, O, S or NRa, f is O, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, and A3 is an optionally substituted C1-C24 alkyl group.

[0056]

[0057] In some implementations, L5 is -(C=O)NRa-, g is 1, and Y1 and Y2 are each independently C and CR\

[0058] 0, S or NRa, f is O, A1 is an optional substituted C1-C24 sodium group, A2 is an optional substituted C1-C24 sodium group, A3 is an optional

[0059]

[0060] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is O, and Y2 is C, CRa, or NR. a A1 is an optional substituted C1-C24 solenoid, A2 is an optional substituted C1-C24 solenoid, A3 is an optional substituted C1-C24 solenoid, R1 is an optional substituted C6-C24 solenoid, R2 is an optional substituted C6-C24 solenoid, R3 is an optional substituted 2(24 solenoid, R4 is an optional substituted C1-C12 solenoid, R5 is an optional substituted C1-C12 solenoid.

[0061] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is C, Y2 is O, C, S or NRa, A1 is an optionally substituted C1-C24 sonic group, A2 is an optionally substituted C1-C24 sonic group, A3 is an optionally substituted C1-C24 sonic group, R1 is an optionally substituted C6-C24 sonic group, R2 is an optionally substituted C6-C24 sonic group, R3 is an optionally substituted C2-C24 sonic group, R4 is an optionally substituted C1-C12 sonic group, and R5 is an optionally substituted C1-C12 sonic group.

[0062] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is S, Y2 is 0 or 6^, A1 is an optionally substituted C1-C24 group, A2 is an optionally substituted C1-C24 group, A3 is an optionally substituted C1-C24 group, R1 is an optionally substituted C6-C24 group, R2 is an optionally substituted C6-C24 group, R3 is an optionally substituted C1-C14 group, R4 is an optionally substituted C1-C12 group, and R5 is an optionally substituted C1-C12 group.

[0063] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is NRa, Y2 is O, NR\C or CRa, and Al is any

[0064]

[0065] C24 alkyl group, R1 is an optional substituted C6-C24 alkyl group, R2 is an optional substituted C6-C24 alkyl group, R3 is an optional substituted C2-C24 alkyl group, R4 is an optional substituted C1-C12 alkyl group, and R5 is an optional substituted C1-C12 alkyl group.

[0066] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is O, Y2 is C, CRa or NRa, Al is an optionally substituted C1-C24 sonic group, A2 is an optionally substituted C1-C24 sonic group, A3 is an optionally substituted C1-C24 sonic group, R1 is an optionally substituted C6-C24 sonic group, R2 is an optionally substituted C6-C24 sonic group, R3 is an optionally substituted C2-C24 sonic group, R4 is an optionally substituted C1-C12 sonic group, and R5 is an optionally substituted C1-C12 sonic group.

[0067] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is C, and Y2 is O, C, S, or NR. a A1 is an optional substituted C1-C24 solenoid, A2 is an optional substituted C1-C24 solenoid, A3 is an optional substituted C1-C24 solenoid, R1 is an optional substituted C6-C24 solenoid, R2 is an optional substituted C6-C24 solenoid, R3 is an optional substituted 2(24 solenoid, R4 is an optional substituted C1-C12 solenoid, R5 is an optional substituted C1-C12 solenoid.

[0068] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is S, Y2 is C or CRa, Ai is an optionally substituted C1-C24 sodium group, A2 is an optionally substituted C1-C24 sodium group, A3 is an optionally substituted C1-C24 sodium group, R1 is an optionally substituted C6-C24 sodium group, R2 is an optionally substituted C6-C24 sodium group, R3 is an optionally substituted C2-C24 sodium group, R4 is an optionally substituted C1-C12 sodium group, and R5 is an optionally substituted C1-C12 sodium group.

[0069] In some embodiments, L5 is a chemical bond, -CH2- or -(C=O)-, Y1 is NRa, Y2 is O, NR\C or CRa, A1 is an optionally substituted C1-C24α group, A2 is an optionally substituted C1-C24α group, A3 is an optionally substituted C1-C24α group, R1 is an optionally substituted C6-C24α group, R2 is an optionally substituted C6-C24α group, R3 is an optionally substituted C2-C24α group, R4 is an optionally substituted C1-C12α group, and R5 is an optionally substituted C1-C12α group.

[0070] In some embodiments, R1 is an alkyl-substituted C6-C24 alkyl group, R2 is an alkyl-substituted C6-C24 alkyl group, R4 is an optionally substituted C1-C12 alkyl group, and R5 is an optionally substituted C1-C12 alkyl group.

[0071] In some implementations, R4 and R5 can be connected in a ring.

[0072] On the other hand, this application provides a lipid carrier comprising the compound described in this application, a pharmaceutically acceptable salt, a stereoisomer, a tautomer, a solvate, a chelate, or a prodrug.

[0073] In some embodiments, the lipid carrier comprises a first lipid compound and a second lipid compound.

[0074] In some embodiments, the first lipid compound in the lipid carrier comprises the compound described in this application, pharmaceutically.

[0075] Acceptable salts, stereoisomers, tautomers, solvates, chelates, or prodrugs, as well as optional other cationic lipids.

[0076] In some embodiments, the second lipid compound in the lipid carrier comprises one or more of anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids.

[0077] In some implementations, the ligand in the lipid carrier includes one or more of antibodies, peptides, and nucleic acid aptamers.

[0078] In some embodiments, the cationic lipids in the lipid carrier comprise DLinDMA, DODMA, DLin-MC2-

[0079] One or more of MPZ, DLin-MC3-DMA, DLin-KC2-DMA, DOTAP, Cl 2-200 > DC-ChoL ALC-0315, ALC-0366, SM102 and DOTMA.

[0080] In some embodiments, the anionic lipid in the lipid carrier comprises one or more of phosphatidylserine, phosphatidylinositol, phosphatidic acid, and phosphatidylglycerol.

[0081] In some embodiments, the phosphatidylglycerol comprises one or more of DPPG, DOPG, and myristoyl phosphatidylglycerol.

[0082] In some embodiments, the phosphatidylserine comprises one or more of DOPS and DPPS. In some embodiments, the phosphatidic acid comprises one or more of DSPA, DPPA, and DOPA. In some embodiments, the neutral lipid comprises DOPE, DSPC, DPPC, DOPC, POPC, POPE, etc.

[0083] Lipids of at least one of DPPE, DMPE, DSPE, DEPC, HSPC and SOPE, or modified with anionic or cationic modifying groups.

[0084] In some implementations, the steroid comprises berberine, cholesterol, or a derivative thereof.

[0085] In some embodiments, the polymer-bound lipids comprise polyethylene glycol-modified lipids.

[0086] In some embodiments, the molar ratio of the compound to the neutral lipid is about 2:1 to 10:

[0087]

[0088] In some implementations, the molar ratio of the compound to the steroid is about 5:1 to 1:1.

[0089] In some embodiments, the molar ratio of the compound to the lipid bound to the polymer is about 100:1 to 20:1.

[0090]

[0091] On the other hand, this application provides a nucleic acid lipid nanocomposition, the composition comprising the compound described in this application, a pharmaceutically acceptable salt, a stereoisomer, a tautomer, a solvate, a chelate or a prodrug, and a therapeutic agent.

[0092] In some embodiments, the nucleic acid lipid nanocomposition comprises the lipid carrier and therapeutic agent described above. In some embodiments, the therapeutic agent comprises a nucleic acid drug.

[0093] In some embodiments, the nucleic acid drug comprises one or more of RNA, DNA, antisense nucleic acid, aptamer, ribozyme, siRNA, mRNA, dsRNA, miRNA, microRNA inhibitor, immunostimulatory nucleic acid, and PNA.

[0094] combination.

[0095] On the other hand, this application provides a cell comprising the said compound, lipid carrier, and / or the said composition. On the other hand, this application provides a kit comprising the said compound, lipid carrier, the said composition, and / or the said cell.

[0096] On the other hand, this application provides a pharmaceutical formulation comprising the said compound, lipid carrier, said composition, said cells and / or said kit, as well as pharmaceutically acceptable excipients, carriers and diluents.

[0097] On the other hand, this application provides a method for delivering nucleic acids, comprising the said compound, lipid carrier, said composition, said cells, said kit and / or said pharmaceutical preparation.

[0098] On the other hand, this application provides a method for administering a therapeutic agent, comprising the compound, a lipid carrier, the composition, the cells, the kit, and / or the pharmaceutical preparation, wherein the therapeutic agent is mixed with the compound, or the therapeutic agent is present in the lipid carrier, the composition, the cells, the kit, and / or the pharmaceutical preparation.

[0099] On the other hand, this application provides the use of the said compound, the said lipid carrier, the said composition, the said cell, the said kit and / or the said pharmaceutical preparation in the preparation of a medicament, and a method for treating and / or preventing a disease or condition.

[0100] Other aspects and advantages of this application will readily be apparent to those skilled in the art from the detailed description below. The detailed description below shows and describes only exemplary embodiments of this application. As those skilled in the art will recognize, the content of this application enables them to make modifications to the disclosed specific embodiments without departing from the spirit and scope of the invention to which this application pertains. Accordingly, the descriptions in the accompanying drawings and specification of this application are merely exemplary and not restrictive.

[0101] Attached Figure Description

[0102] The specific features of the invention involved in this application are shown in the appended claims. The features and advantages of the invention can be better understood by referring to the exemplary embodiments and drawings described in detail below. A brief description of the drawings is as follows:

[0103] Figure 1 shows the PCSK9 gene editing effect in mice after administration of the delivery substance of this application at a dose of 0.5 mpk. Figure 2 shows the PCSK9 gene editing effect in mice after administration of the delivery substance of this application at a dose of 0.125 mpk. Figure 3 shows the PCSK9 gene editing effect in mice after administration of the delivery substance of this application and the control delivery substance at a dose of 0.125 mpk.

[0104] Detailed Implementation

[0105] The following specific embodiments illustrate the implementation of the invention. Those skilled in the art can easily understand other advantages and effects of the invention from the content disclosed in this specification. Before further describing this application, it should be understood that the terminology used herein is for description only and not for limiting specific embodiments.

[0106] Terminology Definition

[0107] In this application, the term "compound" generally refers to a substance having two or more different elements. For example, the compound in this application can be an organic compound, a compound with a molecular weight of less than 500, a compound with a molecular weight of less than 1000, a compound with a molecular weight of more than 1000, or a compound with a molecular weight of more than 10,000 or more than 100,000. In this application, a compound can also refer to a compound linked by chemical bonds. For example, it can be a compound in which one or more molecules with a molecular weight of less than 1000 are linked by chemical bonds to a biological macromolecule, which can be a polysaccharide, protein, nucleic acid, polypeptide, etc. For example, the compound in this application can include a compound in which a protein is linked to one or more molecules with a molecular weight of less than 1000, a compound in which a protein is linked to one or more molecules with a molecular weight of less than 10,000, or a compound in which a protein is linked to one or more molecules with a molecular weight of less than 100,000.

[0108] As described herein, the term "compound" includes various crystalline forms, pharmaceutically acceptable salts, hydrates, or solvates of compounds. The compounds of this application include tautomers, mesosomes, racemates, enantiomers, and / or diastereomers of compounds. In this application, the term "tautomer" (or "tautomer form") refers to structural isomers with different energies that can interconvert through a low energy barrier. If tautomerism is possible (e.g., in solution), chemical equilibrium of the tautomer can be achieved. For example, proton tautomers (or proton transfer tautomers) include (but are not limited to) interconversions via proton transfer, such as keto-enol isomerization, imine-enamine isomerization, amide-imine alcohol isomerization, etc. In this application, the term "mesosome" generally refers to a molecule containing more than one chiral carbon atom but with zero total optical rotation. The terms “racemate”, “race mixture”, or “race mixture” refer to a composition obtained by mixing equimolar amounts of a pair of enantiomers.

[0109] As described herein, the term "stereoisomer" (or "optical isomer") refers to a stable isomer that, due to having at least one chiral element (including a chiral center, chiral axis, chiral plane, etc.), has a perpendicular asymmetric plane, thereby enabling the rotation of plane-polarized light. Because the compounds of this application contain asymmetric centers and other chemical structures that may lead to stereoisomerism, this application also includes these stereoisomers and mixtures thereof. Since the compounds and their salts of this application include asymmetric carbon atoms, they can exist as single stereoisomers, racemates, or mixtures of enantiomers and diastereomers. Typically, these compounds can be prepared as racemic mixtures. However, if desired, such compounds can be prepared or isolated to obtain pure stereoisomers, i.e., single enantiomers or diastereomers, or mixtures enriched with single stereoisomers (purities of 398%, 395%, 393%, 390%, 388%, 385%, or 380%). The single stereoisomer of a compound is synthesized from an optically active starting material containing the desired chiral center, or obtained by preparing a mixture of enantiomers followed by separation or resolution, such as by converting to a mixture of diastereomers followed by separation or recrystallization, chromatographic treatment, using chiral resolving reagents, or direct separation of enantiomers on a chiral chromatographic column. Starting compounds with specific stereochemistry are commercially available or can be prepared according to the methods described below and then resolved using methods well known in the art.

[0110] Unless otherwise stated, all optical isomers of the compounds described in this application are within the scope of this application. In all compounds described in this application, each chiral carbon atom may optionally be in the R configuration or the S configuration, or a mixture of the R and S configurations.

[0111] In this application, the term "pharmaceutically acceptable salt" refers to a salt of a compound that, within the bounds of reasonable medical judgment, is suitable for contact with human and animal tissues without excessive toxicity, irritation, allergic reactions, or other problems or complications, is commensurate with a reasonable benefit / risk ratio, and possesses the desired pharmacological activity. "Pharmaceutically acceptable salt" generally includes, but is not limited to, salts formed by the reaction of compounds of this application with pharmaceutically acceptable inorganic / organic acids or inorganic / organic bases; such salts are also known as acid addition salts or base addition salts. Specifically, "pharmaceuticalally acceptable acid addition salt" refers to a salt formed with an inorganic acid (e.g., but not limited to, hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.) or with an organic acid (e.g., but not limited to, trifluoroacetic acid, citric acid, maleic acid, fumaric acid, succinic acid, tartaric acid, lactic acid, pyruvic acid, oxalic acid, formic acid, acetic acid, benzoic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, etc.). "Pharmaceutical-acceptable base addition salts" refer to salts prepared by adding inorganic bases (such as, but not limited to, sodium hydroxide, potassium hydroxide, calcium hydroxide, molybdenum hydroxide, etc.) or organic bases (such as, but not limited to, diethylamine, triethylamine, ethambutol, trimethylamine, ethanolamine, dicyclohexylamine, choline, etc.) to free acids.

[0112] In this application, the terms "optionally substituted substituent" and "optionally substituted" mean that the reference group may be substituted or not substituted by one or more additional groups selected individually and independently from H, =O, alkyl, alkenyl, cycloalkyl, cycloalkenyl, cyclocycloalkyl, alkylene, alkenyl, cycloalkylene, cycloalkenylene, alicyclic, aliheterocyclic, heterocyclic, aryl, heteroaryl, heterocyclic, hydroxyl, alkoxy, alkyl, hydroxyl, halogen, tanyl, thioresidue, thioalkyl, N-alkylated group, isohydrogen acid, thiocyanate, isothiocyanate, nitro, perhaloalkyl and amino, and their protected derivatives. Non-limiting examples of optional substituents include H, C1-C25 alkyl, C1-C25 alkenyl, halogen-substituted C1-C25 alkenyl, halogen-substituted C1-C25 cycloalkyl, C1-C25 cycloalkyl, C1-C25 cycloalkyl, halogen-substituted C1-C25 alkyl, C1-C25 alkoxy, halogen-substituted C1-C25 alkoxy, C1-C25 alkylene, C1-C25 alkenyl, C1-C25 cycloalkylene, C1-C25 alkylene, halogen-substituted C1-C25 alkenyl, halogen-substituted C1-C25 cycloalkylene, halogen-substituted C1-C25 cycloalkylene, halogen-substituted C1-C25 cycloalkylene, halogen-substituted C1-C25 cycloalkylene.

[0113] Alkenyl, aryl, heteroaryl, cycloalkyl, heterocycloalkyl, 3- to 10-membered heterocyclic, 5- to 10-membered heteroaryl, F, Cl, Br, I, -CN, =O, =N-OH, =N-OR, =NR, -OR-, -OROR-, -C(O)R-, -C(O)OR-, -NRaR-, -OC(O)R-, -ORC(O)R-, -ORC(O)OR-, -OROC(O)R-, -ORS2R-, -OC(O)OR-, -ORC(O)SR-, -ORSC(O)R-, -OROC(O)NR a -> -ORNR a C(O)O-> -ORNR a C(O)-> -ORNR a C(O)NR a -^ -ORS(O)R-, -ORS(O)2R-> -C(O)NHR-, -ORC(O)NHR, -ORNHC(O)R, -ORNHR, -C(O)NR2> -OC(O)NHR, -OC(O)NR2, -SR-, -SRC(O)R-, -SRC(O)OR-, -SROC(O)R-, -SRS2R-, -SROR-, -SRC(O)S-> -SRSC(O) -, -SRNR a C(O)-> -SRC(O)NR a -. -SRNRa c(O)NRa_, -SROC(O)NR a -> -SRNR a C(O)O-> -S(O)R-, -SRS(O)R-, -SRS(O)2R-> -S(0)2R, -NHR-, -NRaR, -NR a RC(O)R, -NR a RC(O)OR. -NR a ROC(O)R> -S(O)2NHR, -S(O)2NRaR, -NHS(O)2NR a R. -NHS(O)2R> -NRS(O)2R, where R 3It is H or C1-C25 alkyl, wherein each R is independently selected from H, C1-C25 alkyl, C1-C25 alkenyl, halogen-substituted C1-C25 alkenyl, halogen-substituted C1-C25 cycloalkyl, C1-C25 bulk, C1-C25 cyclocycloalkyl, C1-C25 cycloalkyl, C1-C25 cycloalkyl, C1-C25 alkoxy, halogen-substituted C1-C25 alkoxy, C1-C25 alkylene, C1-C25 alkenyl, C1-C25 cycloalkylene, C1-C25 alkylene, halogen-substituted C1-C25 alkenyl, halogen-substituted C1-C25 cycloalkyl, halogen-substituted C1-C25 cycloalkylene, halogen-substituted C1-C25 cycloalkylene, aryl, heteroaryl, cycloalkyl, heterocycloalkyl. 3- to 10-membered heterocyclic groups, 5- to 10-membered heteroaryl groups, F, Cl, Br, I. The position and number of these substituents are determined by the well-known valence state restrictions of each group; for example, =0 is a suitable substituent for alkyl groups but not for aryl groups.

[0114] The term “optional” or “optionally” means that the event or situation described below may or may not occur, including both the occurrence and non-occurrence of the event or situation.

[0115] The terms “contain” and “include” are used in their open, non-restrictive sense.

[0116] In this application, the term "main chain," when referring to a specific functional group, typically refers to the carbon chain containing the most carbon atoms, or the longest carbon chain containing functional groups. In some embodiments, when chains of the same length can serve as main chains for the functional groups, the carbon chain with the most branches (or the simplest branches) is selected as its main chain. In some embodiments, the carbon chain serving as the main chain may contain non-carbon atoms.

[0117] In this application, the term "carbinol" generally refers to any group containing only carbon and hydrogen, including saturated, unsaturated, aromatic, straight-chain or branched or cyclic (including polycyclic) groups, and may also include groups having a specified number of carbon atoms (e.g., Ci-Cio represents 1 to 10 carbon atoms). Carbinols include, but are not limited to: alkyl, alkylene, alkenyl, alkenyl, cycloalkyl, cycloalkenyl, cyclocycloalkylene, cycloalkylene, aryl, heteroaryl. Carbinols can be substituted or unsubstituted, substituent or non-substituent; for example, when substituted, the substituent can be substituted at any usable connection point, and the substituent can be independently and optionally selected from alkyl, alkenyl, cycloalkyl, alkoxy, alkylthio, alkylamino, halogen, pteryl, hydroxyl

[0118] It is substituted by one or more of the following substituents: alkyl, nitro, hydroxyl, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocyclic alkoxy, cycloalkylthio, heterocyclic alkylthio, and oxo.

[0119] In this application, the term "alkyl" generally refers to a group formed by removing a hydrogen atom from an alkyl group. Alkyl groups can be substituted or unsubstituted, substituted or non-substituted. The term "alkyl" refers to a straight-chain, branched, or cycloalkyl group, or a combination thereof, having a specified number of carbon atoms (i.e., Ci-Cn refers to 1 to 12 carbons), and can be fully saturated, monounsaturated, or polyunsaturated, and can include divalent and polyvalent groups. Examples of saturated alkyl groups include, but are not limited to, groups such as methyl, ethyl, n-propyl, isopropyl, n-butyl, tert-butyl, isobutyl, sec-butyl, cyclohexyl, (cyclohexyl)methyl, cyclopropylmethyl, and (e.g.) homologs or isomers of n-pentyl, n-hexyl, n-heptyl, n-octyl, etc. Unsaturated alkyl groups are groups having one or more double or triple bonds. Examples of unsaturated alkyl groups include, but are not limited to, vinyl, 2-propenyl, butenyl, 2-isopentenyl, 2-(butadienyl), 2,4-pentadienyl, 3-(1,4-pentadienyl), ethoxyl, 1-propane and 3-propaneyl, 3-butaneyl, and higher homologs and isomers. The alkyl group can be substituted or unsubstituted, substituent or non-substituent. For example, when substituted, the substituent can be substituted at any usable connection point. The substituent can be independently selected from one or more substituents chosen from alkyl, alkenyl, ethoxy, alkoxy, alkylthio, alkylamino, halogen, alkyl, hydroxyl, nitro, hydroxyl, cycloalkyl, heterocyclic, aryl, heteroaryl, cycloalkoxy, heterocyclic alkoxy, cycloalkylthio, heterocyclic alkylthio, and oxo.

[0120] The term "alkylene" generally refers to a group formed by removing two hydrogen atoms from an alkyl group. Alkylenes can be substituted or unsubstituted, substituted or non-substituted. The term "alkylene" refers to a straight-chain, branched, or cycloalkyl group, or a combination thereof, having a specified number of carbon atoms (i.e., C1-C24 means 1 to 24 carbons), and can be fully saturated, monounsaturated, or polyunsaturated. As used herein, non-limiting examples of alkylene groups include methylene, ethylene, n-propylene, isopropylene, n-butylene, isobutylene, secondary butylene, tert-butylene, n-pentylene, isopentylene, hexylene, etc. In this application, the term "cycloalkyl" generally refers to cycloalkyl, polycycloalkyl, and cycloalkenyl groups, and their combinations with alkyl groups, such as cycloalkylalkyl groups. Cycloalkyl groups can be interrupted by 1, 2, or 3 heteroatoms selected from -O-, -NH-, or -S-. However, the term "cycloalkyl" generally refers to a cycloalkyl group that does not contain heteroatomic interruptions. Examples of cycloalkyl groups include cyclopentyl, cyclohexyl, cyclohexenyl, cyclohexylmethyl, and adamantyl. Cycloalkyl groups can be substituted or unsubstituted. The term "cycloalkylene" generally refers to a divalent cycloalkyl group as defined above, which can be substituted or unsubstituted.

[0121] In this application, the term "bulk group" includes unsaturated aliphatic groups that are similar in length to alkyl groups described above, but contain at least one carbon-carbon triple bond. For example, it may include straight-chain cyclohydroxyl groups (e.g., ethoxyl, propoxyl, butoxyl, pentoxyl, hexoxyl, heptoxyl, octoxyl, nonoxyl, decoxyl, etc.), branched bulk groups (e.g., 1-propoxyl), or combinations thereof, and cycloalkyl or cycloalkenyl-substituted bulk groups. The cyclohydroxyl group may be substituted or unsubstituted. The term "cyclohydroxyl group" generally refers to an unsaturated monocyclic, bicyclic, or tricyclic ring having a certain number of carbon atoms and one or more carbon-carbon triple bonds. For example, a C3-8 cyclohydroxyl group refers to a cyclohydroxyl group having 3-8 carbon atoms. The cyclohydroxyl group may be substituted or unsubstituted. In some embodiments, the cyclohydroxyl group has one carbon-carbon triple bond in the ring. In other embodiments, the cyclohydroxyl group has more than one carbon-carbon triple bond in the ring. Representative examples of cyclocyclosporyl groups include, but are not limited to, cyclopropylcyclosporyl, cyclobutylcyclosporyl, cyclopentylcyclosporyl, cyclohexylcyclosporyl, cycloheptylcyclosporyl, cyclooctylcyclosporyl, cyclononylcyclosporyl, and cyclodecylcyclosporyl.

[0122] In this application, the term "alkenyl" generally refers to a partially unsaturated branched or straight-chain aldehyde group having at least one carbon-carbon double bond, the double bond being located at any position on the carbon chain. The atomic orientation around the double bond is either cis (Z) or trans (E) conformation. Alkenyl groups can be substituted or unsubstituted. In this application, the terms "C2-C12 alkenyl," "C2-C24 alkenyl," and "C6-C24 alkenyl" generally refer to alkenyl groups containing at least 2 or at least 6, and at most 12 or 24 carbon atoms, respectively. In this application, non-limiting examples of alkenyl groups include vinyl, propenyl, butenyl, pentenyl, hexenyl, heptenyl, octenyl, nonenyl, decenyl, etc. In this application, the term "alkenylene" refers to a partially unsaturated branched or straight-chain divalent aldehyde group derived from an alkenyl group. Alkenylene groups can be substituted or unsubstituted. As used herein, non-limiting examples of alkenyl groups include vinylene, propenylene, butenylene, pentenylene, hexenylene, heptenylene, octene, nonenylene, decenylene, etc. In this application, the term "cycloalkenyl" includes a monovalent, partially unsaturated, cyclic alkenyl group that may contain a number of carbon atoms and has at least one carbon-carbon double bond. In one embodiment, there is no carbon-carbon triple bond. Cycloalkenyl groups may be substituted or unsubstituted. Exemplary groups include, but are not limited to, cyclobutenyl, cyclopentenyl, 1-methylcyclopentenyl, cyclohexenyl, and cyclohexadienyl. In this application, the term "cycloalkenyl" generally refers to a divalent cycloalkenyl group as defined above, which may be substituted or unsubstituted. Exemplary groups include, but are not limited to, cyclopropenylene, cyclobutenylene, etc.

