A lipid compound and lipid carriers, nucleic acid lipid nanoparticle compositions and pharmaceutical formulations based thereon

Abstract

The present application belongs to the field of gene drug delivery, and particularly relates to a lipid compound, a lipid carrier based on the same, a nucleic acid lipid nanoparticle composition and a pharmaceutical preparation. The compound with the structure of formula (I) can be used to prepare a lipid carrier alone or together with other lipid compounds. The lipid carrier shows pH responsiveness, has a high encapsulation efficiency for nucleic acid drugs, and helps to improve the delivery efficiency of the nucleic acid drugs in vivo. Moreover, the lipid carrier can deliver the nucleic acid drugs to organs in need of enrichment, and has a good application prospect.

Description

Technical Field

[0001] This invention belongs to the field of gene drug delivery, specifically relating to a lipid compound and its lipid carrier, nucleic acid lipid nanoparticle composition and pharmaceutical formulation. Background Technology

[0002] Gene therapy is a hot research topic in modern biomedicine. For example, nucleic acid drugs can be used to prevent cancer, bacterial and viral infections, and treat diseases with genetic causes. Because nucleic acid drugs are easily degraded and have difficulty entering cells, they need to be encapsulated and delivered to target cells using vectors. Therefore, developing safe and efficient delivery vectors is a prerequisite for the clinical application of gene therapy.

[0003] Lipid nanoparticles (LNPs) are currently a research hotspot in the field of non-viral gene vectors. In 2018, the FDA approved the use of LNPs to deliver patisiran (onpattro) for the treatment of hereditary transthyretin amyloidosis, and since then, research on the delivery of nucleic acid drugs using LNP technology has seen explosive growth. In particular, at the end of 2020, the FDA approved the COVID-19 vaccines from Moderna and BioNTech & Pfizer, both of which utilize LNP technology to deliver mRNA drugs for the prevention of the COVID-19 virus.

[0004] LNPs are typically composed of four types of lipid compounds: cationic lipids, neutral lipids, sterols, and amphiphilic lipids. Among them, cationic lipids have the greatest impact on the performance of LNPs, such as affecting the encapsulation efficiency of nucleic acid drugs, the delivery efficiency of nucleic acid drugs in vivo, or cytotoxicity.

[0005] Therefore, there is a need to develop more novel compounds (such as cationic lipid compounds) to provide more options for delivering gene drugs. Summary of the Invention

[0006] The problem the invention aims to solve

[0007] The present invention aims to provide a series of compounds that can be used alone to prepare lipid carriers or co-prepared with other lipid compounds to prepare lipid carriers, thereby improving the delivery efficiency of nucleic acid drugs in vivo and enabling the delivery of nucleic acid drugs to organs that require enrichment.

[0008] The present invention also provides a lipid carrier comprising the above-described compounds.

[0009] The present invention also provides a nucleic acid lipid nanoparticle composition comprising the above-described compound or the above-described lipid carrier.

[0010] The present invention also provides pharmaceutical formulations comprising the above-described compounds, or the above-described lipid carriers, or the above-described nucleic acid lipid nanoparticle compositions.

[0011] Solution for solving the problem

[0012] In a first aspect, the present invention provides compounds of formula (I) or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs thereof.

[0013]

[0014] in:

[0015] X is in:

[0016] Ra and Ra' are each independently hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 groups, each independently selected from hydroxyl, oxo, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 Substitution of aryl or 5-10 heteroaryl groups; or,

[0017] Ra and Ra' are each independent of each other. or,

[0018] Ra and Ra' are connected to each other to form Z;

[0019] Each Z is independently C1-C 24 Alkylene, C1-C 24 Heteroalkyl, C6-C 10 arylene or 5-10 quinone heteroarylene;

[0020] W represents hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 independently selected from C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 Substitution of aryl or 5-10 heteroaryl groups; or,

[0021] W is

[0022] Each of A1, A2, A3, and A4 is independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)-, -C(=O)NR2-, -NR2C(=O)NR2-, -OC(=O)NR2-, -NR2C(=O)O-, or -O(C=O)O-;

[0023] Each R1 is independently C 1- C 24 Alkyl or C 2- C 24 alkenyl;

[0024] Each R2 is independently either hydrogen or C. 1- C 24 alkyl;

[0025] Each of B1, B2, B3 and B4 is independently a C1-C8 alkylene group or a C2-C8 alkenyl group;

[0026] Each m is independently either 0 or 1;

[0027] The heteroalkyl, heteroalkylene, heterocycloalkyl, heteroaryl, and heteroalkylene each independently have 1 to 3 heteroatoms or heteroatom groups, and the heteroatoms or heteroatom groups are each independently N, NH, O, S, S(=O) or S(=O)2.

[0028] In a second aspect, the present invention provides specific examples of compounds of formula (I) above, or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs thereof.

[0029] Thirdly, the present invention provides a lipid carrier comprising the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug.

[0030] Fourthly, the present invention provides a nucleic acid lipid nanoparticle composition comprising the above-mentioned compound or its pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug, or the above-mentioned lipid carrier, and a nucleic acid drug.

[0031] Fifthly, the present invention provides a pharmaceutical formulation comprising the above-mentioned compound or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex or prodrug thereof, or the above-mentioned lipid carrier, or the above-mentioned nucleic acid lipid nanoparticle composition, and a pharmaceutically acceptable excipient, carrier and diluent.

[0032] The effects of the invention

[0033] This invention provides a series of novel compounds of formula (I), which, as a new type of cationic lipid, can be used alone to prepare lipid carriers or co-prepared with other lipid compounds. They exhibit controllable particle size, uniform distribution, monodispersity, and high encapsulation efficiency for negatively charged drugs. Furthermore, due to the presence of a tertiary amine structure, they can exhibit different potentials at different pH levels. Under acidic conditions, they exhibit a positive charge when encapsulating negatively charged drugs, allowing the positively charged lipid carrier to attract the negatively charged drug. They can also exhibit electroneutrality or electronegativity under neutral conditions in vivo, avoiding significant cytotoxicity. The presence of multiple degradable functional groups allows the lipids to facilitate greater release and better expression of nucleic acids in vivo; the presence of multiple degradable functional groups also results in faster lipid metabolism. In addition, this lipid carrier can deliver nucleic acid drugs to organs requiring enrichment.

[0034] Furthermore, the compound has a simple synthetic route, uses inexpensive and readily available raw materials, and has high market potential. Attached Figure Description

[0035] Figure 1 This is an image of mice after intramuscular injection of LNP@mRNA prepared from compound 1 of the present invention.

[0036] Figure 2 An imaging anatomical diagram of mice after intramuscular injection of LNP@mRNA prepared from compound 1 of the present invention.

[0037] Figure 3 This is an imaging image of mice after intravenous injection of LNP@mRNA prepared from compound 8 of the present invention.

[0038] Figure 4 An imaging anatomical diagram of mice intravenously injected with LNP@mRNA prepared from compound 8 of the present invention.

[0039] Figure 5This is an imaging image of mice after intravenous injection of LNP@mRNA prepared from compound 58 of the present invention.

[0040] Figure 6 An imaging anatomical diagram of mice intravenously injected with LNP@mRNA prepared from compound 58 of the present invention.

[0041] Figure 7 The metabolism of LNP@mRNA prepared from compound 1 of the present invention and Dlin-MC3-DMA in mouse liver.

[0042] Figure 8 The metabolism of LNP@mRNA prepared from compound 1 of the present invention and Dlin-MC3-DMA in mouse spleen. Detailed Implementation

[0043] Before further describing the invention, it should be understood that the invention is not limited to the specific embodiments described herein; it should also be understood that the terminology used herein is for description only and not for limiting specific embodiments.

[0044] [Terminology Definition]

[0045] Unless otherwise stated, the following terms have the following meanings:

[0046] The term "pharmaceutically acceptable salt" refers to a salt of the compounds of this invention that is substantially non-toxic to organisms. Pharmaceutically acceptable salts generally include (but are not limited to) salts formed by the reaction of the compounds of this invention with pharmaceutically acceptable inorganic / organic acids or inorganic / organic bases; such salts are also known as acid addition salts or base addition salts. Common inorganic acids include (but are not limited to) hydrochloric acid, hydrobromic acid, sulfuric acid, phosphoric acid, etc.; common organic acids include (but are 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.; common inorganic bases include (but are not limited to) sodium hydroxide, potassium hydroxide, calcium hydroxide, barium hydroxide, etc.; and common organic bases include (but are not limited to) diethylamine, triethylamine, ethylaminobutanol, etc.

[0047] The term "stereoisomer" (or "optical isomer") refers to a stable isomer that has a perpendicular asymmetric plane due to having at least one chiral element (including a chiral center, chiral axis, chiral plane, etc.), thereby enabling the rotation of plane-polarized light. Since the compounds of this invention contain asymmetric centers and other chemical structures that may lead to stereoisomerism, this invention also includes these stereoisomers and mixtures thereof. Because the compounds of this invention and their salts 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 (purity ≥98%, ≥95%, ≥93%, ≥90%, ≥88%, ≥85%, or ≥80%). 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, for example, by converting it into a mixture of diastereomers followed by separation or recrystallization, chromatographic treatment, using chiral resolving reagents, or by direct separation of the 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 by methods well known in the art.

[0048] 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 can be achieved in the tautomer. 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. Unless otherwise stated, all tautomer forms of the compounds of this invention are within the scope of this invention.

[0049] The term "solvent" refers to a substance formed by the combination of a compound of the present invention 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.

[0050] 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.

[0051] The term "non-covalent complex" refers to a compound formed through the interaction of another molecule, where no covalent bond is formed between the two molecules. Complexation can occur, for example, through van der Waals interactions, hydrogen bonding, and electrostatic interactions (also known as ionic bonding).

[0052] The term "prodrug" refers to a derived compound that, when administered to a patient, can directly or indirectly provide the compounds of the present invention. Particularly preferred derived compounds or prodrugs are those that, when administered to a patient, can improve the bioavailability of the compounds of the present invention (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 the present invention are within the scope of the present invention, and various prodrug forms are well known in the art.

[0053] The term "independently" means that at least two groups (or ring systems) in a structure with the same or similar value ranges can have the same or different meanings under specific circumstances. For example, if substituent X and substituent Y are independently hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl, then when substituent X is hydrogen, substituent Y can be hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl; similarly, when substituent Y is hydrogen, substituent X can be hydrogen, halogen, hydroxyl, cyano, alkyl, or aryl.

[0054] 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.

[0055] The terms “contain” and “include” are used in their open, non-restrictive sense.

[0056] The term "alkyl" refers to a monovalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, without unsaturation, and linked to other segments by a single bond, including (but not limited to) methyl, ethyl, propyl, isopropyl, butyl, sec-butyl, isobutyl, and tert-butyl. For example, "C1-C..." 24 "Alkyl" refers to an alkyl group containing 1 to 24 carbon atoms. Specifically, the term "branched C" refers to an alkyl group... 10 -C 15 "Alkyl" refers to a branched alkyl group containing 10 to 15 carbon atoms, including (but not limited to) 1-butylhept-1-yl and 2-butyloct-1-yl.

[0057] The term "alkylene" refers to a divalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, without saturation, and connected to other segments by two single bonds, including (but not limited to) methylene, 1,2-ethylene, 1,3-propylene, and 1,4-butylene. For example, "C1-C..." 24 "Alkylene" refers to an alkylene group containing 1 to 24 carbon atoms.

[0058] The term "heteroalkyl" refers to a monovalent, straight-chain or branched aliphatic group composed of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) that are each independently N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by a single bond. Examples include (but not limited to) 4-oxahepten-1-yl (i.e., 3-(propoxy)propyl-1-yl), 7-oxaoctyl-1-yl (i.e., 6-(methoxy)hexyl-1-yl), 2-oxahexyl-1-yl (i.e., (butoxy)methyl), and 4-aza-4-methylpentan-1-yl (i.e., 3-(dimethylamino)propyl-1-yl), etc. For example, "C1-C 24 "Heteroalkyl" refers to a heteroalkyl group containing 1 to 24 carbon atoms and 1 to 3 heteroatoms (or heterogroups) that are each independently N, NH, O, S, S (=O) or S (=O)2.

[0059] The term "heteroalkylene" refers to a divalent, straight-chain or branched aliphatic group composed of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) each independently being N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by two single bonds, including (but not limited to) 4-aza-4-methyl-1,7-heptene and 3,4-dithia-1,6-hexene, etc. For example, "C1-C 24 "Heteroalkylene" refers to a heteroalkylene compound containing 1 to 24 carbon atoms and 1 to 3 heteroatoms (or heterogroups) that are each independently N, NH, O, S, S (=O) or S (=O)2.

[0060] The term "alkenyl" refers to a monovalent, straight-chain or branched aliphatic group composed only of carbon and hydrogen atoms, containing at least one double bond, and connected to other segments by a single bond, including (but not limited to) vinyl, propenyl, allyl, and other groups. For example, "C2-C..." 24 "Alkenyl" refers to an alkenyl group containing 2 to 24 carbon atoms.

[0061] The term "alkenyl" refers to a divalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, containing at least one double bond, and connected to other segments via two single bonds, including (but not limited to) other segments. For example, "C2-C" 24"Alkenyl" refers to alkenyl groups containing 2 to 24 carbon atoms.

[0062] The term "alkynyl" refers to a monovalent, straight-chain or branched aliphatic group consisting only of carbon and hydrogen atoms, containing at least one triple bond, and linked to other segments by a single bond. This includes (but is not limited to) ethynyl, propynyl, propynyl, and 4-pentyn-1-yl groups. For example, "C2-C..." 24 "Alkyne group" refers to an alkynyl group containing 2 to 24 carbon atoms.

[0063] The term "cycloalkyl" refers to a monovalent monocyclic or polycyclic (e.g., fused, bridged, or spirocyclic) aliphatic group consisting only of carbon and hydrogen atoms and linked to other segments by a single bond, including (but not limited to) cyclopropyl, cyclobutyl, cyclopentyl, and cyclohexyl. For example, "C3-C..." 24 "Alkyl" refers to a cycloalkyl group containing 3 to 24 cyclic atoms.

[0064] The term "heterocyclic alkyl" refers to a monovalent monocyclic or polycyclic (e.g., fused, bridged, or spirocyclic) aliphatic group consisting of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) that are each independently N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by a single bond, including (but not limited to) pyrrolidine-1-yl, piperidin-1-yl, and 4-methylpiperazin-1-yl. For example, "3-24 membered heterocyclic alkyl" refers to a heterocyclic alkyl group containing 3 to 24 cyclic atoms (or heterogroups).

[0065] The term "aryl" refers to a monovalent monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) aromatic group consisting only of carbon and hydrogen atoms and linked to other segments by a single bond, including (but not limited to) phenyl, naphthyl, anthracene, and phenanthrene groups. For example, "C6-C..." 10 "Aryl" refers to an aryl group containing 6 to 10 cyclic atoms.

[0066] The term "arylene" refers to a divalent monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) aromatic group composed only of carbon and hydrogen atoms and linked to other segments by two single bonds, including (but not limited to) 1,4-phenylene, 1,4-naphthylene, 9,10-anthraylene, and 9,10-phenanthylene. For example, "C6-C..." 10 "Aryl" refers to an aryl group containing 6 to 10 cyclic atoms.

[0067] The term "heteroaryl" refers to a monovalent monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) aromatic group composed of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) that are each independently N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by a single bond, including (but not limited to) pyrazolyl (e.g., 1H-imidazol-1-yl), oxazolyl (e.g., oxazol-2-yl), and thiazolyl (e.g., thiazolyl-4-yl). For example, "5-10-membered heteroaryl" refers to a heteroaryl group containing 5 to 10 cyclic atoms (or heterogroups).

[0068] The term "hybrid aryl" refers to a divalent monocyclic or polycyclic (e.g., bicyclic, tricyclic, or tetracyclic) aromatic group composed of a carbon atom, a hydrogen atom, and one to three heteroatoms (or heterogroups) that are independently N, NH, O, S, S(=O), or S(=O)2, and connected to other segments by two single bonds, including (but not limited to) pyrazolyl (e.g., 1,4-imidazolyl), oxazolyl (e.g., 2,4-imidazolyl), and thiazolyl (e.g., 2,5-imidazolyl). For example, "5-10-membered heteroaryl" refers to a heteroaryl group containing 5 to 10 cyclic atoms (or heterogroups).

[0069] The term "hydroxyl group" refers to the -OH group.

[0070] The term "oxo group" refers to the =O group.

[0071] [General Formula Compound]

[0072] This invention provides compounds of formula (I) or pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs thereof.

[0073]

[0074] in:

[0075] X is in:

[0076] Ra and Ra' are each independently hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 groups, each independently selected from hydroxyl, oxo, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 Substitution of aryl or 5-10 heteroaryl groups; or,

[0077] Ra and Ra' are each independent of each other. or,

[0078] Ra and Ra' are connected to each other to form Z;

[0079] Each Z is independently C1-C 24 Alkylene, C1-C 24 Heteroalkyl, C6-C 10 arylene or 5-10 quinone heteroarylene;

[0080] W represents hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 independently selected from C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 Substitution of aryl or 5-10 heteroaryl groups; or,

[0081] W is

[0082] Each of A1, A2, A3, and A4 is independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)-, -C(=O)NR2-, -NR2C(=O)NR2-, -OC(=O)NR2-, -NR2C(=O)O-, or -O(C=O)O-;

[0083] Each R1 is independently C 1- C 24 Alkyl or C 2- C 24 alkenyl;

[0084] Each R2 is independently either hydrogen or C. 1- C 24 alkyl;

[0085] Each of B1, B2, B3 and B4 is independently a C1-C8 alkylene group or a C2-C8 alkenyl group;

[0086] Each m is independently either 0 or 1;

[0087] The heteroalkyl, heteroalkylene, heterocycloalkyl, heteroaryl, and heteroalkylene each independently have 1 to 3 heteroatoms or heteroatom groups, and the heteroatoms or heteroatom groups are each independently N, NH, O, S, S(=O) or S(=O)2.