[0123] In this application, the term "alicyclic group" generally refers to a residue derived from the same or multiple different carbon atoms of an alicyclic ring by removing a hydrogen atom. The term "cycloalkyl group" generally refers to a saturated or partially unsaturated monocyclic or polycyclic cyclic group containing 3 to 24 carbon atoms, 3 to 12 carbon atoms, 3 to 10 carbon atoms, or 3 to 8 carbon atoms. The cycloalkyl group may be substituted or unsubstituted. Non-limiting examples of alicyclic groups include cyclopropane, cyclobutane, cyclopentane, cyclopentenyl, cyclohexane, cyclohexenyl, cyclohexadienyl, cycloheptane, cyclohepttrienyl, cyclooctane, etc.; polycyclic carbocyclic rings may include spirocyclic, fused-ring, and bridged-ring carbocyclic rings. The alicyclic group may be substituted or unsubstituted.

[0124] In this application, the term "partially unsaturated" generally refers to a cyclic structure in which the cyclic molecules contain at least one double or triple bond. The term "partially unsaturated" encompasses cyclic structures with multiple unsaturations, but is not intended to include aromatic or heteroaromatic rings as defined in this application. The term "unsaturated" indicates that a portion has one or more degrees of unsaturation.

[0125] In this application, the term "heteroatom" generally refers to atoms other than carbon and hydrogen, including oxygen, nitrogen, sulfur, phosphorus, chlorine, etc., and also includes any oxidized form of nitrogen or sulfur and any quaternary salt form of basic nitrogen. The term "nitrogen" may include nitrogen that has undergone a per-substitution reaction. Heteroatoms may be substituted or unsubstituted.

[0126] In this application, the term "aliphatic heterocyclic group" generally refers to a stable, non-aromatic 3-8 member monocyclic carbon ring structure, a fused 7-10 member bicyclic heterocyclic structure, or a bridged 6-10 member bicyclic heterocyclic structure. These cyclic structures can be saturated or partially saturated, and in addition to carbon atoms, they contain one or more heteroatoms selected from the group consisting of oxygen, sulfur, and nitrogen. For example, it may contain 1-4 heteroatoms as defined above. When used to refer to atoms in an aliphatic heterocyclic cyclic structure, the term "nitrogen" may include nitrogen that has undergone a persubstitution reaction. For example, an alicyclic group may include a "heterocyclic alkyl group," which can refer to a stable, non-aromatic 3-7 member monocyclic alkyl structure, a fused 7-10 member bicyclic heterocyclic structure, or a bridged 6-10 member bicyclic heterocyclic structure. In addition to carbon atoms, these cyclic structures contain one or more heteroatoms, wherein the heteroatoms can be selected from the group consisting of oxygen, sulfur, and nitrogen. For example, it may contain 1-4 heteroatoms as defined above. Heterocyclic alkyl groups may be substituted or unsubstituted. Alicyclic groups may be substituted or unsubstituted. In this application, the term "aryl" generally refers to a residue derived from the removal of a hydrogen atom from an aromatic ring. The term "aromatic ring" can refer to a 6- to 14-membered all-carbon monocyclic ring or a fused polycyclic ring (i.e., a ring sharing adjacent carbon atom pairs) with a conjugated puff-electron system, and can be 6- to 10-membered, such as benzene and thalassium. The aromatic ring may be fused to a heteroaryl, heterocyclic, or cycloalkyl ring, wherein the ring attached to the parent structure is an aryl ring. The aryl group may be substituted or unsubstituted, and when substituted, the substituent may be one or more of the following groups, independently selected from the group consisting of: alkyl, alkenyl, hydroxyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxyl, nitro, hydroxyl, cycloalkyl, heterocyclic alkyl, aryl, heteroaryl, cycloalkoxy, heterocyclic alkoxy, cycloalkylthio, and heterocyclic alkylthio. The aryl group may be substituted or unsubstituted.

[0127] In this application, the term "heteroaryl" generally refers to a residue derived from the removal of a hydrogen atom from a carbon atom of a heteroaryl ring. The term "heteroaryl ring" refers to a heteroaryl system comprising 1 to 4 heteroatoms and 5 to 14 ring atoms, wherein the heteroatoms may be selected from the group consisting of oxygen, sulfur, and nitrogen. The heteroaryl group may be 5 to 10-membered, and may be 5- or 6-membered, such as quinoline, phenenyl, pyrrolyl, N-alkylpyrrolyl, nifedipine, pyrazinyl, imidyl, tetramethylpyrrolyl, etc. The heteroaryl ring may be fused to an aryl, heterocyclic, or cycloalkyl ring, wherein the ring connected to the parent structure is the heteroaryl ring. The heteroaryl group can be optionally substituted or unsubstituted. When substituted, the substituent can be one or more of the following groups, independently selected from the group consisting of: alkyl, alkenyl, hydroxyl, alkoxy, alkylthio, alkylamino, halogen, sulfhydryl, hydroxynitro, hydroxyl, cycloalkyl, heterocycloalkyl, aryl, heteroaryl, cycloalkoxy, heteroepoxy, cycloalkylthio, and heterocycloalkylthio. The heteroaryl group can be substituted or unsubstituted.

[0128] In this application, the term "heterocyclic group" refers to a monocyclic, bicyclic, tricyclic, or tetracyclic group having a carbon atom and at least one heteroatom in its ring system, and may include fused, bridged, active spirocyclic rings. For example, "4- to 8-membered heterocyclic group" refers to a saturated or partially unsaturated monocyclic or polycyclic heterocyclic group containing 4 to 8 carbon atoms and heteroatom active heteroatom groups for ring formation.

[0129] In this application, as those skilled in the art will know, terms such as “alkyl,” “alkenyl,” and “cycloalkyl” may be preceded by an identifier indicating the number of atoms present in the group under specific conditions, for example, C1-C4 alkyl, C3-C8 cycloalkoxy, C1-C4 alkylamino, etc., where the subscript number following “C” indicates the number of carbon atoms present in the group. For example, C3 alkyl refers to an alkyl group having three carbon atoms (e.g., n-propyl, isopropyl); in Ci-io, the members of the group may have any number of carbon atoms falling within the range of 1-10.

[0130] One or more hydrogen atoms in a group, for example, up to five, or for example, one to three hydrogen atoms, are independently replaced by the corresponding number of substituents. The substituents are only in their possible chemical positions, and those skilled in the art can determine (by experiment or theory) possible or impossible substitutions without much effort. For example, a group having a free amino or hydroxyl group may be unstable when combined with a carbon atom having an unsaturated bond (such as an alkene).

[0131] In this application, the term "neutral lipid" refers to any of a variety of lipid substances that exist in a zwitterionic form that is either uncharged or neutral at a selected pH. At physiological pH, such lipids include, but are not limited to: phosphatidylcholine such as 1,2-distearyl-sn-glycero-3-phosphocholine (DSPC), 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC), 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1,2-dioleoyl-sn-glycero-3-phosphocholine (DOPC), phosphatidylethanolamine such as 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE), sphingomyelin (SM), ceramides, and steroids such as sterols and their derivatives. Neutral lipids can be synthetic or of natural origin. The term "steroid" refers to compounds containing the following carbon skeleton:

[0132]

[0133] The term "solvent" refers to a substance formed by the combination of a compound of this application or a pharmaceutically acceptable salt thereof with at least one solvent molecule through non-covalent intermolecular forces. Common solvates include (but are not limited to) hydrates, ethanol compounds, acetone compounds, etc.

[0134] The term "chelate" refers to a complex with a cyclic structure, obtained through the chelation of two or more ligands with the same metal ion to form a chelate ring.

[0135] The term "prodrug" refers to a derivative compound that, when administered to a patient, can directly or indirectly provide the compound of this application. Particularly preferred derivative compounds or prodrugs are those that, when administered to a patient, can improve the bioavailability of the compound of this application (e.g., facilitate absorption into the bloodstream) or promote the delivery of the parent compound to its site of action (e.g., the lymphatic system). Unless otherwise stated, all prodrug forms of the compounds of this application are within the scope of this application, and various prodrug forms are well known in the art.

[0136] In this application, the term "polymer-bound lipid" refers to a molecule comprising both a lipid moiety and a polymer moiety. An example of a polymer-bound lipid is a polyethylene glycol-modified lipid. The term "polyethylene glycol-modified lipid" refers to a molecule comprising both a lipid moiety and a polyethylene glycol moiety. Polyethylene glycol-modified lipids are known in the art and include 1-(monomethoxy-polyethylene glycol)-2,3-dimyristoylglycerol (PEG-DMG), etc.

[0137] In this application, the term "ligand-bound lipid" refers to a lipid complex formed by the covalent binding of lipids to a specific ligand (such as a protein, small molecule compound, or other biomolecule), thereby endowing liposomes with specific functions, such as improving targeting and reducing side effects. The ligand can be a protein (e.g., a receptor, enzyme), a small molecule (e.g., a drug, hormone, small molecule compound), an antibody, a peptide, a nucleic acid, or other biomolecule, including but not limited to molecules that can bind to lipids and alter their properties.

[0138] In this application, the term "nucleic acid" refers to a polymer containing at least two deoxyribonucleic acids or ribonucleic acids in single-stranded or double-stranded form, and includes DNA, RNA, and their hybrids. DNA may be in the form of antisense molecules, plasmid DNA, cDNA, PCR products, or vectors. RNA may be in the form of small hairpin RNA (shRNA), messenger RNA (mRNA), antisense RNA, miRNA, micRNA, multivalent RNA, Dicer substrate RNA, or viral RNA (vRNA), and combinations thereof. Nucleic acids include nucleic acids containing known nucleotide analogs or modified backbone residues or bonds, which are synthetic, naturally occurring, or non-natural and have similar binding properties to a reference nucleic acid. Examples of such analogs include, but are not limited to, thiophosphates, aminophosphates, methylphosphates, chiral-methylphosphates, 2'-O-methylribonucleic acids, and peptide nucleic acids (PNAs). Unless specifically defined, this term covers nucleic acids containing known analogs of natural nucleotides that have similar binding properties to a reference nucleic acid. Unless otherwise stated, specific nucleic acid sequences also implicitly encompass variants of their conserved modifications (e.g., degenerate codon substitutions), alleles, orthologs, single nucleotide polymorphisms, complementary sequences, and explicitly stated sequences. A “nucleotide” contains the following sugars: deoxyribose (DNA) or ribose (RNA); the base and phosphate group of the nucleotide are linked together by the phosphate group. A “base” includes pyridine and pyridine, further including natural compounds such as adenine, thymic pyridine, cytosine, urosine, myosine, and natural analogs, as well as synthetic derivatives of pyridine and pyridine, including but not limited to modifications that place new reactive groups (such as, but not limited to, amines, alcohols, thiols, compounds, and alkyl halides).

[0139] In this application, the term "delivery" refers to providing an entity to a target. For example, delivering a therapeutic and / or preventative agent to a subject may involve administering a nanoparticle composition comprising the therapeutic and / or preventative agent to the subject (e.g., via intravenous, intramuscular, or subcutaneous routes). Administering a nanoparticle composition to a mammal or mammalian cells may involve contacting one or more cells with the nanoparticle composition. Here, "contact" refers to establishing a physical connection between two or more entities, and methods for in vivo and ex vivo contact between cells and external entities are well-known in the field of biology.

[0140] In this application, the term "administration" means introducing a composition or agent (e.g., nucleic acid) into an individual and includes the simultaneous and sequential introduction of one or more compositions or agents. "Administration" can refer to, for example, treatment, pharmacokinetics, diagnosis, research, placebo, and experimental methods. "Administration" also encompasses in vitro and ex vivo treatment. The introduction of a composition or agent into an individual is by any suitable route, including oral, nasal, pulmonary, extra-gastric (intravenous, intramuscular, intraperitoneal, or subcutaneous), rectal, intralymphatic, intratumoral, or local administration. Administration includes self-administration and administration by another person. Administration can be performed by any suitable route. A suitable route of administration enables the composition or agent to perform its intended function. For example, if the suitable route is intravenous, then the composition or agent is administered by introducing it into the individual's vein.

[0141] In this application, the term “prevention” covers the following situations: (1) suppressing the onset of a disease in a subject or patient who may be at risk and / or susceptible to the disease, but has not yet experienced or exhibited any or all symptoms or signs of the disease; and / or (2) slowing the onset of symptoms or signs of a disease in a subject or patient who may be at risk and / or susceptible to the disease, but has not yet experienced or exhibited any or all symptoms or signs of the disease.

[0142] In this application, the term “treatment” covers the following: (1) suppressing the disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., preventing the further development of the symptoms and / or signs), (2) improving the disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease (e.g., reversing the symptoms and / or signs), and / or (3) achieving any measurable reduction of the disease in a subject or patient who is experiencing or exhibiting symptoms or signs of the disease.

[0143] In this application, the terms “subject” or “patient” mean any organism to which the compounds and / or compositions described in this application may be administered, for example for experimental, diagnostic, preventive and / or therapeutic purposes. Typical subjects include animals (e.g. mammals, such as mice, rats, rabbits, non-human primates and humans) and / or plants.

[0144] In this application, the term "comprising" generally means including the explicitly specified features, but does not exclude other elements.

[0145] "Above" and "below" usually refer to cases that include the number itself.

[0146] In this application, the term "about" generally refers to a variation within a range of 0.5% to 10% above or below a specified value, such as a variation within a range of 0.5%, 1%, 1.5%, 2%, 2.5%, 3%, 3.5%, 4%, 4.5%, 5%, 5.5%, 6%, 6.5%, 7%, 7.5%, 8%, 8.5%, 9%, 9.5%, or 10% above or below a specified value.

[0147] Invention Details

[0148]

[0149] In this configuration, A3 is optionally substituted, Li and L2 are each independently selected from optionally substituted heteroatoms, Gi and G2 are each independently selected from an optionally substituted main chain consisting of 5 to 25 atoms, and G3 and G4 are each independently selected from optionally substituted substituents. For example, Li and L2 are each independently selected from optionally substituted heteroatoms such as N, O, or S. For example, Li and L2 are optionally substituted. For example, when Li and L2 are N, they can be substituted by any of the following groups: H, alkyl, alkenyl, cycloalkyl, cycloalkylene, cycloalkylene, cycloalkylene, cycloalkylene, cycloalkylene, alicyclic, alicyclic, hydroxyl, β-group, halogen, amino, alkyl, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C17 alkyl, C18 alkyl, C19 alkyl, C20 alkyl, C21 alkyl, C22 alkyl, C23 alkyl, C24 alkyl, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C10 alkenyl. C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2 bulk, C3 hydroxyl, C4 hydroxyl, C5 hydroxyl, C6 bulk, C7 hydroxyl, C8 hydroxyl, C9 hydroxyl, C10 bulk, C11 hydroxyl, C12 hydroxyl, C13 hydroxyl, C14 hydroxyl, C15 hydroxyl, C16 hydroxyl, C17 hydroxyl, C18 hydroxyl, C19 hydroxyl, C20 hydroxyl, C21 hydroxyl, C22 hydroxyl, C23 hydroxyl, C24 hydroxyl. For example, Aa is optionally selected from any of the following groups or any combination thereof: alkyl, alkyl, alkenyl, cycloalkyl, alkenyl, cycloalkyl, cycloalkenyl, cyclocycloalkyl, cycloalkenyl, alicyclic, alicyclic.For example, A3 is optionally selected from any of the following groups or any combination thereof: C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene, C10 alkylene, C11 alkylene, C12 alkylene, C13 alkylene, C14 alkylene, C15 alkylene, C16 alkylene, C17 alkylene, C18 alkylene, C19 alkylene, C20 alkylene, C21 alkylene, C22 alkylene, C23 alkylene, C24 alkylene, C1 alkenyl, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5.

[0150] alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C10 alkenyl, C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20

[0151]

[0152] Cycloalkyl, C6 cycloalkylene, C7 cycloalkylene, C8 cycloalkylene, C9 cycloalkylene, C10 cycloalkylene, C11 cycloalkylene, C12 cycloalkylene, C13 cycloalkylene, C14 cycloalkylene, C15 cycloalkylene, C16 cycloalkylene, C17 cycloalkylene, C18 cycloalkylene, C19 cycloalkylene, C20 cycloalkylene, C21 cycloalkylene, C22 cycloalkylene, C23 cycloalkylene, C24 cycloalkylene, C3 cycloalkylene, C4 cycloalkylene, C5 cycloalkylene, C6 cycloalkylene, C7 cycloalkylene, C8 cycloalkylene, C9 cycloalkylene, C10 cycloalkylene, Cu cycloalkylene, C12 cycloalkylene, C13 cycloalkylene, C14 cycloalkylene, C15 cycloalkylene, C16 cycloalkylene, C17 cycloalkylene C18, C19, C20, C21, C22, C23, and C24 cyclic alkenyl groups. In this application, the main chains of G1 and G2 have a length of 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 atoms. For example, G1, G2, G3, and G4 are each independently selected from any of the following groups or any combination thereof: alkyl, alicyclic, alihexacyclic, alkyl, alkoxy, thioalkyl, N-alkylated, alkenyl, and scallopyl.For example, Gi, G2, G3, and G are each independently selected from any of the following groups or any combination thereof: C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C17 alkyl, C18 alkyl, C19 alkyl, C20 alkyl, C21 alkyl, C22 alkyl, C23 alkyl, C24 alkyl, C25 alkyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C8 alkenyl, C9 alkenyl, C10 alkenyl, C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C25 alkenyl. C5 alkoxy, C6 alkoxy, C7 alkoxy, C& alkoxy, C9 alkoxy, C10 alkoxy, C11 alkoxy, C12 alkoxy, C13 alkoxy, C14 alkoxy, C15 alkoxy, C16 alkoxy, C17 alkoxy, C18 alkoxy, C19 alkoxy, C2Q alkoxy, C2L alkoxy, C22 alkoxy, C23 alkoxy, C4 alkoxy, C25 alkoxy, C4 alkoxy, C5 alkoxy, C6 alkoxy, C7 alkoxy, C8 alkoxy, C9 alkoxy, C10 alkoxy, C11 alkoxy, C12 alkoxy, C13 alkoxy, C14 alkoxy, C15 alkoxy, C16 alkoxy, C17 alkoxy, C18 alkoxy, C19 alkoxy, C20 alkoxy, C21 alkoxy, C22 alkoxy, C23 alkoxy, C24 alkoxy. For example, Gi, G2, G3, and G4 may be optionally substituted. For example, at least one atom in Gi, G2, G3, and G4 may be substituted with N, O, or S, and the substituted atom may be optionally substituted. For example, R4 and R5 may be any of the following groups or any combination thereof: alkyl, alkyl, alkenyl, alkylene, alkenylene, cycloalkyl, cycloalkenyl, cycloalkylene, cycloalkenylene, aryl, heteroaryl, alicyclic, aliheterocyclic. For example, R4 and R5 may be any of the following groups or any combination thereof: H, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C& alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C& alkenyl.

[0153] C9-olefin, Cio-olefin

[0154]

[0155] C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, C8 cycloalkyl, C3 cycloalkenyl, C4 cycloalkenyl, C5 cycloalkenyl, C6 cycloalkenyl, C7 cycloalkenyl, C8 cycloalkenyl, C3 cycloalkenyl, C4 cycloalkenyl, C5 cycloalkenyl, C6 cycloalkenyl, C7 cycloalkenyl, C8 cycloalkenyl, 4-membered heterocyclic group, 5-membered heterocyclic group, 6-membered heterocyclic group, 7-membered heterocyclic group, 8-membered heterocyclic group, C6 aryl, C7 aryl, C8 aryl, C9 aryl, C10 aryl, 5-membered heteroaryl, 6-membered heteroaryl, 7-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, 10-membered heteroaryl Monoaryl aryl. For example, R4 and R5 may be optionally substituted. For example, R4 and R5 may be linked together to form a ring. For example, when the above-mentioned groups are substituted, the substituent is any one of the following groups or any combination thereof: H, =O, alkyl, halogen, hydroxyl, amino, alkyl, alkoxy, alkenyl, bulk, alkylene, alkenyl, cycloalkyl, cycloalkenyl, cyclocycloyl, cycloalkylene, alicyclic, alicyclic, C1alkyl, C2alkyl, C3alkyl, C4alkyl, C5alkyl, C6alkyl, C7alkyl, C8alkyl, C9alkyl, C10alkyl, C11alkyl, C12alkyl, C13alkyl, C14alkyl, C15alkyl, C16alkyl, C17alkyl, C18alkyl, C19alkyl, C20alkyl, C21alkyl, C22alkyl, C23alkyl, C24alkyl, C2alkenyl, C3alkenyl, C4alkenyl, C5alkenyl, C6alkenyl, C7alkenyl, C8alkenyl, C9alkenyl, C10alkenyl. C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2 cycloalkyl, C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, C8 cycloalkyl, C9 cycloalkyl, C10 cycloalkyl, C11 cycloalkyl, C12 cycloalkyl, C13 cycloalkyl, C14 cycloalkyl, C15 cycloalkyl, C16 cycloalkyl, C17 cycloalkyl, C18 cycloalkyl, C19 cycloalkyl, C20 cycloalkyl, C21 cycloalkyl, C22 cycloalkyl, C23 cycloalkyl, C24 cycloalkyl, C1-C24 alkylene, C1-C24 alkenyl, C3-C24 cycloalkylene C3-C24 cycloene groups, 4-π to 8-π heterocyclic groups.

[0156] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene; G1 and G2 are optionally substituted C4-C24 alkoxy groups; the above groups are optionally substituted, wherein the substituent is =O or C1-C24 alkyl, and the substitution position is not limited. In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene; G1 and G2 are optionally substituted C4-C24 alkoxy groups, wherein one atom of G1 and G2 is replaced by an O atom; the above groups are optionally substituted, wherein the substituent is =O or C1-C24 alkyl, and the substitution position is not limited.

[0157] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene group; G1 and G2 are optionally substituted C4-C24 alkoxy groups, wherein one atom of Gi and G2 is replaced by an N atom, the N atom being optionally substituted; the above groups are optionally substituted, the substituents being =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0158] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene group; G1 and G2 are optionally substituted C4-C24 alkoxy groups, wherein one atom of Gi and G2 is replaced by an S atom, the S atom being optionally substituted; the above groups are optionally substituted, the substituents being =O or C1-C24 alkyl, and the substitution position is not limited.

[0159] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene group; G1 and G2 are optionally substituted...

[0160] C4-C24 alkoxy, wherein the Gi and Gi atoms are replaced by S atoms; the above groups may optionally be substituted, wherein the substituents are =0 or C1-C24 alkyl, and the substitution position is not limited.

[0161] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene group; G1 and G2 are optionally substituted C4-C24 alkoxy groups, wherein the two atoms Gi and G2 are replaced by N atoms, and the N atoms are optionally substituted; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0162] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene group; G1 and G2 are optionally substituted C4-C24 alkoxy groups, wherein the two atoms Gi and G2 are replaced by N and O atoms, respectively, and the N atom is optionally substituted; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0163] In some embodiments, A3 in formula (I) is an optionally substituted C1-C24 alkylene group; G1 and G2 are optionally substituted C4-C24 alkoxy groups, wherein the two atoms Gi and G2 are replaced by N and O atoms, respectively, and the N atom is optionally substituted; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0164] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl; R4 and R5 are optionally substituted C1-C12 alkyl, or R4 and R5 are linked into a ring; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0165] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl, wherein one atom of G3 and G4 is replaced by an O atom; R4 and R5 are optionally substituted C1-C12 alkyl, or R4 and R5 are linked into a ring; the above groups are optionally substituted, wherein the substituent is =O or C1-C24 alkyl, and the substitution position is not limited.

[0166] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl, wherein one atom of G3 and G4 is replaced by an S atom; R4 and R5 are optionally substituted C1-C12 alkyl, or R4 and R5 are linked into a ring; the above groups are optionally substituted, wherein the substituent is =O or C1-C24 alkyl, and the substitution position is not limited.

[0167] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl, wherein one atom of G3 and G4 is replaced by an N atom, and the N atom is optionally substituted; R4 and R5 are optionally substituted C1-C12 alkyl, or R4 and R5 are linked into a ring; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0168] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl, wherein the two atoms of G3 and G4 are replaced by N atoms, and the N atoms are optionally substituted; R4 and R5 are optionally substituted C1-C12 alkyl, or R4 and R5 are linked into a ring; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl, and the substitution position is not limited.

[0169] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl groups, wherein the two atoms of G3 and G4 are replaced by S atoms; R4 and R5 are optionally substituted C1-C12 alkyl groups, or R4 and R5 are linked into a ring; the above groups are optionally substituted, wherein the substituents are =0 or =

[0170] C1-C24 alkyl groups, with no restrictions on substitution positions.

[0171] In some embodiments, A3 in formula (II) is an optionally substituted C1-C24 alkylene; L1 and L2 are optionally substituted S or N atoms; G3 and G4 are C4-C24 alkyl groups, wherein the two atoms of G3 and G4 are replaced by N and O atoms, respectively, and the N atom is optionally substituted; R4 and R5 are optionally substituted C1-C12 alkyl groups, or R4 and R5 are linked to form a ring; the above groups are optionally substituted, and the substituents are =O, H or C1-C24 alkyl groups, and the substitution position is not limited.