[0088] In one implementation, Ra and Ra' in formula (I) are each independently hydrogen, C1-C 12 Alkyl, C2-C 12 alkenyl, C2-C 12 Alkyne group, C1-C 12 Heteroalkyl, C3-C 12 Cycloalkyl, 3-12 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-3 groups, each independently selected from hydroxyl, oxo, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The group is substituted with an aryl or 5-10 heteroaryl group; preferably, Ra and Ra' are each independently C1-C6 alkyl, C2-C6 alkynyl, or C1-C6 alkyl. 10 Heteroalkyl or 3-10-membered heterocyclic alkyl, wherein the alkyl, ynyl, heteroalkyl, and heterocyclic alkyl are optionally composed of 1-2 groups each independently selected from hydroxyl, oxo, 3-10-membered heterocyclic alkyl, C6-C 10 Substitution of aryl or 5-10 heteroaryl groups.

[0089] In one implementation, each Z in equation (I) is independently C1-C 12 Alkylene, C1-C 12 Heteroalkyl, C6-C 10 Arylene or 5-10 heteroarylene; preferably, each Z is independently C1-C6 alkylene or C1-C6 heteroarylene.

[0090] In one implementation, W in formula (I) represents hydrogen, C1-C 12 Alkyl, C2-C 12 alkenyl, C2-C 12 Alkyne group, C1-C 12 Heteroalkyl, C3-C 12 Cycloalkyl, 3-12 membered heterocycloalkyl, C6-C 10Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-3 independently selected from C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The group is substituted with an aryl or 5-10 heteroaryl group; preferably, W is C1-C. 12 Alkyl or C1-C 12 Heteroalkyl groups, wherein the alkyl and heteroalkyl groups are optionally substituted with one or two groups, each independently selected from 3-10-membered heterocyclic alkyl groups.

[0091] In one implementation, each R1 in equation (I) is independently C. 3- C 24 Alkyl or C 3- C 24 Alkenyl; preferably, each R1 is independently a branched C 3- C 24 Alkyl or branched C 3- C 24 Alkenyl group.

[0092] In one implementation, each R2 in formula (I) is independently hydrogen or C. 1- C6 alkyl; preferably, each R2 is independently hydrogen or C 1- C4 alkyl.

[0093] In one embodiment, each of A1, A2, A3 and A4 in formula (I) is independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)- or -C(=O)NR2-; preferably, each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -O-, -SS-, -NR2C(=O)- or -C(=O)NR2-.

[0094] In one embodiment, each of B1, B2, B3 and B4 in formula (I) is independently C1-C6 alkylene or C2-C6 alkenylene; preferably, each of B1, B2, B3 and B4 is independently C1-C4 alkylene or C2-C4 alkenylene.

[0095] In some embodiments, the compound of formula (I) has a structure as shown in formula (I-1) or formula (I-1'):

[0096]

[0097] in:

[0098] Ra represents hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 groups, each independently selected from hydroxyl, oxo, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The aryl or 5-10 heteroaryl groups are substituted; preferably, Ra is C1-C. 12 Alkyl groups, preferably C1-C6 alkyl groups;

[0099] W represents hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 independently selected from C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The group is substituted with an aryl or 5-10 heteroaryl group; preferably, W is C1-C. 12 Alkyl groups, preferably C1-C6 alkyl groups;

[0100] A1, A2, A3, and A4 are each independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)-, or -C(=O)NR2-; preferably, A1, A2, A3, and A4 are each independently -O(C=O)-, -(C=O)O-, -SS-, -NR2C(=O)-, or -C(=O)NR2-;

[0101] R1 is C1-C 24 Alkyl or C2-C 24 Alkenyl; preferably, R1 is C6-C 20 Alkyl groups, preferably branched C6-C 20 Alkyl groups, more preferably branched C4 groups10 -C 15 alkyl;

[0102] Each R2 is independently either hydrogen or C. 1-12 Alkyl group; preferably, each R2 is independently hydrogen;

[0103] B1, B2, B3 and B4 are each independently C1-C8 alkylene or C2-C8 alkenylene; preferably, B1, B2, B3 and B4 are each independently C1-C6 alkylene, more preferably C1-C4 alkylene.

[0104] The heteroalkyl, heterocycloalkyl, and heteroaryl groups each independently have 1 to 3 heteroatoms or heterogroups, and each heteroatom or heterogroup is independently N, NH, O, S, S(=O) or S(=O)2.

[0105] In some embodiments, the compound of formula (I) has a structure as shown in formula (I-2) or formula (I-2'):

[0106]

[0107] in:

[0108] Ra represents hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 groups, each independently selected from hydroxyl, oxo, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The aryl or 5-10 heteroaryl groups are substituted; preferably, Ra is C1-C. 12 Alkyl, C2-C 12 Alkyne group, C1-C 12 Heteroalkyl, C3-C 12 Cycloalkyl or 3-12-membered heterocycloalkyl, wherein the alkyl, alkynyl, heteroalkyl, cycloalkyl, and heterocycloalkyl are optionally composed of 1-4 groups each independently selected from hydroxyl, oxo, C3-C8 cycloalkyl, 3-8-membered heterocycloalkyl, C6-C 10The group is substituted with an aryl or 5-10-membered heteroaryl group; more preferably, Ra is a C1-C6 alkyl, C2-C6 ynyl, C1-C6 heteroalkyl, C3-C6 cycloalkyl, or 3-6-membered heterocyclic alkyl, wherein the alkyl, ynyl, heteroalkyl, cycloalkyl, and heterocyclic alkyl are optionally replaced by 1-2 groups each independently selected from hydroxyl, oxo, C3-C8 cycloalkyl, 3-8-membered heterocyclic alkyl, C6-C6 cycloalkyl, C3 ... 10 Substitution of aryl or 5-10 heteroaryl groups;

[0109] Each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)- or -C(=O)NR2-; preferably, each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -O-, -SS-, -NR2C(=O)- or -C(=O)NR2-;

[0110] Each R1 is independently C1-C 24 Alkyl or C2-C 24 Alkenyl; preferably, each R1 is independently C6-C. 20 Alkyl groups, preferably branched C6-C 20 Alkyl groups, more preferably branched C4 groups 10 -C 15 alkyl;

[0111] Each R2 is independently either hydrogen or C. 1- C 12 Alkyl group; preferably, each R2 is independently hydrogen;

[0112] Each of B1, B2, B3 and B4 is independently C1-C8 alkylene or C2-C8 alkenylene; preferably, each of B1, B2, B3 and B4 is independently C1-C6 alkylene, more preferably C1-C4 alkylene.

[0113] The heteroalkyl, heterocycloalkyl, and heteroaryl groups each independently have 1 to 3 heteroatoms or heterogroups, and each heteroatom or heterogroup is independently N, NH, O, S, S(=O) or S(=O)2.

[0114] In some embodiments, the compound of formula (I) has a structure as shown in formula (I-3) or formula (I-3'):

[0115]

[0116] in:

[0117] Each Z is independently C1-C 24 Alkylene, C1-C 24 Heteroalkyl, C6-C 10 Arene-aryl or 5-10 heteroaryl; preferably, each Z is independently C1-C 12 Alkylene, preferably C1-C6 alkylene;

[0118] W represents hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, ynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 independently selected from C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The group is substituted with an aryl or 5-10 heteroaryl group; preferably, W is C1-C. 12 Alkyl or C1-C 12 The alkyl group and the heteroalkyl group are optionally substituted with 1 to 4 groups, each independently selected from C3-C8 cycloalkyl or 3-8-membered heteroalkyl; more preferably, W is a C1-C8 alkyl or C1-C8 heteroalkyl group, the alkyl group and the heteroalkyl group are optionally substituted with 1 to 2 groups, each independently selected from C3-C8 cycloalkyl or 3-8-membered heteroalkyl.

[0119] A1, A2, A3, and A4 are each independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)-, or -C(=O)NR2-; preferably, A1, A2, A3, and A4 are each independently -SS-, -NR2C(=O)-, or -C(=O)NR2-;

[0120] R1 is C 1- C 24 Alkyl or C 2- C 24 Alkenyl; preferably, R1 is C6-C 20 Alkyl groups, preferably branched C6-C 20 Alkyl groups, more preferably branched C4 groups 10 -C 15 alkyl;

[0121] Each R2 is independently either hydrogen or C. 1-12Alkyl group; preferably, each R2 is independently hydrogen;

[0122] B1, B2, B3 and B4 are each independently C1-C8 alkylene or C2-C8 alkenylene; preferably, B1, B2, B3 and B4 are each independently C1-C6 alkylene, more preferably C1-C4 alkylene.

[0123] The heteroalkyl, heteroalkylene, heterocycloalkyl, heteroaryl, and heteroalkylene each independently have 1 to 3 heteroatoms or heteroatom groups, and the heteroatoms or heteroatom groups are each independently N, NH, O, S, S(=O) or S(=O)2.

[0124] In some embodiments, the compound of formula (I) has a structure as shown in formula (I-4) or formula (I-4'):

[0125]

[0126] in:

[0127] Ra and Ra' are each independently hydrogen, C1-C 24 Alkyl, C2-C 24 alkenyl, C2-C 24 Alkyne group, C1-C 24 Heteroalkyl, C3-C 24 Cycloalkyl, 3-24 membered heterocycloalkyl, C6-C 10 Aryl or 5-10 heteroaryl, wherein the alkyl, alkenyl, alkynyl, heteroalkyl, cycloalkyl, heterocycloalkyl, aryl, and heteroaryl groups are optionally composed of 1-4 groups, each independently selected from hydroxyl, oxo, C1-C6 alkyl, C1-C6 heteroalkyl, C3-C 10 Cycloalkyl, 3-10 membered heterocycloalkyl, C6-C 10 The group is substituted with an aryl or 5-10 heteroaryl group; preferably, Ra and Ra' are each independently C1-C. 12 Alkyl group, wherein the alkyl group is optionally substituted with 1-4 hydroxyl groups; more preferably, Ra and Ra' are each independently C1-C6 alkyl groups, wherein the alkyl group is optionally substituted with 1-2 hydroxyl groups;

[0128] Z is C1-C 24 Alkylene, C1-C 24 Heteroalkyl, C6-C 10 arylene or 5-10 heteroarylene; preferably, Z is C1-C 12 Alkylene or C1-C 12 Heteroalkylene, preferably C1-C6 alkylene or C1-C6 heteroalkylene;

[0129] Each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)- or -C(=O)NR2-; preferably, each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -SS-, -NR2C(=O)- or -C(=O)NR2-;

[0130] Each R1 is independently C1-C 24 Alkyl or C2-C 24 Alkenyl; preferably, each R1 is independently C6-C. 20 Alkyl groups, preferably branched C6-C 20 Alkyl groups, more preferably branched C4 groups 10 -C 15 alkyl;

[0131] Each R2 is independently either hydrogen or C. 1- C 12 Alkyl group; preferably, each R2 is independently hydrogen;

[0132] Each of B1, B2, B3 and B4 is independently C1-C8 alkylene or C2-C8 alkenylene; preferably, each of B1, B2, B3 and B4 is independently C1-C6 alkylene, more preferably C1-C4 alkylene.

[0133] The heteroalkyl, heteroalkylene, heterocycloalkyl, heteroaryl, and heteroalkylene each independently have 1 to 3 heteroatoms or heteroatom groups, and the heteroatoms or heteroatom groups are each independently N, NH, O, S, S(=O) or S(=O)2.

[0134] In some embodiments, the compound of formula (I) has a structure as shown in formula (I-5) or formula (I-5'):

[0135]

[0136] in:

[0137] Z is C1-C 24 Alkylene, C1-C 24 Heteroalkyl, C6-C 10 arylene or 5-10 heteroarylene; preferably, Z is C1-C 12 Alkylene or C1-C 12 Heteroalkylene, preferably C1-C8 alkylene or C1-C8 heteroalkylene;

[0138] Each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -C(=O)-, -O-, -S(O)-, -SS-, -C(=O)S-, -SC(=O)-, -NR2C(=O)- or -C(=O)NR2-; preferably, each of A1, A2, A3 and A4 is independently -O(C=O)-, -(C=O)O-, -O-, -SS-, -NR2C(=O)- or -C(=O)NR2-;

[0139] Each R1 is independently C1-C 24 Alkyl or C2-C 24 Alkenyl; preferably, each R1 is independently C6-C. 20 Alkyl groups, preferably branched C6-C 20 Alkyl groups, more preferably branched C4 groups 10 -C 15 alkyl;

[0140] Each R2 is independently either hydrogen or C. 1- C 12 Alkyl group; preferably, each R2 is independently hydrogen;

[0141] Each of B1, B2, B3 and B4 is independently C1-C8 alkylene or C2-C8 alkenylene; preferably, each of B1, B2, B3 and B4 is independently C1-C6 alkylene, more preferably C1-C4 alkylene.

[0142] The heteroalkyl and heteroaryl groups each independently have 1 to 3 heteroatoms or heterogroups, wherein each heteroatom or heterogroup is independently N, NH, O, S, S(=O) or S(=O)2.

[0143] In some specific implementation plans, each Each excerpt is independently selected from the following:

[0144]

[0145] In some specific implementation schemes, each Z is independently selected from the following fragments:

[0146]

[0147] In some specific implementation schemes, X is selected from the following excerpt:

[0148]

[0149] [Specific compound]

[0150] This invention provides a series of specific compounds falling within the scope of the general formula, including (but not limited to):

[0151]

[0152]

[0153]

[0154]

[0155]

[0156]

[0157]

[0158]

[0159]

[0160]

[0161] [Lipid carrier]

[0162] This invention provides a lipid carrier comprising any of the aforementioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs. This type of lipid carrier exhibits high encapsulation efficiency for nucleic acid drugs, significantly improving the in vivo delivery efficiency of nucleic acid drugs.

[0163] In some embodiments, the lipid carrier comprises a first lipid compound and a second lipid compound, wherein the first lipid compound comprises any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs and optionally cationic lipids, and the second lipid compound comprises one or more of anionic lipids, neutral lipids, sterols and amphiphilic lipids.

[0164] In some specific implementations, the first lipid compound is any of the above-mentioned compounds or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or prodrug.

[0165] In some other specific embodiments, the first lipid compound is any of the above-mentioned compounds or a pharmaceutically acceptable salt, stereoisomer, tautomer, solvate, chelate, non-covalent complex, or a combination of a prodrug and a cationic lipid.

[0166] In some specific implementations, the second lipid compound is a combination of neutral lipids, sterols, and amphiphilic lipids.

[0167] In other specific embodiments, the second lipid compound is a combination of anionic lipids, neutral lipids, sterols, and amphiphilic lipids.

[0168] In some specific embodiments, the above-mentioned cationic lipids include (but are not limited to) one or more combinations of DLinDMA, DODMA, DLin-MC2-MPZ, DLin-KC2-DMA, DOTAP, C12-200, DC-Chol and DOTMA, with DLin-KC2-DMA and DOTAP being preferred.

[0169] In some specific embodiments, the above-mentioned anionic lipids include (but are not limited to) one or more combinations of phosphatidylserine, phosphatidylinositol, phosphatidic acid, phosphatidylglycerol, DOPG, DOPS and myristoyl phosphatidylglycerol, preferably DOPG and DOPS.

[0170] In some specific embodiments, the aforementioned neutral lipids include (but are not limited to) at least one of DOPE, DSPC, DPPC, DOPC, DPPG, POPC, POPE, DPPE, DMPE, DSPE, and SOPE, or lipids modified with anionic or cationic modifying groups, preferably DSPC. The anionic or cationic modifying groups are not limited.

[0171] In some specific embodiments, the aforementioned amphiphilic lipids include (but are not limited to) one or more combinations of PEG-DMG, PEG-c-DMG, PEG-C14, PEG-c-DMA, PEG-DSPE, PEG-PE, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, Tween-20, Tween-80, PEG-DPG, PEG-s-DMG, DAA, PEG-c-DOMG, and GalNAc-PEG-DSG, preferably PEG-DMG and Tween-80.

[0172] In some specific implementations, in the lipid carrier, the molar ratio of the first lipid compound, anionic lipid, neutral lipid, sterol, and amphiphilic lipid is (20–65):(0–20):(5–25):(25–55):(0.3–15); exemplaryly, the molar ratio can be 20:20:5:50:5, 30:5:25:30:10, 20:5:5:55:15, 65:0:9.7:25:0.3, etc.; wherein, in the first lipid compound, the molar ratio of any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes, or prodrugs and cationic lipids is (1–10):(0–10); exemplaryly, the molar ratio can be 1:1, 1:2, 1:5, 1:7.5, 1:10, 2:1, 5:1, 7.5:1, 10:1, etc.