[0172]

[0173] The compound comprises the structure (I), wherein A3 is a C3 alkylene group, G1 and G2 are C17 alkoxy groups substituted with a alkyl group and a C6 alkyl group, and one atom in G1 and G2 is replaced by an O atom, and the molecular formulas of Gi and G2 are as follows:

[0174] O(CH2)6OC(=O)CH(C6H13)C8H17.

[0175] In this application, the compound comprises structure (III):

[0176]

[0177] Wherein, L1 and L2 are each independently selected from optionally substituted heteroatoms; L5 is a chemical bond or an optionally substituted substituent; g is 0 or 1; Y1 and Y2 are each independently selected from optionally substituted heteroatoms or carbon atoms; E is -O-, -S(o)b-, or -SS-, where b is 0, 1, or 2; f is 0 or 1; A1, A2, and A3 are each independently selected from optionally substituted substituents; R3 is an optionally substituted alkyl group or r

[0178] *N, R; Ri and R2 are each independently selected from optionally substituted substituents; R4 and R5 are each independently selected from H or optionally substituted arugula, or R4 and R5 are linked to form a ring. For example, Li and L2 are each independently selected from optionally substituted heteroatoms such as N, O, S. For example, Li and L2 are optionally substituted. For example, when Li and L2 are N, they can be substituted by any of the following groups: H, alkyl, alkenyl, cycloalkyl, cycloalkylene, cycloalkylene, cycloalkylene, cycloalkylene, cycloalkylene, alicyclic, alihexacyclic, hydroxyl, hydroxyl, halogen, amino, alkyl, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C17 alkyl, C18 alkyl, C19 alkyl, C20 alkyl, C21 alkyl, C22 alkyl, C23 alkyl, C24 alkyl, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C2 alkenyl, C9 alkenyl, C10 alkenyl. C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2 ketyl, C3 ketyl, C4 ketyl, C5 ketyl, C6 ketyl, C7 ketyl, C8 ketyl, C9 ketyl, C10 ketyl, C11 bulkyl, C12 ketyl, C13 bulkyl, C14 ketyl, C15 bulkyl, C16 ketyl, C17 bulkyl, C18 bulkyl, C19 ketyl, C20 ketyl, C21 ketyl, C22 bulkyl, C23 ketyl, C24 bulkyl. For example, Y1 and Y2 are each independently substituted with optional N, O, S, Atoms such as C. For example, Yi and Yz are optionally substituted.For example, when Yi and Y2 are C, they can be substituted by any of the following groups: H, =O, alkyl, alkenyl, alkylene, alkenylene, cycloalkyl, cycloalkenyl, cycloalkylene, cycloalkylene, cycloalkylene, alicyclic, alicyclic, C1alkyl, C2alkyl, C3alkyl, C4alkyl, C5alkyl, C6alkyl, C7alkyl, C8alkyl, C9alkyl, C10alkyl, C11alkyl, C12alkyl, C13alkyl, C14alkyl, C15alkyl, C16alkyl, C17alkyl, C18alkyl, C19alkyl, C20alkyl, C11alkyl, C22alkyl, C23alkyl, C24alkyl, C2alkenyl, C3alkenyl, C4alkenyl, C5alkenyl, C6alkenyl, C7alkenyl, C8alkenyl, C9alkenyl, C10alkenyl, C11alkenyl, C12alkenyl. C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2.

[0179]

[0180] Or any combination thereof: alkyl, alkyl, alkenyl, alkylene, alkenylene, cycloalkyl, cycloalkenyl, cycloalkylene, cycloalkylene, cycloalkenylene, aryl, heteroaryl, alicyclic, alicyclic, C2alkyl, C3alkyl, C4alkyl, C5alkyl, C6alkyl, C7alkyl, C10alkyl, C11alkyl, C12alkyl, C13alkyl, C14alkyl, C15alkyl, C16alkyl, C17alkyl, C1Xalkyl, C19alkyl, C20alkyl, C21alkyl, C22alkyl, C13alkyl, C24alkyl, C2alkenyl, C3alkenyl, C4alkenyl, C5alkenyl, C6alkenyl, C7alkenyl, C10alkenyl, C11alkenyl, C12alkenyl, C13alkenyl C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2 bulk, C3 bulk, C4 bulk, C5 bulk, C6 bulk, C7 bulk, C2 bulk, C9 bulk, C10 bulk, C11 bulk, C12 bulk, C13 bulk, C14 alkenyl, C15 bulk, C16 bulk, C17 bulk, C18 alkenyl, C19 bulk, C20 bulk, C21 bulk, C12R4 bulk, C23 bulk, C24 bulk, AJ, R5. For example, when R4 and R5 are connected, they can form a ring. For example, R4 and R5

[0181]

[0182] Alkyl, alkenyl, bulk, alkylene, alkenylene, cycloalkyl, cycloalkenyl, cyclobulb, cycloalkylene, cycloalkylene, cycloalkylene, aryl, heteroaryl, alicyclic, alicyclic, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C& alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, G alkenyl, C8 alkenyl, C9 alkenyl, C10 alkenyl, C11 alkenyl, C12 alkenyl, C2 fusible, C3 fusible, C4 fusible, C5 fusible, C6 fusible, C7 fusible, C8 fusible, C9 fusible, C10 fusible, Cu fusible, C12 fusible, C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, C8 cycloalkyl, C3 cycloalkenyl, C4 cycloalkenyl, C5 cycloalkenyl, C6 cycloalkenyl, C7 cycloalkenyl, C8 cycloalkenyl, C3 cyclobutane, C4 cyclobutane, C5 cyclobutane, C6 cyclobutane, G cyclobutane, & cyclobutane, 4-membered heterocyclic, 5-membered heterocyclic, 6-membered heterocyclic, 7-membered heterocyclic, 8-membered heterocyclic, C6 aryl, C7 aryl, C8 aryl, C9 aryl, C10 aryl, 5-membered heteroaryl, 6-membered heteroaryl, 7-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, 10-membered heteroaryl. For example, R4 and R5 may optionally be substituted. For example, R4 and R5 may be linked into a ring.For example, AH, A2, and A3 can be any of the following groups or any combination thereof: alkyl, alkyl, alkenyl, alkylene, alkylene, cycloalkyl, cycloalkenyl, cyclobulb, cycloalkylene, cycloalkylene, aryl, heteroaryl, alicyclic, aliheterocyclic, C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene, C10 alkylene, C11 alkylene, C12 alkylene, C13 alkylene, C14 alkylene, C15 alkylene, C16 alkylene, C17 alkylene, C18 alkylene, C19 alkylene, C20 alkylene, C21 alkylene, C22 alkylene, C23 alkylene, C24 alkylene, C1 alkenyl, C2 alkenyl, C3 alkenyl, C4 alkenyl. C5-alkyl, C6-alkyl, C7-alkyl, C&-alkyl, C9-alkyl, C10-alkyl, C11-alkyl, C12-alkyl, C13-alkyl, C14-alkyl, C15-alkyl, C16-alkyl, C17-alkyl, C18-alkyl, C19-alkyl, C20-alkyl, C21-alkyl, C22-alkyl, C23-alkyl, C24-alkyl, C3-cycloalkyl, C4-cycloalkyl, C5-cycloalkyl, C6-cycloalkyl, C7-cycloalkyl, C8-cycloalkyl, C9-cycloalkyl, C10-cycloalkyl, C11-cycloalkyl, C12-cycloalkyl, C13-cycloalkyl, C14-cycloalkyl, C15-cycloalkyl, C16-cycloalkyl, C17-cycloalkyl, C18-cycloalkyl C19 cycloalkyl, C20 cycloalkyl, C21 cycloalkyl, C22 cycloalkyl, C23 cycloalkyl, C24 cycloalkyl, C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, C7 cycloalkyl, C8 cycloalkyl, C9 cycloalkyl, C10 cycloalkyl, C11 cycloalkyl, C12 cycloalkyl, C13 cycloalkyl, C14 cycloalkyl, C15 cycloalkyl, C16 cycloalkyl, C17 cycloalkyl, C18 cycloalkyl, C19 cycloalkyl, C20 cycloalkyl, C21 cycloalkyl, C22 cycloalkyl, C23 cycloalkyl, C24 cycloalkyl. For example, A1, A2, and A3 may optionally be substituted.For example, when the above-mentioned groups are substituted, the substituent is any one of the following groups or any combination thereof: H, =O, alkyl, halogen, alkyl group, hydroxyl group, amino group, alkyl group, alkoxy group, alkenyl group, bulk group, alkylene group, alkenyl group, cycloalkyl group, cycloalkenyl group, cycloalkyl group, cycloalkylene group, alicyclic group, alicyclic heterocyclic group, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C17 alkyl, C18 alkyl, C19 alkyl, C20 alkyl, C21 alkyl, C22 alkyl, C23 alkyl, C24 alkyl, C2 alkenyl, C3 alkenyl, C4 alkenyl, C5 alkenyl, C6 alkenyl, C7 alkenyl, C11 alkenyl, C9 alkenyl, C10 alkenyl. C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, G alkyl, & alkyl, C9 alkyl, C10 alkyl, Cu alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C17 alkyl, C18 alkyl, C19 bulk alkyl, C20 alkyl, C21 bulk alkyl, C22 alkyl, C23 bulk alkyl, C24 bulk alkyl, C1-C24 alkylene, C1-C24 alkenyl, C3-C24 cycloalkylene C3-C24 cycloene group, 4- to 8-membered heterocyclic group.

[0183] In some embodiments, Li and L2 in formula (III) are O atoms; L5 is -CH2-; A1, A2, and A3 are optionally substituted C1-C24 alkylene groups; g is 1; Yi and Y2 are optionally substituted C or O atoms; f % 0; R1, R2, and R3 are optionally substituted C2-C24 alkyl groups; R4 and R5 are optionally substituted C1-C12 alkyl groups; the above groups are optionally substituted, and the substituents are H, C1-C24 alkyl, or hydroxyl. For example, in some more specific embodiments, R3 is J. For example, in some further more specific embodiments, R4 and R5 can be linked into a ring. For example, in one embodiment of formula (III), L1 and L2 are S atoms. For example, in some embodiments, Yi and Y2 are optionally substituted C or S atoms. For example, in some embodiments, Yi and Y2 are optionally substituted N or C atoms. For example, in some embodiments, Yi and Y2 are optionally substituted N or O atoms.

[0184] In some more specific embodiments, Li and L2 in formula (III) may also be N and S atoms, wherein the N atom may be substituted with H or a C1-C24 alkyl group. In some even more specific embodiments, L5 in formula (III) may also be a chemical bond, -(C=O)-, -(C=O)O-, -O(C=O)-, -(C=O)S-, or -(C=O)NRa-, wherein Ra is H or a C1-C12 alkyl group.

[0185] In other embodiments, Li and L2 in formula (III) are O atoms; L5 is -CH2-; Ai, A2, and A3 are optionally substituted C1-C24 alkylene groups; g is 0; f is 1; E is -O-, -S(O>-, or -SS-, where b is 0, 1, or 2; Ri, R2, and R3 are optionally substituted C2-C24 alkyl groups; R4 and R5 are optionally substituted C1-C12 alkyl groups; The substituents are H, C1-C24 alkyl groups, or hydroxyl groups. For example, in some more specific embodiments, R4 and R5 can be linked into a ring.

[0186]

[0187] In this context, L1 and L2 are S atoms. For example, in some embodiments, Y1 and Y2 are optionally substituted C or S atoms. For example, in some embodiments, Y1 and Y2 are optionally substituted N or C atoms. For example, in some embodiments, Y1 and Y2 are optionally substituted N or O atoms.

[0188] In some more specific embodiments, Li and L2 in formula (III) can also be N and S atoms, wherein the N atom can be

[0189] It can be substituted with H or C1-C24 alkyl groups. In some further, more specific embodiments, L5 in formula (III) can also be a chemical bond, -(C=O)-, -(C=O)O-, -O(C=O)-, -(C=O)S- or -(C=O)NRa-, where R a J% H or C1-C12 alkyl.

[0190] For example, in some embodiments, the structure of the compound is as follows (as described in structure EAB-039 provided in this application):

[0191]

[0192] The structure (III) is defined as follows: Li and L2 are 0 atoms, L5 is -(C=0)-, * is a non-substituted C8 alkyl group, A3 is a non-substituted C3 alkylene group, the N atom attached to A3 is connected to its attached R4 and R5 groups to form a non-substituted 5-membered ring, Ai and A2 are non-substituted C5 alkylene groups, g is 1, Yi is a C atom, γ2 is a 0 atom, f % 0, Ri and R2 are C8 alkyl groups substituted with C4 alkyl groups.

[0193] In this application, the compound comprises either structure (Illa) or structure (Illb):

[0194]

[0195] Li is selected from optionally substituted heteroatoms; L5 is a chemical bond or an optionally substituted substituent; Yi and Y2 are each independently optionally substituted heteroatoms or carbon atoms; Ai, A2, and A3 are each independently selected from optionally substituted substituents; R3 is an optionally substituted alkyl group; Ri and R2 are each independently selected from optionally substituted substituents; R4 and R5 are each independently selected from H or optionally substituted substituents, or R4 and R5 are linked to form a ring. For example, Li and L2 are each independently selected from optionally substituted N, O, S, etc. heteroatoms. For example, Yi and Y2 are each independently substituted N, O, S, C, etc. atoms. For example, L5 is a chemical bond, -CH2-, -(C=O)-, -(C=O)S-, -(C=O)O-, or -(C=O)NRa-, where Ra is H or a C1-C12 alkyl group. For example, R3 can be any of the following groups or any combination thereof: alkyl, alkyl, alkenyl, cycloalkyl, alkylene, alkenylene, cycloalkyl, cycloalkenyl, cyclocycloalkylene, cycloalkenylene, aryl, heteroaryl, alicyclic, aliheterocyclic, C2 alkyl

[0196] Alkenyl, C3alkyl, C4alkyl, C5alkyl, C6alkyl, C7alkyl, C8alkyl, C9alkyl, C10alkyl, C11alkyl, C12alkyl, C13alkyl, C14alkyl, C15alkyl, C16alkyl, C17alkyl, C18alkyl, C19alkyl, C20alkyl, C21alkyl, C22alkyl, C23alkyl, C24alkyl, C2alkenyl, C3alkenyl, C4alkenyl, C5alkenyl, C6alkenyl, C7alkenyl, C&alkenyl, C9alkenyl, C10alkenyl, C11alkenyl, C12alkenyl, C13alkenyl, C14alkenyl, C15alkenyl, C16alkenyl, C17alkenyl, C18alkenyl, C19alkenyl, C20alkenyl, C21alkenyl, C22alkenyl, C23alkenyl, C24alkenyl, C2alkenyl, C3alkenyl C4 cooking base, C5 cooking base, C6 cooking base, C7 cooking base, C8 cooking base, C9 cooking base, C10 cooking base, C11 cooking base, C12 cooking base, C13 cooking base, C14 cooking base, C15 cooking base, C16 cooking base, C17 cooking base, C18 cooking base, C19 block base, C20 cooking base, C21 block base, C22 block base, C23 block base, C24 block base. For example, R3 may be optionally replaced.For example, A1, A2, and A3 can be any of the following groups or any combination thereof: alkyl, alkyl, alkenyl, bulk, alkylene, alkenyl, cycloalkyl, cycloalkenyl, cycloalkylene, cycloalkylene, aryl, heteroaryl, alicyclic, aliheterocyclic, C1 alkylene, C2 alkylene, C3 alkylene, C4 alkylene, C5 alkylene, C6 alkylene, C7 alkylene, C8 alkylene, C9 alkylene, C10 alkylene, C11 alkylene, C12 alkylene, C13 alkylene, C14 alkylene, C15 alkylene, C16 alkylene, C17 alkylene, C18 alkylene, C19 alkylene, C20 alkylene, C21 alkylene, C22 alkylene, C13 alkylene, C24 alkylene, C1 alkenyl, C2 alkenyl, C3 alkenyl, C4 alkenyl. C5-alkyl, C6-alkyl, C7-alkyl, &-alkyl, C9-alkyl, Cio-alkyl, CH-alkyl, Cu-alkyl, CH-alkyl, C14-alkyl, C15-alkyl, C16-alkyl, C17-alkyl, C18-alkyl, C19-alkyl, C20-alkyl, C21-alkyl, C22-alkyl, C23-alkyl, C24-alkyl, C3-alkyl-alkyl, C4-alkyl-alkyl, C5-alkyl-alkyl, C6-alkyl-alkyl, C7-alkyl-alkyl, C8-alkyl-alkyl, C9-alkyl-alkyl, C10-alkyl-alkyl, C11-alkyl-alkyl, C12-alkyl-alkyl, C13-alkyl-alkyl, C14-alkyl-alkyl, C15-alkyl-alkyl, C16-alkyl-alkyl, C17-alkyl-alkyl, C18-alkyl-alkyl, C19-alkyl-alkyl C20 cycloalkylene, C21 cycloalkylene, C22 cycloalkylene, C23 cycloalkylene, C24 cycloalkylene, C3 cycloalkylene, C4 cycloalkylene, C5 cycloalkylene, C6 cycloalkylene, C7 cycloalkylene, C8 cycloalkylene, C9 cycloalkylene, C10 cycloalkylene, C11 cycloalkylene, C12 cycloalkylene, C13 cycloalkylene, C14 cycloalkylene, C15 cycloalkylene, C16 cycloalkylene, C17 cycloalkylene, C18 cycloalkylene, C19 cycloalkylene, C20 cycloalkylene, C21 cycloalkylene, C22 cycloalkylene, C23 cycloalkylene, C24 cycloalkylene. For example, A1, A2, and A3 may optionally be substituted.For example, R4 and R5 can be any of the following groups or any combination thereof: alkyl, alkyl, alkenyl, alkylene, alkenylene, cycloalkyl, cycloalkenyl, cycloalkylene, cycloalkylene, cycloalkylene, aryl, heteroaryl, alicyclic, alicyclic, H, C1alkyl, C2alkyl, C3alkyl, C4alkyl, C5alkyl, C6alkyl, C7alkyl, C8alkyl, C9alkyl, C10alkyl, C11alkyl, C12alkyl, C2alkenyl, C3alkenyl, C4alkenyl, C5alkenyl, C6alkenyl, C7alkenyl, C8alkenyl, C9alkenyl, C10alkenyl, C11alkenyl, C12alkenyl, C2 bulkyl, C3 bulkyl, C4 bulkyl, C5 bulkyl, C6alkenyl, C7 bulkyl, Cx alkyl, C9 alkyl, C10 bulkyl, Cu alkyl, C12 bulk, C3 cycloalkyl, C4 cycloalkyl, C5 cycloalkyl, C6 cycloalkyl, G cycloalkyl, C8 cycloalkyl, C3 cycloalkenyl, C4 cycloalkenyl, C5 cycloalkenyl, C6 cycloalkenyl, C7.

[0197] Cycloalkenyl, C8 cycloalkenyl, C3 cyclocyclic, C4 cyclocyclic, C5 cyclocyclic, C6 cyclocyclic, C7 cyclocyclic, C8 cyclocyclic, 4-membered heterocyclic, 5-membered heterocyclic, 6-membered heterocyclic, 7-membered heterocyclic, 8-membered heterocyclic, C6 aryl, C7 aryl, C8 aryl, C9 aryl, C10 aryl, 5-membered heteroaryl, 6-membered heteroaryl, 7-membered heteroaryl, 8-membered heteroaryl, 9-membered heteroaryl, 10-membered heteroaryl. For example, R4 and R5 may optionally be substituted. For example, R4 and R5 may be linked to form a ring. For example, when the above-mentioned groups are substituted, the substituent is any one of the following groups or any combination thereof: H, =O, benzoyl, halogen, thallyl, hydroxyl, amino, alkyl, alkoxy, alkenyl, cycloalkyl, alkenyl, cycloalkyl, cycloalkylene, cycloalkylene, cycloalkylene, alicyclic, alicyclic, C1 alkyl, C2 alkyl, C3 alkyl, C4 alkyl, C5 alkyl, C6 alkyl, C7 alkyl, C8 alkyl, C9 alkyl, C10 alkyl, C11 alkyl, C12 alkyl, C13 alkyl, C14 alkyl, C15 alkyl, C16 alkyl, C17 alkyl, C18 alkyl, C19 alkyl, C20 alkyl, C21 alkyl, C22 alkyl, C23 alkyl.

[0198]

[0199] Alkenyl, C11 alkenyl, C12 alkenyl, C13 alkenyl, C14 alkenyl, C15 alkenyl, C16 alkenyl, C17 alkenyl, C18 alkenyl, C19 alkenyl, C20 alkenyl, C21 alkenyl, C22 alkenyl, C23 alkenyl, C24 alkenyl, C2 hydroxyl, C3 hydroxyl, C4 hydroxyl, C5 hydroxyl, C6 hydroxyl, C7 hydroxyl, C8 hydroxyl, C9 hydroxyl, C10 bulk, C11 bulk, C12 hydroxyl, C13 bulk, C14 hydroxyl, C15 hydroxyl, C16 hydroxyl, C17 hydroxyl, C18 hydroxyl, C19 hydroxyl, C20 bulk, C21 hydroxyl, C22 hydroxyl, C23 hydroxyl, C24 hydroxyl, C1-C24 alkylene, C1-C24 alkenyl C3-C24 cycloalkylene, C3-C24 cycloolefin, 4- to 8-membered heterocyclic groups.

[0200] In some embodiments, L5 in formula (Illa) is -CH2- or -(C=O)-; A1, A2, and A3 are optionally substituted Ci-C24 alkylene or C1-C24 alkenylene; Y1 and Y2 are O, C, or -CRJ, wherein Ra is H or C1-C12 alkyl; Ri, R2, and Ra are optionally substituted C2-C24 alkyl; R4 and R5 are optionally substituted H or C1-C12 alkyl; the above groups are optionally substituted, wherein the substituents are H, C1-C24 alkyl, or hydroxyl. For example, in some more specific embodiments, R4 and R5 may be linked into a ring. For example, in some further more specific embodiments, R4 and R5 are optionally substituted 4- to 8-membered heterocyclic groups or 5- to 10-membered heteroaryl groups.

[0201] In some embodiments, L5 in formula (Illb) is -CH2- or -(C=O)-; A1, A2, and A3 are optionally substituted Ci-C24 alkylene or C1-C24 alkenylene; Y1 and Y2 are O, C, or -CRa-, wherein Ra is H or C1-C12 alkyl; R1 and R2 are optionally substituted C2-C24 alkyl; R4 and R5 are optionally substituted H or C1-C12 alkyl; the above groups are optionally substituted, wherein the substituents are H, C1-C24 alkyl, or hydroxyl. For example, in some more specific embodiments, R4 and R5 may be linked into a ring. For example, in some further more specific embodiments, R4 and R5 are optionally substituted 4- to 8-membered heterocyclic groups or 5- to 10-membered heteroaryl groups.

[0202] On the other hand, this application provides a compound, or a pharmaceutically acceptable salt, stereoisomer, or solvent compound thereof, said compound being selected from any of the structures listed in the following table:

[0203]

[0204]

[0205]

[0206]

[0207]

[0208]

[0209]

[0210] It should be understood that any embodiment of a compound comprising structures (I), (II), (III), or (Illa) to (Illb) as shown above, and any particular substituents and / or variables of the compound structures (I), (II), (III), or (Illa) to (Illb) as shown above, may be independently combined with other embodiments and / or substituents and / or variables of compounds comprising structures (I), (II), (III), or (Illa) to (Illb) to form embodiments of this application not specifically shown above. Furthermore, if a list of substituents and / or variables of any particular R group, L group, G group, A group, Y group, E group, or variable b, g, or f is listed in a particular embodiment and / or claim, it should be understood that each individual substituent and / or variable may be removed from the particular embodiment and / or claim, and the remaining list of substituents and / or variables will be considered within the scope of this application. It should be understood that this application also includes all permutations and combinations permitted under all selections of the substituents involved in the above embodiments.

[0211] It should be understood that, in this specification, the combination of substituents and / or variables in the described formula is permitted only if such contribution results in a stable compound.

[0212] Lipid carrier

[0213] On the other hand, this application provides a lipid carrier comprising one or more of the compounds described in this application. In some embodiments, the lipid carrier comprises a first lipid compound and a second lipid compound. In some embodiments, the first lipid compound comprises one or more of the compounds described in this application, plus optionally other cationic lipids. In some embodiments, the second lipid compound comprises one or more excipients selected from anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids. For example, a composition of lipid nanoparticles (LNPs) is provided, the LNPs comprising any one of the compounds described in this application, and one or more excipients selected from anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids. In other embodiments, the lipid carrier also comprises other pharmaceutically acceptable excipients and / or carriers.