[0173] In some more specific embodiments, in the lipid carrier, the molar ratio of the first lipid compound, anionic lipid, neutral lipid, sterol and amphiphilic lipid is (20-55):(0-13):(5-25):(25-51.5):(0.5-15); wherein, in the first lipid compound, the molar ratio of any of the above compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs and cationic lipids is (3-4):(0-5).

[0174] [Nucleic Acid Nanoparticle Composition]

[0175] The present invention provides a nucleic acid nanoparticle composition comprising any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs or the above-mentioned lipid carriers, as well as a nucleic acid drug.

[0176] In some implementations, the aforementioned nucleic acid drugs include (but are not limited to) one or more combinations of DNA, siRNA, mRNA, dsRNA, antisense nucleic acid, antisense oligonucleotide, microRNA, antisense microRNA, antagomir, microRNA inhibitors, microRNA activators, and immunostimulatory nucleic acids.

[0177] In some specific implementation schemes, the mass ratio of the above-mentioned nucleic acid drug to any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs is 1:(3-40).

[0178] In some other specific implementations, the mass ratio of the above-mentioned nucleic acid drug to the above-mentioned lipid carrier is 1:(3-40).

[0179] For example, the above mass ratio can be 1:3, 1:5, 1:10, 1:15, 1:20, 1:30, etc.

[0180] [Pharmaceutical Preparations]

[0181] The present invention provides a pharmaceutical formulation comprising any of the above-mentioned compounds or their pharmaceutically acceptable salts, stereoisomers, tautomers, solvates, chelates, non-covalent complexes or prodrugs, or the above-mentioned lipid carriers, or the above-mentioned nucleic acid lipid nanoparticle compositions, and pharmaceutically acceptable excipients, carriers and diluents.

[0182] In some embodiments, the particle size of the above-mentioned pharmaceutical preparation is 30 to 500 nm; for example, the particle size can be 30 nm, 50 nm, 100 nm, 150 nm, 250 nm, 350 nm, 500 nm, etc.

[0183] In some specific implementation schemes, the encapsulation rate of the nucleic acid drug in the above-mentioned drug formulation is greater than 50%; for example, the encapsulation rate can be 55%, 60%, 65%, 70%, 75%, 79%, 80%, 85%, 89%, 90%, 93%, 95%, etc.

[0184] [Preparation Method]

[0185] Unless otherwise specified, the experimental methods described in the following examples are conventional methods; unless otherwise specified, the reagents and materials are commercially available.

[0186] In this invention, the "equivalent (eq)" ratio refers to the molar ratio of the solvent or the drug.

[0187] In this invention, "appropriate amount" means that the amount of solvent or reagent added can be adjusted within a large range and has little impact on the synthesis result, and no specific limitation is required.

[0188] In the following examples, all solvents and reagents used were of analytical or chemical purity; all solvents were redistilled before use; and all anhydrous solvents were processed according to standard or literature methods.

[0189] Example 1: Synthesis of Compound 1

[0190] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0191] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0192] Intermediate product 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 1.

[0193] 1 H-NMR (400MHz, CDCl3): δ4.17-4.06(m,16h),4.01-3.99(m,8H),2.81-2.78(m,8H),2.48-2.45(m, 12H),2.41-2.31(m,18H),2.21(s,3H),1.92-1.83(m,4H),1.77-1.21(m,96H),0.98-0.87(m,24H).

[0194] Example 2: Synthesis of Compound 2

[0195] 2-Butyloctanoic acid (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 1,3-propanediol (3.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0196] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then succinic acid (3.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0197] Intermediate 2 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 3.

[0198] Intermediate product 3 (5.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 2.

[0199] 1 H-NMR (400MHz, CDCl3): δ4.23-4.12(m,32H),2.81-2.78(m,8H),2.70-2.64(m,16h),2.55-2.51(m ,16h),2.39-2.34(m,4H),2.28(s,3H),2.04-2.00(m,8H),1.80-1.19(m,84H),0.93-0.89(m,24H).

[0200] Example 3: Synthesis of Compound 3

[0201] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctylamine (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0202] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0203] Intermediate 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 3.

[0204] 1 H-NMR (400MHz, CDCl3): δ4.16-4.09(m,16h),3.98-3.89(m,8H),3.42-3.36(m,8H) ,2.69-2.11(m,35H),1.98-1.91(m,4H),1.78-1.19(m,100H),0.93-0.89(m,24H).

[0205] Example 4: Synthesis of Compound 4

[0206] 2-Butyloctanoic acid (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 3-aminopropanol (3.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0207] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then succinic acid (3.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0208] Intermediate 2 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 3.

[0209] Intermediate 3 (5.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 4.

[0210] 1H-NMR (400MHz, CDCl3): δ4.16-4.09(m,24H),3.98-3.89(m,8H),3.21-3.18(m,8H) ,2.77-2.74(m,16h),2.69-2.11(m,13H),1.78-1.19(m,92H),0.93-0.89(m,24H).

[0211] Example 5: Synthesis of Compound 5

[0212] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0213] 2-(hydroxymethoxy)ethanol (5.0 eq) was dissolved in an appropriate amount of anhydrous dichloromethane. Acryloyl chloride (1.0 eq) was slowly added at 0 °C, and the mixture was stirred for 10 min. Then, triethylamine (1.0 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 2.

[0214] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then intermediate 2 (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 3.

[0215] Intermediate 3 (5.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 5.

[0216] 1 H-NMR (400MHz, CDCl3): δ6.18-6.14(m,8H),4.18-4.14(m,16h),3.76-3.63(m,16h) ),2.49-2.16(m,35H),1.94-1.89(m,4H),1.79-1.17(m,82H),0.93-0.89(m,24H).

[0217] Example 6: Synthesis of Compound 6

[0218] Cystamine dihydrochloride (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0219] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0220] Intermediate 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 6.

[0221] 1 H-NMR (400MHz, CDCl3): δ3.65-3.51(m,24H), 2.83-2.79(m,16h), 2.49-2.21(m,23H), 1.56-1.17(m,68H), 0.93-0.88(m,24H).

[0222] Example 7: Synthesis of Compound 10

[0223] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0224] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0225] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and 1-(3-aminopropyl)imidazole (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 10.

[0226] 1 H-NMR (400MHz, CDCl3): δ7.63(s,1H),7.13(s,1H),6.94(s,1H),4.23-4.24(m,2H),3.65-3. 52(m,12H),2.83-2.79(m,8H),2.45-2.27(m,10H),1.48-1.17(m,32H),0.93-0.88(m,12H).

[0227] Example 8: Synthesis of Compound 11

[0228] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0229] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0230] Intermediate product 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and ethanolamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The product was then concentrated and purified to obtain compound 11.

[0231] 1 H-NMR (400MHz, CDCl3): δ4.46-4.19(m,12h),3.98-3.89(m,4H),3.42-3.36(m,2H),2.69-2.11(m,16H),1.78-1.19(m,48H),0.93-0.89(m,12H).

[0232] Example 9: Synthesis of Compound 12

[0233] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0234] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0235] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and ethanolamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 12.

[0236] 1 H-NMR (400MHz, CDCl3): δ3.65-3.41(m,14H), 2.83-2.80(m,8H), 2.61-2.24(m,8H), 1.48-1.17(m,32H), 0.93-0.88(m,12H).

[0237] Example 10: Synthesis of Compound 15

[0238] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0239] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0240] Intermediate 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and 4-aminobutanol (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 15.

[0241] 1 H-NMR (400MHz, CDCl3): δ4.46-4.19(m,12h),3.98-3.89(m,4H),3.58-3.49(m,2H ),3.02-2.95(m,2H),2.69-2.11(m,14H),1.78-1.19(m,52H),0.93-0.89(m,12H).

[0242] Example 11: Synthesis of Compound 21

[0243] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0244] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0245] Intermediate product 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and 2-(4-methylpiperazin-1-yl)ethylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 21.

[0246] 1 H-NMR (400MHz, CDCl3): δ4.46-4.19(m,12h), 3.98-3.89(m,4H), 2.62-2.01(m,29H), 1.78-1.19(m,48H), 0.93-0.89(m,12H).

[0247] Example 12: Synthesis of Compound 22

[0248] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0249] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0250] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N,N'-bis(2-hydroxyethyl)ethylenediamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 22.

[0251] 1 H-NMR (400MHz, CDCl3): δ4.24-4.04(m,12H),3.79-3.78(m,4H),3.04-2.98(m,4H ),2.59-2.24(m,20H),1.94-1.88(m,2H),1.59-1.21(m,48H),0.89-0.88(m,12H).

[0252] Example 13: Synthesis of Compound 26

[0253] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0254] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0255] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N-propylbutanamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 26.

[0256] 1 H-NMR (400MHz, CDCl3): δ3.65-3.41(m,6H),3.04-2.99(m,2H),2.84-2.76(m,4H) ),2.50-2.44(m,4H),2.24-2.20(m,1H),1.49-1.11(m,22H),0.89-0.88(m,12H).