[0214] In some embodiments, the composition comprises any one or more of the compounds described in this application, and one or more excipients selected from anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids, wherein the molar ratio of the compound to the neutral lipid is from about 2:1 to about 10:1. For example, a composition is provided comprising any one or more of the compounds described in this application, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids, wherein the compounds have a molar ratio to the neutral lipids in the following numerical ranges: about 2:1 to about 3:1, about 2:1 to about 4:1, about 2:1 to about 5:1, about 2:1 to about 6:1, about 2:1 to about 7:1, about 2:1 to about 8:1, about 2:1 to about 9:1, about 2:1 to about 10:1, about 3:1 to about 4:1, about 3:1 to about 5:1, about 3:1 to about 6:1, about 3:1 to about 7:1, about 3:1 to about 8:1, about 3:1 to about 9:1, about 3:1 to about 10:1, about 4:1 to about 5:1, about 4:1 to about 6:1. Approximately 4:1 to approximately 7:1, Approximately 4:1 to approximately 8:1, Approximately 4:1 to approximately 9:1, Approximately 4:1 to approximately 10:1, Approximately 5:1 to approximately 6:1, Approximately 5:1 to approximately 7:1, Approximately 5:1 to approximately 8:1, Approximately 5:1 to approximately 9:1, Approximately 5:1 to approximately 10:1, Approximately 6:1 to approximately 7:1, Approximately 6:1 to approximately 8:1, Approximately 6:1 to approximately 9:1, Approximately 6:1 to approximately 10:1, Approximately 7:1 to approximately 8:1, Approximately 7:1 to approximately 9:1, Approximately 7:1 to approximately 10:1, Approximately 8:1 to approximately 9:1, Approximately 8:1 to approximately 10:1, Approximately 9:1 to approximately 10:1 =

[0215] In some embodiments, the composition comprises any one or more of the compounds described in this application, and one or more excipients selected from anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids, wherein the molar ratio of the compound to the steroid is from about 5:1 to about 1:1. For example, a composition comprising any one or more of the compounds described in this application, a therapeutic agent, and one or more lipids selected from neutral lipids, steroids, and polymer-bound lipids is provided.

[0216] A composition of excipients for lipids bound to ligands, wherein the compound has a molar ratio to the neutral lipid in the following numerical ranges: about 5:1 to about 4:1, about 5:1 to about 3:1, about 5:1 to about 2:1, about 5:1 to about 1:1, about 4:1 to about 3:1, about 4:1 to about 2:1, about 4:1 to about 1:1, about 3:1 to about 2:1, about 3:1 to about 1:1, about 2:1 to about 1:1.

[0217] In some embodiments, the composition comprises any one or more of the compounds described in this application, and one or more excipients selected from anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids. In some embodiments, the composition comprises any one or more of the compounds described in this application, a therapeutic agent, and one or more excipients selected from neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids, wherein the compound and the polymer-bound lipid have a molar ratio in the following numerical ranges: about 100:1 to about 90:1, about 100:1 to about 80:1, about 100:1 to about 70:1, about 100:1 to about 60:1, about 100:1 to about 50:1, about 100:1 to about 40:1, about 100:1 to about 30:1, about 100:1 to about 20:1, about 90:1 to about 80:1, about 90:1 to about 70:1, about 90:1 to about 60:1, about 90:1 to about 50:1, about 90:1 to about 40:1, about 90:1 From approximately 30:1, approximately 90:1 to approximately 20:1, approximately 80:1 to approximately 70:1, approximately 80:1 to approximately 60:1, approximately 80:1 to approximately 50:1, approximately 80:1 to approximately 40:1, approximately 80:1 to approximately 30:1, approximately 80:1 to approximately 20:1, approximately 70:1 to approximately 60:1, approximately 70:1 to approximately 50:1, approximately 70:1 to approximately 40:1, approximately 70:1 to approximately 30:1, approximately 70:1 to approximately 20:1, approximately 60:1 to approximately 50:1, approximately 60:1 to approximately 40:1, approximately 60:1 to approximately 30:1, approximately 60:1 to approximately 20:1, approximately 50:1 to approximately 40:1, approximately 50:1 to approximately 30:1, approximately 50:1 From approximately 20:1, from approximately 40:1 to approximately 30:1, from approximately 40:1 to approximately 20:1, from approximately 30:1 to approximately 20:1.

[0218] In this application, the diameter of the lipid carrier is about 500 nm or less. For example, the composition has diameters of the following values: about 500 nm, about 450 nm, about 400 nm, about 350 nm, about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 100 nm, about 75 nm, about 50 nm, and about 25 nm.

[0219] Composition

[0220] On the other hand, this application provides a composition comprising any one or more of the compounds described in this application and a therapeutic agent. In some embodiments, the composition comprises any one of the lipid carriers described in this application and a therapeutic agent. In some embodiments, the composition comprises any one or more of the compounds described in this application, a therapeutic agent, and a lipid carrier. In other embodiments, the composition also comprises other pharmaceutically acceptable excipients and / or carriers.

[0221] In this application, the therapeutic agent comprises one or more nucleic acids. It should be understood that the disclosure of this application is not limited to the specific nucleic acids disclosed in this specification, and is not limited in scope to any particular source, sequence, or type of nucleic acid. However, those skilled in the art can easily identify related homologues from various other nucleic acid sources, including nucleic acids from non-human species. The nucleic acids intended for use in the disclosure of this application may comprise sequences based on naturally occurring sequences. In some embodiments, the therapeutic agent comprises one or more nucleic acids, and the nucleic acids comprise one or more modified nucleotides (e.g.,

[0222] Compounds containing one or more glycosidically modified ribosomals are intended to possess desired properties. In some embodiments, the therapeutic agent comprises one or more nucleic acids, which are oligonucleotides comprising modified ribosomals (e.g., ribosomals comprising modified sugars, modified ribosomal bases, modified bonds, and / or modified ribosomals at other positions such as the 3,3 position of the sugar and / or the 5,5 position of the 5,5 position of the 5,5 position of the 5,5 position of the sugar on the 3,5-terminal ribosomal), and the oligonucleotides obtained by selecting a particular modification may possess desired characteristics.

[0223] Cells and reagent kits

[0224] On the other hand, this application provides a cell comprising the compounds, lipid carriers, and / or compositions described in this application. For example, the cell is a carrier cell comprising a negatively charged cell membrane. For example, the LNP containing the compound described in this application is protonated at a suitable pH, thereby aggregating with the cell membrane of the carrier cell through electrostatic interaction. For example, the compounds, lipid carriers, and / or compositions described in this application are protonated at a suitable pH, causing the cell membrane of the carrier cell to contain the compounds and / or compositions through endocytosis or membrane permeation.

[0225] On the other hand, this application provides a kit comprising the compounds, lipid carriers, compositions, and / or cells described in this application. For example, the kit includes at least one vial, tube, flask, bottle, syringe, or other suitable container into which the components can be placed. For example, if more than one component is present in the kit, the kit will also include a second, third, or other additional container into which additional components can be placed separately. For example, various combinations of components can be contained in the containers. For example, all INP components are combined in a single container. For example, some or all LNP components are provided in separate containers. In some embodiments, the kit will also include packaging for accommodating the various containers. For example, the kit includes instructions for using the kit components. For example, the instructions may be in physical paper form and / or machine-readable electronic form.

[0226] pharmaceutical preparations

[0227] On the other hand, this application provides a pharmaceutical formulation comprising the compound described in this application, the lipid carrier described in this application, the composition described in this application, the cells described in this application, and / or the kit described in this application, as well as pharmaceutically acceptable excipients, carriers, and diluents. In some embodiments, the particle size of the above-mentioned pharmaceutical formulation is 30-500 nm, for example, the pharmaceutical formulation has a particle size of about 500 nm, about 450 nm, about 400 nm, about 350 nm, about 300 nm, about 250 nm, about 200 nm, about 150 nm, about 100 nm, about 75 nm, about 50 nm, and about 30 nm. In some implementation schemes, the encapsulation rate of the nucleic acid drug in the aforementioned drug formulation is greater than 50%. For example, the encapsulation rate can be 55%, 60%, 65%, 70%, 75%, 79%, 80%, 85%, 88%, 90%, 92%, 93%, 96%, 97%, or 99%.

[0228] 100%, etc.

[0229] Method of administering therapeutic agents

[0230] On the other hand, this application provides a method for administering a therapeutic agent, the method comprising providing the compound described in this application, the lipocarrier described in this application, the composition described in this application, the cells described in this application, the kit described in this application, and / or the pharmaceutical formulation described in this application. In some embodiments, the method includes mixing the therapeutic agent described in this application with the compound described in this application. In other embodiments, the method includes placing the therapeutic agent described in this application in the lipocarrier described in this application, in the composition described in this application, in the cells described in this application, in the kit described in this application, and / or in the pharmaceutical formulation described in this application.

[0231] Methods for delivering nucleic acids

[0232] On the other hand, this application provides a method for delivering nucleic acids, the method comprising providing the compound described in this application, the lipid carrier described in this application, the composition described in this application, the cells described in this application, the kit described in this application, and / or the pharmaceutical preparation described in this application. In some embodiments, the method comprises mixing the nucleic acid described in this application with the compound described in this application. In other embodiments, the method comprises placing the nucleic acid described in this application in the lipid carrier described in this application, in the composition described in this application, in the cells described in this application, in the kit described in this application, and / or the pharmaceutical preparation described in this application. For example, the method for delivering nucleic acids comprises systemic delivery of the compound described in this application, the lipid carrier described in this application, the composition described in this application, the cells described in this application, the kit described in this application, and / or the pharmaceutical preparation described in this application, so as to widely expose it to most of the body, which can be performed by any means known in the art, including but not limited to intravenous, intra-arterial, subcutaneous, and intraperitoneal delivery. For example, the method for delivering nucleic acids comprises local delivery of the compounds, lipid carriers, compositions, cells, kits, and / or pharmaceutical formulations described in this application to a target site within the organism, such delivery may be performed by, for example, direct injection into a disease site (e.g., a tumor or site of inflammation) or a target organ (e.g., the liver, heart, kidney, etc.). For example, the local delivery includes local application or injection techniques, including but not limited to intramuscular, subcutaneous, or intradermal injection. For example, the local delivery does not exclude systemic pharmacological effects.

[0233] Methods of treating and / or preventing diseases or conditions

[0234] On the other hand, this application provides a pharmaceutical composition comprising the compounds described in this application, the lipid carriers described in this application, the composition described in this application, the cells described in this application, the kits described in this application, and / or the pharmaceutical formulations described in this application. In some embodiments, the pharmaceutical composition further comprises pharmaceutically acceptable excipients. In some embodiments, the pharmaceutical composition is used for treating and / or preventing diseases.

[0235] On the other hand, this application provides the use of the compounds, lipid carriers, compositions, cells, kits, and / or pharmaceutical preparations described in this application for the preparation of drugs for the treatment or prevention of diseases.

[0236] On the other hand, this application provides the compounds, lipid carriers, compositions, cells, kits, and / or pharmaceutical preparations described in this application for the treatment or prevention of diseases.

[0237] On the other hand, this application provides a method for treating and / or preventing disease, comprising administering to a subject or patient an appropriate dose of the compound described in this application, the lipid carrier described in this application, the composition described in this application, the cells described in this application, the kit described in this application, and / or the pharmaceutical preparation described in this application.

[0238] In this application, the disease includes cancer.

[0239] In this application, the disease includes diseases related to protein or enzyme deficiency.

[0240] In this application, the diseases described include those associated with a specific miRNA or a group or more miRNAs. The examples below are not intended to be limited by any theory; they are merely illustrative of the compounds, preparation methods, and uses of this application, and are not intended to limit the scope of the invention.

[0241] Example

[0242] Example 1: Preparation of the compound

[0243] Synthesis of Intermediate 1

[0244] 1. Synthesis of Compound 2

[0245] Compound 1 (5 g, 28.59 mmol) was dissolved in 50 mL of LMF solution, and TBDPSC1 (8.77 g) and imidazole (4.52 g) were added sequentially. The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was confirmed to be complete by TLC, it was quenched with water, extracted with ethyl acetate to obtain the organic phase, washed with saturated brine, dried and concentrated, and then purified by silica gel column chromatography to obtain compound 2 (10.8 g, 95.3%).

[0246]

[0247] 1H NMR (400 MHz, CDC13) 87.70 - 7.63 (m, 4H), 7.44 - 7.34 (m, 6H), 3.69 - 3.57 (m, 5H), 2.29 (d, J = 7.6 Hz, 2H), 1.67 - 1.51 (m, 4H), 1.36 - 1.22 (m, 8H), 1.04 (s, 9H) O

[0248] 2. Synthesis of Compound 3

[0249] Compound 2 (3 g, 7 mmol) and potassium carbonate (2.55 g) were added to a mixed solvent of 35 mL methanol and 17 mL water, and heated to 85°C. The mixture was stirred for 2 hours until the reaction solution became clear. After confirming complete consumption of the starting materials by TLC, the mixture was cooled to room temperature, concentrated under reduced pressure, and the pH was adjusted to 2-3. Then, 20 mL of brine was added for dilution, and the mixture was extracted with ethyl acetate to obtain the organic phase. The organic phase was dried and concentrated to obtain crude compound 3 (2.88 g, 100%), which was used directly in the next step.

[0250]

[0251] 3. Synthesis of Compound 5

[0252] Compound 3 (288 g, 7 mmol), EDCI (268 g), DMAP (342 mg), DIEA (4.52 g), and compound 4 (1.96 g, 10.5 mmol) were sequentially dissolved in 50 mL of dichloromethane solution. The mixture was stirred overnight at room temperature under nitrogen protection. After TLC confirmation of complete reaction, the reaction was quenched with water, extracted with ethyl acetate to obtain the organic phase, washed with saturated brine, dried, concentrated, and then purified by silica gel column chromatography to obtain compound 5 (3.3 g, 81%).

[0253]

[0254] ¹H NMR (400 MHz, CDCl₃) 87.69–7.63 (m, 4H), 7.45–7.33 (m, 6H), 3.97 (d, J = 6.0 Hz, 2H), 3.64 (t, J = 6.4 Hz, 2H), 2.29 (t, J = 7.6 Hz, 2H), 1.66–1.48 (m, 5H), 1.37–1.20 (m, 24H), 1.04 (s, 9H), 0.92–0.84 (m, 6H).

[0255] 4. Synthesis of Compound 6

[0256] Compound 5 (3.3 g, 5.67 mmol) was dissolved in 30 mL of tetrahydrofuran solution, and then TBAF (11.4 mL, 11.4 mmol, 1 minTHF) was added. The mixture was stirred at room temperature for 2 hours. After the reaction was confirmed to be complete by TLC, the organic phase was extracted with ethyl acetate, washed with saturated brine, dried and concentrated, and then purified by silica gel column chromatography to obtain compound 6 (2 g, 100%).

[0257]

[0258] 1HNMR (400 MHz, CDC13) 53.97 (d, J = 6.0 Hz, 2H), 3.64 (t, J = 6.8 Hz, 2H), 2.30 (t, J = 7.6 Hz, 2H), 1.68 - 1.53 (m, 5H), 1.37 - 1.21 (m, 24H), 0.92- 0.78 (m, 6H).

[0259] 5. Synthesis of Compound 8

[0260] At 0°C, compound 6 (2 g, 5.84 mmol), compound 7 (243 mg, 2.34 mmol), and DCC (1.45 g, 7 mmol) were dissolved sequentially in 20 mL of dichloromethane solution. The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was confirmed to be complete by TLC, the mixture was filtered, washed with dichloromethane, concentrated, and purified by silica gel column chromatography to obtain compound 8 (1.8 g, 100%).

[0261]

[0262] 1H NMR (400 MHz, CDC13) 54.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.36 (s, 2H),

[0263] 2.30 (t, J = 7.6 Hz, 4H), 1.68 - 1.59 (m, 10H), 1.43 - 1.15 (m, 48H), 0.97 - 0.75 (m, 12H) O

[0264] 6. Synthesis of Intermediate 1

[0265] Compound 8 (3.7 g, 4.9 mmol) and p-toluenesulfonic acid (1.27 g, 7.35 mmol) were dissolved sequentially in 50 mL of acetone solvent.

[0266] NBS (875 mg, 4.9 mmol) was added to the solution under nitrogen protection and stirred at 70°C for 5 hours under nitrogen protection. After the reaction was confirmed to be complete by LCMS, the solution was filtered, washed with ethyl acetate, concentrated, and purified by silica gel column chromatography to obtain intermediate 1 (3 g, 74%).

[0267]

[0268] 1H NMR (400 MHz, CDC13) 54.83 (s, 1H), 4.21 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.30 (t, J = 7.6 Hz, 4H), 1.72 - 1.60 (m, 10H), 1.35 - 1.20 (m, 48H), 0.96 - 0.77 (m, 12H).

[0269] Synthesis of EAB-001

[0270] 7. Synthesis of Compound 11

[0271] Compound 9 (3.79 g, 37.8 mmol) and compound 10 (5 mL, 39.7 mmol) were dissolved in 140 mL of methanol solution and stirred at room temperature for 3 hours. Sodium borohydride (2.15 g, 56.8 mmol) was then slowly added, and stirring continued at room temperature for 1 hour. After TLC confirmation of complete reaction, 50 mL of 1 M NaOH was added for quenching, followed by dilution with 100 mL of water. The mixture was extracted with ethyl acetate to obtain the organic phase. After drying and concentrating the organic phase, silica gel column chromatography was performed to obtain compound 11 (3 g, 42.6%).

[0272]

[0273] 8. Synthesis of compound EAB-001

[0274] Intermediate 1 (300 mg, 0.36 mmol), compound 11 (134 mg, 0.72 mmol), and triethylamine (181.8 mg, 1.8 mmol) were sequentially dissolved in 10 mL of chloroform solution. The mixture was stirred overnight at 70°C under nitrogen protection. After LC-MS confirmation of complete reaction, 20 mL of water was added to quench the reaction, followed by dichloromethane chromatography to obtain the organic phase. The organic phase was dried, concentrated, and then purified by silica gel column chromatography to give a yellow oily compound EAB-001 (31.29 mg, 25%).

[0275]

[0276] 1H NMR (400 MHz, CDC13) 54.22 (s, 1H), 4.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.00 - 2.32 (m, 12H), 2.29 (t, J = 7.6 Hz, 4H), 1.65 - 1.59 (m, 12H), 1.46 - 1.42 (m, 2H), 1.29 (d, J = 12.7 Hz, 54H), 0.92 to 0.85 (m, 15H)»

[0277] Synthesis of EAB-002

[0278] 9. Synthesis of Compound 13

[0279] Compound 12 (2.0 g, 15.626 mmol) and compound 10 (1.674 g, 16.407 mmol) were dissolved in 70 mL of methanol solution and stirred at room temperature for 3 hours. Then, sodium borohydride (890 mg, 23.439 mmol) was slowly added, and stirring was continued at room temperature for 1 hour. After the reaction was confirmed to be complete by TLC, 30 mL of 1 M NaOH was added to quench the reaction, followed by dilution with 70 mL of water. The organic phase was extracted with ethyl acetate, dried, concentrated, and then purified by silica gel column chromatography to obtain compound 13 (1.2 g, 35.9%).

[0280] Floating; — 』、、

[0281]

[0282] Eighth of the 1000s to Miwen (a type of Chinese document)

[0283]

[0284] 1HNMR (400 MHz, CDC13) 52.65 (t, J = 7.2 Hz, 2H), 2.59 (t, J = 7.2 Hz, 2H), 2.34 to 2.29 (m, 2H), 2.22 (s, 6H), 1.98 (s, 1H), 1.72 - 1.60 (m, 2H), 1.54 - 1.43 (m, 2H), 1.29 (t, J = 9.1 Hz, 10H), 0.91 - 0.83 (m, 3H)o

[0285] 10. Synthesis of compound EAB-002

[0286] Intermediate 1 (200 mg, 0.24 mmol), compound 13 (80 mg, 0.36 mmol), and triethylamine (122 mg, 1.20 mmol) were dissolved sequentially in 10 mL of chloroform solution. The mixture was stirred overnight at 7CTC under nitrogen protection. After the reaction was confirmed to be complete by LCMS, the mixture was quenched with water, extracted with dichloromethane, and the organic phase was dried, concentrated, and then purified by silica gel column chromatography to obtain a yellow oily compound EAB-002 (30.02 mg, 12.9%).

[0287]

[0288] 1H NMR (400 MHz, CDC13) 84.22 (s, 1H), 4.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.75 -2.60 (m, 4H), 2.59-2.11 (m, 12H), 1.65 - 1.58 (m, 12H), 1.47- 1.42 (m, 2H), 1.33 - 1.23 (m, 58H), 0.92 -0.84 (m, 15H)o

[0289] Synthesis of EAB-003

[0290] 11. Synthesis of Compound 15

[0291] Compound 14 (2 g, 12.82 mmol) and compound 10 (1.37 g, 13.46 mmol) were dissolved in 70 mL of methanol solution and stirred at room temperature for 3 hours. Sodium borohydride (731 mg, 19.23 mmol) was then slowly added, and stirring continued at room temperature for 1 hour. After TLC confirmation of complete reaction, 1 M NaOH (20 mL) was added to quench the reaction. The mixture was diluted with water and then ethyl acetate was used to obtain the organic phase. After drying and concentrating the organic phase, silica gel column chromatography was performed to obtain compound 15 (950 mg, 30.62%).

[0292]

[0293] 12. Synthesis of compound EAB-003

[0294] Compound 15 (300 mg, 0.36 mmol), intermediate 1 (174 mg, 0.72 mmol), and triethylamine (181.8 mg, 1.8 mmol) were dissolved sequentially in 10 mL of chloroform solution. The mixture was stirred overnight at 7 CTC under nitrogen protection. After the reaction was confirmed to be complete by LC-MS, water was added to quench the reaction. The organic phase was then extracted with dichloromethane. After drying and concentrating the organic phase, silica gel column chromatography was performed to obtain a yellow oily compound EAB-003 (64.06 mg, 17.88%).

[0295]

[0296] 1H NMR (400 MHz, CDC13) 54.22 (s, 1H), 4.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.90 - 2.32 (m, 12H), 2.29 (t, J = 7.6 Hz, 4H), 1.86- 1.74 (m, 2H), 1.67- 1.59 (m, 12H), 1.45 - 1.41 (m, 2H), 1.32 - 1.25 (m, 60H), 0.92 - 0.85 (m, 15H).

[0297] Synthesis of intermediate 2

[0298] Intermediate 1 (1.8 g, 2.163 mmol), compound 10 (662 mg, 6.490 mmol), and triethylamine (1.092 g, 10.815 mmol) were sequentially dissolved in 20 mL of chloroform. The mixture was stirred at 70°C for 16 hours under nitrogen protection. After LC-MS confirmation of complete reaction, water was added to quench the reaction. The organic phase was then extracted with dichloromethane, dried, concentrated, and purified by silica gel column chromatography to obtain a yellow oily intermediate 2 (750 mg, 40.6%).

[0299]

[0300] 1H NMR (400 MHz, CDC13) 54.15 (t, J = 6.8 Hz, 4H), 4.04 (s, 1H), 3.97 (d, J = 6.0 Hz, 4H), 2.65 (t, J = 7.2 Hz, 2H), 2.43 - 2.36 (m, 2H), 2.33 - 2.25 (m, 10H), 1.74 - 1.58 (m, 12H), 1.33 - 1.23 (m, 48H), 0.92 - 0.85 (m, 12H).

[0301] Synthesis of EAB-004

[0302] Intermediate 2 (250 mg, 0.293 mmol) was first dissolved in 5 mL of methanol solution, and then compound 16 (162 mg) was added sequentially.

[0303] Add 0.879 mg (1.465 mmol) and 2 drops of acetic acid, stir overnight at room temperature under nitrogen protection, then add sodium borohydride (93 mg, 1.465 mmol), stir for another 2 hours at room temperature. After the product is detected by LCMS, extract with water and dichloromethane to obtain the organic phase. After drying and concentrating the organic phase, purify by preparative liquid chromatography to obtain the colorless oily compound EAB-004 (30.38 mg, 10.2%).

[0304]

[0305] 1H NMR (400 MHz, CDC13) 54.21 (s, 1H), 4.16 - 4.10 (m, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.30 - 3.06 (m, 2H), 2.82 - 2.75 (m, 6H), 2.65 - 2.59 (m, 2H), 2.30 (t, J = 7.6 Hz, 4H), 2.07-1.87 (m, 2H), 1.64-1.59 (m, 12H), 1.47-1.40 (m, 4H), 1.31-1.25 (m, 64H), 0.91 - 0.87 (m, 15H) O

[0306] Synthesis of EAB-005

[0307] Intermediate 2 (200 mg, 0.234 mmol), triethylamine (71 mg, 0.703 mmol), and compound 17 (124 mg, 0.703 mmol) were sequentially dissolved in 5 mL of dichloromethane solution and stirred overnight at room temperature. After the reaction was confirmed to be complete by LCMS, water was added to quench the reaction, and then dichloromethane was used to obtain the organic phase. After drying and concentrating the organic phase, it was purified by silica gel column chromatography and preparative liquid chromatography to obtain the colorless oily compound EAB-005 (29.67 mg, 12.7%).

[0308]

[0309] 1H NMR (400 MHz, CDC13) 8 5.26 - 5.02 (m, 1H), 4.27 - 4.12 (m, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.56 - 3.36 (m, 2H), 2.77 - 2.01 (m, 14H), 1.70 - 1.58 (m, 14H), 1.34 - 1.24 (m, 58H), 0.91 -0.85 (m, 15H)o

[0310] Synthesis of EAB-006

[0311] Intermediate 2 (150 mg, 0.176 mmol), triethylamine (54 mg, 0.528 mmol), and compound 18 were sequentially dissolved in 3 mL of dichloromethane solution and stirred overnight at room temperature. After the reaction was confirmed to be complete by LCMS, water was added to quench the reaction, and then dichloromethane was used to obtain the organic phase. After drying and concentrating the organic phase, it was purified by preparative liquid chromatography to obtain the colorless oily compound EAB-006 (17.77 mg, 9.9%).