[0257] Example 14: Synthesis of Compound 28

[0258] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0259] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0260] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N,N'-bis(2-hydroxyethyl)ethylenediamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was concentrated and purified to obtain compound 28.

[0261] 1 H-NMR (400MHz, CDCl3): δ3.65-3.41(m,16h), 2.83-2.80(m,8H), 2.61-2.24(m,14H), 1.48-1.17(m,32H), 0.93-0.88(m,12H).

[0262] Example 15: Synthesis of Compound 30

[0263] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0264] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0265] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N-isopropyl-N-(2-(piperazin-1-yl)ethyl)prop-2-amine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was concentrated and purified to obtain compound 30.

[0266] 1 H-NMR (400MHz, CDCl3): δ3.65-3.61(m,2H),3.53-3.50(m,4H),2.84-2.69(m,6H) ,2.53-2.51(m,2H),2.37-2.23(m,13H),1.49-1.14(m,16h),1.01-0.88(m,18H).

[0267] Example 16: Synthesis of Compound 32

[0268] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0269] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0270] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N-(3-aminopropyl)diethanolamine (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was then concentrated and purified to obtain compound 32.

[0271] 1 H-NMR (400MHz, CDCl3): δ4.61-4.58(m,4H),4.24-4.04(m,12H),3.79-3.78(m,4H) ),2.48-2.24(m,16h),1.94-1.88(m,2H),1.59-1.21(m,50H),0.89-0.88(m,12H).

[0272] Example 17: Synthesis of Compound 36

[0273] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0274] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0275] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and (4-aminopiperazin-1-yl)methanol (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was concentrated and purified to obtain compound 36.

[0276] 1H-NMR (400MHz, CDCl3): δ4.61-4.58(m,2H),4.24-4.04(m,12H),2.94-2.85(m,4H) ),2.64-2.24(m,20H),1.94-1.88(m,2H),1.59-1.21(m,48H),0.89-0.88(m,12H).

[0277] Example 18: Synthesis of Compound 38

[0278] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0279] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0280] Intermediate 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and cystamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 38.

[0281] 1 H-NMR (400MHz, CDCl3): δ4.48-4.14(m,24H), 3.79-3.73(m,8H), 2.76-2.24(m,32H), 1.94-1.88(m,4H), 1.59-1.21(m,96H), 0.89-0.88(m,24H).

[0282] Example 19: Synthesis of Compound 41

[0283] Adipic acid (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctyl alcohol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate product 1.

[0284] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0285] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and 4-(aminomethoxy)butanol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was concentrated and purified to obtain compound 41.

[0286] 1 H-NMR (400MHz, CDCl3): δ4.48-4.04(m,14H),3.76-3.61(m,6H),3.42-3.33(m,2H) ),2.51-2.24(m,12H),1.94-1.88(m,2H),1.59-1.21(m,52H),0.89-0.88(m,12H).

[0287] Example 20: Synthesis of Compound 43

[0288] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0289] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0290] Intermediate 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and N-(3-aminopropyl)diethanolamine (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was concentrated and purified to obtain compound 43.

[0291] 1 H-NMR (400MHz, CDCl3): δ4.63-4.60(m,4H),3.65-3.51(m,12H),2.83-2.79(m,8H),2.49-2.21(m,10H),1.56-1.17(m,34H),0.93-0.88(m,12H).

[0292] Example 21: Synthesis of Compound 45

[0293] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0294] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0295] Intermediate product 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N was added. 1 N 3 -Dimethylpropyl-1,3-diamine (1.0 eq) was added and stirred at room temperature for 16 h. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to give compound 45.

[0296] 1 H-NMR (400MHz, CDCl3): δ3.65-3.51(m,12H), 2.83-2.79(m,8H), 2.49-2.21(m,16h), 1.56-1.17(m,34H), 0.93-0.88(m,12H).

[0297] Example 22: Synthesis of Compound 47

[0298] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0299] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0300] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and (4-aminopiperazine)-1-methanol (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was concentrated and purified to obtain compound 47. 1 H-NMR (400MHz, CDCl3): δ4.92-4.89(m,2H),3.54-3.48(m,8H),2.83-2.79(m,12H ),2.62-2.55(m,8H),2.34-2.27(m,6H),1.56-1.17(m,32H),0.93-0.88(m,12H).

[0301] Example 23: Synthesis of Compound 49

[0302] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0303] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0304] Intermediate 2 (5.0 eq) was dissolved in an appropriate amount of methanol, and cystamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 49.

[0305] 1 H-NMR (400MHz, CDCl3): δ3.65-3.51(m,26H),2.83-2.79(m,16h),2.64-2.48(m,16h),2.64-2.48(m,4H),1.56-1.17(m,64H),0.93-0.88(m,24H).

[0306] Example 24: Synthesis of Compound 53

[0307] 2-tert-Butoxycarbonylaminoethanethiol (1.0 eq) was dissolved in an appropriate amount of dichloromethane and methanol (V / V = 1 / 1). 2-Mercaptoethanol (10.0 eq) was slowly added while stirring for 4 h. Iodine (0.1 eq) was then dissolved in methanol and slowly added to the reaction mixture. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC until complete. The product was concentrated and purified to obtain intermediate product 1.

[0308] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0309] Intermediate 2 (1.0 eq) was dissolved in an appropriate amount of ethyl acetate, and a 4M hydrochloric acid-ethyl acetate solution (6V) was added. The mixture was stirred at room temperature for 2 h. The reaction was monitored by TLC until complete. The product was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 3.

[0310] Intermediate product 3 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 4.

[0311] Intermediate 4 (6.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was concentrated and purified to obtain compound 53.

[0312] 1 H-NMR (400MHz, CDCl3): δ3.95-3.91(m,8H),3.86-3.83(m,8H),3.55-3.50(m,8H),2.84- 2.76(m,16H),2.50-2.44(m,20H),2.15(s,3H),1.49-1.11(m,68H),0.89-0.88(m,24H).

[0313] Example 25: Synthesis of Compound 54

[0314] 2-Hydroxyethyl disulfide (5.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0315] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0316] Intermediate 2 (6.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was concentrated and purified to obtain compound 54.

[0317] 1 H-NMR (400MHz, CDCl3): δ4.43-4.36(m,8H),3.96-3.89(m,16H),2.84-2.76(m,16H),2.50-2.14(m,23H),1.49-1.11(m,68H),0.89-0.88(m,24H).

[0318] Example 26: Synthesis of Compound 55

[0319] 2-tert-Butoxycarbonylaminoethanethiol (1.0 eq) was dissolved in an appropriate amount of dichloromethane and methanol (V / V = 1 / 1), and 2-mercaptoethanol (10.0 eq) was added. The mixture was stirred for 10 min, and then 2-mercaptoethanol (1.1 eq) was slowly added. The mixture was stirred for 4 h. Iodine (0.1 eq) was dissolved in methanol and slowly added to the reaction mixture. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until it was complete. The mixture was concentrated and purified to obtain intermediate product 1.

[0320] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.5 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, filtered, concentrated, and purified to obtain intermediate 2.

[0321] Intermediate product 2 (1.0 eq) was dissolved in an appropriate amount of ethyl acetate, and a 4M hydrochloric acid-ethyl acetate solution (6V) was added. The mixture was stirred at room temperature for 2 h. The reaction of the starting material was monitored by TLC until complete. The product was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 3.

[0322] Intermediate product 3 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 4.

[0323] Intermediate 4 (6.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was concentrated and purified to obtain compound 55.

[0324] 1 H-NMR (400MHz, CDCl3): δ4.43-4.36(m,8H),3.67-3.52(m,16H),2.84-2.76(m,16H),2.50-2.14(m,23H),1.49-1.11(m,68H),0.89-0.88(m,24H).

[0325] Example 27: Synthesis of Compound 56

[0326] 2-Mercaptoethanol (1.0 eq) was dissolved in an appropriate amount of tetrahydrofuran, and PPh3 (1.5 eq) and CBr4 (1.5 eq) were added. The mixture was stirred at room temperature for 2 h. The reaction of the starting material was monitored by TLC until complete. The pH was adjusted to 3-4, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, concentrated, and purified to obtain intermediate 1.

[0327] 2-Butyloctanol (1.0 eq) was dissolved in an appropriate amount of DMF, and intermediate 1 (2.0 eq) and potassium carbonate (3.0 eq) were added. After the addition was complete, the mixture was stirred at 90 °C for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0328] Intermediate product 2 (1.0 eq) was dissolved in an appropriate amount of dichloromethane and methanol (V / V = 1 / 1), and mercaptoethylamine (10.0 eq) was added. The mixture was stirred for 4 h. Then, iodine (0.1 eq) was dissolved in methanol and slowly added to the reaction mixture. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until it was complete. The mixture was then concentrated and purified to obtain intermediate product 3.

[0329] Intermediate product 3 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 4.

[0330] Intermediate 4 (6.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction was monitored by TLC until the starting material was completely reacted. The mixture was concentrated and purified to obtain compound 56.

[0331] 1 H-NMR (400MHz, CDCl3): δ3.67-3.62(m,16H), 3.51-3.21(m,16H), 2.84-2.76(m,16H), 2.50-2.14(m,19H), 1.68-1.11(m,72H), 0.89-0.88(m,24H).

[0332] Example 28: Synthesis of Compound 57

[0333] 2-Hydroxyethyl disulfide (1.0 eq) was dissolved in an appropriate amount of dichloromethane. Acryloyl chloride (1.2 eq) was slowly added at 0 °C, and the mixture was stirred for 10 min. Then, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0334] 2-Butyloctanol (1.0 eq) was dissolved in an appropriate amount of tetrahydrofuran, and PPh3 (1.5 eq) and CBr4 (1.5 eq) were added. The mixture was stirred at room temperature for 2 h. The reaction of the starting material was monitored by TLC until complete. The pH was adjusted to 3-4, and the mixture was extracted with ethyl acetate. The combined organic layers were washed with brine, dried over anhydrous sodium sulfate, concentrated, and purified to obtain intermediate 2.

[0335] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of DMF, and intermediate 2 (2.0 eq) and potassium carbonate (3.0 eq) were added. After the addition was complete, the mixture was stirred at 90 °C for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 3.

[0336] Intermediate 3 (6.0 eq) was dissolved in an appropriate amount of methanol, and N,N-bis(3-aminopropyl)methylamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 57.

[0337] 1 H-NMR (400MHz, CDCl3): δ3.95-3.91(m,8H),3.86-3.83(m,16H),3.55-3.20(m,8H),2.84 -2.76(m,16H),2.50-2.44(m,16H),2.15(s,3H),1.49-1.11(m,72H),0.89-0.88(m,24H).

[0338] Example 29: Synthesis of Compound 58

[0339] 2-Butyloctanoic acid (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 1,3-propanediol (3.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0340] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then succinic acid (3.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0341] Intermediate 2 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and DMAP (0.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 4-hydroxybutyl acrylate (1.2 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 3.

[0342] Intermediate product 3 (3.0 eq) was dissolved in an appropriate amount of methanol, and ethanolamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The product was then concentrated and purified to obtain compound 58.

[0343] 1 H-NMR (400MHz, CDCl3): δ4.22-4.16(m,16H),3.89-3.85(m,4H),3.56-3.52(m,2H),2.81-2 .78(m,8H),2.57-2.46(m,6H),2.11-1.98(m,6H),1.69-1.19(m,40H),0.93-0.89(m,12H).

[0344] Example 30: Synthesis of Compound 59

[0345] Cystamine disulfate (2.0 eq) was dissolved in an appropriate amount of dichloromethane, and HOBT (1.5 eq), EDCI (1.5 eq), and triethylamine (3.0 eq) were added. The mixture was stirred for 10 min, and then 2-butyloctanoic acid (1.0 eq) was added. After the addition was complete, the mixture was stirred at room temperature for 16 h. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate product 1.

[0346] Intermediate 1 (1.0 eq) was dissolved in an appropriate amount of dichloromethane, and acryloyl chloride (1.2 eq) was slowly added at 0 °C. After stirring for 10 min, triethylamine (1.2 eq) was added, and the mixture was stirred at room temperature for 16 h after the addition was complete. The reaction of the starting material was monitored by TLC until complete. The mixture was extracted twice with ethyl acetate, and the organic phase was collected. Then, anhydrous sodium sulfate was added for drying, and the mixture was filtered, concentrated, and purified to obtain intermediate 2.

[0347] Intermediate 2 (3.0 eq) was dissolved in an appropriate amount of methanol, and N,N-diisopropylethylenediamine (1.0 eq) was added. The mixture was stirred at room temperature for 16 h after the addition was complete. The reaction was monitored by TLC until the starting material was fully reacted. The mixture was then concentrated and purified to obtain compound 59.

[0348] 1 H-NMR (400MHz, CDCl3): δ3.64-3.48(m,12H), 2.83-2.22(m,20H), 1.56-1.17(m,32H), 1.11-0.98(m,12H), 0.93-0.88(m,12H).

[0349] Example 31

[0350] Compounds 1, 2, 6, 8, 11, 15, 21, 38, 54, 55, 58, and 59 were dissolved in ethanol (concentration of 24.4 mg / mL based on total lipid weight) along with cholesterol, DSPC (distearylphosphatidylcholine), and PEG-DMG (polyethylene glycol dimyristate) in a molar ratio of 50:38.5:10:1.5 (where the equivalent of compounds 1, 2, 6, 8, 11, 15, 21, 38, 54, 55, 58, or 59 is 50, the equivalent of cholesterol is 38.5, the equivalent of DSPC is 10, and the equivalent of PEG-DMG is 1.5). Luciferase mRNA was dissolved in pH [missing information - likely a pH value]. In a 10 mM citrate buffer solution at 4.0 g / mL (drug concentration 0.276 mg / mL), the two solutions were mixed rapidly using microfluidic technology at a volume ratio of 1:3 (where the ethanol equivalent was 1 and the aqueous solution equivalent was 3). The buffer environment was then replaced with PBS at pH 7.4 using dialysis or tangential flow technology to remove the ethanol, thus preparing multiple groups of LNP@mRNA.

[0351] The particle size, PDI, and encapsulation efficiency of each LNP@mRNA were tested, and the results are shown in Table 1.

[0352] Table 1. Particle size, PDI, Zeta, and encapsulation efficiency of each LNP@mRNA

[0353] compound Particle size (nm) PDI Zeta(mV) Encapsulation efficiency (%) 1 149 0.11 -6.6 92 2 204 0.07 -16.2 82 6 136 0.10 -8.1 91 8 357 0.20 -21.0 83 11 53 0.05 -4.7 87 15 85 0.11 -6.4 94 21 80 0.09 -6.1 96 38 95 0.08 -14 89 54 119 0.07 -4.1 92 55 121 0.04 -2.2 94 58 211 0.07 -7.2 87 59 73 0.16 -1.0 81

[0354] The results showed that the LNP@mRNA prepared from the compounds listed in the table above had an encapsulation efficiency of over 80% for mRNA, and the LNP@mRNA prepared from compounds 11, 15, 21, 38, and 59 in combination with the other three lipids had smaller particle sizes. Furthermore, the LNPs prepared from these compounds all exhibited a negative charge under neutral conditions, indicating good biocompatibility. Therefore, the compounds provided by this invention demonstrate a high encapsulation efficiency for nucleic acid drugs, and their use as carriers can improve the in vivo delivery efficiency of nucleic acid drugs.

[0355] Each LNP@mRNA was injected into mice via tail vein, and the fluorescence intensity and organ distribution in the mice were tested 6 hours later. Figure 1 This is an image of mice after intramuscular injection of LNP@mRNA prepared from compound 1. Figure 2 Anatomical imaging of mice after intramuscular injection of LNP@mRNA prepared from compound 1, combined with... Figure 1 and Figure 2 It can be seen that mRNA is mainly expressed in muscle, liver and spleen. Figure 3 This is an image of mice after intravenous injection of LNP@mRNA prepared from compound 8. Figure 4 Anatomical imaging of mice after intravenous injection of LNP@mRNA prepared from compound 8, combined with... Figure 3 and Figure 4 It can be seen that all mRNA is expressed in the spleen. Figure 5 This is an image of mice after intravenous injection of LNP@mRNA prepared from compound 58. Figure 6 Anatomical imaging of mice after intravenous injection of LNP@mRNA prepared from compound 58, combined with... Figure 5 and Figure 6 It can be seen that mRNA is mainly expressed in the spleen, with a small amount expressed in the liver. Therefore, the above-mentioned lipid compounds with specific structures can be selected as lipid carriers according to the organs where nucleic acid drugs need to be enriched.

[0356] Example 32

[0357] Compound 1 and the marketed cationic lipid DLin-MC3-DMA (abbreviated as MC3) were dissolved in ethanol (concentration of 24.4 mg / mL based on total lipid weight) with cholesterol, DSPC (distearate phosphatidylcholine), and PEG-DMG (polyethylene glycol dimyristate) at a molar ratio of 50:38.5:10:1.5 (where the equivalent of compound 1 or MC3 is 50, the equivalent of cholesterol is 38.5, the equivalent of DSPC is 10, and the equivalent of PEG-DMG is 1.5). Luciferase mRNA was dissolved in a 10 mM citrate buffer solution at pH 4.0 (drug concentration of 0.276 mg / mL) at a volume ratio of 1:3 (where the equivalent of the ethanol solution is 1, and the equivalent of the aqueous solution is 3). The two phases were rapidly mixed using microfluidic technology, and the buffer environment was replaced with a pH of 1 using dialysis or tangential flow. The LNP@mRNA was prepared by PBS at 7.4°C to remove ethanol.