[0312]

[0313] ¹H NMR (400 MHz, CDCl₃) 55.30–5.00 (m, ¹H), 4.30–4.20 (m, 2H), 4.20–4.14 (m, 2H), 3.97 (d, J = 5.6 Hz, 4H), 3.59–3.45 (m, 2H), 3.25–3.02 (m, 2H), 2.81 (s, 6H), 2.39–2.20 (m, 8H), 1.71–1.61 (m, ¹²H), 1.32–1.25 (m, ⁶²H), 0.90–0.86 (m, ¹⁵H)

[0314] Synthesis of EAB-007

[0315] 13. Synthesis of Compound 20

[0316] Compound 19 (200 mg, 0.935 mmol) was dissolved in 5 mL of dichloromethane solution, then cooled to 0°C. Under nitrogen protection, 2 drops of DMF and oxaloyl chloride (0.48 mL, 5.6 mmol) were added sequentially. The mixture was stirred at 0°C for 1 hour. After TLC confirmed the reaction was complete, the mixture was concentrated under reduced pressure to obtain a crude product of a pale yellow oily compound 2 (205 mg, 100%), which was used directly in the next step.

[0317]

[0318] 14. Synthesis of EAB-007

[0319] Under nitrogen protection, compound 20 (205 mg, 0.879 mmol) was dissolved in 10 mL of dichloromethane solution, followed by the sequential addition of intermediate 2 (250 mg, 0.293 mmol) and triethylamine (148 mg, 1.466 mmol). The mixture was stirred overnight at room temperature. After LCMS confirmed the reaction was complete, water was added to quench the reaction, followed by dichloromethane extraction to obtain the organic phase. The organic phase was dried and concentrated, and then purified by silica gel column chromatography and preparative liquid chromatography to obtain a pale yellow oily compound EAB-007 (32.97 mg, 10.7%).

[0320]

[0321] 1H NMR (400 MHz, CDC13) 5 5.22 - 5.04 (m, 1H), 4.25 - 4.10 (m, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.44 (t, J = 7.2 Hz, 2H), 2.85 - 1.95 (m, 14H), 1.69- 1.57 (m, 14H), 1.34- 1.23 (m, 66H), 0.92 - 0.85 (m, 15H) =

[0322] Synthesis of intermediate 3

[0323] 15. Synthesis of Compound 23

[0324] Compound 21 (6.91 g, 58.59 mmol), DCC (4.425 g, 21.48 mmol), DMAP (2.86 g, 23.43 mmol), and compound 22 (5 g, 19.53 mmol) were successively dissolved in 50 mL of dichloromethane solution. The mixtures were stirred overnight at room temperature under nitrogen protection. After TLC confirmation of complete reaction, water was added to quench the reaction, and the organic phase was obtained by ethyl acetate chromatography. The organic phase was washed with saturated brine, dried, concentrated, and then subjected to silica gel column chromatography to obtain compound 23 (2.1 g, 30.2%).

[0325]

[0326] 1HNMR (400 MHz, CDC13) 54.07 (t, J = 6.8 Hz, 2H), 3.64 (t, J = 6.4 Hz, 2H), 2.36 - 2.25 (m, 1H), 1.65 - 1.55 (m, 6H), 1.47 - 1.37 (m, 6H), 1.31 - 1.2 (m, 20H), 0.87 (t, J = 6.8 Hz, 6H) O

[0327] 16. Synthesis of Compound 24

[0328] (At TC, compound 23 (1.027 g, 2.88 mmol), compound 7 (100 mg, 15.38 mmol), DMAP (140.77 mg, 1.153 mmol), and DCC (594.2 mg, 2.884 mmol) were dissolved sequentially in 10 mL of dichloromethane solution. The mixture was slowly heated to room temperature and stirred overnight under nitrogen protection. After the reaction was confirmed to be complete by TLC, the mixture was filtered and washed with dichloromethane. The filtrate was concentrated under reduced pressure and then purified by silica gel column chromatography to obtain compound 24 (389 mg, 51.86%).)

[0329]

[0330] 1H NMR (400 MHz, CDC13) 64.14 (t, J = 6.8 Hz, 4H), 4.06 (t, J = 6.8 Hz, 4H), 3.37 (s, 2H), 2.36 - 2.27 (m, 2H), 1.68 - 1.60 (m, 8H), 1.48 - 1.34 (m, 16H), 1.30 - 1.20 (m, 40H), 0.89 - 0.85 (m, 12H) O

[0331] 17. Synthesis of Intermediate 3

[0332] Compound 24 (389 mg, 0.498 mmol) and p-toluenesulfonic acid (129 mg, 0.748 mmol) were dissolved sequentially in 10 mL of ethanocyanin solution. NBS (89 mg, 0.498 mmol) was added under nitrogen protection, and the mixture was stirred at 70°C for 5 hours under nitrogen protection. After TLC confirmation of complete reaction, the mixture was filtered, washed with ethyl acetate, and the filtrate was concentrated under reduced pressure. Silica gel column chromatography yielded a colorless oily intermediate 3 (390 mg, crude).

[0333]

[0334] ¹H NMR (400 MHz, CDCl₃) 54.82 (s, ¹H), 4.22 (t, J = 6.8 Hz, 4H), 4.05 (t, J = 6.8 Hz, 4H), 2.33–2.28 (m, 2H), 1.68–1.60 (m, 8H), 1.45–1.37 (m, 16H), 1.27–1.23 (m, 40H), 0.88–0.85 (m, 12H).

[0335] Synthesis of EAB-009

[0336] Intermediate 3 (100 mg, 0.116 mmol), potassium carbonate (48.3 mg, 0.349 mmol), and compound 10 (20.3 mg, 0.174 mmol) were dissolved sequentially in 5 mL of acetylene solution. The mixture was stirred overnight at 70°C under nitrogen protection. After the reaction was confirmed to be complete by LC-MS, water was added to quench the reaction. The organic phase was then extracted with dichloromethane. After drying and concentrating the organic phase under reduced pressure, it was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-009 (34.54 mg, 33.11%).

[0337]

[0338] 1H NMR (400 MHz, CDC13) 54.15 (t, J = 6.8 Hz, 4H), 4.10 (s, 1H), 4.06 (t, J = 6.8 Hz, 4H), 3.19-3.05 (m, 2H), 2.85 -2.68 (m, 8H), 2.44 (s, 3H), 2.35 -2.25 (m, 2H), 1.97- 1.88 (m, 2H), 1.67 - 1.56 (m, 12H), 1.45 - 1.36 (m, 12H), 1.28 - 1.21 (m, 40H), 0.90- 0.84 (m, 12H)o

[0339] Synthesis of EAB-010

[0340] 18. Synthesis of Compound 25

[0341] Intermediate 3 (1 g, 1.16 mmol), compound 10 (357 mg, 3.49 mmol), and triethylamine (588 g, 5.81 mmol) were sequentially dissolved in 30 mL of chloroform solution and stirred overnight at 70°C under nitrogen protection. After the reaction was confirmed to be complete by LC-MS, water was added to quench the reaction, and then dichloromethane was used to obtain the organic phase. The organic phase was dried, concentrated under reduced pressure, and purified by silica gel column chromatography and preparative liquid chromatography to obtain compound 25 (645 mg, 62.94%).

[0342]

[0343] 19. Synthesis of EAB-010

[0344] Compound 25 (250 mg, 0.284 mmol) was dissolved in 5 mL of methanol solution. Compound 12 (109 mg, 0.851 mmol) and 2 drops of acetic acid were added sequentially. The mixture was stirred overnight at room temperature under nitrogen protection. Then, sodium borohydride (90 mg, 1.42 mmol) was added, and stirring continued for 2 hours at room temperature. After LCMS confirmed product formation, 20 mL of water was added for quenching.

[0345] The organic phase was obtained by dichloromethane extraction, dried and concentrated under reduced pressure, and then purified by preparative liquid chromatography to give a colorless oily compound EAB-010 (21.51 mg, 7.6%). O

[0346]

[0347] 1H NMR (400 MHz, CDC13) 54.22 (s, 1H), 4.14 (t, J = 6.8 Hz, 4H), 4.06 (t, J = 6.8 Hz, 4H),

[0348] 3.35 - 3.00 (m, 2H), 2.81 - 2.57 (m, 8H), 2.34 - 2.27 (m, 2H), 2.07 - 1.83 (m, 2H), 1.69 - 1.58 (m,

[0349] 12H), 1.45 — 1.36 (m, 14H), 1.30 — 1.21 (m, 52H), 0.90 — 0.85 (m, 15H) O

[0350] Synthesis of EAB-013

[0351] Intermediate 2 (200 mg, 0.234 mmol), triethylamine (71 mg, 0.703 mmol), and compound 26 (123 mg, 0.469 mmol) were sequentially added to 10 mL of dichloromethane solution. The mixture was stirred overnight at room temperature under nitrogen protection. After LC-MS confirmation of complete reaction, 20 mL of water was added to quench the reaction. The mixture was then extracted with dichloromethane to obtain the organic phase. The organic phase was dried, concentrated under reduced pressure, and purified by silica gel column chromatography and preparative liquid chromatography to obtain a colorless oily compound EAB-013 (71.74 mg, 28.4%).

[0352]

[0353] 1H NMR (400 MHz, CDC13) 8 5.24 - 5 05 (m, 1H), 4.24 - 4.10 (m, 4H), 3 97 (d, J = 6.0 Hz,

[0354] 4H), 3.53–3.39 (m, 2H), 2.75–2.08 (m, 14H), 1.90–1.56 (m, 18H), 1.32–1.24 (m, 66H), 0.91–0.85 (m, 15H)

[0355] Synthesis of EAB-014

[0356] 20. Synthesis of Compound 28

[0357] Compound 27 (500 mg, 2.208 mmol) and compound 10 (237 mg, 2.319 mmol) were dissolved in 10 mL of methanol and 10 mL of 1,2-dichloroethane solution. The mixture was stirred at room temperature for 3 hours under nitrogen protection. Then, sodium borohydride (126 mg, 3.312 mmol) was slowly added, and the mixture was stirred at room temperature for another hour. After the reaction was confirmed to be complete by TLC, 30 mL of 1 M NaOH was added to quench the reaction. The mixture was then diluted with 50 mL of water and ethyl acetate to obtain the organic phase. After drying and concentrating the organic phase under reduced pressure, the mixture was subjected to silica gel column chromatography to obtain compound 28 (260 mg, 39.4%).

[0358]

[0359] 21. Synthesis of EAB-014

[0360] Intermediate 1 (200 mg, 0.24 mmol), compound 28 (113 mg, 0.36 mmol), and triethylamine (122 mg, 1.20 mmol) were sequentially added to 10 mL of chloroform solution and stirred overnight at 70°C under nitrogen protection. After the reaction was confirmed to be complete by LC-MS, 20 mL of water was added to quench the reaction, and then dichloromethane was used to obtain the organic phase. The organic phase was dried, concentrated under reduced pressure, and purified by silica gel column chromatography and preparative liquid chromatography to obtain a yellow oily compound EAB-014 (15.41 mg, 6.0%).

[0361]

[0362] ¹H NMR (400 MHz, CDCl₃) 54.27 (s, ¹H), 4.20 - 4.08 (m, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.41 - 3.08 (m, 2H), 3.01 - 2.59 (m, 10H), 2.30 (t, J = 7.6 Hz, 4H), 2.24 - 2.00 (m, 2H), 1.66 - 1.59 (m, 12H), 1.32 - 1.24 (m, 72H), 0.91 - 0.86 (m, 15H).

[0363] Synthesis of intermediate 4

[0364] 22. Synthesis of Compound 30

[0365] Compound 29 (10 g, 62.42 mmol) was dissolved in 100 mL of DMF solution, followed by the addition of TBDPSC1 (20.59 g, 74.90 mmol) and imidazole (10.62 g, 156.04 mmol). The mixture was stirred overnight at room temperature under nitrogen protection. After TLC confirmed the reaction was complete, 100 mL of water was added to quench the reaction. The organic phase was then obtained by ethyl acetate chromatography. The organic phase was washed with saturated brine, dried, concentrated under reduced pressure, and then purified by silica gel column chromatography to obtain compound 30 (23 g, 92%).

[0366]

[0367] 1H NMR (400 MHz, CDC13) 57.70 - 7.63 (m, 4H), 7.42 - 7.36 (m, 6H), 3.69 - 3.60 (m, 5H), 2.29 (t, J = 7.6 Hz, 2H), 1.63 - 1.54 (m, 4H), 1.40 - 1.24 (m, 4H), 1.07 - 1.00 (m, 9H).

[0368] 23. Synthesis of Compound 31

[0369] Compound 30 (27 g, 67.8 mmol) was added to a mixed solvent of 60 mL methanol and 30 mL water, and potassium carbonate (22 g, 169.5 mmol) was added. The mixture was heated to 85°C and stirred for 12 hours until the reaction solution became clear. After confirming complete consumption of the starting material by TLC, the reaction solution was cooled to room temperature and concentrated under reduced pressure. The pH was adjusted to 2-3, and then diluted with 50 mL brine. The organic phase was extracted with ethyl acetate. After drying and concentrating the organic phase under reduced pressure, crude compound 31 (23 g, 88%) was obtained. This crude product was used directly in the next step.

[0370]

[0371] 24. Synthesis of Compound 32

[0372] Compound 31 (22.9 g, 59.64 mmol), EDCI (22.9 g, 119.28 mmol), DMAP (3.6 g, 29.8 mmol), DIEA (38.46 g, 298.2 mmol), and compound 4 (16.63 g, 89.45 mmol) were sequentially added to 150 mL of dichloromethane solution. The mixture was stirred overnight at room temperature under nitrogen protection. After TLC confirmation of complete reaction, 50 mL of water was added to quench the reaction. The organic phase was then obtained by ethyl acetate chromatography. The organic phase was washed with saturated brine, dried, concentrated under reduced pressure, and then subjected to silica gel column chromatography to obtain compound 32 (24.4 g, 74.11%).

[0373]

[0374] 1H NMR (400 MHz, CDC13) 57.69 - 7.64 (m, 4H), 7.44 - 7.35 (m, 6H), 3.97 (d, J = 5.6 Hz, 2H), 3.65 (t, J = 6.4 Hz, 2H), 2.29 (t, J = 7.6 Hz, 2H), 1.65 - 1.52 (m, 5H), 1.37 - 1.25 (m, 20H), 1.05 (d, J = 2.8 Hz, 9H), 0.91 - 0.87 (m, 6H).

[0375] 25. Synthesis of Compound 33

[0376] Compound 32 (12 g, 21.74 mmol) was added to 100 mL of tetrahydrofuran solution, followed by the addition of TBAF (44 mL, 43.48 mmol, 1 M in THF). The mixture was stirred at room temperature for 16 hours. After TLC confirmation of complete reaction, 100 mL of ethyl acetate was added for dilution. The organic phase was washed with saturated brine, dried, concentrated under reduced pressure, and then purified by silica gel column chromatography to obtain compound 33 (6 g, 87.90%).

[0377]

[0378] 1HNMR (400 MHz, CDC13) 53.96 (d, J = 5 6 Hz, 2H), 3 63 (t, J = 6.8 Hz, 2H), 2.30 (t, J = 7.6 Hz, 2H), 1.66- 1.53 (m, 5H), 1.40 - 1.34 (m, 4H), 1.31 - 1.23 (m, 16H), 0.93 - 0.84 (m, 6H).

[0379] 26. Synthesis of Compound 34

[0380] At TC, compound 33 (11.6 g, 38.46 mmol), compound 7 (1.6 g, 15.38 mmol), and DCC (9.5 g, 46.14 mmol) were added sequentially to 150 mL of dichloromethane solution. The mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The TLC plate was then...

[0381] After confirming the reaction was complete, the mixture was filtered. The residue was washed with dichloromethane, and the filtrate was concentrated under reduced pressure. The resulting product was then subjected to silica gel column chromatography to obtain compound 34 (9 g, 83.97%).

[0382]

[0383] 1H NMR (400 MHz, CDC13) 54.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 5.6 Hz, 4H), 3.35 (s, 2H), 2.30 (t, J = 7.6 Hz, 4H), 1.68 - 1.57 (m, 10H), 1.40- 1.33 (m, 8H), 1.31 - 1.23 (m, 32H), 0.94- 0.84 (m, 12H) O

[0384] 27. Synthesis of Intermediate 4

[0385] Compound 34 (3.7 g, 5.31 mmol) and p-toluenesulfonic acid (1.27 g, 7.96 mmol) were added sequentially to 100 mL of ethyl acetate solution. NBS (900 mg, 5.04 mmol) was added under nitrogen protection, and the mixture was stirred at 70°C for 5 hours under nitrogen protection. After the reaction was confirmed to be complete by LC-MS, the mixture was filtered, the residue was washed with ethyl acetate, and the filtrate was concentrated under reduced pressure. Silica gel column chromatography was then used to obtain a colorless oily intermediate, intermediate 4 (2.8 g, 67.98%).

[0386]

[0387] 1H NMR (400 MHz, CDC13) 84.82 (s, 1H), 4.21 (t, J = 6.4 Hz, 4H), 3.97 (d, J = 5.6 Hz, 4H), 2.30 (t, J = 7.6 Hz, 4H), 1.72 - 1.59 (m, 10H), 1.42 - 1.34 (m, 8H), 1.31 - 1.25 (s, 32H), 0.91 - 0.85 (m, 12H)o

[0388] Synthesis of intermediate 5

[0389] Intermediate 4 (2.5 g, 3.22 mmol), compound 10 (985 mg, 9.66 mmol), and triethylamine (1.626 g, 16.1 mmol) were sequentially added to 30 mL of chloroform solution. The mixture was stirred overnight at 70°C under nitrogen protection. After LC-MS confirmation of complete reaction, 20 mL of water was added to quench the reaction. The mixture was then extracted with dichloromethane to obtain the organic phase. The organic phase was dried, concentrated under reduced pressure, and purified by silica gel column chromatography and preparative liquid chromatography to obtain a yellow oily intermediate 5 (1.1 g, 42.9%).

[0390]

[0391] 1H NMR (400 MHz, CDC13) 54.15 (t, J = 6.8 Hz, 4H), 4.04 (s, 1H), 3.96 (d, J = 5.6 Hz, 4H), 2.67 (t, J = 6.8 Hz, 2H), 2.56 - 2.47 (m, 2H), 2.34 (s, 6H), 2.30 (t, J = 7.6 Hz, 4H), 1.80 - 1.72 (m,

[0392] 2H), 1.67 - 1.57 (m, 10H), 1.39 - 1.33 (m, 8H), 1.31- 1.31 (m, 32H), 0.91-0.86 (m, 12H) O

[0393] Synthesis of EAB4H5

[0394] Intermediate 5 (200 mg, 0.25 mmol) and triethylamine (76 mg, 0.75 mmol) were added to 10 mL of dichloromethane solution. Compound 37 (74 mg, 0.5 mmol) was added under TC. The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was confirmed to be complete by LCMS, 20 mL of water was added to quench the reaction. The organic phase was then obtained by dichloromethane extraction. After drying and concentrating the organic phase under reduced pressure, it was purified by strip-forming liquid chromatography to obtain a colorless oily compound EAB-015 (66.49 mg, 29.14%).

[0395]

[0396] 1HNMR (4OO MHz, CDC13) 55.14 (s, 1H), 4.21 - 4.10 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.45 - 3.39 (m, 2H), 2.43 - 2.38 (m, 2H), 2.35 - 2.27 (m, 6H), 2.22 (s, 6H), 1.77 -

[0397]

[0398] 2H), 1.67 - 1.55 (m, 14H), 1.38 - 1.33 (m, 8H), 1.31 - 1.25 (m, 36H), 0.90 -0.85 (m, 15H) O

[0399] Synthesis of EABW16

[0400] Intermediate 5 (200 mg, 0.251 mmol), triethylamine (77 mg, 0.753 mmol), and compound 17 (89 mg, 0.502 mmol) were sequentially added to 10 mL of dichloromethane solution. The mixture was stirred overnight at room temperature. After the reaction was confirmed to be complete by LC-MS, 20 mL of water was added to quench the reaction. The mixture was then extracted with dichloromethane to obtain the organic phase. The organic phase was dried, concentrated under reduced pressure, and purified by silica gel column chromatography and HPLC to obtain a colorless oily compound EAB-016 (97.61 mg, 41.5%).

[0401]

[0402] 1HNMR (4OO MHz, CDC13) 55.15 (s, 1H), 4.25 - 4.08 (m, 4H), 3.97 (d, J = 6.0 Hz, 4H), 3.43 (t, J = 7.6 Hz, 2H), 2.49- 2.10 (m, 14H), 1.70 - 1.56 (m, 14H), 1.37- 1.24 (m, 50H), 0.92-0.85 (m, 15H) O

[0403] Synthesis of EAB-017

[0404] Intermediate 5 (200 mg, 0.25 mmol) and triethylamine (76 mg, 0.75 mmol) were added to 10 mL of dichloromethane solution, and compound 18 (103 mg, 0.5 mmol) was added at 0°C. The mixture was stirred overnight at room temperature under nitrogen protection, and the reaction was confirmed by LC-MS.

[0405] After complete extraction, 20 mL of water was added for quenching, followed by extraction with dichloromethane to obtain the organic phase. After drying and concentrating the organic phase under reduced pressure, it was purified by preparative liquid chromatography to obtain the colorless oily compound EAB-017 (70.53 mg, 29 12 %).

[0406]

[0407] ¹H NMR (400 MHz, CDCl₃) 55.14 (s, 1H), 4.22–4.11 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.46–3.38 (m, 2H), 2.43–2.36 (m, 2H), 2.33–2.27 (m, 6H), 2.23 (s, 6H), 1.77–1.71 (m, 2H), 1.68–1.56 (m, 12H), 1.37–1.24 (m, 54H), 0.92–0.85 (m, 15H).

[0408] Synthesis of EAB-018

[0409] Under nitrogen protection, intermediate 5 (300 mg, 0.376 mmol) and triethylamine (190 mg, 1.88 mmol) were added sequentially to 15 mL of dichloromethane solution. The reaction mixture was cooled to 0.3 °C, and then compound 20 (176 mg, 0.752 mmol) was added. The mixture was slowly heated to room temperature and stirred overnight. After LC-MS confirmation of complete reaction, 20 mL of water was added to quench the reaction. The organic phase was extracted with dichloromethane, dried, concentrated under reduced pressure, and purified by silica gel column chromatography and preparative liquid chromatography to obtain a colorless oily compound EAB-018 (79.98 mg, 21.4%).

[0410]

[0411] 1H NMR (400 MHz, CDC13) 55.15 (s, 1H), 4.27 - 4.08 (m, 4H), 3.97 (d, J = 5.6 Hz, 4H), 3.43 (t, J = 7.6 Hz, 2H), 2.50 -2.12 (m, 14H), 1.74 - 1.57 (m, 14H), 1.38 - 1.24 (m, 58H), 0.92-0.85 (m, 15H)o

[0412] Synthesis of EAB-019

[0413] 28. Synthesis of Compound 39

[0414] Compound 38 (2.0 g, 17.515 mmol) and compound 10 (1.876 g, 18.391 mmol) were dissolved in 70 mL of methanol solution and stirred at room temperature for 3 hours. Sodium borohydride (1 g, 26.273 mmol) was then slowly added, and stirring continued at room temperature for 1 hour. After TLC confirmation of complete reaction, 50 mL of 1 M NaOH was added to quench the reaction. The mixture was then diluted with 150 mL of water, extracted with ethyl acetate to obtain the organic phase, dried, concentrated under reduced pressure, and purified by silica gel column chromatography to obtain compound 39 (1.8 g, 51.4%).

[0415]

[0416] baggage

[0417] 1H NMR (400 MHz, CDC13) 52.65 (t, J = 7.2 Hz, 2H), 2.62 - 2.57 (m, 2H), 2.34 - 2.29 (m, 2H), 2.22 (s, 6H), 1.89 (s, 1H), 1.71 - 1.63 (m, 2H), 1.55 - 1.42 (m, 2H), 1.34 - 1.25 (m, 8H), 0.91 -0.85 (m, 3H) O

[0418] 29. Synthesis of EAB-019

[0419] Intermediate 4 (250 mg, 0.322 mmol), compound 39 (100 mg, 0.483 mmol), and triethylamine (163 mg, 1.610 mmol) were sequentially added to 15 mL of chloroform solution. The mixture was stirred overnight at 70°C under nitrogen protection. After the reaction was confirmed to be complete by LC-MS, 20 mL of water was added to quench the reaction. The organic phase was obtained by dichloromethane extraction. After drying and concentrating the organic phase under reduced pressure, it was purified by silica gel column chromatography and preparative liquid chromatography to obtain a colorless oily compound EAB-019 (57.85 mg, 20.0%).

[0420]

[0421] 1H NMR (400 MHz, CDC13) 84.23 (s, 1H), 4.14 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.77 - 2.58 (m, 4H), 2.43 - 2.18 (m, 12H), 1.69 - 1.58 (m, 12H), 1.46 - 1.25 (m, 5 OH), 0.92 - 0.85 (m, 15H)o

[0422] Synthesis of EAB-020

[0423] 30. Synthesis of Compound 41

[0424] Compound 40 (2.0 g, 14.06 mmol) and compound 10 (1.506 g, 14.734 mmol) were dissolved in 70 mL of methanol solution and stirred at room temperature for 3 hours. Then, sodium borohydride (800 mg, 21.09 mmol) was slowly added, and stirring was continued at room temperature for 1 hour. After the reaction was confirmed to be complete by TLC, 50 mL of 1 M NaOH was added to quench the reaction. Then, water (150 mL) was added for dilution, and the organic phase was obtained with ethyl acetate. After drying and concentrating the organic phase under reduced pressure, the residue was purified by silica gel column chromatography (eluent: 0%–15% MeOH in DCM with 1% NH4OH) to obtain compound 41 (1.5 g, 46.8%).