[0358] Each group of LNP@mRNA was injected into mice via tail vein, with 100 μg of mRNA per mouse. The lipid metabolism in the liver and spleen of the mice was then tested. Figure 7 The metabolism of LNP@mRNA in mouse liver was prepared by combining compound 1, MC3, and three other lipids. It can be seen that MC3 reached its peak at 6 hours after LNP@mRNA injection and maintained a high lipid level after 48 hours. In contrast, compound 1 reached its peak at 2 hours after LNP@mRNA injection, and its peak level was much lower than that of MC3 throughout the 48 hours. Therefore, it can be concluded that compound 1 has a faster metabolic rate in the liver than MC3. Figure 8 The metabolism of LNP@mRNA prepared from compound 1, MC3, and three other lipids in the spleen of mice was observed. MC3 reached its peak level 12 hours after LNP@mRNA injection and maintained a high lipid level even after 48 hours. In contrast, compound 1 reached its peak level 2 hours after LNP@mRNA injection, and its peak lipid level was significantly lower than that of compound 1 throughout the 48-hour period. This indicates that compound 1 has a faster metabolic rate in the spleen compared to MC3. Furthermore, the lipids accumulated in the liver and spleen at lower levels, exhibiting a faster metabolic rate, higher biocompatibility, and lower toxicity compared to commercially available cationic lipids.

[0359] Example 33

[0360] Compound 55 was dissolved in ethanol (24.4 mg / mL, total lipid weight) in a molar ratio of 30:20:38.5:10:1.5 with DOTAP ((2,3-dioleopropyl)trimethylammonium chloride), cholesterol, DSPC, and PEG-DMG (where the equivalent of compound 55 was 30, the equivalent of DOTAP was 20, the equivalent of cholesterol was 38.5, the equivalent of DSPC was 10, and the equivalent of PEG-DMG was 1.5). Luciferase mRNA was dissolved in 50 mM citrate buffered saline solution at pH 4.0 (drug concentration 0.276 mg / mL) at a volume ratio of 1:3 (where the equivalent of the ethanol solution was 1 and the equivalent of the aqueous solution was 3). The two phases were rapidly mixed using microfluidic technology, and the buffer environment was replaced with PBS at pH 7.4 using dialysis or tangential flow technology to prepare LNP@mRNA. Adding sucrose as a cryoprotectant yields a nucleic acid lipid nanoparticle drug formulation.

[0361] Example 34

[0362] Compound 56 was dissolved in ethanol (24.4 mg / mL, total lipid weight) with DOTAP, DOPS (dioleoylphosphatidylserine), cholesterol, DSPC, and PEG-DMG (total 15 mg) in a molar ratio of 20:25:15:25:5:10 (where the equivalent of compound 56 is 20, the equivalent of DOTAP is 25, the equivalent of DOPS is 15, the equivalent of cholesterol is 25, the equivalent of DSPC is 5, and the equivalent of PEG-DMG is 10). Luciferase mRNA (5 mg) was dissolved in 50 mM citrate buffered saline solution at pH 4.0 (drug concentration 0.276 mg / mL) in a volume ratio of 1:3 (where the equivalent of the ethanol solution is 1 and the equivalent of the aqueous solution is 3). The two phases were rapidly mixed using microfluidic technology, and the buffer environment was replaced with PBS at pH 7.4 using dialysis or tangential flow technology to prepare LNP@mRNA. Adding sucrose as a cryoprotectant yields a nucleic acid lipid nanoparticle drug formulation.

[0363] Example 35

[0364] Compound 2 was dissolved in ethanol (concentration of 24.4 mg / mL based on total lipid weight) with DLin-KC2-DMA (CAS No.: 1190197-97-7), DOPG (dioleoylphosphatidylglycerol), cholesterol, DSPC, and Tween-80 (total 30 mg) in a molar ratio of 15:5:3:51.5:25:0.5 (where the equivalent of compound 2 is 15, the equivalent of DLin-KC2-DMA is 5, the equivalent of DOPG is 3, the equivalent of cholesterol is 51.5, the equivalent of DSPC is 25, and the equivalent of Tween-80 is 0.5). Luciferase mRNA (1 mg) was dissolved in pH [a solution not specified in the original text]. LNP@mRNA was prepared by rapidly mixing the two phases (drug concentration 0.276 mg / mL) in a 50 mM citrate buffer solution at a volume ratio of 1:3 (where the ethanol equivalent was 1 and the aqueous equivalent was 3). The buffer environment was then replaced with PBS at pH 7.4 using dialysis or tangential flow techniques. Sucrose was added as a cryoprotectant to obtain the nucleic acid lipid nanoparticle drug formulation.

[0365] It should be noted that although specific examples have been used to illustrate the technical solutions of the present invention, those skilled in the art will understand that the present invention is not limited thereto. Various embodiments of the present invention have been described above. These descriptions are exemplary and not exhaustive, and are not limited to the disclosed embodiments. Many modifications and variations will be apparent to those skilled in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen to best explain the principles, practical applications, or technical improvements to the embodiments in the market, or to enable others skilled in the art to understand the embodiments disclosed herein.

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, in: X is or ,in: When X is In this case, Ra is a C1-C6 alkyl group, wherein the alkyl group is substituted with one hydroxyl group; When X is At that time, Ra and Ra' were each independent of each other. Z is ; W is ; Each R1 is an independent branch of C. 10 -C 15 alkyl; Each Each independently .

2. The compound according to claim 1 or a pharmaceutically acceptable salt thereof, characterized in that, X is selected from the following excerpt: 。 3. The following compounds or their pharmaceutically acceptable salts: 。 4. A lipid carrier comprising the compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof.

5. The lipid carrier according to claim 4, characterized in that, The lipid carrier comprises a first lipid compound and a second lipid compound, wherein the first lipid compound comprises a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof and optionally a cationic lipid, and the second lipid compound comprises neutral lipids, sterols, amphiphilic lipids and optionally anionic lipids; In the lipid carrier, the molar ratio of the first lipid compound, the anionic lipid, the neutral lipid, the sterol, and the amphiphilic lipid is (20~65):(0~20):(5~25):(25~55):(0.3~15). In the first lipid compound, the molar ratio of the compound or its pharmaceutically acceptable salt according to any one of claims 1 to 3 to the cationic lipid is (1 to 10): (0 to 10).

6. The lipid carrier according to claim 5, characterized in that, In the lipid carrier, The cationic lipids include one or more of DLinDMA, DODMA, DLin-MC2-MPZ, DLin-KC2-DMA, DOTAP, C12-200, DC-Chol and DOTMA. The anionic lipids include one or more of phosphatidylserine, phosphatidylinositol, phosphatidic acid, phosphatidylglycerol, DPPG, DOPG, DOPS and myristoyl phosphatidylglycerol; The neutral lipids include at least one of DOPE, DSPC, DPPC, DOPC, POPC, POPE, DPPE, DMPE, DSPE and SOPE, or lipids modified with anionic or cationic modifying groups. The amphiphilic lipids include one or more of PEG-DMG, PEG-c-DMG, PEG-C14, PEG-c-DMA, PEG-DSPE, PEG-PE, PEG-modified ceramides, PEG-modified dialkylamines, PEG-modified diacylglycerols, Tween-20, Tween-80, PEG-DPG, PEG-s-DMG, DAA, PEG-c-DOMG, and GalNAc-PEG-DSG.

7. A nucleic acid lipid nanoparticle composition comprising a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, or a lipid carrier according to any one of claims 4 to 6, and a nucleic acid drug.

8. The nucleic acid lipid nanoparticle composition according to claim 7, characterized in that, The nucleic acid drug includes one or more of the following: DNA, siRNA, mRNA, dsRNA, antisense nucleic acid, microRNA, antisense microRNA, antagomir, microRNA inhibitor, microRNA activator, and immunostimulatory nucleic acid.

9. The nucleic acid lipid nanoparticle composition according to claim 7, characterized in that, The mass ratio of the nucleic acid drug to the compound or its pharmaceutically acceptable salt according to any one of claims 1 to 3 is 1:(3 to 40); or, the mass ratio of the nucleic acid drug to the lipid carrier according to any one of claims 4 to 6 is 1:(3 to 40).

10. A pharmaceutical formulation comprising a compound according to any one of claims 1 to 3 or a pharmaceutically acceptable salt thereof, or a lipid carrier according to any one of claims 4 to 6, or a nucleic acid lipid nanoparticle composition according to any one of claims 7 to 9, and a pharmaceutically acceptable excipient, carrier, and diluent.

11. The pharmaceutical preparation according to claim 10, characterized in that, The particle size of the pharmaceutical preparation is 30~500 nm.

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