[0425]

[0426] ¹H NMR (400 MHz, CDCl₃) 52.67 (t, J = 7.2 Hz, 2H), 2.63 - 2.57 (m, 2H), 2.33 (t, J = 7.2 Hz, 2H), 2.23 (s, 6H), 2.11 - 2.01 (m, 1H), 1.73 - 1.63 (m, 2H), 1.54–1.45 (m, 2H), 1.32–1.24 (m,

[0427] 12H), 0.91 - 0.85 (m, 3H) =

[0428] 31. Synthesis of EAB-020

[0429] Intermediate 4 (250 mg, 0.322 mmol), compound 41 (110 mg, 0.483 mmol), and triethylamine (163 mg, 1.610 mmol) were added sequentially to a chloroform (15 mL) solution. The mixture was stirred overnight at 70°C under nitrogen protection. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction. The mixture was extracted with dichloromethane (20 mL x 3), and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–12% MeOH in DCM) and preparative liquid chromatography to obtain a colorless oily compound EAB-020 (49.94 mg, 16.8%).

[0430]

[0431] 1H NMR (400 MHz, CDC13) 54.23 (s, 1H), 4.14 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H),

[0432] 2.76 - 2.61 (m, 4H), 2.46–2.08 (m, 12H), 1.68–1.60 (m, 12H), 1.45–1.25 (m, 54H), 0.93 - 0.84

[0433] (m, 15H)o

[0434] Synthesis of EAB-021

[0435] 32. Synthesis of Compound 43

[0436] Compound 42 (2.0 g, 11.745 mmol) and compound 10 (1.258 g, 12.332 mmol) were dissolved in methanol (70 mL) and stirred at room temperature for 3 hours. Sodium borohydride (670 mg, 17.618 mmol) was then slowly added, and stirring continued at room temperature for 1 hour. After the reaction was complete as monitored by TLC, 1 M NaOH (50 mL) was added to quench the reaction. The mixture was then diluted with water (150 mL) and extracted with ethyl acetate (100 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–15% MeOH in DCM with 1% NH4OH) to give compound 43 (1.7 g, 56.5%).

[0437]

[0438] ¹H NMR (400 MHz, CDCl₃) s 3.42 (s, ¹H), 2.71 (t, J = 7.0 Hz, ²H), 2.66–2.60 (m, ²H), 2.34

[0439] (t, J = 7.0 Hz, 2H), 2.22 (s, 6H), 1.76 - 1.66 (m, 2H), 1.57 - 1.46 (m, 2H), 1.30 - 1.23 (m, 16H), 0.89

[0440] - 0.84 (m, 3H).

[0441] 33. Synthesis of EAB-021

[0442] Intermediate 4 (250 mg, 0.322 mmol), compound 43 (124 mg, 0.483 mmol), and triethylamine (163 mg, 1.610 mmol) were added sequentially to a chloroform (15 mL) solution and stirred overnight at 70°C under nitrogen protection. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction, and the mixture was collected with dichloromethane (20 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%~15% MeOH in DCM) and preparative liquid chromatography to obtain a colorless oily compound EAB-021 (56.90 mg, 18.6%).

[0443]

[0444] ¹H NMR (400 MHz, CDCl₃) 54.23 (s, ¹H), 4.14 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.74–2.61 (m, 4H), 2.42–2.12 (m, ¹²H), 1.68–1.59 (m, ¹²H), 1.46–1.25 (m, ⁵⁸H), 0.92–0.85 (m, ¹⁵H)

[0445] Synthesis of EAB-022

[0446] 34. Synthesis of Compound 45

[0447] Compound 44 (2.0 g, 10.08 mmol) and compound 10 (1.080 g, 10.588 mmol) were dissolved in methanol (50 mL) and 1,2-dichloroethane (20 mL). The mixture was stirred at room temperature for 3 hours, and then sodium borohydride (575 mg, 15.12 mmol) was slowly added. The mixture was stirred at room temperature for another 1 hour. After the reaction was confirmed to be complete by TLC, 1 M NaOH (50 mL) was added to quench the reaction mixture. The mixture was then diluted with water (150 mL), extracted with ethyl acetate (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–15% MeOH in DCM with 1% NH4OH) to obtain compound 45 (1.4 g, 48.8%).

[0448]

[0449] LCMS: Rt: 0.856 min; MS m / z (ELSD): 285.3[M+H]+;

[0450] 1HNMR (400 MHz, CDC13) 53.04 (s, 1H), 2.71 (t, J = 7.2 Hz, 2H), 2.67- 2.59 (m, 2H), 2.35 (t, J = 7.2 Hz, 2H), 2.23 (s, 6H), 1.77- 1.66 (m, 2H), 1.57- 1.47 (m, 2H), 1.31 - 1.24 (m, 20H), 0.90 - 0.85 (m, 3H).

[0451] 35. Synthesis of EAB-022

[0452] Intermediate 4 (300 mg, 0.387 mmol), compound 45 (160 mg, 0.560 mmol), and triethylamine (196 mg, 1.935 mmol) were used.

[0453] The solution was added sequentially to a chloroform (15 mL) solution and stirred overnight at 70°C under nitrogen protection. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction. The mixture was then extracted with dichloromethane (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%~15% MeOH in DCM) and preparative liquid chromatography to obtain a colorless oily compound EAB-022 (40.47 mg, 10.7%).

[0454]

[0455] 1H NMR (400 MHz, CDC13) 54.22 (s, 1H), 4.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.71 (t, J =7.2 Hz, 2H), 2.66 - 2.60 (m, 2H), 2.46 - 2.22 (m, 10H), 1.81 - 1.51 (m, 16H), 1.47 - 1.41 (m, 2H), 1.39 - 1.33 (m, 8H), 1.31 - 1.24 (m, 50H), 0.91 -0.85 (m, 15H)o

[0456] Synthesis of intermediate 6

[0457] 36. Synthesis of Compound 47

[0458] Compound 46 (1 g, 7.575 mmol) was dissolved in dichloromethane (20 mL), and TBDPSC1 (2.5 g, 9.09 mmol) and imidazole (773 mg, 11.364 mmol) were added sequentially. The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was complete as monitored by TLC, water (50 mL) was added to quench the reaction. The mixture was then extracted with dichloromethane (50 mL x 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to obtain compound 47 (2.2 g, 78.5%).

[0459]

[0460] ¹H NMR (400 MHz, CDCl₃) 87.69–7.63 (m, 4H), 7.45–7.34 (m, 6H), 3.71–3.60 (m, 5H), 2.31 (t, J = 7.6 Hz, 2H), 1.77–1.67 (m, 2H), 1.63–1.56 (m, 2H), 1.04 (s, 9H).

[0461] 37. Synthesis of Compound 48

[0462] Compound 47 (2.2 g, 5.946 mmol) was added to a mixture of methanol (20 mL) and water (10 mL), followed by potassium carbonate (2.462 g, 17.838 mmol). The mixture was then heated to 70°C and stirred for 12 hours until the reaction solution became clear. After the starting material was completely consumed as monitored by TLC, the reaction solution was cooled to room temperature and concentrated under reduced pressure. The resulting residue was adjusted to pH 2-3 with 2 MHCl, diluted with 50 mL of brine, extracted with ethyl acetate (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain crude compound 48 (2.0 g, 94.5%), which was used directly in the next step.

[0463]

[0464] 38. Synthesis of Compound 49

[0465] Compound 48 (2.0 g, 5.618 mmol), EDCI (2.16 g, 11.236 mmol), DMAP (275 mg, 2.247 mmol), DIEA (3.62 g, 28.09 mmol), and compound 4 (1.57 g, 8.427 mmol) were sequentially added to a solution of dichloromethane (30 mL). The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was complete as monitored by TLC, water (50 mL) was added to quench the reaction. The mixture was then collected with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine (100 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to obtain compound 49 (900 mg, 30.6%).

[0466]

[0467] 1H NMR (400 MHz, CDC13) 57.70 - 7.63 (m, 4H), 7.46 - 7.33 (m, 6H), 3.97 (d, J = 5.8 Hz, 2H), 3.66 (t, J = 6.2 Hz, 2H), 2.31 (t, J = 7.6 Hz, 2H), 1.77 - 1.69 (m, 2H), 1.62 - 1.58 (m, 2H), 1.30 - 1.24 (m, 16H), 1.04 (s, 9H), 0.91 - 0.85 (m, 6H) =

[0468] 39. Synthesis of Compound 50

[0469] Compound 49 (13.5 g, 21.74 mmol) was added to a tetrahydrofuran (100 mL) solution, followed by the addition of TBAF (44 mL, 43.48 mmol, 1 M in THF). The mixture was stirred at room temperature for 16 hours. After the reaction was complete as monitored by TLC, ethyl acetate (100 mL) was added to dilute the mixture. The organic phase was washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–30% EA in PE) to obtain compound 50 (4.8 g, 65.14%).

[0470]

[0471] 1HNMR (400 MHz, CDC13) 53.98 (d, J = 5.8 Hz, 2H), 3.65 (t, J = 6.0 Hz, 2H), 2.35 (t, J = 7.2

[0472] Hz, 2H), 1.77- 1.67 (m, 2H), 1.64 - 1.59 (m, 4H), 1.27 (m, 16H), 0.91 -0.84 (m, 6H) O

[0473] 40. Synthesis of Compound 51

[0474] (At TC, compound 50 (4.8 g, 16.77 mmol), compound 7 (698 mg, 6.71 mmol), and DCC (4.15 g,

[0475] 20.13 mmol) was added sequentially to a 100 mL solution of dichloromethane, and then the mixture was slowly heated to room temperature and stirred overnight under nitrogen protection. After the reaction was complete as monitored by TLC, the mixture was filtered, the residue was washed with dichloromethane, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to obtain compound 51 (3.7 g, 86.13%).

[0476]

[0477] 1H NMR (400 MHz, CDC13) 54.16 (t, J = 5.4 Hz, 4H), 3.97 (d, J = 5.8 Hz, 4H), 3.37 (s, 2H), 2.34 (t, J = 6.3 Hz, 4H), 1.72 - 1.67 (m, 8H), 1.60 (m, 2H), 1.27 (m, 32H), 0.90 - 0.86 (m, 12H).

[0478] 41. Synthesis of Intermediate 6

[0479] Compound 51 (2.89 g, 4.51 mmol), NBS (803 mg, 4.51 mmol), and TosOH (1.17 g, 6.77 mmol) were added sequentially to a chloroform (30 mL) solution. The mixture was stirred at 70°C for 4 hours under nitrogen protection. After the reaction was complete as monitored by TLC, water (20 mL) was added to quench the reaction. The mixture was extracted with ethyl acetate (20 mL x 3), and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) and preparative liquid chromatography to obtain a yellow oily intermediate 6 (2 g, 49.48%).

[0480]

[0481] 1H NMR (400 MHz, CDC13) 54.83 (s, 1H), 4.24 (t, J = 6.0 Hz, 4H), 3.97 (d, J = 6.0 Hz, 4H), 2.35 (t, J = 6.8 Hz, 4H), 1.76 to 1.69 (m, 8H), 1.60 (m, 2H), 1.27 (m, 32H), 0.90 - 0.87 (m, 12H)=

[0482] Synthesis of intermediate 7

[0483] Intermediate 6 (2 g, 2.78 mmol), compound 10 (568 mg, 5.56 mmol), and triethylamine (1.41 g, 13.89 mmol) were added sequentially to a chloroform (30 mL) solution. The mixture was stirred overnight at 70°C under nitrogen protection. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction. The mixture was extracted with dichloromethane (20 mL x 3), and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% MeOH in DCM) and preparative liquid chromatography to obtain a yellow oily intermediate 7 (630 mg, 30.60%).

[0484]

[0485] Synthesis of EAB-023

[0486] Intermediate 7 (150 mg, 0.20 mmol) and triethylamine (61.44 mg, 0.61 mmol) were added to a solution of dichloromethane (10 mL). Compound 52 (37 mg, 0.31 mmol) was added at 0°C. The mixture was stirred overnight at room temperature under nitrogen protection. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction. The mixture was then collected in dichloromethane (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-023 (53.42 mg, 31.98%).

[0487]

[0488] 1H NMR (400 MHz, CDC13) 55.08 (s, 1H), 4.33 - 4.14 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.44 (t, J = 7.6 Hz, 2H), 2.73 - 1.98 (m, 14H), 1.89 - 1.58 (m, 14H), 1.38 - 1.22 (m, 34H), 0.95 -0.82 (m, 15H)o

[0489] Synthesis of EAB-024

[0490] Intermediate 7 (150 mg, 0.20 mmol) and triethylamine (61.44 mg, 0.61 mmol) were added to a dichloromethane (10 mL) solution. Compound 37 (6 mg, 0.41 mmol) was added under nitrogen protection. The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction. The mixture was extracted with dichloromethane (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-024 (50.45 mg, 29.21%).

[0491]

[0492] 1H NMR (400 MHz, CDC13) 55.08 (s, 1H), 4.35 - 4.10 (m, 4H), 3.96 (d, J = 6.0 Hz, 4H), 3.43 (t, J = 7.6 Hz, 2H), 2.56 -2.21 (m, 14H), 1.84- 1.75 (m, 2H), 1.70- 1.55 (m, 10H), 1.34 - 1.22 (m, 40H), 0.91 -0.86 (m, 15H).

[0493] Synthesis of EAB-025

[0494] Intermediate 7 (150 mg, 0.20 mmol) and triethylamine (61.44 mg, 0.61 mmol) were added to a 10 mL solution of dichloromethane. Compound 17 (54 mg, 0.31 mmol) was added under nitrogen protection at 0°C. The mixture was stirred overnight at room temperature. After the reaction was complete as monitored by LC-MS, water (20 mL) was added to quench the reaction. The mixture was extracted with dichloromethane (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-025 (62.25 mg).

[0495] mg, 34.90%)o

[0496]

[0497] 1H NMR (400 MHz, CDC13) 55.07 (s, 1H), 4.29 - 4.10 (m, 4H), 3.96 (d, J = 6.0 Hz, 4H), 3.49 - 3.35 (m, 2H), 2.54 - 2.13 (m, 14H), 1.76 - 1.58 (m, 12H), 1.40 - 1.14 (m, 44H), 0.95 - 0.82 (m, 15H)o

[0498] Synthesis of EAB-026

[0499] Intermediate 7 (150 mg, 0.20 mmol) and triethylamine (61.44 mg, 0.61 mmol) were added to a solution of dichloromethane (10 mL). Compound 18 (82 mg, 0.41 mmol) was added at 0°C, and the mixture was stirred overnight at room temperature. After the reaction was complete as monitored by LCMS, water (20 mL) was added to quench the reaction. The mixture was extracted with dichloromethane (20 mL x 3), and the organic phase was dried over anhydrous sodium sulfate. The mixture was filtered, concentrated under reduced pressure, and the residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-026 (49.86 mg, 27.09%).

[0500]

[0501] ¹H NMR (400 MHz, CDCl₃) 55.12 (s, ¹H), 4.31–4.14 (m, 4H), 3.96 (d, J = 5.8 Hz, 4H), 3.52–3.38 (m, 2H), 3.00–2.18 (m, ¹⁴H), 1.98–1.55 (m, ¹⁶H), 1.33–1.23 (m, ¹⁴H), 0.93–0.86 (m, ¹⁵H)

[0502] Synthesis of EAB-027

[0503] Intermediate 6 (300 mg, 0.417 mmol), triethylamine (0.29 mL, 2.086 mmol), and compound 53 (215.29 mg, 1.25 mmol) were sequentially added to a chloroform (10 mL) solution and stirred overnight at 65 °C. After the reaction was complete as monitored by LCMS, water (10 mL) was added to quench the reaction, and the mixture was extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-027 (17.62 mg, 5.2%).

[0504]

[0505] 1H NMR (400 MHz, CDC13) 54.24 (s, 1H), 4.19 - 4.14 (m, 4H), 3.97 (d, J = 5.6 Hz, 4H), 2.70

[0506] (t, J = 7.2 Hz, 2H), 2.67–2.62 (m, 2H), 2.36–2.31 (m, 4H), 2.31–2.25 (m, 2H), 2.21 (s, 6H), 1.72–1.66 (m, 8H), 1.48–1.41 (m, 2H), 1.34–1.26 (m, 40H), 0.91–0.86 (m, 15H).

[0507] Synthesis of EAB-028

[0508] Intermediate 6 (300 mg, 0.417 mmol), triethylamine (0.29 mL, 2.086 mmol), and compound 39 (250.35 mg, 1.25 mmol) were added sequentially to a chloroform (10 mL) solution and stirred overnight at 65 °C. After the reaction was complete as monitored by LCMS, water (10 mL) was added to quench the reaction. The mixture was extracted with dichloromethane (10 mL), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-028 (44.04 mg, 12.58%).

[0509]

[0510] 1H NMR (400 MHz, CDC13) 54.23 (s, 1H), 4.18 - 4.14 (m, 4H), 3.97 (d, J = 5.6 Hz, 4H), 2.73 (t, J = 6.7 Hz, 2H), 2.66 -2.61 (m, 2H),2.60 - 2.06 (m, 12H), 1.81 - 1.59 (m, 10H), 1.46 - 1.40 (m, 2H), 1.36 - 1.18 (m, 42H), 0.91 - 0.86 (m, 15H).

[0511] Synthesis of EAB-029

[0512] Intermediate 6 (300 mg, 0.417 mmol), triethylamine (0.29 mL, 2.086 mmol), and compound 41 (285.39 mg, 1.25 mmol) were added sequentially to a chloroform (10 mL) solution and stirred overnight at 65 °C. After the reaction was complete as monitored by LCMS, water (10 mL) was added to quench the reaction, and the mixture was collected with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAB-029 (17.62 mg, 15.17%).

[0513]

[0514] 1H NMR (400 MHz, CDC13) 54.23 (s, 1H), 4.18 - 4.13 (m, 4H), 3.97 (d, J = 5.8 Hz, 4H), 2.70 (t, J = 7.2 Hz, 2H), 2.67 - 2.62 (m, 2H), 2.38 - 2.28 (m, 6H), 2.23 (s, 6H), 1.72 - 1.67 (m, 8H), 1.66 - 1.63 (m, 2H), 1.47 - 1.39 (m, 2H), 1.31 - 1.24 (m, 46H), 0.91 - 0.86 (m, 15H).

[0515] Synthesis of EAB-031

[0516]

[0517] 42. Synthesis of Compound 55

[0518] Intermediate 4 (500 mg, 0.64 mmol) and compound 54 (112.4 mg, 0.96 mmol) were added sequentially to a chloroform (10 mL) solution, and triethylamine (0.27 mL, 1.93 mmol) was added dropwise at room temperature. The reaction mixture was stirred overnight at 65 °C. The reaction was monitored for completeness by TLC (DCM:MEOH = 10:1). The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (30 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOH in DCM) and preparative liquid chromatography to give a pale yellow oily compound 55 (340 mg, 64.9%). O

[0519] 43. Synthesis of EAB-031

[0520] Compound 55 (200 mg, 0.246 mmol), triethylamine (74.96 mg, 0.74 mmol), and compound 17 (86.9 mg, 0.37 mmol) were added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound EAB-031 (26.5 mg, 11.28%).

[0521] L H NMR (400 MHz, CDC13) 55.23 (s, 1H), 4.23 -4.11 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.43 - 3.33 (m, 2H), 2.58 -2.13 (m, 14H), 1.67 - 1.57 (m, 19H), 1.39 - 1.34 (m, 8H), 1.28 (s, 44H), 0.92 - 0.85 (m, 15H).

[0522]

[0523] Intermediate 4 (500 mg, 0.64 mmol) and compound 56 (112.4 mg, 0.96 mmol) were added sequentially to chloroform (10 mL).

[0524] Triethylamine (0.27 mL, 1.93 mmol) was added dropwise to the solution at room temperature, and the reaction mixture was stirred overnight at 65 °C. The reaction was monitored for completeness by TLC (DCM:MEOH = 10:1). The reaction mixture was quenched with water (20 mL) and collected with dichloromethane (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOH in DCM) and preparative liquid chromatography to give a pale yellow oily compound 57 (209 mg, 39.21%).

[0525] 45. Synthesis of EAB-032

[0526] Compound 57 (200 mg, 0.246 mmol), triethylamine (74.96 mg, 0.74 mmol), and compound 17 (86.9 mg, 0.37 mmol) were added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound EAB-032 (26.94 mg, 11.5%).

[0527] 1H NMR (400 MHz, CDC13) 55.22 (s, 1H), 4.21 - 4.12 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.38 - 3.32 (m, 2H), 2.85 - 2.50 (m, 8H), 2.40 - 2.33 (m, 2H), 2.32 - 2.27 (m, 4H), 1.68 - 1.59 (m, 14H), 1.38 - 1.33 (m, 10H), 1.32 - 1.23 (m, 44H), 0.92 - 0.86 (m, 15H).

[0528] Synthesis of EAB-033

[0529]

[0530] 46. ​​Synthesis of compound 3a

[0531] Compound 1a (2 g, 7.14 mmol) and dimethylamine (35 mL, 71.4 mmol, 2.0 M) were added sequentially to a methanol (20 mL) solution, and the reaction mixture was reacted at 45°C for 4 h. The reaction was monitored for completeness by TLC (DCM:MEOH = 10:1). The mixture was concentrated under reduced pressure, and water (20 mL) was added to the reaction mixture. Extraction was performed using ethyl acetate (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a crude, pale yellow oily compound 2a (2.3 g), which was used directly in the next reaction step.

[0532] Compound 2a (500 mg, 2.04 mmol) was added to a solution of dioxane hydrochloride (10 mL), and the reaction mixture was allowed to react overnight at room temperature. The reaction was monitored for completion by TLC (DCM:MEOH = 10:1). The mixture was concentrated under reduced pressure, and water (10 mL) was added to the reaction mixture.

[0533] Wash with ethyl acetate (10 mL x 3). Collect the aqueous phase and freeze-dry to give a white solid (300 mg), which is used directly in the next reaction.

[0534] 47. Synthesis of Compound 58

[0535] Intermediate 4 (500 mg, 0.64 mmol) and compound 3a (139.5 mg, 0.96 mmol) were added sequentially to a chloroform (10 mL) solution, followed by the dropwise addition of triethylamine (0.27 mL, 1.93 mmol) at room temperature. The reaction mixture was stirred overnight at 65°C. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHmDCM) and preparative liquid chromatography to give a pale yellow oily compound 58 (270 mg, 48.81%).

[0536] 48. EAB-033 Synthesis

[0537] Compound 58 (270 mg, 0.321 mmol), triethylamine (0.134 mL, 0.96 mmol), and compound 17 (113.28 mg, 0.64 mmol) were added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (20 mL), and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound EAB-033 (32.15 mg, 10.2%).

[0538] ¹H NMR (400 MHz, CDCl₃) 55.22 (s, 1H), 4.19–4.13 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.37–3.30 (m, 2H), 2.94–2.62 (m, 8H), 2.39–2.33 (m, 2H), 2.32–2.27 (m, 4H), 1.87–1.75 (m, 2H), 1.67–1.61 (m, 12H), 1.36–1.26 (m, 56H), 0.91–0.87 (m, 15H).

[0539] Synthesis of EAB-034

[0540]

[0541] 49. Synthesis of Compound 60

[0542] Intermediate 4 (500 mg, 0.64 mmol) and compound 59 (126 mg, 0.96 mmol) were added sequentially to a chloroform (10 mL) solution, followed by the dropwise addition of triethylamine (0.27 mL, 1.93 mmol) at room temperature. The reaction mixture was stirred overnight at 65 °C. The reaction was monitored for completeness by TLC (DCM:MEOH = 10:1). The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOH in DCM) and preparative liquid chromatography to give a pale yellow oily compound 60 (370 mg, 69.42%).

[0543] 50. Synthesis of EAB-034

[0544] Compound 60 (200 mg, 0.242 mmol), triethylamine (73.59 mg, 0.727 mmol), and compound 17 (85.33 mg, 0.484 mmol) were added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound EAB-034 (56.21 mg, 2402%).

[0545] 1H NMR (400 MHz, CDC13) 5 5.35 - 5.00 (m, 1H), 4.28 - 4.10 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.50 - 3.37 (m, 2H), 3.17 - 2.92 (m, 4H), 2.43 -2.20 (m, 6H), 2.10 - 1.95 (m, 2H), 1.82 - 1.47 (m, 14H), 1.43 - 1.16 (m, 56H), 0.97 - 0.80 (m, 15H).

[0546] Synthesis of intermediate 8

[0547]

[0548] 51. Synthesis of Compound 62

[0549] Compound 31 (22.6 g, 58.85 mmol), EDCI (2256 g, 117.7 mmol), DMAP (2.87 g, 23.54 mmol), DIEA (30.36 g, 235.41 mmol), and compound 61 (17.65 g, 88.28 mmol) were added sequentially to a solution of dichloromethane (50 mL). The reaction mixture was then stirred overnight at room temperature under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 30:1). The reaction was quenched by adding water (50 mL) to the reaction mixture, and then collected with ethyl acetate (50 mL x 3). The organic phase was washed with saturated brine (100 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was separated and purified by silica gel column chromatography (eluent: 0%–5% EA in PE) to obtain a colorless and transparent oily product 62 (30.6 g, 91.86%).

[0550] 52. Synthesis of Compound 63

[0551] Compound 62 (30.6 g, 54.06 mmol) was added to a tetrahydrofuran (30 mL) solution, followed by the addition of TBAF (108.127 mL, 108.127 mmol, 1 min THF). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored for completion by TLC (PE / EA = 7 / 1). The reaction mixture was diluted with ethyl acetate (150 mL), the organic phase was washed with saturated brine (150 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was subjected to silica gel column chromatography (eluent: 0%–30% EA in PE).

[0552] After separation and purification, a yellow oily product 63 (15.6 g, 87.97%) was obtained.

[0553] 53. Synthesis of Compound 64

[0554] At 0°C, compound 63 (10 g, 30.48 mmol), compound 7 (1.268 g, 12.19 mmol), and DCC (7.536 g, 36.58 mmol) were added sequentially to a solution of dichloromethane (100 mL). The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 64 (1.66 g, 7.5%).

[0555] 54. Synthesis of Compound 65

[0556] Compound 64 (1.66 g, 2.28 mmol) and p-toluenesulfonic acid (590.73 mg, 3.43 mmol) were added sequentially to an ethyl acetate (20 mL) solution, followed by the addition of NBS (407.6 mg, 2.28 mmol) under nitrogen protection. The reaction mixture was stirred at 70°C for 4 hours under nitrogen protection. The reaction was monitored for completeness by LC-MS. The reaction mixture was filtered, and the residue was washed with ethyl acetate. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 65 (1.56 g, 84.8%).

[0557] 55. Synthesis of Intermediate 8

[0558] Compound 65 (1.56 g, 1.94 mmol) and compound 10 (301 mg, 2.91 mmol) were dissolved in chloroform (10 mL), and triethylamine (896.19 mg, 5.82 mmol) was added dropwise to the reaction mixture, which was stirred overnight at 65°C. The reaction was monitored by LC-MS until complete. The mixture was then diluted with water (30 mL) and extracted with dichloromethane (30 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% MeOHin DCM) to give a pale yellow oily intermediate 8 (1.38 g, 86 1%).

[0559] LCMS: Rt: 1.614 min; MS m / z (ELSD): 827.5[M+H]+;

[0560] Synthesis of EAB-036

[0561]

[0562] Intermediate 8 (300 mg, 0.36 mmol), compound 66 (134 mg, 0.72 mmol), and triethylamine (181.8 mg, 1.8 mmol) were added sequentially to a chloroform (10 mL) solution. The reaction mixture was stirred overnight at 70 °C under nitrogen protection. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (15 mL) and extracted with dichloromethane (15 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% MeOHin DCM) and preparative liquid chromatography to give a yellow oily compound EAB-036 (102.36 mg, 29.59%).

[0563] 1HNMR (400 MHz, CDC13) 55.14 (s, 1H), 4.90 - 4.82 (m, 2H), 4.20 -4.11 (m, 4H), 3.42 (t, J = 7.6 Hz, 2H), 2.43 - 2.38 (m, 2H), 2.30 - 2.24 (m, 6H), 2.23 - 2.16 (m, 6H), 1.75 - 1.70 (m, 2H), 1.66 - 1.61 (m, 6H), 1.53 - 1.47 (m, 8H), 1.36 - 1.24 (m, 52H), 0.89- 0.85 (m, 15H).

[0564] Synthesis of EAB-037

[0565]

[0566] Intermediate 8 (300 mg, 0.36 mmol), triethylamine (55.09 mg, 0.54 mmol), and compound 17 (96.54 mg, 0.54 mmol) were added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (15 mL) and extracted with dichloromethane (15 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound EAB-037 (129.97 mg, 11.28%).

[0567] ¹H NMR (400 MHz, CDCl₃) 5.14 (s, ¹H), 4.93–4.79 (m, 2H), 4.21–4.09 (m, 4H), 3.45–3.37 (m, 2H), 2.44–2.36 (m, 2H), 2.33–2.18 (m, ¹²H), 1.77–1.70 (m, 2H), 1.65–1.61 (m, 6H), 1.54–1.47 (m, 8H), 1.37–1.21 (m, 54H), 0.90–0.85 (m, ¹⁵H).

[0568] Synthesis of EAB-039

[0569]

[0570] 56. Synthesis of Compound 68

[0571] Intermediate 4 (500 mg, 0.64 mmol) and compound 67 (124 mg, 0.96 mmol) were added sequentially to a chloroform (10 mL) solution, followed by the dropwise addition of triethylamine (0.27 mL, 1.93 mmol) at room temperature. The reaction mixture was stirred overnight at 65°C. The reaction was monitored for completeness by LC-MS. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound 68 (190 mg, 35.73%).

[0572] 57. Synthesis of EAB-039

[0573] Compound 68 (100 mg, 0.121 mmol), triethylamine (18.4 mg, 0.182 mmol), and compound 17 (32.26 mg, 0.121 mmol) were used.

[0574] 0.182 mmol) was added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature. The reaction was monitored for completeness by LCMS. The reaction solution was quenched with water (20 mL), and extracted with dichloromethane (10 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–8% MeOHin DCM) and preparative liquid chromatography to give a pale yellow oily compound EAB-039 (15.6 mg, 13.3%).

[0575] 1HNMR (4OO MHz, CDC13) 55.13 (s, 1H), 4.19 - 4.11 (m, 4H), 3.96 (d, J = 5.6 Hz, 4H), 3.44 (t, J = 7.2 Hz, 2H), 2.68 - 2.43 (m, 6H), 2.43 - 2.37 (m, 2H), 2.29 (t, J = 7.2 Hz, 4H), 1.69 — 1.58 (m, 14H), 1.37- 1.25 (m, 54H), 0.91 — 0.85 (m, 15H).

[0576] Synthesis of EAB-040

[0577] 58. Synthesis of Compound 71

[0578]

[0579] Compound 69 (5.15 g, 39.06 mmol), compound 70 (10 g, 39.06 mmol), and DCC (12 g, 58.59 mmol) were added sequentially to a 100 mL solution of dichloromethane at 0°C. The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 71 (5.5 g, 47.09%).

[0580] 59. Synthesis of Compound 72

[0581]

[0582] Compound 71 (5.5 g, 14.86 mmol), compound 7 (650 mg, 6.25 mmol), and DCC (5.1 g, 24.75 mmol) were added sequentially to a solution of dichloromethane (100 mL) at 0°C. The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 72 (4.7 g, 92.95%).

[0583] 60. Synthesis of Compound 73

[0584]

[0585] Compound 72 (4.7 g, 5.8 mmol) and p-toluenesulfonic acid (1.498 g, 8.71 mmol) were sequentially added to an ethyl acetate (50 mL) solution, followed by the addition of NBS (1.034 g, 5.8 mmol) under nitrogen protection. The reaction mixture was stirred at 70 °C for 4 hours under nitrogen protection. The reaction was monitored for completeness by LC-MS. The reaction mixture was filtered, and the residue was washed with ethyl acetate. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 73 (4.4 g, 85.48%).

[0586] 61. Synthesis of Compound 74

[0587]

[0588] Compound 73 (2 g, 2.25 mmol) was added to a solution of chloroform (20 mL), followed by compound 10 (344.98 mg, 3.38 mmol) and TEA (683 mg, 6.76 mmol). The reaction mixture was incubated overnight at 65°C. The reaction was monitored for completeness by LCMS. The reaction mixture was diluted with dichloromethane (50 mL), and the organic phase was washed with saturated brine (30 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–10% MEOH in DCM) to give a yellow oily product 74 (1.15 g, 56%).

[0589] 62. Synthesis of EAB-040

[0590]

[0591] Compound 74 (500 mg, 0.55 mmol), triethylamine (83.33 mg, 0.825 mmol) and compound 17 (146.03 mg, 0.825 mmol) were added sequentially to a solution of dichloromethane (10 mL), and the reaction mixture was stirred overnight at room temperature under nitrogen protection.

[0592] The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (20 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% MeOH in DCM) and preparative liquid chromatography to give a yellow oily compound EAB-040 (79.64 mg, 13.8%).

[0593] LCMS: Rt: 1.165 min; MS m / z (ELSD): 1049.9[M+H]+;

[0594] CAD: 98.06% purity. Rt:14.947min

[0595] ¹H NMR (400 MHz, CDCl₃) 55.15 (s, ¹H), 4.20–4.12 (m, 4H), 4.05 (t, J = 6.8 Hz, 4H), 3.46–3.38 (m, 2H), 2.43–2.38 (m, 2H), 2.33–2.27 (m, 4H), 2.24–2.18 (m, 6H), 1.78–1.70 (m, 2H), 1.67–1.55 (m, 16H), 1.48–1.35 (m, 10H), 1.29–1.22 (m, 54H), 0.89–0.85 (m, 15H).

[0596] Synthesis of EAB-041

[0597]

[0598] Compound 75 (3.7 g, 39.06 mmol), compound 70 (10 g, 39.06 mmol), and DCC (12 g, 58.59 mmol) were added sequentially to a solution of dichloromethane (100 mL) at 0°C. The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 76 (6.7 g, 49.68%).

[0599] 64. Synthesis of Compound 77

[0600] At 0°C, compound 76 (6.7 g, 20.42 mmol), compound 7 (849.75 mg, 8.17 mmol), and DCC (5.04 g, 24.51 mmol) were added sequentially to a solution of dichloromethane (100 mL). The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 77 (5.3 g, 89.47%).

[0601] 65. Synthesis of Compound 78

[0602] Compound 77 (5.3 g, 7.31 mmol) and p-toluenesulfonic acid (1.886 g, 10.96 mmol) were added sequentially to an ethyl acetate (50 mL) solution, followed by the addition of NBS (1.3 g, 7.31 mmol) under nitrogen protection. The reaction mixture was stirred at 70°C for 4 hours under nitrogen protection. The reaction was monitored for completeness by LC-MS. The reaction mixture was filtered, and the residue was washed with ethyl acetate. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 78 (4.5 g, 76.75%).

[0603] 66. Synthesis of Compound 79

[0604] Compound 78 (2 g, 2.49 mmol) was added to a chloroform (20 mL) solution, followed by compound 10 (381.5 mg, 3.74 mmol).

[0605] 7.48 mmol) and TEA (755.6 mg, 7.48 mmol). The reaction mixture was incubated overnight at 65°C. The reaction was monitored for completeness by LCM S. The reaction solution was diluted with dichloromethane (50 mL), the organic phase was washed with saturated brine (10 mL x 3), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% MEOH in DCM) to give a yellow oily product compound 79 (926 mg, 45%).

[0606] 67. Synthesis of EAB-041

[0607] Compound 79 (300 mg, 0.36 mmol), triethylamine (55.09 mg, 0.54 mmol), and compound 917 (96.54 mg, 0.54 mmol) were sequentially added to a solution of dichloromethane (10 mL). The reaction mixture was stirred overnight at room temperature under nitrogen protection. The reaction was monitored for completeness by LCMS. The reaction mixture was quenched with water (20 mL) and extracted with dichloromethane (20 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% MeOH in DCM) and preparative liquid chromatography to give a yellow oily compound EAB-041 (166.67 mg, 47.49%).

[0608] 1H NMR (400 MHz, CDC13) 55.22 - 5.06 (m, 1H), 4.30 -4.16 (m, 4H), 4.11 - 4.05 (m, 4H), 3.53 - 3.41 (m, 2H), 3.07 - 2.72 (m, 4H), 2.68 - 2.54 (m, 4H), 2.38 - 2.21 (m, 4H), 2.09 - 1.96 (m, 2H), 1.78 - 1.67 (m, 10H), 1.60 - 1.54 (m, 4H), 1.46 - 1.39 (m, 4H), 1.32 - 1.21 (m, 5 OH), 0.91 - 0.83 (m, 15H).

[0609] EAB-043

[0610]

[0611] 68. Synthesis of Compound 81

[0612] Compound 80 (10 g, 68.49 mmol) was dissolved in DMF (100 mL), followed by the addition of TBDPSC1 (22.6 g, 82.19 mmol) and imidazole (10.6 g, 171.23 mmol). The reaction mixture was then stirred overnight at room temperature under nitrogen protection. The reaction was monitored for completion by TLC (PE:EA = 10:1). The reaction was quenched by adding water (100 mL) and then extracted with ethyl acetate (100 mL x 3). The organic phases were combined, washed with saturated brine (200 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–5% EA in PE) to obtain a colorless and transparent [residue / solution].

[0613] Oily product 81 (23.8 g, 90.48%).

[0614] 69. Synthesis of Compound 82

[0615] Compound 81 (23.8 g, 61.97 mmol) was added to a mixed solvent of methanol (200 mL) and water (100 mL), followed by potassium carbonate (25.66 g, 185.93 mmol). The reaction mixture was heated to 70 °C and stirred for 2 hours until the reaction solution became clear. The complete consumption of the starting material was monitored by TLC (PE:EA = 5:1). The reaction solution was cooled to room temperature and then concentrated directly under reduced pressure. The resulting residue was adjusted to pH 2–3 with 2 MHCl, diluted with brine (100 mL), and extracted with ethyl acetate (100 mL x 3). The combined organic phases were dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure to give a pale yellow oily product 82 (24 g, 100%), which was used directly in the next step.

[0616] 70. Synthesis of Compound 84

[0617] Compound 82 (5 g, 13.51 mmol), EDCI (5.18 g, 27.02 mmol), DMAP (329.7 mg, 2.7 mmol), DIEA (4.36 g, 33.78 mmol), and compound 83 (2.92 g, 20.27 mmol) were added sequentially to a solution of dichloromethane (50 mL). The reaction mixture was then stirred overnight at room temperature under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 30:1). The reaction was quenched by adding water (50 mL) and then extracted with ethyl acetate (50 mL x 3). Combine the organic phases, wash with saturated brine (100 mL x 3), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate and purify the residue by silica gel column chromatography (eluent: 0%–5% EA in PE) to obtain a colorless and transparent oily product 84 (5.2 g, 77.58%).

[0618] 71. Synthesis of Compound 85

[0619] Compound 84 (5.2 g, 10.48 mmol) was added to a tetrahydrofuran (50 mL) solution, followed by the addition of TBAF (20.97 mL, 20.97 mmol, 1 min THF). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored for completeness by TLC (PE / EA = 7 / 1). The reaction mixture was diluted with ethyl acetate (50 mL), washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–30% EA in PE) to give a yellow oily product 85 (2.3 g, 85.03%).

[0620] 72. Synthesis of Compound 87

[0621] At 0°C, compounds 85 (2.2 g, 8.52 mmol), 86 (909.56 mg, 5.68 mmol), and DCC (3.513 g, 17.05 mmol) were added sequentially to a 100 mL solution of dichloromethane. The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 87 (1.24 g, 54.56%).

[0622] 73. Synthesis of Compound 88

[0623] Compound 87 (1.24 g, 3.1 mmol) was added to a solution of dioxane hydrochloride (4 M, 15 mL). The reaction mixture was then stirred overnight at room temperature under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was directly concentrated under reduced pressure.

[0624] The crude product was condensed to obtain 88 (970 mg, 90.96%) of a colorless oily product, which was used directly in the next step.

[0625] 74. Synthesis of Compound 94

[0626] Compound 82 (5 g, 13.51 mmol), EDCI (5.16 g, 27.02 mmol), DMAP (329.7 mg, 2.7 mmol), DIEA (5.87 g, 33.78 mmol), and compound 93 (5.18 g, 20.27 mmol) were added sequentially to a solution of dichloromethane (50 mL). The reaction mixture was then stirred overnight at room temperature under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 30:1). The reaction was quenched by adding water (50 mL) and then extracted with ethyl acetate (50 mL x 3). Combine the organic phases, wash with saturated brine (100 mL x 3), dry with anhydrous sodium sulfate, filter, concentrate the filtrate under reduced pressure, and separate and purify the residue by silica gel column chromatography (eluent: 0%~5% EA in PE) to obtain a colorless and transparent oily product 94 (5.8 g, 70.47%).

[0627] 75. Synthesis of Compound 89

[0628] Compound 94 (5.8 g, 9.52 mmol) was added to a tetrahydrofuran (50 mL) solution, followed by the addition of TBAF (19.04 mL, 19.04 mmol, 1 min THF). The reaction mixture was stirred at room temperature for 2 hours. The reaction was monitored for completeness by TLC (PE / EA = 7 / 1). The reaction mixture was diluted with ethyl acetate (50 mL), washed with saturated brine (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–30% EA in PE) to give a yellow oily product 89 (2.3 g, 85.03%).

[0629] 76. Synthesis of Compound 90

[0630] Compound 88 (418.38 g, 1.21 mmol), compound 89 (450 mg, 1.21 mmol), and DCC (751.62 mg, 1.82 mmol) were added sequentially to a solution of dichloromethane (10 mL) at 0°C. The reaction mixture was then slowly heated to room temperature and stirred overnight under nitrogen protection. The reaction was monitored for completeness by TLC (PE:EA = 10:1). The reaction mixture was filtered, and the residue was washed with dichloromethane. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 90 (700 mg, 82.69%).

[0631] 77. Synthesis of Compound 91

[0632] Compound 90 (700 mg, 1.004 mmol) and p-toluenesulfonic acid (259.11 mg, 1.5 mmol) were added sequentially to an ethyl acetate (20 mL) solution, followed by the addition of NBS (178.76 mg, 1.004 mmol) under nitrogen protection. The reaction mixture was stirred at 70 °C for 4 hours under nitrogen protection. The reaction was monitored for completeness by LC-MS. The reaction mixture was filtered, and the residue was washed with ethyl acetate. The filtrate was then concentrated under reduced pressure, and the residue was purified by silica gel column chromatography (eluent: 0%–10% EA in PE) to give a colorless oily product 91 (496 mg, 63.72%).

[0633] 78. Synthesis of Compound 92

[0634] Compound 91 (496 mg, 0.64 mmol) and compound 10 (97.92 mg, 0.96 mmol) were dissolved in chloroform (10 mL), and then triethylamine (193.92 mg, 1.92 mmol) was added dropwise to the reaction mixture, which was stirred overnight at 65°C. The reaction was monitored by LCMS until complete. The reaction was then quenched with water (10 mL) and collected with dichloromethane (10 mL x 3). The organic phases were combined and purified with anhydrous sulfur.

[0635] The sodium sulfate was dried, filtered, and the filtrate was concentrated under reduced pressure. The residue was separated and purified by silica gel column chromatography (eluent: 0%–10% MeOH in DCM) to give a pale yellow oily product 92 (150 mg, 29.4%).

[0636] 79. Synthesis of EAB-043

[0637] Compound 92 (150 mg, 0.188 mmol), compound 17 (49.69 mg, 0.282 mmol), and triethylamine (38.02 mg, 0.564 mmol) were added sequentially to a solution of dichloromethane (10 mL). The reaction mixture was stirred overnight at room temperature under nitrogen protection. The reaction was monitored by LCMS until complete. The reaction mixture was quenched with water (10 mL) and extracted with dichloromethane (10 mL x 3). The organic phases were combined, dried over anhydrous sodium sulfate, filtered, and the filtrate was concentrated under reduced pressure. The residue was purified by silica gel column chromatography (eluent: 0%–10% MeOH in DCM) and preparative liquid chromatography to give a yellow oily compound EAB-043 (33.07 mg, 18.75%).

[0638] Synthesis of EAC-001

[0639] Intermediate 3 (400 mg, 0.46 mmol), triethylamine (235 mg, 2.33 mmol), and compound 95 (148 mg, 0.93 mmol) were sequentially added to a 10 mL solution of chloroform. The mixture was stirred overnight at 70 °C under nitrogen protection. After the reaction was complete as monitored by LCMS, the reaction mixture was directly concentrated under reduced pressure. The resulting residue was purified by preparative liquid chromatography to obtain a colorless oily compound, EAC-001 (31.24 mg, 7.16%).

[0640]

[0641] 1H NMR (400 MHz, CDC13) 54.46 (s, 1H), 4.15 (t, J = 6.8 Hz, 4H), 4.06 (t, J = 6.8 Hz, 4H), 2.88 (t, J = 7.2 Hz, 4H), 2.51 -2.39 (m, 4H), 2.34 - 2.22 (m, 14H), 1.72 - 1.49 (m, 12H), 1.48- 1.33 (m, 12H), 1.32 — 1.20 (m, 40H), 0.87 (t, J = 6.8 Hz, 12H).

[0642] Synthesis of EAC-004

[0643] Intermediate 3 (300 mg, 0.35 mmol), triethylamine (176 mg, 1.74 mmol), and compound 96 (130 mg, 0.69 mmol) were added sequentially to a chloroform (10 mL) solution. The mixture was stirred overnight at 70°C under nitrogen protection. The reaction was monitored by LC-MS until complete. After the reaction was complete, the reaction mixture was concentrated under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAC-004 (44.83 mg, 13.30%).

[0644]

[0645] 1H NMR (400 MHz, CDC13) 54.24 (s, 1H), 4.14 (t, J = 6.8 Hz, 4H), 4.06 (t, J = 6.8 Hz, 4H), 2.71 (t, J = 7.2 Hz, 4H), 2.34 - 2.27 (m, 6H), 2.26 - 2.19 (m, 12H), 1.68 - 1.53 (m, 16H), 1.44- 1.35 (m, 12H), 1.30 - 1.21 (s, 40H), 0.87 (t, J = 6.6 Hz, 12H).

[0646] Synthesis of EAC-007

[0647] 1. Synthesis of Compound 99

[0648] Compound 97 (300 mg, 1.58 mmol) was dissolved in FA (5 mL) solution and stirred at room temperature for 3 hours. The mixture was then concentrated under reduced pressure. The resulting residue and compound 98 (184 g, 1.58 mmol) were dissolved in methanol (10 mL) solution and stirred at room temperature for 3 hours. Sodium borohydride (294 mg, 4.74 mmol) was then slowly added, and stirring continued at room temperature for 1 hour. After the reaction was complete as monitored by LCMS, 1 M NaOH (10 mL) was added to quench the reaction. The mixture was then diluted with water (50 mL) and extracted with ethyl acetate (50 mL). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 99 (250 mg, 73.5%). The crude product was used directly in the next step.

[0649]

[0650] 2. Synthesis of EAC-007

[0651] Intermediate 3 (500 mg, 0.582 mmol), triethylamine (0.4 mL, 2.91 mmol), and compound 99 (250 mg, 1.163 mmol) were sequentially added to a chloroform (10 mL) solution. The mixture was stirred overnight at 7 CTC under nitrogen protection, and the reaction was monitored for completeness by LC-MS. After the reaction was complete, the reaction mixture was directly concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to give a colorless oily compound EAC-007 (7.65 mg, 1.32%).

[0652]

[0653] ¹H NMR (400 MHz, CDCl₃) 54.19 (s, ¹H), 4.17–4.11 (m, 4H), 4.06 (t, J = 6.8 Hz, 4H), 3.05–2.35 (m, 20H), 2.36–2.27 (m, 2H), 1.62–1.60 (m, 24H), 1.39–1.36 (m, 8H), 1.27–1.24 (m, 40H), 0.89–0.84 (m, 12H).

[0654] Synthesis of EAC-010

[0655] 3. Synthesis of Compound 101

[0656] Compound 97 (200 mg, 1.06 mmol) was dissolved in FA (5 mL) solution and stirred at room temperature for 3 hours, then concentrated under reduced pressure. The resulting residue and compound 100 (93 mg, 1.06 mmol) were dissolved in methanol (10 mL) solution and stirred at room temperature for 3 hours. Sodium borohydride (203 mg, 3.18 mmol) was then slowly added, and stirring was continued at room temperature for 1 hour. The reaction was monitored for completeness by LCMS. The reaction was quenched by adding 1 M NaOH (10 mL), then diluted with water (50 mL), extracted with ethyl acetate (50 mL x 3), dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to obtain compound 101 (200 mg, 100%). The crude product was used directly in the next step.

[0657]

[0658] 4. Synthesis of EAC-010

[0659] Intermediate 3 (460 mg, 0.534 mmol), triethylamine (0.37 mL, 2.67 mmol), and compound 101 (150 mg, 0.802 mmol) were sequentially added to a 10 mL solution of chloroform. The mixture was stirred overnight at 70°C under nitrogen protection, and the reaction was monitored for completeness by LC-MS. After the reaction was complete, the reaction mixture was directly concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to give a colorless oily compound EAC-010 (44.92 mg, 8.7%).

[0660]

[0661] 1H NMR (400 MHz, CDC13) 84.33 (s, 1H), 4.15 (t, J = 6.8 Hz, 4H), 4.06 (t, J = 6.8 Hz, 4H), 2.90 -2.82 (m, 2H), 2.72 (t, J = 7.2 Hz, 2H), 2.62 -2.17 (m, 18H), 1.70 - 1.51 (m, 20H), 1.40 - 1.36 (m, 8H), 1.28 - 1.22 (m, 40H), 0.89 - 0.85 (m, 12H).

[0662] Synthesis of EAGOU

[0663] 5. Synthesis of Compound 104

[0664] Compound 102 (500 mg, 3.85 mmol), potassium carbonate (2.65 g, 19.23 mmol), and compound 103 (1.41 g, 5.77 mmol) were added sequentially to an acetylene (20 mL) solution. The mixture was stirred overnight at 70°C under nitrogen protection, and the reaction was monitored for completeness by LC-MS. After the reaction was complete, the reaction mixture was filtered, and the filtrate was directly concentrated under reduced pressure to give compound 104 (720 mg, 87%). The crude product was used directly in the next step.

[0665]

[0666] 6. Synthesis of EAC-011

[0667] Intermediate 3 (500 mg, 0.580 mmol), triethylamine (0.3 mL, 1.74 mmol), and compound 104 (250 mg, 1.16 mmol) were sequentially added to a 15 mL solution of chloroform. The mixture was stirred overnight at 7 CTC under nitrogen protection, and the reaction was monitored by LC-MS until complete. After the reaction was complete, the reaction mixture was directly concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to give a colorless oily compound EAC-011 (8.50 mg, 1.47%).

[0668]

[0669] ¹H NMR (400 MHz, CDCl₃) 84.21 (s, ¹H), 4.14 (dd, J = 6.8, 4.8 Hz, 4H), 4.06 (t, J = 6.8 Hz, 4H), 2.76–2.70 (m, 2H), 2.67 (d, J = 7.2 Hz, 2H), 2.65–2.20 (m, 18H), 1.63–1.53 (m, 24H), 1.40–1.36 (m, 8H), 1.27–1.23 (m, 40H), 0.87 (t, J = 6.4 Hz, 12H).

[0670] Synthesis of EAC-012

[0671] 7. Synthesis of Compound 36

[0672] Compound 97 (400 mg, 2.11 mmol) was dissolved in FA (5 mL) solution and stirred at room temperature for 3 hours, then concentrated under reduced pressure. The resulting residue and compound 35 (305 mg, 2.11 mmol) were dissolved in methanol (10 mL) solution and stirred at room temperature for 3 hours. Sodium borohydride (398 mg, 6.33 mmol) was then slowly added, and stirring was continued at room temperature for 1 hour. The reaction was monitored by LCMS to ensure complete reaction. The reaction was quenched by adding 1 M NaOH (10 mL), then diluted with water (50 mL), and collected with ethyl acetate (50 mL x 3). The organic phase was dried over anhydrous sodium sulfate, filtered, and concentrated under reduced pressure to give a pale yellow oily product 36 (350 mg, 68.3%). The crude product was used directly in the next step.

[0673] L

[0674]

[0675] 8. Synthesis of EAC-012

[0676] Intermediate 3 (500 mg, 0.580 mmol), triethylamine (0.3 mL, 1.74 mmol), and compound 36 (300 mg, 1.16 mmol) were sequentially added to a chloroform (10 mL) solution and stirred overnight at 70°C under nitrogen protection. The reaction was monitored by LC-MS until complete. After the reaction was complete, the reaction mixture was directly concentrated under reduced pressure, and the resulting residue was purified by preparative liquid chromatography to give a colorless oily compound EAC-012 (12.27 mg, 2%).

[0677]

[0678] ¹H NMR (400 MHz, CDCl₃) 54.19 (s, ¹H), 4.17–4.10 (m, 4H), 4.06 (t, J = 6.8 Hz, 4H), 2.83–2.26 (m, 22H), 1.66–1.55 (m, 12H), 1.47–1.32 (m, 20H), 1.31–1.20 (m, 44H), 0.90–0.85 (m, 12H)

[0679] Synthesis of EAC-013

[0680] Intermediate 4 (300 mg, 0.387 mmol), triethylamine (0.27 mL, 1.937 mmol), and compound 96 (217.4 mg, 1.162 mmol) were added sequentially to a chloroform (10 mL) solution and stirred overnight at 65 °C. The reaction was monitored by LC-MS until complete. The reaction mixture was quenched with water (10 mL), extracted with dichloromethane (10 mL), dried over anhydrous sodium sulfate, filtered, concentrated under reduced pressure, and the residue was purified by preparative liquid chromatography to obtain a colorless oily compound EAC-013 (23.97 mg, 7.01%).

[0681]

[0682] 1H NMR (400 MHz, CDC13) 54.24 (s, 1H), 4.13 (t, J = 6.8 Hz, 4H), 3.97 (d, J = 6.9 Hz, 4H),

[0683] 2.71 (t, J = 7.2 Hz, 4H), 2.36-2.27 (m, 8H), 2.27 - 2.17 (m, 12H), 1.86 - 1.75 (m, 4H), 1.67 - 1.60 (m, 12H), 1.37- 1.34 (m, 4H), 1.31 - 1.22 (m, 34H), 0.92 - 0.85 (m, 12H).

[0684] Comparison of the synthesis of lipids Comuound 200 and 201

[0685]

[0686] Example 2 Preparation and characterization of lipid nanoparticles

[0687] The cationic lipid compound of Example 1 or dilinoleylmethyl-4-dimethylaminobutyrate (DLin-MC3-DMA, usually abbreviated as MC3) was dissolved in ethanol with DSPC, cholesterol and DMG-PEG2000 in a molar ratio of 50:10:38.5:1.5 to prepare ethanol lipid solutions.

[0688] Epigenetic editing tool mRNA was diluted in 10 mM citrate buffer (pH = 4) to obtain an aqueous solution of mRNA. Liposomes were prepared by mixing the ethanol lipid solution and the aqueous mRNA solution at a volume ratio of 1:3 using a microfluidic nanoparticle synthesizer (Macro & Nano) with an N / P ratio of 6. The ethanol was removed by overnight dialyzing with 20 mM Tris as the dialysate for 12–24 h to obtain the LNP formulation encapsulating the epigenetic editing tool mRNA.

[0689] The size, polydispersity index (PDI), and encapsulation efficiency of each LNP formulation were determined using the Malvern Zetasizer Nano ZS (Malvern UK) and the Quant-it Ribogreen RNA Quantification Kit (ThermoFisher Scientific, UK). The test results are shown in Table 1.

[0690] Table 1 shows the particle size, PDI, and encapsulation efficiency of each LNP formulation.

[0691]

[0692]

[0693]

[0694] The results showed that the LNP formulations prepared from the lipid compounds in Example 1 had an encapsulation rate of more than 88% for mRNA, and the encapsulation rates of compounds EAC-007, EAC-010, and EAC-011 could reach 100%. This demonstrates that the compounds provided in this application have a high encapsulation rate for nucleic acid drugs, which can successfully transport nucleic acid molecules into cells for expression. Using these as carriers can improve the delivery efficiency of nucleic acid drugs in vivo, thereby exerting corresponding biological activities.

[0695] Example 3 Evaluation of the effect of PCSK9 gene editing in mice

[0696] After PCSK9 is generated in liver cells, it leads to the degradation of low-density lipoprotein receptor (LDL-R) on the surface of liver cells, thereby hindering the liver from effectively clearing cholesterol from the blood and ultimately causing elevated blood lipid levels. By reducing the expression of PCSK9, the binding of PCSK9 to LDL-R can be reduced, thereby preventing the degradation of LDL-R on the surface of liver cells. This results in more LDL-R on the liver surface, thus more effectively clearing LDL-Co from the blood. This application provides a method for delivering the epigenetic editing tool EPIREG-mRNA (nucleic acid sequence shown in the table below) to the liver. Lipid nanoparticles prepared in Example 2 were injected into 8-10 week old male C57BL / 6J mice via tail vein at a dose of 0.5 mpk or 0.125 mpk, and blood was collected from the mice 3 days after administration. The expression level of PCSK9 protein in blood was detected using the Mouse Proprotein Convertase 9 / PCSK9 Quantikine ELISA Kit (R&D systems) and compared with the PBS group to obtain the relative expression level of PCSK9 in different drug administration groups. The experimental results are shown in Figures 1 and 2.

[0697] The results showed that, compared with the control group MC3-formed LNP and PBS group, the LNP prepared by the compound provided in this application had a better silencing effect on the PCSK9 gene, reducing the expression level of the PCSK9 gene by 50%. Specifically, EAB-004 showed a better silencing effect on the PCSK9 gene.

[0698] At a dose of 0.5 mpk, the inhibition rate of the PCSK9 gene reached 95%. The evaluation data of this embodiment can prove that the LNP prepared by the compound provided in this application can successfully deliver nucleic acid molecules into cells and express them, and can be used as a nucleic acid delivery system in the field of liver-targeted gene therapy.

[0699] The nucleic acid sequence of this embodiment

[0700]

[0701]

[0702]

[0703]

[0704]

[0705] Example 4: Comparative Evaluation of PCSK9 Gene Editing Effects in Mice

[0706] The PCSK9-silencing gene-editing tool EPIREG mRNA sequence (SEQ ID NO: 1) was encapsulated into LNPs prepared from cationic lipid molecules (Compound 200 and 201) with aminopropylene glycol-linked head and tail groups, as described in Example 2. The LNPs were administered via tail vein injection to 8-10 week old male C57BL / 6J mice at a dose of 0.125 mpk, serving as a control group. Similarly, the experimental group was administered via tail vein injection to 8-10 week old male C57BL / 6J mice at a dose of 0.125 mpk, with partial injection of the lipid nanoparticles prepared in Example 2. Blood samples were collected from the mice 3 days after administration. The expression level of PCSK9 protein in the blood was detected using the Mouse Proprotein Convertase 9 / PCSK9 Quantikine ELISA Kit (R&D systems).

[0707] As shown in Figure 3, the LNP prepared by the cationic lipid molecule EAB-016 provided in this application has a better silencing effect on the PCSK9 gene. When administered at a dose of 0.125 mpk, the inhibition effect on the PCSK9 gene can reach 90%, while the inhibition effect of the control group is only 20%. The data of this embodiment can prove that the efficiency of the cationic lipid in delivering nucleic acid molecules is better than that of the cationic lipid with aminopropylene glycol connecting the head and tail groups.

[0708] The foregoing detailed description of this application is intended to enable those skilled in the art to understand and implement its contents, but it should not be construed as limiting the scope of protection of this application. All equivalent modifications made in accordance with the spirit and essence of this application are subject to legal interpretation.

[0709] Any changes or modifications should be covered within the scope of protection of this application.

Claims

Claims 1. Compounds of formula (I), Or its stereoisomers, tautomers, or pharmaceutically acceptable salts, wherein: The A3 is optionally substituted, and the Gi and G2 are each independently selected from the optional substituted main chain consisting of 5 to 25 atoms.

2. The compound according to claim 1, or its stereoisomers, tautomers, or pharmaceutically acceptable salts, characterized in that, The compound comprises the following structure (II). Among them, the Li and L2 are each independently selected from the heteroatoms that are substituted by choice; A3 is an optional substituent; R4 and * are each independently selected from H or optional substituents, or R4 and R5 are linked together to form a ring; G3 and G4 are each independently selected from optional substituents.

3. The compound according to any one of claims 1-2, or its stereoisomers, tautomers, or pharmaceutically acceptable salts, characterized in that, The compound contains the following structure (III). (III), wherein, the Li and L2 are each independently selected from the heteroatoms that are substituted by choice; L5 is a chemical bond or an optional substituent; g is 0 or 1; Yl and Y2 are each independently a heteroatom or carbon atom with optional substitution; E is -0-, -S(O)b-, or -SS-, where b is 0, 1, or 2; f is 1; R4 and R5 are each independently selected from H or optional substituents, or R4 and R5 are linked together to form a ring.

4. The compound according to any one of claims 2 or 3, wherein the Li and L2 are each independently -O-, -S-, -N-, or -NR-. a - where Ra is H or Ci-Cu alkyl.

5. The compound according to any one of claims 3-4, wherein L5 is a chemical bond, -CH2-, -(C=O)-, -(C=O)S-, -(C=O)O- or -(C=O)NRa-, wherein Ra is H or a C1-C12 alkyl group.

6. The compound according to any one of claims 3-5, wherein Yi and Y2 are each independently C, O, S, N, CR, or NR. 3 , where R? is H or C1-C12 alkyl.

7. The compound according to any one of claims 1-6, or its stereoisomers, tautomers, or pharmaceutically acceptable salts, characterized in that, The compound contains the following structure (Illa). Among them, the L5 is -CH2- or -(C=0)-; Yi and Y2 are each independently -0-, -C- or -CR?-, where Ra is H or C1-C12 alkyl; Ai, A2, and A3 are each independently selected from any substituted substituents; R3 is an optional substituted alkyl group; Ri and R2 are each independently selected from optional substituents; R4 and * are each independently selected from H or optional substituents, or R4 and R5 are linked together to form a ring.

8. The compound according to any one of claims 1-7, or its stereoisomers, tautomers, or pharmaceutically acceptable salts, characterized in that, The compound contains the following structure (Illb). Among them, the L5 is -CH2- or -(C=0)-; Y1 and Y2 are each independently -0-, -C- or -CRa-, where Ra is H or C1-C12 alkyl; Ai, A2, and A3 are each independently selected from any substituted substituents; Ri and R2 are each independently selected from optional substituents; R4 and * are each independently selected from H or optional substituents, or R4 and R5 are linked together to form a ring.

9. The compound according to any one of claims 1-8, wherein Ai, A2 and A3 are each independently an optionally substituted alkyl, alicyclic, and / or heterocyclic group.

10. The compound according to any one of claims 1-9, wherein each of Ai, A2 and A3 is independently an optionally substituted alkyl, alkenyl, and / or alkyl group.

11. The compound according to any one of claims 1-10, wherein each of Ai, A2 and A3 is independently an optionally substituted cycloalkyl, cycloalkenyl, and / or cyclocyclobenzyl group.

12. The compound according to any one of claims 1-11, wherein Ai, A2 and A3 are each independently independently a optionally substituted C1-C24 alkylene, C1-C24 alkenylene, C3-C24 cycloalkylene and / or C3-C24 cycloalkenylene.

13. The compound according to any one of claims 2-12, wherein R1, R2, R4 and * are each independently an optionally substituted alkyl, alicyclic, alicyclic, heterocyclic, aryl, and / or heteroaryl group.

14. The compound according to any one of claims 2-13, wherein Ri, *, R4 and * are each independently an optionally substituted alkyl, alkenyl, and / or alkyl group.

15. The compound according to any one of claims 2-14, wherein Ri, R2, R4 and Rs are each independently an optionally substituted cycloalkyl, cycloalkenyl, and / or cyclocyclobenzyl group.

16. The compound according to any one of claims 3-15, wherein R1 and R2 are each independently independently a optionally substituted C6-C24 alkyl, C6-C24 alkenyl, and / or C6-C24 chloroalkyl.

17. The compound according to any one of claims 3-16, wherein R3 is an optionally substituted C2-C24 alkyl, C2-C24 alkenyl, and / or C2-C24 chloroalkyl.

18. The compound according to any one of claims 2-17, wherein R4 and * are each independently substituted Ci-C12 alkyl, C2-C12 alkenyl, C2-C12 cycloalkyl, C3-C8 cycloalkyl, C3(8 cycloalkenyl, C3(8 cycloalkyl, 4 to 8-membered heterocyclic, Ce-C10 aryl and / or 5 to 10-membered heteroaryl.

19. The compound according to any one of claims 1-18, wherein Gi, G2, G3 and G4 are each independently substituted with an optional alkyl, alkoxy, thioalkyl, alicyclic and / or heterocyclic group consisting of 5 to 25 atoms.

20. The compound according to any one of claims 1-19, wherein Gi, G2, G3 and G4 are each independently an optionally substituted alkyl, alkenyl, and / or bulk group consisting of 5 to 25 atoms.

21. The compound according to any one of claims 1-20, wherein one, two, or three atoms of Gi, G2, G3, and G4 are each independently replaced by N, O, or S, wherein, When replaced by an N atom, the N atom can be replaced by R?, where Ra is H or a C1-C12 alkyl group.

22. The compound according to any one of claims 3-21, wherein the Li and L2 are each independently -O-, -S-, or -NR-. a When g is 1, Yi and Y2 are each independently C, O, S, NR, or CR. a B 3 It is H or C1-C12 alkyl.

23. The compound according to any one of claims 3-21, wherein L5 is -(C=O)O-, g is 0, f is 1, E is -O-, -S(O)b-, or -SS-, wherein b is 0, 1, or 2, A1 is an optionally substituted C1-C24 sodium group, A2 is an optionally substituted C1-C24 sodium group, A3 is an optionally substituted C1-C24 sodium group, R1 is an optionally substituted C6-C24 sodium group, R2 is an optionally substituted C6-C24 sodium group, R3 is an optionally substituted C2-C24 sodium group, or R4 is an optionally substituted C1-C12 sodium group. R The sodium group, R5 is an optional C1-C12 sodium group. 5 24. The compound according to any one of claims 3-21, wherein L5 is -(C=O)S-, g is 0, f is 1, E is -O-, -S(O)b-, or -SS-, wherein b is 0, 1, or 2, Ai is an optionally substituted C1-C24 alkyl group, and A2 is an optionally substituted group.

25. The compound according to any one of claims 3-21, wherein L5 is -(C=O)NRa-, g is 0, f is 1, E is -O-, -S(O)b- or -SS-, wherein b is 0, 1 or 2, Ai is an optionally substituted C1-C24 alkyl group, and A2 is an optionally substituted C1-C24 alkyl group.

26. The compound according to any one of claims 3-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, and g is...

27. The compound according to any one of claims 3-21, wherein L5 is -(C=O)O-, g is 1, Y1 and Y2 are each independently C, C, O, S or NRa, f is 0, A1 is an optionally substituted C1-C24 alkyl group, and A2 is an optionally substituted group.

28. The compound according to any one of claims 3-21, wherein L5 is -(C=O)S-, g is 1, Yi and Y2 are each independently C, C, O, S or NRa, f is 0, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, A3 is an optionally substituted C1-C24 alkyl group, R1 is an optionally substituted C6-C24 alkyl group, and R2 is an optionally substituted C6-C24 alkyl group.

29. The compound according to any one of claims 3-21, wherein L5 is -(C=O)NRa-, g is 1, Yi and Y2 are each independently C, C, O, S or NRa, f is 0, Ai is an optionally substituted C1-C24 alkyl group, and A2 is an optionally substituted C1-C24 alkyl group. C12 sodium group, R5 is an optional C1-C12 sodium group. lyN , Dagger R5 30. The compound according to any one of claims 7, 9-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, Yi is O, Y2 is NRa, C or CRa, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, A3 is an optionally substituted C1-C24 alkyl group, R1 is an optionally substituted C6-C24 alkyl group, R2 is an optionally substituted C6-C24 alkyl group, R3 is an optionally substituted C2-C24 alkyl group, R4 is an optionally substituted C1-C12 alkyl group, and R5 is an optionally substituted C1-C12 hydroxyl group.

31. The compound according to any one of claims 7, 9-21, wherein L5 is a chemical bond, -CHz- or -(C=O)-, Yi is C, Y2 is O, C, S or NRa, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, A3 is an optionally substituted C1-C24 alkyl group, R1 is an optionally substituted C6-C24 alkyl group, R2 is an optionally substituted C6-C24 alkyl group, R3 is an optionally substituted C2-C24 alkyl group, R4 is an optionally substituted C1-C12 alkyl group, and R5 is an optionally substituted C1-C12 alkyl group.

32. The compound according to any one of claims 7, 9-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, Yi is S, Y2 is C or CRa, A1 is an optionally substituted C1-C24 sodium group, A2 is an optionally substituted C1-C24 sodium group, A3 is an optionally substituted C1-C24 sodium group, R1 is an optionally substituted C6-C24 sodium group, R2 is an optionally substituted C6-C24 sodium group, R3 is an optionally substituted C2-C24 sodium group, R4 is an optionally substituted C1-C12 sodium group, and R5 is an optionally substituted C1-C12 sodium group.

33. The compound according to any one of claims 7, 9-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, Yi is NRa, Y2 is O, NR\C or CRa, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, A3 is an optionally substituted C1-C24 alkyl group, R1 is an optionally substituted C6-C24 alkyl group, R2 is an optionally substituted C6-C24 alkyl group, R3 is an optionally substituted C2-C24 alkyl group, R4 is an optionally substituted C1-C12 alkyl group, and R5 is an optionally substituted C1-C12 alkyl group.

34. The compound according to any one of claims 8-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, Yi is O, Y2 is NRa, c or CRa, A1 is an optionally substituted C1-C24 sodium group, A2 is an optionally substituted C1-C24 sodium group, A3 is an optionally substituted C1-C24 sodium group, R1 is an optionally substituted C6-C24 sodium group, R2 is an optionally substituted C6-C24 sodium group, R4 is an optionally substituted C1-C12 sodium group, and R5 is an optionally substituted C1-C12 sodium group.

35. The compound according to any one of claims 8-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, Yi is C, Y2 is O, C, S or NRa, A1 is an optionally substituted C1-C24 sodium group, A2 is an optionally substituted C1-C24 sodium group, A3 is an optionally substituted C1-C24 sodium group, R1 is an optionally substituted C6-C24 sodium group, R2 is an optionally substituted C6-C24 sodium group, R4 is an optionally substituted C1-C12 sodium group, and R5 is an optionally substituted C1-C12 sodium group.

36. The compound according to any one of claims 8-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, Yi is S, Y2 is C or CRa, A1 is an optionally substituted C1-C24 alkyl group, A2 is an optionally substituted C1-C24 alkyl group, A3 is an optionally substituted C1-C24 alkyl group, R1 is an optionally substituted C6-C24 alkyl group, R2 is an optionally substituted C6-C24 alkyl group, R4 is an optionally substituted C1-C12 alkyl group, and R5 is an optionally substituted C1-C12 alkyl group.

37. The compound according to any one of claims 8-21, wherein L5 is a chemical bond, -CH2- or -(C=O)-, and Yi is NR. a Y2 is 0, NR, C, or CR?, Ai is an optional substituted C1-C24 sodium group, A2 is an optional substituted Ci-C24 sodium group, A3 is an optional substituted C1-C24 sodium group, R1 is an optional substituted C6-C24 sodium group, R2 is an optional substituted C6-C24 sodium group, R4 is an optional substituted C1-C12 sodium group, and R5 is an optional substituted C1-C12 hydroxyl group.

38. The compound according to any one of claims 2-37, wherein R4 and * are linked in a ring.

39. The compound according to any one of claims 1-38, or a pharmaceutically acceptable salt, stereoisomer, or solvent compound thereof, wherein the compound is selected from the group consisting of:

40. A lipid carrier comprising any one of the compounds of claims 1-39 or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate or prodrug thereof.

41. The lipid carrier according to claim 40, characterized in that, The lipid carrier comprises a first lipid compound and a second lipid compound.

42. The lipid carrier according to any one of claims 40-41, wherein the first lipid compound comprises the compound according to any one of claims 1-38 or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate or prodrug, and optionally other cationic lipids.

43. The lipid carrier according to any one of claims 40-42, wherein the second lipid compound comprises one or more of anionic lipids, neutral lipids, steroids, polymer-bound lipids, and ligand-bound lipids.

44. The lipid carrier according to any one of claims 40-43, wherein the ligand comprises one or more of antibodies, peptides, and nucleic acid aptamers.

45. The lipid carrier according to any one of claims 40-44, wherein the cationic lipid comprises DLinDMA, One or more of DODMA, DLin-MC2-MPZ, DLin-MC3-DMA, DLin-KC2-DMA, DOTAP, C12-200, DC-Chok ALC-0315, ALC-0366, SM102 and DOTMA.

46. ​​The lipid carrier according to any one of claims 40-45, wherein the anionic lipid comprises one or more of phosphatidylserine, phosphatidylinositol, phosphatidic acid, and phosphatidylglycerol.

47. The lipid carrier according to any one of claims 40-46, wherein the phosphatidylglycerol comprises one or more of DPPG, DOPG and myristoylphosphatidylglycerol.

48. The lipid carrier according to any one of claims 40-47, wherein the phosphatidylserine comprises one or more of DOPS and DPPS.

49. The lipid carrier according to any one of claims 40-48, wherein the phosphatidic acid comprises one or more of DSPA, DPPA, and DOPA.

50. The lipid carrier according to any one of claims 40-49, wherein the neutral lipid comprises at least one of DOPE, DSPC, DPPC, DOPC, POPC, POPE, DPPE, DMPE, DSPE, DEPC, HSPC and SOPE, or a lipid modified with an anionic or cationic modifying group.

51. The lipid carrier according to any one of claims 40-50, wherein the steroid comprises berberine, cholesterol, or a derivative thereof.

52. The lipid carrier according to any one of claims 40-51, wherein the polymer-bound lipid comprises polyethylene glycol-modified lipid.

53. The lipid carrier according to any one of claims 40-52, wherein the molar ratio of said compound to said neutral lipid is about 10:1 to 2:

10.

54. The lipid carrier according to any one of claims 40-53, wherein the molar ratio of said compound to steroid is about 5:1 to 1:

10.

55. The lipid carrier according to any one of claims 40-54, wherein the molar ratio of the compound to the polymer-bound lipid is about 100:1 to 20:

10.

56. A nucleic acid lipid nanoparticle composition, said composition comprising any one of the compounds of claims 1-39 or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate or prodrug, and therapeutic agent thereof.

57. A nucleic acid lipid nanoparticle composition, said composition comprising the lipid carrier and therapeutic agent as described in any one of claims 40-55.

58. The composition according to any one of claims 56-57, wherein the therapeutic agent comprises a nucleic acid drug.

59. The composition according to any one of claims 56-58, wherein the nucleic acid drug comprises one or more of RNA, DNA, antisense nucleic acid, aptamer, ribozyme, siRNA, mRNA, dsRNA, miRNA, microRNA inhibitor, immunostimulatory nucleic acid, and PNA.

60. A cell comprising a compound as described in any one of claims 1-39, a lipid carrier as described in any one of claims 40-55, and / or a composition as described in any one of claims 56-59.

61. A kit comprising a compound as described in any one of claims 1-39, a lipid carrier as described in any one of claims 40-55, a composition as described in any one of claims 56-59, and / or cells as described in claim 60.

62. A pharmaceutical formulation comprising a compound as described in any one of claims 1-39, a lipid carrier as described in any one of claims 40-55, and / or a composition as described in any one of claims 56-59, cells as described in claim 60, and / or a kit as described in claim 61, and pharmaceutically acceptable excipients, carriers, and diluents.

63. A method for delivering nucleic acids, the method comprising a compound as described in any one of claims 1-39, a lipid carrier as described in any one of claims 40-55, and / or a composition as described in any one of claims 56-59, a cell as described in claim 60, a kit as described in claim 61, and / or a pharmaceutical formulation as described in claim 62.

64. A method of producing a therapeutic agent, the method comprising a compound as described in any one of claims 1-39, a lipid carrier as described in any one of claims 40-55, and / or a composition as described in any one of claims 56-59, cells as described in claim 60, a kit as described in claim 61, and / or a pharmaceutical formulation as described in claim 62.

65. A method for treating and / or preventing a disease or condition, the method comprising a compound as described in any one of claims 1-39, a lipid carrier as described in any one of claims 40-55, and / or a composition as described in any one of claims 56-59, a cell as described in claim 60, a kit as described in claim 61, and / or a pharmaceutical preparation as described in claim 62.

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